Background

Metabolic reprogramming exists within tumor cells and tumor microenvironment (TME) in breast cancer (BC), but little is known about how the immunometabolic interplay of BC evolves during treatment. Using temporal proteogenomic profiling, we studied in-depth BC immunometabolism and its potential therapeutic vulnerabilities.

Methods

BC tissue (pre/on/post neoadjuvant chemotherapy; NAC) was longitudinally collected from the PROMIX trial (NCT00957125) of NAC (n=150 patients) in HER2-negative BC and analyzed by: bulk RNA microarray (n=122), single nucleus RNA-seq (snRNA-seq) (n=8), whole-exome sequencing (WES) (n=20), and mass spectrometry-based proteomics (n=29), including bulk/single-cell immunometabolic phenotype/cluster deconvolution, protein correlation network and clonal evolution analyses.

Results

Baseline and dynamic change of immunometabolic phenotype based on bulk gene expression profiling suggested tumors with hot TME or downregulation of tricarboxylic acid (TCA) cycle, amino acid or nucleotide metabolism were associated with higher pathologic complete response in multivariable analysis. BC proteomes showed TCA cycle-related protein module was starkly elevated within cold tumors, and vice versa. Potential drug targets (FASN, LDHB, LDHA, IDH2, MDH2) in metabolic pathways regulating the TME were revealed through unbiased proteogenomic differential abundance analyses (cold vs hot). Fewer subclones were identified in hot (28.3%) tumors than cold (43.1%) tumors (P<.001), which were more likely to have accelerated growth relative to their parent if included known metabolic drivers (SDHA, CACNA1D, ACSL3, ATIC, MED12). Metabolic flexibility of breast epithelial cells was dissected by five snRNA-based metabolic clusters (C): C0 (normal and tumor cells, lowest global metabolic activity); C1 (OXPHOS and glycolysis, cold tumor exclusively); C2 (OXPHOS but mutually with glycolysis); C3 (glutathione); C4 (Notch signaling).

Conclusions

This longitudinal proteogenomic study shows that interaction of tumor intrinsic metabolic states and TME is associated with treatment outcome, shedding light on the importance to target tumor metabolism for immunoregulation.

Clinical trial identification

NCT00957125; September 8, 2016.

Legal entity responsible for the study

The authors.

Funding

Roche Sweden.

Disclosure

T. Foukakis: Financial Interests, Institutional, Other, contracted research: Pfizer, Roche; Financial Interests, Personal, Advisory Board: Affibody, Novartis, Pfizer, Roche, Exact Sciences, Veracyte; Financial Interests, Personal, Royalties: UpToDate; Non-Financial Interests, Personal, Speaker’s Bureau: Pfizer. All other authors have declared no conflicts of interest.

Background

Neoadjuvant atezolizumab plus docetaxel, trastuzumab and pertuzumab therapy showed favorable efficacy and safety profiles in HER2-positive early breast cancer (EBC) in NEO-PATH phase 2 study, which provided evidence of combining immunotherapy and anti-HER2 treatment as a potential new therapeutic option. We evaluated potential genomic biomarkers in NEO-PATH trial.

Methods

Tumor tissue were collected before neoadjuvant chemotherapy from all enrolled patients and at surgery in patients who did not achieve pathologic complete response (pCR). Targeted sequencing using FoundationOne CDx^TM and whole transcriptome sequencing (WTS) were performed. The association between genomic features and pCR achievement was analyzed by Fisher's exact test for the enrichment test by mutations and by differential gene expression analysis using edgeR package. A p-value or false discovery rate (FDR) less than 0.05 were considered as statistically significant.

Results

Targeted sequencing result were obtained from 63 patients(T1) and 15 patients(T3), respectively. Pre-treatment RAD21 amplification (p=0.0002), MYC amplification (p=0.0095) and MYC pathway mutations (p=0.0095) were more frequently detected in non-pCR group (n=23) compared to pCR group (n=40), and co-amplification of ERBB2 and MYC were enriched in non-pCR group (p=0.01687). When genomic mutations were compared before and after neoadjuvant chemotherapy, ERBB2 amplification (p= 0.0026) and CDK12 amplification (p=0.0006) were enriched in pre-treatment tumors and PTPRO mutation (p=0.0229) was enriched in residual tumor tissue. WTS result were obtained from 62 patients(T1) and 10 patients(T3), respectively. Luminal subtypes were enriched in non-pCR group (p = 0.0558). In patients with non-pCR, LRP1B (FDR=4.04e-04), ESR1 (FDR=1.07e-4) and RAD21 (FDR=2.05e-03) were highly expressed and ESR1, luminal and MYC related genesets showed higher expression (FDR<0.05).

Conclusions

Luminal subtype and MYC amplification were associated with non-pCR in patients with HER2-positive EBC who were treated with immunotherapy and anti-HER2 treatment combination. Further evaluation of the predictive role of these mutations are warranted.

Clinical trial identification

NCT03881878.

Legal entity responsible for the study

Yeon Hee Park.

Funding

This work was supported by a grant from the Ministry of Health and Welfare, Republic of Korea (HA17C0055) and by the National R&D Program for Cancer Control, Ministry of Health and Welfare, Republic of Korea (1720150). The study drugs were provided by Roche (trastuzumab, pertuzumab, and atezolizumab), Sanofi (docetaxel), and Dong-A ST (tripegfilgrastim).

Disclosure

H.K. Ahn: Non-Financial Interests, Institutional, Principal Investigator: AstraZeneca, Roche, Boehringer Ingelheim, MSD, Pfizer, Eli Lilly; Financial Interests, Personal, Invited Speaker: MSD Boehringer Ingelheim. S. Im: Financial Interests, Personal, Invited Speaker: Merck, AstraZeneca, Eisai, GSK, Hanmi, Eli Lilly, MSD, Pfizer, Novartis; Financial Interests, Institutional, Research Grant: Roche, AstraZeneca, Pfizer, Dae Woong; K.H. Jung: Non-Financial Interests, Personal, Advisory Board: Roche, AstraZeneca, Novartis, BiXink, Takeda. Y.H. Park: Financial Interests, Personal, Advisory Board: AstraZeneca, Pfizer, Roche, Novartis, MSD, Daiichi Sankyo; Financial Interests, Personal, Invited Speaker: AstraZeneca, Pfizer, Roche, Novartis, MSD; Financial Interests, Institutional, Other, Research Grant: AstraZeneca, Pfizer, Roche, MSD; Non-Financial Interests, Principal Investigator: AstraZeneca, Pfizer, Novartis, MSD, Lilly, Roche, Daiichi Sankyo. All other authors have declared no conflicts of interest.

Background

Pelareorep (pela) is an intravenously delivered non-engineered oncolytic reovirus demonstrating anti-tumor activity through innate and adaptive immune responses. Previous data from the AWARE-1 study demonstrated that addition of atezolizumab (atezo) to pela increased the CelTIL score (study’s primary endpoint) by more than 30% in 60% of HR+/HER2- early breast cancer (BC) patients. CelTIL is a composite measure of tumor cellularity and tumor-infiltrating lymphocytes (TILs) and has been associated with a better prognosis in BC. The increase in CelTIL score observed in AWARE-1 was accompanied by PD-L1 upregulation. Here we report additional translational research results from AWARE-1.

Methods

HR+/HER2- early BC patients were enrolled in two cohorts: Cohort 1 (C1)– pela + letrozole (n=10); and Cohort 2 (C2)– pela + letrozole + atezo (n=10). Pela was administered on days 1, 2 and 8, 9, and atezo was given on day 3. Tumor biopsies collected pre-treatment and on days 3 and 21 were examined by immunohistochemistry (IHC), T cell receptor sequencing (TCR-seq), digital spatial profiling (DSP), and the PAM50 assay. Blood samples collected at these timepoints were evaluated by TCR-seq and flow cytometry.

Results

IHC showed a significant increase (p-value=0.04) in caspase 3 staining in all patients from day3 to day 21 that reached significance in C2. PAM50 analysis indicated a conversion of baseline disease from luminal B to luminal A in both cohorts, with 100% of patients converting to luminal A in C2. TCR-seq showed a decrease in pre- vs. post-treatment blood T cell diversity in both cohorts, which was significant (p-value=0.01) in C2. An association between decreased post-treatment T cell diversity and pre- vs. post-treatment increases in TILs was observed in both cohorts, reaching significance in C2 (p-value=0.01). DSP showed an increase in activated T cells but not exhausted T cells in both cohorts.

Conclusions

These data show that pela induces an inflamed tumor phenotype and demonstrate its synergy with atezo. Moreover, these data support pela’s immune-based mechanism of action and suggest that combining pela with atezo may improve outcomes in BC.

Clinical trial identification

EudraCT number: 2018-003345-42.

Legal entity responsible for the study

Oncolytics Biotech Inc.

Funding

Oncolytics Biotech Inc.

Disclosure

J. Gavila Gregori: Financial Interests, Personal, Other: AstraZeneca; Financial Interests, Personal, Other: Pfizer; Financial Interests, Personal, Other: Novartis; Financial Interests, Personal, Other: Roche. H. Loghmani: Financial Interests, Personal, Full or part-time Employment: Oncolytics Bioteh Inc. L.M. Manso Sanchez: Financial Interests, Personal, Advisory Board: AstraZeneca; Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Advisory Board: Novartis; Financial Interests, Personal, Advisory Board: Tesaro; Financial Interests, Personal, Advisory Board: Pfizer. T. Pascual: Financial Interests, Personal, Invited Speaker: Pfizer; Financial Interests, Personal, Invited Speaker: AstraZeneca; Financial Interests, Personal, Invited Speaker: Lilly. M. Juan: Financial Interests, Personal, Advisory Role: Grifols. A. Prat: Financial Interests, Personal, Invited Speaker: Roche; Financial Interests, Institutional, Other, Contracted research:: Roche; Financial Interests, Personal, Invited Speaker, Lecture fees: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Novartis; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Novartis; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Amgen; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Amgen; Financial Interests, Personal, Invited Speaker, Lecture fees: BMS; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: BMS; Financial Interests, Personal, Invited Speaker, Lecture fees: NanoString; Financial Interests, Institutional, Other, Contracted research: NanoString; Financial Interests, Institutional, Invited Speaker, Lecture fees: NanoString; Financial Interests, Institutional, Other, Contracted research: Novartis; Financial Interests, Institutional, Other, Contracted research: Roche; Financial Interests, Institutional, Other, Contracted research: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Contracted research; Financial Interests, Personal, Invited Speaker, Lecture fees: Daiichi Sankyo; Financial Interests, Institutional, Invited Speaker, Clinical trials: Daiichi Sankyo; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Puma; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Oncolytics; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: MSD; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Guardan Health; Financial Interests, Personal, Expert Testimony, Advisory role/consultancy: Peptomyc; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Lilly; Financial Interests, Institutional, Other, Clinical trials: Lilly; Financial Interests, Institutional, Other, Contracted research: Boehringer; Financial Interests, Institutional, Other, Contracted research: Sysmex Europa GmbH; Financial Interests, Institutional, Other, Contracted research: Medica Scientia inno. Research; Financial Interests, Institutional, Other, Contracted research: Celgene; Financial Interests, Institutional, Other, Contracted research: Astellas; Financial Interests, Institutional, Other, Clinical trials: Roche; Financial Interests, Institutional, Other, Clinical trials: AMgen; Financial Interests, Personal, Invited Speaker, Leadership role: Reveal Genomics, SL.; Financial Interests, Institutional, Other, Clinical trials: Boehringer; Non-Financial Interests, Institutional, Other, Leadership roles: Patronage committee: SOLTI Foundation; Non-Financial Interests, Institutional, Other, Leadership roles: Patronage committee: Actitud Frente al Cáncer Foundation. T.C. Heineman: Financial Interests, Personal, Officer, Chief Medical Officer: Oncolytics Biotech; Financial Interests, Personal, Stocks/Shares: Oncolytics Biotech. All other authors have declared no conflicts of interest.

Background

The RNA modification have been correlated with patient’s survival in various cancers. However, whether RNA modification genes are associated with molecular classification and potential application in breast cancer patients’ prognostication and prediction of disease-free survival (DFS) is largely unknown. Here we aim to develop and validate of a deep learning RNA modification model to predict DFS in patients with breast cancer, and further to analysis the association of the RNA modification model with immune microenvironment.

Methods

In this study, a total of 2,236 non-metastatic breast cancer patients were included. We built the DeepSurv in TensorFlow, a deep-learning survival neural network-based model on 2,149 non-metastatic breast cancer patients' data with 112 RNA modification genes in training cohort from The Cancer Genome Atlas (TCGA) and NCBI Gene Expression Omnibus (GEO). The algorithm was externally validated on an independent test cohort, comprising 87 non-metastatic breast cancer patients in Sun Yat-sen Memorial Hospital of Sun Yat-sen University. The primary end point was DFS.

Results

In the training cohort, we established a deep-learning survival neural network model showed promising results to predict DFS risk score in breast cancer patients; patients with low-risk vs high-risk score had statistically significantly longer DFS [hazard ratio (HR) = 14.88, 95% CI = 11.96–18.53, P < 0.005]. The RNA modification risk score was associated with DFS (AUCs for 2-, 3-, 5-, and 7-year survival were 0.92, 0.93, 0.93 and 0.94, respectively) superior molecular subtype, tumor, node, and metastasis staging system. Similarly, in the test cohort, patients with low-risk vs high-risk score had statistically significantly longer DFS (P < 0.005), the AUCs for 2- and 3-year survival were 0.89, and 0.89, respectively. The RNA modification model was validated across various subgroups. Further, this study identified that model obviously related to multiple tumor immune microenvironment characteristics.

Conclusions

This study developed and validated novel deep learning survival neural network model showed reliable individual DFS information in prognostic evaluation and treatment recommendation in patients with breast cancer.

Legal entity responsible for the study

Beijing Normal University-Hong Kong Baptist University United International College, Hong Kong Baptist University.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

Background

When triple-negative breast cancer (TNBC) patients have residual disease after neoadjuvant chemotherapy (NACT), they have a high risk of metastatic relapse. With immune infiltrate in TNBC being prognostic and predictive of response to treatment, our aim was to develop an immunologic transcriptomic signature using post NACT samples to predict relapse more accurately.

Methods

We identified 115 samples of residual tumors from post-NACT TNBC patients. We profiled the expression of 770 genes related to cancer microenvironment using the NanoString PanCancer IO360 panel to develop a prognostic transcriptomic signature, and to describe the immune microenvironments of the residual tumors.

Results

Thirty-eight (33%) patients experienced metastatic relapse. Hierarchical clustering separated patients into 5 clusters with distinct prognosis based on pathways linked to immune activation, epithelial to mesenchymal transition and cell cycle. Clusters enriched in immune activation had a significantly better prognosis. We selected eight immune-related genes (BLK, GZMM, CXCR6, LILRA1, SPIB, CCL4, CXCR4, SLAMF7) to develop a prognostic transcriptomic signature. We also developed an extended signature comprising 55 genes with a high correlation score (≥ 0.8) with at least one gene from the 8-gene signature. A high 8-gene signature score or extended signature score was associated with a significantly better distant-relapse free interval in our cohort (HR 0.18 [95%CI 0.09-0.35], P < 0.001 and HR 0.27 [95%CI 0.14-0.51], P < 0.001, respectively). Both signatures were tested on TNBC patients from METABRIC cohort (n = 251) and were associated with a better disease-specific survival (8-gene signature: HR 0.53 [95%CI 0.35-0.8], P < 0.01 and extended signature: HR 0.43 [95%CI 0.2- 0.64], P < 0.001). In our cohort, the 8-gene signature as well as the RCB score were validated as independent prognostic factors in multivariate analysis.

Conclusions

Lack of immune activation after NACT is associated with a high risk of distant relapse. We propose a prognostic signature based on immune infiltrate that could help select patients in need of adjuvant therapies.

Legal entity responsible for the study

Gaëtan MacGrogan.

Funding

Comité Prévention Dépistage des Cancers.

Disclosure

M. Arnedos: Financial Interests, Personal, Invited Speaker: Novartis; Financial Interests, Personal, Advisory Board: Puma, AstraZeneca, AbbVie; Financial Interests, Institutional, Other, Masterclass: Roche, Genomic Health; Financial Interests, Institutional, Other, Preceptorship: Novartis; Financial Interests, Institutional, Invited Speaker: Novartis, Puma. H. Bonnefoi: Financial Interests, Personal, Advisory Board: AstraZeneca; Financial Interests, Institutional, Invited Speaker: Bayer. All other authors have declared no conflicts of interest.

Background

CanAssist Breast (CAB) is an IHC based prognostic test that predicts risk of distant recurrence in hormone receptor (HR)-positive, HER2-negative early breast cancer in 5 years from diagnosis. CAB has been validated extensively on retrospective cohorts from India, USA and Europe showing its comparable performance across these divergent cohorts. The current study for the first time demonstrates the recurrence risk predictions up to 10 years in a sub-cohort of a prospective clinical trial, TEAM (Tamoxifen, Exemestane Adjuvant Multinational).

Methods

CAB has been assessed on 490 Dutch patient samples enrolled in the TEAM trial from 22 centers. TEAM is a large, international clinical trial that recruited 9766 post-menopausal women, randomized for the use of hormonal therapy, sequential (2-3 years Tamoxifen + 3-2 years exemestane) or exemestane alone (5 years) and patients were followed up for at least 10 years (median=10.4, range: 0.93-15.45). The team performing CAB were blinded to clinical outcomes. The recurrence risk predictions by CAB were compared with patient outcomes by the LUMC team. The performance of CAB was estimated with Kaplan-Meier survival analysis and hazard ratios (HR).

Results

Our study cohort had 67% patients with node-positive disease, 55% had tumors greater than 2cm (T2) and 28% had poorly differentiated tumors (G3). 78% of the cohort was treated with hormonal therapy alone, either exemestane alone or sequential therapy for a period of 5 years. CAB stratified 68% of the total cohort as low risk with a DRFI (distant relapse free interval) of 86% and 32% as high risk at 10 years. In the node-positive sub-cohort, the risk proportions were 63:37 with a low risk DMFI of 88% [HR-2.84 (1.66-4.88, p<0.0001)] at 10 years and 93% at 5 years slightly higher to DMFI of post-menopausal women with RS 0-25 who participated in RxPONDER trial. The 10 year low risk DMFI was 83% (HR: 1.78, 0.9-3.3, p=0.05) in sequential and 90% in exemestane arm (HR: 3.3, 1.7-6.5, p<0.0001). Only CAB high risk patients showed a chemotherapy benefit with an improved DMFI of 14%.

Conclusions

Data from a randomised trial show the usefulness of CAB for long term (10 year) recurrence risk predictions in early stage HR-positive, HER2-negative breast cancer.

Clinical trial identification

NCT00279448, NCT00032136, and NCT00036270; Netherlands Trial Registry NTR267.

Legal entity responsible for the study

Leiden University Medical Center.

Funding

Oncostem Diagnostics, Pfizer Oncology.

Disclosure

M. Bakre: Financial Interests, Personal and Institutional, Ownership Interest: OncoStem Diagnostics Pvt Ltd. A. Gunda, B.A. Savitha, C. Prakash, P. Shrivastava, T. Kaur, M. Siraganahalli Eshwaraiah: Financial Interests, Personal, Full or part-time Employment: OncoStem Diagnostics Pvt Ltd. P. Kuppen: Financial Interests, Institutional, Research Grant, from OncoStem Diagnostics Pvt Ltd.: Leiden University Medical Center. All other authors have declared no conflicts of interest.

Background

A possible association between changes in tumor variant allele fraction (VAF) from baseline to cycle 2/ day 1 (C2D1) using the 74-gene Guardant360 assay and progression free survival (PFS) was observed in 45 patients (pts) with ABC treated with a CDK4/6i + ET. Here, we tested the same methodology in an independent dataset and explored the association between overall survival (OS), intrinsic subtype (IS) and ctDNA dynamics.

Methods

Baseline and C2D1 plasma samples were obtained from 113 pts with HR+/HER2-negative ABC treated with CDK4/6i + ET in an international, multicentric observational study. The first 45 pts were used to derive the mean VAF ratio (mVAFR: mean of the log VAFR for all mutations) methodology associated with PFS (NPJ Breast Cancer 2021). mVAFR was calculated for each alteration with a VAF ≥0.4% at any time point. Pts with VAFs <0.4% at all timepoints were considered to have ctDNA-low disease. Molecular response was defined by the mVAFR. The main objective was to assess the association of mVAFR with PFS in the 68-pt independent validation cohort. We also explored the association of mVAFR with OS (n=113) and with IS (n=54). PFS and OS uni- and multivariable cox models were performed adjusting for clinical variables.

Results

With a median follow up (mFUP) of 16.6 months (m), pts with medium or high mVAFR had shorter median PFS than pts with low mVAFR and ctDNA-low disease (11.2 vs 25.0m; adjusted hazard ratio [aHR]=2.85, 95% confidence interval [CI] 1.2-6.7, p=0.017). mVAFR as a continuous variable was also found associated with PFS (HR=2.0, 95% CI 1.0–4.1, p=0.047). In the 113-pt combined cohort, with a mFUP of 19.4m, pts with medium or high mVAFR had shorter OS than pts with either low mVAFR or ctDNA-low disease (31.3m vs not reached; aHR=10.9, 95% CI 3.7-31.6, p<0.001). mVAFR as a continuous variable was also found associated with OS (aHR=4.1, 95% CI 1.7-9.5, p<0.001). Finally, mVAFR was associated with IS (p=0.022).

Conclusions

Early ctDNA dynamics after CDK4/6i + ET can identify different pt populations. The absence of molecular response identifies pts with poor outcome, and future studies should focus on improving treatment options for this population.

Legal entity responsible for the study

The authors.

Funding

AstraZeneca.

Disclosure

O. Martinez Saez: Financial Interests, Personal, Invited Speaker: Novartis; Financial Interests, Personal, Invited Speaker: Eisai; Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Other, Travel expenses: Roche; Financial Interests, Personal, Other, Medical advisory: Reveal Genomics. F. Brasó-Maristany: Financial Interests, Personal, Invited Speaker: Reveal Genomics. T. Pascual: Financial Interests, Personal, Invited Speaker: Pfizer; Financial Interests, Personal, Invited Speaker: AstraZeneca; Financial Interests, Personal, Invited Speaker: Lilly. M. Munoz: Financial Interests, Personal, Expert Testimony: Roche; Financial Interests, Personal, Expert Testimony: Novartis; Financial Interests, Personal, Other, International conference travel grants: Roche; Financial Interests, Personal, Other, International conference travel grants: Pfizer; Financial Interests, Personal, Other, International conference travel grants: Lilly; Financial Interests, Personal, Expert Testimony: Eisai; Financial Interests, Personal, Advisory Board: Pierre Fabre. I. Faull: Financial Interests, Member of the Board of Directors: Guardant Health. J. Odegaard: Financial Interests, Personal, Member of the Board of Directors: Guardant Health. G. Patel: Financial Interests, Personal, Member of the Board of Directors: AstraZeneca. R. McEwen: Financial Interests, Personal, Full or part-time Employment: AstraZeneca. D. Carroll: Financial Interests, Personal, Full or part-time Employment: AstraZeneca. E.M. Ciruelos: Financial Interests, Personal, Other, Speakers Bureau. Educational activities: Roche; Financial Interests, Personal, Invited Speaker, Symposia and Educational activities: Roche; Financial Interests, Personal, Advisory Board, Non-permanent advisor: Roche; Financial Interests, Personal, Other, Speakers Bureau. Educational activities: Lilly; Financial Interests, Personal, Invited Speaker, Symposia and Education: Lilly; Financial Interests, Personal, Advisory Board, Non-permanent advisor: Lilly; Financial Interests, Personal, Advisory Board, Non-permanent advisor: Novartis; Financial Interests, Personal, Invited Speaker, Educational activities: Pfizer; Financial Interests, Personal, Advisory Board, Non-permanent advisor: Pfizer; Financial Interests, Personal, Advisory Board, Non-permanent advisor: AstraZeneca; Financial Interests, Personal, Advisory Board, Non-permanent advisor: Daiichi Sankyo; Financial Interests, Personal, Advisory Board, Non-permanent advisor: MSD; Financial Interests, Institutional, Funding, PI for Patricia 2 trial (sponsor: SOLTI Group): Pfizer; Financial Interests, Institutional, Funding, PI for Prometeo 2 trial (sponsor: SOLTI Group): Pfizer; Financial Interests, Institutional, Funding, PI for TATEN trial (sponsor: SOLTI Group): MSD; Financial Interests, Institutional, Funding, PI for NEREA trial (sponsor: SOLTI Group): Puma; Financial Interests, Institutional, Funding, PI for ATREZZO trial (sponsor: SOLTI Group): Roche; Non-Financial Interests, Invited Speaker, Non-profit organization dedicated to breast cancer research: SOLTI Cooperative Group; Non-Financial Interests, Advisory Role, Scientific Evaluator at ISCIII (Spanish Government Academic Research Platform): Instituto de Salud Carlos III. A. Prat: Financial Interests, Personal, Invited Speaker: Roche; Financial Interests, Institutional, Other, Contracted research:: Roche; Financial Interests, Personal, Invited Speaker, Lecture fees: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Novartis; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Novartis; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Amgen; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Amgen; Financial Interests, Personal, Invited Speaker, Lecture fees: BMS; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: BMS; Financial Interests, Personal, Invited Speaker, Lecture fees: NanoString; Financial Interests, Institutional, Other, Contracted research: NanoString; Financial Interests, Institutional, Invited Speaker, Lecture fees: NanoString; Financial Interests, Institutional, Other, Contracted research: Novartis; Financial Interests, Institutional, Other, Contracted research: Roche; Financial Interests, Institutional, Other, Contracted research: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Contracted research; Financial Interests, Personal, Invited Speaker, Lecture fees: Daiichi Sankyo; Financial Interests, Institutional, Invited Speaker, Clinical trials: Daiichi Sankyo; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Puma; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Oncolytics; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: MSD; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Guardan Health; Financial Interests, Personal, Expert Testimony, Advisory role/consultancy: Peptomyc; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Lilly; Financial Interests, Institutional, Other, Clinical trials: Lilly; Financial Interests, Institutional, Other, Contracted research: Boehringer; Financial Interests, Institutional, Other, Contracted research: Sysmex Europa GmbH; Financial Interests, Institutional, Other, Contracted research: Medica Scientia inno. Research; Financial Interests, Institutional, Other, Contracted research: Celgene; Financial Interests, Institutional, Other, Contracted research: Astellas; Financial Interests, Institutional, Other, Clinical trials: Roche; Financial Interests, Institutional, Other, Clinical trials: Amgen; Financial Interests, Personal, Invited Speaker, Leadership role: Reveal Genomics, SL.; Financial Interests, Institutional, Other, Clinical trials: Boehringer; Non-Financial Interests, Institutional, Other, Leadership roles: Patronage committee: SOLTI Foundation; Non-Financial Interests, Institutional, Other, Leadership roles: Patronage committee: Actitud Frente al Cáncer Foundation. All other authors have declared no conflicts of interest.

Background

The aim of this study was to evaluate real-world use of ODX RS in early breast cancer (BC) pN₀/N₁ patients and its impact on adjuvant CT decisions.

Methods

LISE is an ongoing observational study comprehensively evaluating BC characteristics, management practices and outcomes in 3,122 patients diagnosed between January 2010 and December 2021 at Courlancy Cancer Institute (Reims, France).

Results

Among 1,160 eligible HR+/HER2- patients from an interim analysis (N=1,626), ODX testing was performed in 231 patients (19.9%). Pathological tumor size distribution was as follows: pT_1a 0.9%, pT_1b 12.8%, pT_1c 58.0%, pT₂ 27.9%, pT_3-4 0.5%. 78.4% of cases were pN₀, 8.6% were pN_1mic and 13.0% pN₁. Scarff-Bloom-Richardson (SBR) grading was as follows: SBR I 16.0%, SBR II 68.5%, SBR III 15.5%. Luminal A and Luminal B HER2- subtypes accounted for 31.1% and 68.8% of cases, respectively. Multivariate analysis revealed highest ODX use in pT_1c or pT₂ tumors, with Ki-67 ≥20% and SBR I-II grading. RS distribution and its impact on adjuvant CT decisions are reported in the table. Table: 11P

Conclusions

ODX testing demonstrated major changes in therapeutic choices. In pN₀ and pN_1mic-N₁ patients, CT indication was decreased by 69.4% and 75.4%, respectively. Our real-world clinical practice results describe the ‘grey area’ population requiring ODX prescription and confirm the utility of ODX to guide management and avoid overtreatment.

Legal entity responsible for the study

Bruno Cutuli, Institut du Cancer Courlancy, Reims, France.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

Background

RAD51C and RAD51D are two paralogs of RAD51, essential for the repair of DNA breaks by homologous recombination (HRR). Germline pathogenic variants (PV) in these genes have been associated with HRR deficiency (HRD) and antitumor response to DNA damaging agents, including PARP inhibitors. The evaluation of RAD51 nuclear foci provides a functional measure of HRD, whereas genomic HRD captures accumulated tumor genomic instability. Biallelic inactivation of RAD51C has been associated with genomic HRD. We aimed to evaluate functional and genomic HRD in patients with primary breast and ovarian cancer (BC/OC) and germline PV in RAD51C/D (gRAD51C/D) to help understand the impact of these alterations in HRR status and provide evidence for therapeutic decision-making.

Methods

51 primary and untreated FFPE tumor samples were obtained from gRAD51C/D high-grade OC (n=26), ER- BC (n=14) and ER+ BC (n=11) patients, included in the Spanish Hereditary Cancer-SEOM registry. An immunofluorescence-based assay was used to evaluate RAD51 and functional HRD was defined as RAD51 score ≤10%. The genomic instability score (GIS), tumor HRR-gene mutation calling and gene-specific LOH (gsLOH) status was obtained with the Myriad myChoice CDx assay and analysis via the Myriad review algorithm.

Results

A successful RAD51 score was obtained in 40/51 (78%) and GIS in 27/29 (93%). The prevalence of HRD by the RAD51 test in gRAD51C was 12/30 (40%) and in gRAD51D was 8/10 (80%), compared to >90% in gBRCA1/2 and gPALB2 BC. Genomic HRD was 11/18 (61%) in gRAD51C samples and 8/9 (89%) in gRAD51D tumors. RAD51 scores and GIS were concordant in 19/21 (91%) cases. Of these, all HRD tumors by the RAD51 test (12/21) were also HRD by GIS and presented gsLOH. HRR proficiency by RAD51 (9/21) was consistent with low GIS, except for two OC cases with high GIS, gRAD51C mutation and gsLOH. Interestingly, all ER+ BC cases (n=5) were HRR proficient by RAD51 and GIS, and lacked gsLOH.

Conclusions

RAD51 scores and GIS are highly concordant in gRAD51C/D BC/OC and reveal a lower prevalence of HRD than expected, primarily in gRAD51C tumors. HRP was predominant in gRAD51C ER+ BC, which did not exhibit biallelic inactivation.

Legal entity responsible for the study

The authors.

Funding

Asociación Española Contra el Cáncer (AECC) LaCaixa Foundation (CaixaImpulse grant) Generalitat de Catalunya (AGAUR-Producte and PERIS) Instituto de Salud Carlos III (ISCIII) Fondo Europeo de Desarrollo Regional (FEDER).

Disclosure

A. Llop-Guevara: Other, Personal and Institutional, Proprietary Information, Patent WO2019122411A1: Methods based on the detection of RAD51 foci in tumor cells: None. M. Romey: Financial Interests, Institutional, Funding: Myriad Genetics. P.G. Nuciforo: Financial Interests, Institutional, Advisory Board: Bayer; Financial Interests, Institutional, Advisory Board: MSD Oncology; Financial Interests, Institutional, Invited Speaker: Novartis; Other, Institutional, Other, Consultant: Targos Molecular Pathology GmbH. C. Denkert: Financial Interests, Personal, Advisory Board: MSD Oncology; Financial Interests, Personal, Advisory Board: Daiichi Sankyo; Financial Interests, Personal, Advisory Board: Molecular Health; Financial Interests, Personal, Invited Speaker: AstraZeneca; Financial Interests, Personal, Invited Speaker: Merck; Financial Interests, Personal, Ownership Interest, Cofounder and shareholder of Sividon Diagnostics until 2016: Sividon Diagnostic; Financial Interests, Personal, Invited Speaker: VmScope digital pathology software; Financial Interests, Institutional, Funding: Roche; Financial Interests, Institutional, Research Grant: Myriad. V. Serra Elizalde: Financial Interests, Institutional, Invited Speaker: AstraZeneca; Financial Interests, Personal, Other, WO2019122411A1: Methods based on the detection of rad51 foci in tumor cells: TBD; Financial Interests, Institutional, Research Grant, Testing various novel targeted agents in patient-derived tumour models: AstraZeneca. J. Balmaña: Financial Interests, Personal, Advisory Board: AstraZeneca; Financial Interests, Personal, Advisory Board: Pfizer; Financial Interests, Institutional, Other, Steering Committee Member: AstraZeneca; Financial Interests, Institutional, Principal Investigator, Local PI in clinical trials: MedSir; Financial Interests, Institutional, Principal Investigator, Local PI in clinical trials: Pfizer; Other, Personal, Proprietary Information, Patent WO2019122411A1: Methods based on the detection of RAD51 foci in tumor cells: None. All other authors have declared no conflicts of interest.

Background

The effect of tertiary lymphoid structures (TLSs) in patients with breast cancer (BC) remains controversial. The objective of current study is to investigate the clinicopathological and prognostic significance of TLSs in BC. Given unique difficulties for detecting and quantifying TLSs, a TLS-associated gene signature based on The Cancer Genome Atlas (TCGA) BC cohort was used to validate and supplement our results. Our study was registered in the PROSPERO (registration number: CRD42022302921).

Methods

Ectronic platforms (PubMed, Web of Science, EMBASE, the Cochrane Library, NKI and Wangfang) were searched systematically to identify relevant researches as of January 11, 2022. We calculated combined odds ratios (ORs) with 95% confidence intervals (CIs) to determine the relationship between clinicopathological parameters and TLSs. The pooled hazard ratios (HRs) and 95% CIs were also calculated to evaluate the prognostic significance of TLSs. The TLS signature based on TCGA BC cohort was applied to validate and supplement our results.

Results

15 studies with 3898 patients were eligible for enrollment in our study. The combined analysis indicated that the presence of TLSs was related to improved disease-free survival (DFS) (HR= 0.61, 95% CI: 0.41–0.90, p< 0.05) and overall survival (OS) (HR= 1.66, 95% CI: 1.26-2.20, p < 0.001). Additionally, the presence of TLSs was positively correlated with early tumor TNM stage and high tumor-infiltrating lymphocytes. TLS presence was positively related to human epidermal growth factor receptor 2 (HER-2) and Ki-67 but inversely correlated with the status of estrogen and progesterone receptor. Simultaneously, our study found that tumor immune microenvironment was more favorable in the high TLS signature group than in the low TLS signature group. Consistently, BC patients in the high TLS signature group exhibited better survival outcomes compared to those in the low TLS signature group, suggesting TLSs were favorable prognostic biomarkers.

Conclusions

TLS presence was strongly related to clinicopathological features and prognosis in patients with BC, which has the potential to act as an effective biomarker for the clinical assessments and predicting prognosis of BC.

Legal entity responsible for the study

The authors.

Funding

The National Natural Science Foundation of China.

Disclosure

All authors have declared no conflicts of interest.

Background

Changes occurring in host-associated microbial communities (i.e. microbiota) may modulate responses to checkpoint blockade immunotherapy. We previously showed that anti-programmed cell death 1 protein P added to microtubule-targeting chemotherapy E has an encouraging antitumor activity in HR+ MBC pts regardless of programmed death-ligand 1 status. The CALADRIO study assessed the impact of gut and oral microbiota on clinical outcome of pts from the KELLY trial.

Methods

The phase 2 KELLY trial investigated the efficacy and safety of P plus E in 44 pts with pre-treated, HR+, HER2-negative, locally recurrent inoperable or MBC (NCT03222856). Fecal and saliva samples were prospectively collected at baseline (BL), after 3 cycles, and end of treatment from a subset of pts. Tumor response was grouped into clinical benefit (CB; complete or partial responses, or stable disease [SD] ≥24 weeks) and no CB (SD <24 weeks or progressive disease) as per RECIST 1.1. Shotgun metagenomic and 16S rRNA gene sequencing were used to characterize fecal and saliva microbiota profiles, respectively. Microbiota data were analyzed using MEGAHIT, LEfSe, Wilcoxon ranked sum, and PERMANOVA methods.

Results

A total of 58 fecal and 68 saliva samples were collected. Overall P+E did not cause significant gut or oral microbiota perturbations, indicating any drug-related microbial toxicity. Across all pts, dominant gut microbiota genera included Bacteroides and Faecalibacterium, with a common oral microbe, Prevotella, also present. LeFSe analysis suggested CB was driven in part by gut-associated Bacteroides fragilis and oral-associated Streptococcus with an abundance ≥50%. Pts experiencing CB had gut and oral microbiota richness at BL and a decrease over treatment potentially related to the antibiotic usage. Several typical oral microbes in both saliva and fecal samples were also observed, suggesting a potential translocation along the oral-gut axis.

Conclusions

These preliminary findings suggest potential avenues for downstream microbiota pts stratification before commencement of treatment. Further investigation is required in larger cohorts.

Clinical trial identification

NCT03222856.

Legal entity responsible for the study

MEDSIR.

Funding

MEDSIR.

Disclosure

M. Gion Cortes: Financial Interests, Personal, Sponsor/Funding: Roche; Financial Interests, Personal, Sponsor/Funding: Pfizer. J.M. Perez Garcia: Non-Financial Interests, Personal, Advisory Role: Roche; Non-Financial Interests, Personal, Advisory Role: Lilly; Non-Financial Interests, Personal, Advisory Role: Daiichi Sankyo; Financial Interests, Personal, Sponsor/Funding: Roche. A. Prat: Non-Financial Interests, Personal, Full or part-time Employment: Novoartis; Financial Interests, Personal, Stocks/Shares: Reveal Genomics; Financial Interests, Personal, Sponsor/Funding: Pfizer, Novartis, Roche, MSD Oncology, Lilly, Daiichi Sankyo, Amgen, Guardant Health; Non-Financial Interests, Personal, Advisory Role: Amgen, Roche, Novartis, Pfizer, Bristol Myers Squibb, Boehringer, Puma, Oncolytics Biotech, Daiichi Sankyo, AbbVie, AstraZeneca, NanoString Technologies (to the Institution); Financial Interests, Institutional, Funding: Roche, Novartis, Incyte, Puma Biotechnology; Financial Interests, Personal, Ownership Interest: Patents:- PCT/EP2016/080056, - WO/2018/096191, US 63/023785, HER2DX (filing); Financial Interests, Personal, Sponsor/Funding: Daiichi Sankyo; Other, Personal, Other: Oncolytics, Peptomyc S.L. A. Llombart Cussac: Non-Financial Interests, Personal, Leadership Role: Eisai, Celgene, Lilly, Pfizer, Roche, Novartis, MSD;; Financial Interests, Personal, Stocks/Shares: MEDSIR, Initia-Research; Non-Financial Interests, Institutional, Advisory Role: Lilly, Roche, Pfizer, Novartis, Pierre Fabre, GenomicHealth, GSK; Non-Financial Interests, Personal, Speaker’s Bureau: Lilly, AstraZeneca, MSD; Financial Interests, Personal, Funding: Roche, Foundation Medicine, Pierre Fabre, Agendia; Financial Interests, Personal, Sponsor/Funding: Roche, Lilly, Novartis, Pfizer, AstraZeneca. M. Mancino: Financial Interests, Personal and Institutional, Full or part-time Employment: MEDSIR. J. Cortés: Non-Financial Interests, Personal, Advisory Role: Roche, Celgene, Cellestia, AstraZeneca, Biothera Pharmaceutical, Merus, Seattle Genetics, Daiichi Sankyo, Erytech, Athenex, Polyphor, Lilly, Servier, Merck Sharp&Dohme, GSK, Leuko, Bioasis, Clovis Oncology.; Financial Interests, Personal, Sponsor/Funding: Roche, Novartis, Celgene, Eisai, Pfizer, Samsung Bioepis, Lilly, Merck Sharp&Dohme, Daiichi Sankyo.; Financial Interests, Personal and Institutional, Funding: Roche, Ariad pharmaceuticals, AstraZeneca, Baxalta GMBH/Servier Affaires, Bayer healthcare, Eisai, F.Hoffman-La Roche, Guardanth health, Merck Sharp&Dohme, Pfizer, Piqur Therapeutics, Puma C, Queen Mary University of London.; Financial Interests, Personal, Stocks/Shares: MEDSIR. A. Malfettone: Financial Interests, Personal and Institutional, Full or part-time Employment: MEDSIR. All other authors have declared no conflicts of interest.

Background

Breast cancer (BC) constitutes the most common female malignancy in the world and is the main cause of cancer-associated mortality. Personalized classification of histopathological subtypes is crucial to estimate a patient’s prognosis and administer appropriate treatment strategies. To date, the immunohistochemistry-based (IHC) classification scheme in use is still too broad therefore individuals of the same IHC tumor subtype may not benefit from the same treatment regimen. Indeed, characterizing the molecular profiles unique to each of the BC subtypes allows treatments to be tailored to these profiles in a more specific way than using a standalone IHC status.

Methods

To address this problem we have developed the SubType Classifier that combines whole genome sequencing (WGS) data and machine learning algorithms supporting molecular-based breast cancer subtype classification. For this purpose, we used WGS data of 1039 BC patients derived from 3 different databases (ICGC, TCGA, HMF), further divided into training (n=802) and hold-out (n=237) datasets. Training cohort has been labeled by the IHC-based subtype estimation, where HER2 status was corrected by the ERBB2 copy number variations (Wojtaszewska M. et al, 2021). The hold-out set was composed of samples with estrogen and/or progesterone receptors and HER2 status confirmed by IHC results.

Results

The CatBoost Classifier achieved the best performance on ∼1000 copy number variation (CNV)-based genomic features selected as the biologically most relevant. Validation on the hold-out dataset has provided >0.9 discriminative power by area under receiver operating characteristic (AUROC) curve analysis. Within the prediction of three TNBC, ER+HER2- and ER+HER2+ subtypes, the precision ranged from 0.69 to 0.90 and recall from 0.67 to 0.93. However, the classification of ER-HER2+ class remains a challenge due to the high power of ERBB2-correlated genes and high similarity to other ER+HER2+ class (0.3 precision and ∼0.2, recall).

Conclusions

Results obtained by the SubType Classifier confirmed that the use of comprehensive genomic data supports the IHC-based canonical breast cancer subtype classification, providing deeper insights about tumor biology.

Legal entity responsible for the study

MNM Diagnostics Sp. z o.o.

Funding

Has not received any funding.

Disclosure

M. Piernik, A. Wozna: Financial Interests, Personal, Stocks/Shares: MNM Diagnostics Sp. z o.o. P. Zawadzki: Financial Interests, Personal and Institutional, Ownership Interest: MNM Diagnostics Sp. z o.o. All other authors have declared no conflicts of interest.

Background

Early prediction of treatment response is crucial for the optimal treatment of advanced breast cancer. We aimed to explore whether monitoring early changes in plasma human epidermal growth factor receptor 2 (HER2) levels using digital PCR (dPCR) could predict the treatment response in advanced breast cancer.

Methods

This was a multicenter, prospective, noninterventional clinical study of patients with advanced breast cancer. All enrolled patients underwent blood testing to measure the HER2 levels by digital PCR before treatment initiation and once every 3 weeks during the study. The primary endpoints were (1) the diagnostic value of dPCR for detecting HER2 status in the blood and (2) the relevance of potential changes in the plasma HER2 level at 3 weeks from baseline for predicting treatment response.

Results

Overall, 85 patients were enrolled between October 9^th, 2018, and January 23^rd, 2020. dPCR had a specificity of 91.67% (95% CI: 80.61% to 97.43%) for detecting HER2 amplification, and the area under the receiver operating characteristic (ROC) curve was 0.84 (P<0.0001). A clinically relevant specificity threshold of approximately 90%, which was equivalent to a ≥15% decrease in the plasma HER2 ratio at 3 weeks from baseline, showed a positive predictive value of 97.37% (95% CI: 77.11% to 98.65%) in terms of predicting clinical benefit. Patients whose plasma HER2 ratio was reduced by ≥15% had a longer median progression-free survival (PFS) than those whose ratio was reduced by <15% (9.20 months vs. 4.50 months, P=0.0008).

Conclusions

Early changes in the plasma HER2 ratio may predict the treatment response in patients with advanced breast cancer and could facilitate optimal treatment selection.

Clinical trial identification

NCT03947736.

Legal entity responsible for the study

The authors.

Funding

This study was supported by the National Natural Science Foundation of China (81672634, 82172650), Beijing Medical Award Foundation (YXJL-2020-0941-0763).

Disclosure

All authors have declared no conflicts of interest.

Background

Clinically, breast cancer (BC) is divided into subgroups based on the assessment of estrogen receptor (ER), progesterone receptor, and human epidermal growth factor receptor 2 (HER2) status. HER2-/ER+ tumors constitute the luminal subgroup. By using the gene expression profiling assay PAM50, luminal tumors can be stratified into the intrinsic molecular subtypes Luminal A (LumA), Luminal B (LumB) and HER2-enriched (LumHER2). The study at hand aims to characterize the LumHER2 subtype based on clinicopathological and transcriptomic features to elucidate what cases constitute the subgroup and to identify drivers of disease that may be used to improve treatment decisions.

Methods

The analyses were based on mRNA-sequencing and clinical review data from 6660 cases of the Sweden Cancerome Analysis Network - Breast study, of which 4413 were clinically HER2-/ER+ (LumHER2: 79, LumA: 2856, LumB: 1249). Approaches included comparative statistics of clinicopathological variables, survival analyses based on recurrence-free intervals as well as differential gene expression, metagene, and single-gene expression analyses.

Results

The LumHER2 subtype is associated with larger and higher graded tumors than the LumA subtype. In patients that received endocrine therapy (E) or both endocrine and chemotherapy (EC), LumHER2 tumors were associated with a higher risk of relapse than LumA and LumB tumors, independent of age, tumor size, and lymph node status (E: LumA - HR 0.31, 95% CI 0.13-0.79; LumB - HR 0.41, 95% CI 0.17-0.98; EC: LumA - HR 0.39, 95% CI 0.16-0.99; LumB - HR 0.37, 95% CI 0.15-0.90). Metagene patterns indicated LumHER2 tumors to have a higher immune response and lower steroid response than the other luminal subtypes with similar proliferative characteristics to LumB tumors. ESR1 was less expressed in LumHER2 tumors, while ERBB2 expression was similar between all subtypes.

Conclusions

LumHER2 breast cancer is associated with a faster disease recurrence than other luminal subtypes, regardless of received treatment. The subgroup is not predominantly constituted by clinically misclassified cases and less ER-dependent than other luminal subtypes, providing a possible explanation for the comparatively poor response to endocrine therapy.

Legal entity responsible for the study

Lund University.

Funding

Breast and Lung Cancer Research Group, Faculty of Medicine, Department of Clinical Sciences, Lund University.

Disclosure

All authors have declared no conflicts of interest.

Background

The late occurrence of isolated local recurrence (LR) of luminal BC calls into question whether these are true LR or new primaries. We evaluated the concordance of the mutation profiles between matched primary tumors (PT) and LR.

Methods

We collected data from women with LR of luminal BC treated at Gustave Roussy and Trieste University Hospital between 1992 and 2018. Following a quality control, targeted NGS was performed on the IonTorrent S5 NGS platform with the Oncomine Tumor Mutation Load Assay covering 409 critical oncogenes or tumor suppressor genes to analyze PT and LR samples. For the bioinformatic analysis, we retained the matched samples that passed quality check and had a total mean depth coverage ≥ 300X. Variants were filtered on their allele frequency ≥ 5% and a read count ≥ 5X, and on the basis of their genomic effect and annotation.

Results

Among the 131 eligible patients, 82 matched PT and LR were sequenced and analyzed. The table summarizes patient, PT and LR characteristics. Median time until LR was 6.0 years. After a median follow-up of 4.6 years following LR, median DDFS and OS were 8.1 and 8.7 years, respectively. In the PT, the main genes with more than 20% of variants were KMT2D, ATRX, MTOR, ATM and KMT2A. In the LR, the main genes with more than 10% of variants were KMT2D, NOTCH2, PIK3CA, and KMT2A. The main gene with recurrent mutation present in both PT and LR was PIK3CA (4.8%). This analysis showed that 17 cases (20%) shared the same variants between PT and LR. Table: 18P

Patient and tumor characteristics

Conclusions

Although we found a wide range of molecular alterations in LR, a minority of patients had concordant mutational profiles between PT and LR. These findings, if confirmed in larger studies, may better guide the choice of therapy after LR excision.

Legal entity responsible for the study

The authors.

Funding

Has not received any funding.

Disclosure

M. Lacroix-Triki: Financial Interests, Advisory Board: Myriad genetics; Financial Interests, Advisory Board: Exact sciences; Financial Interests, Advisory Board: Rroche diagnostics; Financial Interests, Advisory Board: AstraZeneca; Financial Interests, Advisory Board: Daiichi Sankyo; Financial Interests, Advisory Board: MSD; Financial Interests, Advisory Board: Roche. E. Rouleau: Financial Interests, Invited Speaker: BMS; Financial Interests, Invited Speaker: AstraZeneca; Financial Interests, Invited Speaker: Roche; Financial Interests, Invited Speaker: Clovis; Financial Interests, Invited Speaker: GSK. L. Lacroix: Financial Interests, Advisory Role: Adept Field Solutions; Financial Interests, Advisory Role: AstraZeneca; Financial Interests, Advisory Role: Bayer; Financial Interests, Advisory Role: Boehringer; Financial Interests, Advisory Role: BMS; Financial Interests, Advisory Role: Lilly; Financial Interests, Advisory Role: Icomed; Financial Interests, Advisory Role: Genomic Health; Financial Interests, Advisory Role: Medimmune; Financial Interests, Advisory Role: Novartis; Financial Interests, Advisory Role: Pfizer; Financial Interests, Advisory Role: QualWorld; Financial Interests, Advisory Role: Taiho Oncology; Financial Interests, Advisory Role: Roche; Financial Interests, Advisory Role: Thermofisher; Financial Interests, Advisory Role: VelaDx; Financial Interests, Sponsor/Funding: Abbott; Financial Interests, Sponsor/Funding: Amgen; Financial Interests, Sponsor/Funding: AstraZeneca; Financial Interests, Sponsor/Funding: Beckman Coulter; Financial Interests, Sponsor/Funding: Bayer; Financial Interests, Sponsor/Funding: Boeringer; Financial Interests, Sponsor/Funding: BMS; Financial Interests, Sponsor/Funding: Illumina; Financial Interests, Sponsor/Funding: Genomic Health; Financial Interests, Sponsor/Funding: Guardant health; Financial Interests, Sponsor/Funding: Lilly; Financial Interests, Sponsor/Funding: Medimmune; Financial Interests, Sponsor/Funding: Myriad; Financial Interests, Sponsor/Funding: Novartis; Financial Interests, Sponsor/Funding: Pfizer; Financial Interests, Sponsor/Funding: Roche; Financial Interests, Sponsor/Funding: Siemens Healthineer; Financial Interests, Sponsor/Funding: Taiho Oncology; Financial Interests, Sponsor/Funding: Thermofisher; Financial Interests, Sponsor/Funding: VelaDx. F. André: Financial Interests, Institutional, Research Grant: AstraZeneca; Financial Interests, Institutional, Research Grant: Lilly; Financial Interests, Institutional, Research Grant: Novartis; Financial Interests, Institutional, Research Grant: Pfizer; Financial Interests, Institutional, Research Grant: Roche; Financial Interests, Institutional, Research Grant: Daiichi; Other, Founder: Pegacsy. B. Pistilli: Financial Interests, Institutional, Advisory Role: AstraZeneca; Financial Interests, Personal, Advisory Role: Myriad; Financial Interests, Personal, Advisory Role: Pierre Fabre; Financial Interests, Institutional, Advisory Role: Pfizer; Financial Interests, Institutional, Invited Speaker: Daiichi Sankyo; Financial Interests, Institutional, Invited Speaker: Novartis; Financial Interests, Institutional, Invited Speaker: Puma; Financial Interests, Personal, Sponsor/Funding: AstraZeneca; Financial Interests, Personal, Sponsor/Funding: Pierre Fabre; Financial Interests, Personal, Sponsor/Funding: MSD; Financial Interests, Institutional, Advisory Board: Novartis; Financial Interests, Institutional, Advisory Board: AstraZeneca; Financial Interests, Institutional, Advisory Board: Daiichi Sankyo. All other authors have declared no conflicts of interest.

Background

Breast cancer is one of the leading causes of death worldwide. Tumor vascularity and immune disturbance are one of the hallmarks of cancer and related with patients’ survival. Here we report how tumor vascularity assessed by Hounsfield Units(HU) on patients’ computed tomography(CT) and hosts’ immunologic environment represented by serum neutrophil to lymphocyte ratio(NLR) played and affected patients’ survival.

Methods

Materials and methods: Female patients with breast cancer who got preoperative treatment from 2008 to 2018 at Wonju Severance Hospital, Wonju, Korea, were included. Clinicopathological data were collected. Maximum HU of tumor and aorta on CT and their ratio(TAR) were calculated. Peripheral complete blood cell counts were collected and NLR were calculated. Patients were categorized according to the cutoff values retrieved from the receiver operating characteristic curve and Kaplan-Meier curves were generated to compare overall and distant disease free survival.

Results

The final cohort included 740 patients with mean age, 54.3±11.3 (22-89) years old. TAR was 0.347±0.108(range, 0.062-1.114) and NLR was 2.29 ±1.53 (range, 0.61 – 10.47). Cut-off values for TAR and NLR were 0.27 and 1.61, respectively. Both high TAR and NLR groups showed significantly worse recurrence free-interval (p<0.001, p=0.002 respectively), distant recurrence free-interval (p<0.001, p=0.034, respectively) and overall survival(p=0.002, p=0.007 respectively). Further analysis was done on TAR and NLR combined groups and only patients with high TAR and NLR showed worse prognosis than others. Any group with just one worse factor, either TAR or NLR, did not show worse prognosis than patients with higher value in TAR and NLR(p<0.001).

Conclusions

Increased HU on CT representing tumor aspect and NLR reflecting immune status of hosts influenced patients survival with significance. This impact was magnified when patients had increased value of both factors, suggesting a close relationship between tumor vascularity and patients’ immune cells.

Legal entity responsible for the study

The authors.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

Background

Immune checkpoint inhibition (ICI) is an established treatment in PDL1-positive metastatic triple-negative (TN) breast cancer (BC). However, the immune landscape of breast cancer brain metastasis (BCBM) remains poorly defined and BCBM still represents an area of unmet clinical need.

Methods

The tumour infiltrating lymphocytes (TILs) and the mRNA levels of 770 immune-related genes (NanoString nCounter Immuno-oncology IO360 panel) were assessed in primary BCs and BCBMs. The prognostic role of ARG2 transcript and protein expression (immunohistochemistry) and its association with outcome (Kaplan-Meier Log-rank survival analysis) and T-cell depletion (Wilcoxon signed-rank t-test) was determined.

Results

A significant reduction of TILs was identified in the BCBMs in comparison to primary BCs. 11.5% of primary BCs had a high immune-infiltrate (hot), 46.2% were altered (immunosuppressed/excluded) and 34.6% were cold (no/low immune infiltrate), whereas only 3.8% of BCBMs were hot, 23.1% altered and 73.1% cold. Compared to the primary BCs, we identified 112 downregulated genes in BCBM [false-discovery rate (FDR)<0.01, log2 fold-change (FC)>1.5], involved in matrix remodelling-metastasis, cytokine-chemokine signalling, lymphoid compartment, antigen presentation and immune cell adhesion-migration. Four genes, ARG2, SOX2, EGF, NCAM1, were upregulated in BCBM (FDR<0.01), compared to the primary BCs. High mRNA expression of ARG2 in primary BCs was associated with a worse distant metastasis-free survival (DMFS, p=0.038), while ARG2 protein expression was associated with a worse breast-brain metastasis-free (BMFS, p=0.027) and overall survival (p=0.019). There was a significant difference between the transcript levels of ARG2 and cytotoxic cells, T-cells and CD8 T-cells in primary BCs (p<0.0001 for all comparisons), with higher transcript levels of T-cells and lower ARG2. In BCBM, the significant transcript difference remained between ARG2, Cytotoxic cells and T-cells but not CD8 T-cells (p=0.0014, p=0.021 and p=0.3219 respectively) with higher ARG2 and lower T-cell transcripts.

Conclusions

This study highlights the immunological differences between primary BCs and BCBMs and the potential importance of ARG2 expression in T-cell depletion and as a prognostic biomarker in metastatic BC.

Legal entity responsible for the study

Breast Cancer group, University of Liverpool.

Funding

Infrastructure support from the Liverpool Experimental Cancer Medicine Centre (Grant Reference: C18616/A25153), the Clattterbridge Cancer Charity and the Liverpool University Hospitals NHS Foundation Trust. The study has also emanated from research supported in part by a Grant from Science Foundation Ireland under Grant number [20/FFP-P/8597 (DV)], Breast Cancer NOW Fellowship [Aug 2019 SF1310 (DV)] and Science Foundation Ireland Future for Frontiers Award, [19/FFP/6443 (LY)].

Disclosure

All authors have declared no conflicts of interest.

Background

The use of neoadjuvant therapy (NAT) for operable breast cancer (BC) has progressively increased over time and it is today recommended by major guidelines. The achievement of a pathological complete response (pCR) is associated with an improved outcome. As a consequence new strategies are required to early identify patients who will not respond. In this persepctive, metabolomics may represent an innovative technology to identify host related factors correlated with outcome. In this research we evaluate the use metabolomics analysis coupled to artificial intelligence to predict treatment outcome for BC patients undergoing NAT.

Methods

Untargeted metabolomics analysis was performed on serum samples from 66 operable BC patients treated with NAT. Small molecules were extracted from serum, derivatized and then analyzed using bi-dimensional gas chromatography/mass spectrometer (GCxGC-MS). The metabolomics profiling was then evaluated according to response to therapy (pCR versus residual disease). A machine learning approach was implemented with Boruta features selection algorithm combined with genetical algorithm as classifier, on a training set including 28 plasma samples and externally validated on the remaing 12 samples.

Results

Among the 66 enrolled patients, 27 (41%) were HER2 +, 23 (35%) and 16 (24%) were luminal B. Overall, 52 patients have received surgery so far. pCR was achieved in 29 patients (56%) and residual disease in 23 (44%). A total of 670 small molecules were quantified by untargeted metabolomics analysis; 77 of these resulted differentially expressed (p <0.05 and fold change > 1.3) between patients achieving a pCR or residual disease. A prediction model, combining metabolomic signatures and machine learning, was implemented on 40 metabolomic profiles. With this approach we were able to correctly identify the type of response with an accuracy of 98%.

Conclusions

By using this omic approach, we were able to identify a metabolomic signature correlated with the type of response to NAT. Among differentially expressed molecules, we identified several fatty acids, amino acids and small molecules that could be targeted by selected dietary supplements. Updated analysis of the different biological BC subtypes will be presented.

Legal entity responsible for the study

The authors.

Funding

AIRC.

Disclosure

All authors have declared no conflicts of interest.

Background

Establishing biobanking in cancer research is important for health research infrastructure to collect, store, process, and distribute high-quality human biological samples and associated data. The Biobank for Translational Medicine (B4MED) Unit at the European Institute of Oncology (IEO) is a landmark in this field. The aim of this analysis was to retrospectively examine the evolution of biobank activity for breast cancer research during the 10 years since the B4MED foundation.

Methods

All B4MED activities are controlled and tracked by Nautilus Laboratory Information Management System, a management system integrated with IEO medical records database. The files of B4MED were interrogated for the number of samples associated with patients treated for breast cancer at IEO, including their storage and use since the beginning of B4MED. All clinicopathologic characteristics, including histological subtype, grade, and stage, were retrieved into a pseudo-anonymized database. To extract the data, we performed a query with the following keywords: Breast, Carcinoma, and Primary.

Results

A total of 3,858 breast cancers and integrated clinicopathologic data were collected in our biobank (2012-2022). The samples, stored at -80°C, included tumor (n=3,858), matched non-neoplastic tissues (n=2,101; 55%), and biofluids (n=3,423; 89%), i.e. blood, serum, and plasma. For 1,774 (45%) patients both the normal tissue and blood samples were stored. Subtypes included 3,188 (82.6%) invasive carcinoma of no special type, 428 (11.1%) lobular, 134 (3.5%) mixed ductal-lobular, and 84 (2.2%) special types. Grade, stage, and molecular subtypes were available for all patients. Biomarkers were carefully annotated (e.g. we were able to retrieve 1,478 HER2-low breast cancers samples). All patients signed the Scientific Research Participation Agreement.

Conclusions

B4MED at IEO is an invaluable source of biomaterials that in the last decade led to remarkable scientific innovations related to new biomarkers and novel drugs, for increasingly personalized treatment strategies designed for breast cancer patients. The next challenge is to integrate digital and molecular pathology data to take advantage of machine learning protocols for next-generation biobanking.

Legal entity responsible for the study

The authors.

Funding

European Institute of Oncology and Italian Ministries of Health and Research.

Disclosure

All authors have declared no conflicts of interest.

Background

Immune checkpoint inhibitors exhibit limited response rates in patients with triple-negative breast cancer (TNBC), suggesting that additional immune escape mechanisms may exist. Here, we aimed to observe the function shifts of immune-related genes across various tumor microenvironments and identify previously undescribed regulators of TNBC immunity.

Methods

We performed two-step customized in vivo CRISPR screens targeting disease-related immune genes using different mouse models with multi-dimensional immune deficiency characteristics. Functional screening and transcriptomics analysis identified Lgals2 as a candidate regulator in TNBC involving immune escape. The distribution of tumor-infiltrating immune cells were analyzed through flow cytometry and single-cell RNA sequencing of tumors isolated from mouse xenograft models. We further conducted in vitro co-culture experiments to investigate the crosstalk between tumor-associated macrophages and TNBC cells mediated by Lgals2. Blockade of LGALS2 using an inhibitory antibody were also applied to assess its clinical significance in TNBC.

Results

The in vivo CRISPR screens characterized gene functions in the different tumor microenvironments and recovered canonical immunotherapy targets. Lgals2 was consistently correlated with immune escape in two-step screens and its expression was significantly higher in tumor tissues from TNBC patients compared with normal tissues. In addition, deletion of Lgals2 in tumor cells have no effect on tumor proliferation in vitro, but attenuated tumor growth in vivo. We further demonstrated that tumor cell-intrinsic Lgals2 resulted in the immunosuppressive phenotype of the TNBC microenvironment and induced the significantly increased number of tumor-associated macrophages in tumor tissues. Mechanistic studies revealed that Lgasl2 facilitated M2-like polarization and proliferation of macrophages through CSF1/CSF1R axis. Moreover, anti-LGALS2 antibody arrested tumor growth and reversed the immune suppression.

Conclusions

Lgals2 plays a critical role in the immunosuppressive nature of the TNBC microenvironment. We provide a theoretical basis for LGALS2 as a potential immunotherapy target in TNBC.

Legal entity responsible for the study

Fudan University Shanghai Cancer Center.

Funding

National Natural Science Foundation of China.

Disclosure

All authors have declared no conflicts of interest.

Background

Intermediate biopsies during neoadjuvant treatment might offer opportunities for prediction of pathological complete response (pCR) and patient outcome, which could be a basis for de-escalation of treatment. However, their relevance for clinical decisions is not clear.

Methods

We evaluated intermediate biopsies that were taken during neoadjuvant treatment from 297 patients with invasive breast cancer from three prospective, randomized trials (GeparQuattro, -Quinto, -Sixto). We evaluated the presence of invasive breast cancer, tumor-infiltrating lymphocytes (TILs) and Ki-67 expression and compared the results to pre-treatment samples. We explored the association of residual cancer in the intermediate biopsies as well as dynamics in Ki-67 and TILs with pCR rates and disease-free survival.

Results

Of the 297 tumors, 87 (29%) samples were triple-negative (TNBC), 138 (46%) HR+/HER2- and 72 (24%) HER2+. We found invasive tumor cells in 70% of biopsies, with significant differences between the subtypes (HR+/HER2-: 84%; TNBC: 62%, HER2+: 51%; p<0.001). In the complete cohort, 8% of patients with invasive tumor cells in intermediate biopsies achieved a pCR after completion of treatment (pCR rate in patients with residual tumor: TNBC: 19%, HR+/HER2: 3%; HER2+: 11%). If no residual cancer was present, pCR rate was 50% in the complete cohort (TNBC: 48%, HR+/HER2-: 27%; HER2+: 66%). An increase in TILs from baseline biopsy to intermediate biopsy was associated with a higher probability of pCR in the overall study cohort (p=0.001). A similar or increased Ki-67 was associated with a low probability of pCR in the overall study cohort (p=0.004) and with a shorter disease-free survival in patients with TNBC (p=0.04).

Conclusions

Intermediate biopsies can identify patients that are unlikely to achieve a pCR after therapy. After further validation, this may be useful for adaptation of treatment. For translational biomarker research, intermediate biopsies are useful tool to study mechanisms of therapy resistance and biomarker discovery. Additional studies will be needed to demonstrate if standardized sampling procedures can further improve response prediction through intermediate biopsies.

Legal entity responsible for the study

German Breast Group.

Funding

Has not received any funding.

Disclosure

M. Untch: Non-Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Amgen GmbH; Non-Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: AstraZeneca; Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: BMS; Non-Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Celgene GmbH; Non-Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Daiichi Sankyo; Non-Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Eisai; Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Lilly Deutschland; Non-Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Lilly Int.; Non-Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: MSD Merck; Non-Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Myriad Genetics; Non-Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Pfizer GmbH; Non-Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Roche Pharma AG; Non-Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Sanofi Aventis Deutschland GmbH; Non-Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Novartis; Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Pierre Fabre; Non-Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Clovis Oncology; Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Seatlle Genetics; Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Seagen; Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: GSK; Financial Interests, Personal and Institutional, Other, All fees to the institution/employer: Gilead. T. Karn: Other, Personal, Other, Patent pending: EP18209672. M. van Mackelenbergh: Financial Interests, Institutional, Other, Honoraria: Amgen; Financial Interests, Institutional, Other, Honoraria: AstraZeneca; Financial Interests, Institutional, Other, Honoraria: Novartis; Financial Interests, Institutional, Other, Honoraria: Pfizer; Financial Interests, Institutional, Other, Honoraria: Pierre Fabre; Financial Interests, Institutional, Other, Honoraria: Lilly; Financial Interests, Institutional, Other, Honoraria: Genomic Health; Financial Interests, Institutional, Other, Honoraria: Molecular Health; Financial Interests, Institutional, Other, Honoraria: Roche; Financial Interests, Institutional, Other, Honoraria: Gilead; Financial Interests, Institutional, Other, Honoraria: Seagen; Financial Interests, Institutional, Other, Honoraria: GSK. J. Huober: Financial Interests, Personal, Other, personal fees: Gilead; Financial Interests, Personal, Other, personal fees: Seagen; Financial Interests, Personal and Institutional, Research Grant, Grant, personal fees: Lilly; Financial Interests, Personal and Institutional, Research Grant, Grant, Travel, personal fees: Novartis; Financial Interests, Personal, Other, Travel, personal fees, Other: Pfizer; Financial Interests, Personal, Other, Travel, personal fees, Other: Daiichi; Financial Interests, Personal, Other, personal fees: Roche; Financial Interests, Personal, Other, personal fees: MSD; Financial Interests, Personal, Other, personal fees: AbbVie; Financial Interests, Personal, Other, personal fees: Eisai; Financial Interests, Institutional, Research Grant, Grant: Hexal; Financial Interests, Institutional, Research Grant, Grant, Travel, Other: BMS. W.D. Schmitt: Financial Interests, Personal, Other, Honoraria: AstraZeneca; Financial Interests, Personal, Other, Honoraria: GlaxoSmithKline; Financial Interests, Personal, Funding: Myriad Genetics. F. Marmé: Financial Interests, Personal, Other, Personal Fees: Roche/Genentech; Financial Interests, Personal, Other, Personal Fees: AstraZeneca; Financial Interests, Personal, Other, Personal Fees: Pfizer; Financial Interests, Personal, Other, Personal Fees: Tesaro; Financial Interests, Personal, Other, Personal Fees: Novartis; Financial Interests, Personal, Other, Personal Fees: Amgen; Financial Interests, Personal, Other, Personal Fees: PharmaMar; Financial Interests, Personal, Other, Personal Fees: Genomic Health; Financial Interests, Personal, Other, Personal Fees: CureVac; Financial Interests, Personal, Other, Personal Fees: EISAI; Financial Interests, Personal, Other, Persona Fees: BMS; Financial Interests, Personal, Other, Personal Fees: Clovis Oncology; Financial Interests, Personal, Other, Personal Fees: GSK; Financial Interests, Personal, Other, Personal Fees: MSD; Financial Interests, Personal, Other, Personal Fees: Seagen; Financial Interests, Personal, Other, Personal Fees: Myriad Genetics; Financial Interests, Personal, Other, Personal Fees: Pierre Fabre; Financial Interests, Personal and Institutional, Research Grant, Personal Fees: Gilead/Immunomedics; Financial Interests, Personal, Other, Persona fees: Janssen-Cilag. E. Stickeler: Financial Interests, Personal, Other, personal fees: Roche; Financial Interests, Personal, Other, personal fees: Gilead; Financial Interests, Personal, Other, personal fees: MSD; Financial Interests, Personal, Other, personal fees: Lilly; Financial Interests, Personal, Other, personal fees: Pfizer; Financial Interests, Personal, Other, personal fees: Seagen; Financial Interests, Personal, Other, personal fees: PharmaMar. H. Tesch: Financial Interests, Institutional, Other, Consulting fees, honoraria for lectures, presentations: Novartis; Financial Interests, Institutional, Other, Consulting fees, honoraria for lectures, presentations: Roche; Financial Interests, Institutional, Other, Consulting fees, honoraria for lectures, presentations: GSK; Financial Interests, Institutional, Other, Consulting fees, honoraria for lectures, presentations: Seagen; Financial Interests, Institutional, Other, Consulting fees, honoraria for lectures, presentations: Pfizer; Financial Interests, Institutional, Other, Consulting fees, honoraria for lectures, presentations: Lilly; Financial Interests, Institutional, Other, Consulting fees, honoraria for lectures, presentations: AstraZeneca; Financial Interests, Institutional, Other, Consulting fees, honoraria for lectures, presentations: Daiichi; Financial Interests, Institutional, Other, Consulting fees, honoraria for lectures, presentations: Exact Science. P.A. Fasching: Financial Interests, Personal, Advisory Board, Advisory Board, Invited Speaker: Novartis; Financial Interests, Institutional, Research Grant, Grant, Institutional Funding: BioNTech; Financial Interests, Personal and Institutional, Advisory Board, Advisory Board, Invited Speaker, Research Grant: Pfizer; Financial Interests, Personal, Advisory Board, Advisory Board, Invited Speaker: Daiichi Sankyo; Financial Interests, Personal, Advisory Board, Advisory Board, Invited Speaker: AstraZeneca; Financial Interests, Personal, Advisory Board, Advisory Board, Invited Speaker: Eisai; Financial Interests, Personal, Advisory Board, Advisory Board, Invited Speaker: Merck Sharp & Dohme; Financial Interests, Institutional, Research Grant, Grant, Institutional Funding: Cepheid; Financial Interests, Personal, Advisory Board, Advisory Board, Invited Speaker: Lilly; Financial Interests, Personal, Advisory Board, Advisory Board: Pierre Fabre; Financial Interests, Personal, Advisory Board, Advisory Board, Invited Speaker: Seagen; Financial Interests, Personal, Advisory Board, Advisory Board, Invited Speaker: Roche; Financial Interests, Personal, Advisory Board, Advisory Board: Hexal; Financial Interests, Personal, Advisory Board, Advisory Board: Agendia; Financial Interests, Personal, Advisory Board, Advisory Board: Sanofi Aventis; Financial Interests, Personal, Advisory Board, Advisory Board, Invited Speaker: Gilead. A. Schneeweiss: Financial Interests, Personal and Institutional, Research Grant, Research Grant, Travel expenses, Honoraria: BMS; Financial Interests, Personal and Institutional, Research Grant, Research Grant, Expert testimony, Travel expenses, Honoraria: Roche; Financial Interests, Institutional, Research Grant, Research Grant: AbbVie; Financial Interests, Institutional, Research Grant, Research Grant: Molecular Partner; Financial Interests, Personal, Expert Testimony, Expert testimony, Honoraria: AstraZeneca; Financial Interests, Personal, Other, Honoraria, Travel expenses: Pfizer; Financial Interests, Personal, Other, Honoraria: Novartis; Financial Interests, Personal, Other, Honoraria: MSD; Financial Interests, Personal, Other, Honoraria: Tesaro; Financial Interests, Personal, Other, Honoraria: Lilly; Financial Interests, Personal, Other, Honoraria: Seagen; Financial Interests, Personal, Other, Honoraria: Gilead. V. Müller: Financial Interests, Personal, Invited Speaker, speaker and consultancy honoraria: Amgen; Financial Interests, Personal, Invited Speaker, speaker honoraria: AstraZeneca; Financial Interests, Personal, Invited Speaker, speaker and consultancy honoraria: Daiichi Sankyo; Financial Interests, Personal, Invited Speaker, speaker and consultancy honoraria: Eisai; Financial Interests, Personal, Invited Speaker, speaker honoraria: Pfizer; Financial Interests, Personal, Invited Speaker, speaker and consultancy honoraria: MSD; Financial Interests, Personal and Institutional, Invited Speaker, speaker and consultancy honoraria, Institutional research support: Novartis; Financial Interests, Personal and Institutional, Invited Speaker, speaker and consultancy honoraria, Institutional research support: Roche; Financial Interests, Personal, Invited Speaker, speaker honoraria: Teva; Financial Interests, Personal and Institutional, Invited Speaker, speaker honoraria, Institutional research support: Seagen; Financial Interests, Personal, Invited Speaker, speaker and consultancy honoraria: GSK; Financial Interests, Personal, Invited Speaker, speaker and consultancy honoraria: Gilead; Financial Interests, Personal, Advisory Role, consultancy honoraria: Genomic Health; Financial Interests, Personal, Advisory Role, consultancy honoraria: Hexal; Financial Interests, Personal, Advisory Role, consultancy honoraria: Pierre Fabre; Financial Interests, Personal, Advisory Role, consultancy honoraria: ClinSol; Financial Interests, Personal, Advisory Role, consultancy honoraria: Lilly; Financial Interests, Institutional, Other, Institutional research support: Genentech. S. Loibl: Financial Interests, Institutional, Research Grant, Grant, Other: AbbVie; Financial Interests, Institutional, Advisory Board, honorarium for Ad Board, Other: Amgen; Financial Interests, Institutional, Research Grant, Grant, Other: AstraZeneca; Financial Interests, Institutional, Advisory Board, honorarium for Ad Board, Other: Bayer; Financial Interests, Institutional, Advisory Board, honorarium for Ad Boards, Other: BMS; Financial Interests, Institutional, Research Grant, Grant, Other: Celgene; Non-Financial Interests, Institutional, Research Grant, Grant, Medical Writing, Other: Daiichi Sankyo; Financial Interests, Institutional, Advisory Board, honorarium for Ad Board, Other: Eirgenix; Financial Interests, Institutional, Advisory Board, honorarium for Ad Board, Other: GSK; Non-Financial Interests, Institutional, Research Grant, Grant, Medical Writing, Other: Immunomedics/Gilead; Financial Interests, Institutional, Advisory Board, honorarium for Ad Boards, Other: Lilly; Financial Interests, Institutional, Advisory Board, honorarium for Ad Board, Other: Merck; Non-Financial Interests, Institutional, Research Grant, Grant, Medical Writing, Other: Novartis; Non-Financial Interests, Institutional, Research Grant, Grant, Medical Writing, Other: Pfizer; Financial Interests, Institutional, Advisory Board, honorarium for Ad Board & Lecture, Other: Pierre Fabre; Financial Interests, Institutional, Advisory Board, honorarium for Ad Board & Lecture, Other: prIME/Medscape; Non-Financial Interests, Institutional, Advisory Board, honorarium for Ad Board, Medical Writing, Other: Puma; Financial Interests, Institutional, Advisory Board, honorarium for Lecture, Other: Samsung; Non-Financial Interests, Institutional, Advisory Board, honorarium for Ad Board, Medical Writing, Other: Seagen; Non-Financial Interests, Institutional, Advisory Board, honorarium for Ad Boards & Lectures, Medical Writing, Other: Roche; Other, Institutional, Other, Patent Pending, Immunsignature in TNBC: EP14153692.0; Other, Institutional, Other, Patent Pending, Signature for CDK 4/6 Inhibitor: EP21152186.9; Other, Institutional, Other, Patent Issued, Predicting response to an Anti-HER2 containing therapy: EP15702464.7; Other, Institutional, Other, Patent Pending, GeparNuevo: EP19808852.8; Other, Institutional, Royalties, Patent Issued, Royalties, VM Scope GmbH: Digital Ki67 Evaluator. C. Denkert: Financial Interests, Personal, Ownership Interest, Stock and Other Ownership Interests: Sividon Diagnostics; Financial Interests, Personal, Advisory Role, Consulting or Advisory Role: MSD Oncology; Financial Interests, Personal, Advisory Role, Consulting or Advisory Role: Daiichi Sankyo; Financial Interests, Personal, Advisory Role, Consulting or Advisory Role: Molecular Health; Financial Interests, Personal, Advisory Role, Consulting or Advisory Role: AstraZeneca; Financial Interests, Personal, Advisory Role, Consulting or Advisory Role: Merck; Financial Interests, Personal and Institutional, Advisory Role, Consulting or Advisory Role, Research Funding: Roche; Financial Interests, Personal, Advisory Role, Consulting or Advisory Role: Lilly; Financial Interests, Institutional, Research Grant, Research Funding: Myriad Genetics; Other, Personal, Royalties, Patents, Royalties, Other Intellectual Property: VMScope digital pathology software; Other, Personal, Royalties, Patents, Royalties, Other Intellectual Property: WO2015114146A1; Other, Personal, Royalties, Patents, Royalties, Other Intellectual Property: WO2010076322A1; Other, Personal, Royalties, Patents, Royalties, Other Intellectual Property: WO2020109570A1. All other authors have declared no conflicts of interest.

Background

The 27-gene HER2DX prognostic assay has been evaluated in >1300 patients (pts) with early-stage HER2+ breast cancer (BC) (EBioMedicine 2022). Here, we explored the association of the research-based HER2DX with a list of 485 genes linked to relapse-free survival (RFS) in the N9831 pivotal trial (JCO 2015).

Methods

N9831 randomized 1282 pts with early-stage HER2+ BC to adjuvant multi-agent chemotherapy (CT) (arm A), 1-year of trastuzumab starting concomitantly with CT (arm B) and 1-year of trastuzumab after CT (arm C). In JCO 2015, 485 genes were significantly (p<0.01) associated with RFS in 849 pts in arms B/C using DASL technology. Here, the primary objective was to identify available genes from the HER2DX gene list associated with RFS in arms B/C. A representation factor and the probability of finding a gene overlap were calculated. The secondary objective was to compare the expression of the N9831 485 prognostic genes in research-based HER2DX low- vs high-risk groups in 376 HER2+ BCs from METABRIC and TCGA datasets. Differentially expressed genes between both risk groups were identified (false discovery rate<5%).

Results

Five genes (CD27, CD79A, IL2RG, LAX1 and PIM2) were found in both prognostic gene lists (N9831 and HER2DX) and all belong to the HER2DX IGG signature (5 of 11 genes available; 45.5%). In N9831, the RFS hazard ratio estimate for each gene was 0.84 (CD27; p=0.002), 0.78 (CD79A; p=0.005), 0.81 (IL2RG; p=0.002), 0.85 (LAX1; p=0.008) and 0.80 (PIM2; p=0.007). The overlap between the 2 gene lists was not due to chance (enrichment factor=10.7; p<0.001). Of the genes associated with better prognosis in N9831 (n=330), 96.8% in METABRIC and 96.9% in TCGA were significantly more expressed in the research-based HER2DX low-risk group compared to the high-risk group. Of the genes associated with worse prognosis in N9831 (n=155), 66.6% in METABRIC and 35.6% in TCGA were significantly more expressed in the research-based HER2DX high-risk group compared to low-risk group.

Conclusions

The biology captured by HER2DX is significantly associated with prognostic genes found in pts treated with adjuvant trastuzumab-based CT in the N9831 trial. This result reinforces the prognostic ability of HER2DX in early-stage HER2+ BC.

Legal entity responsible for the study

The authors.

Funding

Has not received any funding.

Disclosure

F. Brasó-Maristany: Financial Interests, Personal, Invited Speaker: Reveal Genomics. O. Martinez Saez: Financial Interests, Personal, Invited Speaker: Novartis; Financial Interests, Personal, Invited Speaker: Eisai; Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Other, Travel expenses: Roche. L. Pare Brunet: Financial Interests, Personal, Full or part-time Employment: Reveal Genomics. M. Marín-Aguilera: Financial Interests, Personal, Full or part-time Employment: Reveal Genomics. P.F. Conte: Financial Interests, Personal, Expert Testimony, expert testimony for drug approval with AIFA: Roche; Financial Interests, Personal, Advisory Board, advisory board for alpelisb trial: Novartis; Financial Interests, Personal, Invited Speaker: Eli Lilly; Financial Interests, Personal, Invited Speaker: AstraZeneca; Financial Interests, Institutional, Research Grant: Merch Kga; Financial Interests, Institutional, Research Grant: Roche; Financial Interests, Institutional, Research Grant: Novartis. V. Guarneri: Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Invited Speaker: Eli Lilly; Financial Interests, Personal, Advisory Board: Eli Lilly; Financial Interests, Personal, Advisory Board: Novartis; Financial Interests, Personal, Invited Speaker: Novartis; Financial Interests, Personal, Advisory Board: MSD; Financial Interests, Personal, Advisory Board: Gilead; Financial Interests, Institutional, Invited Speaker: Eli Lilly; Financial Interests, Institutional, Invited Speaker: Roche; Financial Interests, Institutional, Invited Speaker: BMS; Financial Interests, Institutional, Invited Speaker: Novartis; Financial Interests, Institutional, Invited Speaker: AstraZeneca; Financial Interests, Institutional, Invited Speaker: MSD; Financial Interests, Institutional, Invited Speaker: Synton Biopharmaceuticals; Financial Interests, Institutional, Invited Speaker: Merck. T. Pascual: Financial Interests, Personal, Invited Speaker: Pfizer; Financial Interests, Personal, Invited Speaker: AstraZeneca; Financial Interests, Personal, Invited Speaker: Lilly. A. Vivancos: Financial Interests, Personal, Other, Consultant: Sysmex; Financial Interests, Personal, Advisory Board: Novartis; Financial Interests, Personal, Advisory Board: Merck; Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Advisory Board: Bristol Meyers Squibb; Financial Interests, Personal, Advisory Board: Guardant Health; Financial Interests, Personal, Advisory Board: Bayer; Financial Interests, Personal, Other, Technology Transfer DX Field: Ferrer; Financial Interests, Institutional, Research Grant, Preclinical Research Grant: Bristol Meyers Squibb; Financial Interests, Institutional, Research Grant, Preclinical Research Grant: Novartis; Financial Interests, Institutional, Research Grant, Preclinical Research Grant: Debio; Financial Interests, Institutional, Research Grant, Preclinical Research Grant: SYSMEX; Financial Interests, Institutional, Research Grant, Preclinical Research Grant: Cellestia Biotech; Financial Interests, Institutional, Research Grant, Preclinical Research Grant: Roche; Financial Interests, Institutional, Research Grant, Preclinical Research Grant: Chittern. J. Parker: Financial Interests, Personal, Stocks/Shares: Reveal Genomics; Financial Interests, Personal, Advisory Board: Reveal Genomics. P. Villagrasa Gonzalez: Financial Interests, Personal, Stocks/Shares: Reveal Genomics; Financial Interests, Personal, Officer: Reveal Genomics. S.M. Tolaney: Financial Interests, Personal, Advisory Board, Ad Board Participant/Consultant: AstraZeneca; Financial Interests, Personal, Advisory Board, Ad Board Participant/Consultant: Eli Lilly; Financial Interests, Personal, Advisory Board, Ad board participant/Consultant: Pfizer; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Novartis; Financial Interests, Personal, Advisory Board, Ad Board participant/consultant: Merck; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Gilead; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Genentech/Roche; Financial Interests, Personal, Other, Consultant: Nektar; Financial Interests, Personal, Other, Consultant: NanoString; Financial Interests, Personal, Advisory Board, Ad board participant: Bristol Myers Squibb; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Eisai; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Sanofi; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Seagen; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Daiichi Sankyo; Financial Interests, Personal, Advisory Board, Ad board participant/consultant/DSMC: Odonate; Financial Interests, Personal, Other, Steering Committee Member/Consultant: CytomX; Financial Interests, Personal, Other, Consultant: Athenex; Financial Interests, Personal, Advisory Board, Ad board participant: OncoPep; Financial Interests, Personal, Advisory Board, Ad board participant: OncoSec; Financial Interests, Personal, Advisory Board, Ad board participant: Certara; Financial Interests, Personal, Advisory Board, Ad board participant: Mersana Therapeutics; Financial Interests, Personal, Advisory Board, Ad board participant: Ellipses Pharma; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: 4D Pharma; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Puma; Financial Interests, Personal, Invited Speaker, Invited speaker for pharma supported educational activity: Chugai Pharmaceuticals; Financial Interests, Personal, Advisory Board, Advisory board participant: G1 Therapeutics; Financial Interests, Personal, Other, Consultant: Blueprint Medicines; Financial Interests, Personal, Advisory Board, Advisory Board Participation: Zymeworks; Financial Interests, Personal, Advisory Board, Advisory Board participation: Zentalis; Financial Interests, Personal, Advisory Board, Advisory board participation: Reveal Genomics; Financial Interests, Personal, Advisory Board, Advisory Board participation: OncXerna; Financial Interests, Institutional, Funding: AstraZeneca; Financial Interests, Institutional, Funding: Eli Lilly; Financial Interests, Institutional, Funding: Pfizer; Financial Interests, Institutional, Funding: Sanofi; Financial Interests, Institutional, Funding: Seagen; Financial Interests, Institutional, Funding: Odonate; Financial Interests, Personal and Institutional, Invited Speaker: CytomX; Financial Interests, Institutional, Funding: Cyclacel; Financial Interests, Institutional, Funding: Exelixis; Financial Interests, Institutional, Funding: Gilead; Financial Interests, Institutional, Funding: Bristol Myers Squibb; Financial Interests, Institutional, Funding: Eisai; Financial Interests, Institutional, Funding: Merck; Financial Interests, Institutional, Funding: Novartis; Financial Interests, Institutional, Funding: Nektar; Financial Interests, Institutional, Funding: Genentech/Roche. L.A. Carey: Financial Interests, Institutional, Funding, research funding: Syndax; Financial Interests, Institutional, Funding, research funding: Immunomedics; Financial Interests, Institutional, Funding, research funding: Novartis; Financial Interests, Institutional, Funding, research funding: NanoString Technologies; Financial Interests, Institutional, Funding, research funding: AbbVie; Financial Interests, Institutional, Funding, research funding: Seattle Genetics; Financial Interests, Institutional, Funding, research funding: Veracyte; Non-Financial Interests, Advisory Role, uncompensated relationship - all monies go to UNC. Dr. Carey does not have any signatory authority over any UNC account: sanofi; Non-Financial Interests, Advisory Role, uncompensated relationship - all monies go to UNC. Dr. Carey does not have any signatory authority over any UNC account: Novartis; Non-Financial Interests, Advisory Role, uncompensated relationship - all monies go to UNC. Dr. Carey does not have any signatory authority over any UNC account: G1 Therapeutics; Non-Financial Interests, Advisory Role, uncompensated relationship - all monies go to UNC. Dr. Carey does not have any signatory authority over any UNC account: Genentech/Roche; Non-Financial Interests, Advisory Role, uncompensated relationship - all monies go to UNC. Dr. Carey does not have any signatory authority over any UNC account: GlaxoSmithKline; Non-Financial Interests, Advisory Role, uncompensated relationship - all monies go to UNC. Dr. Carey does not have any signatory authority over any UNC account: AstraZeneca/ Daiichi Sankyo; Non-Financial Interests, Advisory Role, uncompensated relationship - all monies go to UNC. Dr. Carey does not have any signatory authority over any UNC account: Apitude Health; Non-Financial Interests, Advisory Role, uncompensated relationship - all monies go to UNC. Dr. Carey does not have any signatory authority over any UNC account: Exact Sciences. C.M. Perou: Financial Interests, Personal, Stocks/Shares: Reveal Genomics; Financial Interests, Personal, Advisory Role: Reveal Genomics; Financial Interests, Personal, Stocks/Shares: BioClassifier LLC; Financial Interests, Personal, Advisory Role: BioClassifier LLC. A. Prat: Financial Interests, Personal, Invited Speaker: Roche; Financial Interests, Institutional, Other, Contracted research:: Roche; Financial Interests, Personal, Invited Speaker, Lecture fees: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Novartis; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Novartis; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Amgen; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Amgen; Financial Interests, Personal, Invited Speaker, Lecture fees: BMS; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: BMS; Financial Interests, Personal, Invited Speaker, Lecture fees: NanoString; Financial Interests, Institutional, Other, Contracted research: NanoString; Financial Interests, Institutional, Invited Speaker, Lecture fees: NanoString; Financial Interests, Institutional, Other, Contracted research: Novartis; Financial Interests, Institutional, Other, Contracted research: Roche; Financial Interests, Institutional, Other, Contracted research: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Contracted research; Financial Interests, Personal, Invited Speaker, Lecture fees: Daiichi Sankyo; Financial Interests, Institutional, Invited Speaker, Clinical trials: Daiichi Sankyo; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Puma; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Oncolytics; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: MSD; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Guardan Health; Financial Interests, Personal, Expert Testimony, Advisory role/consultancy: Peptomyc; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Lilly; Financial Interests, Institutional, Other, Clinical trials: Lilly; Financial Interests, Institutional, Other, Contracted research: Boehringer; Financial Interests, Institutional, Other, Contracted research: Sysmex Europa GmbH; Financial Interests, Institutional, Other, Contracted research: Medica Scientia inno. Research; Financial Interests, Institutional, Other, Contracted research: Celgene; Financial Interests, Institutional, Other, Contracted research: Astellas; Financial Interests, Institutional, Other, Clinical trials: Roche; Financial Interests, Institutional, Other, Clinical trials: AMgen; Financial Interests, Personal, Invited Speaker, Leadership role: Reveal Genomics, SL.; Financial Interests, Institutional, Other, Clinical trials: Boehringer; Non-Financial Interests, Institutional, Other, Leadership roles: Patronage committee: SOLTI Foundation; Non-Financial Interests, Institutional, Other, Leadership roles: Patronage committee: Actitud Frente al Cáncer Foundation. All other authors have declared no conflicts of interest.

Background

In patients with triple-negative breast cancer (TNBC) undergoing neoadjuvant chemotherapy (NACT), higher tumor-infiltrating lymphocytes (TILs) strongly correlate with increased rates of pathologic complete response (pCR) and improved survival. The aim of the present work is to investigate the changes of TIL levels from baseline biopsy to residual disease (RD) in TNBC patients failing to achieve pCR after standard NACT with anthracycline-taxane (A-T) +/- carboplatin (Cb).

Methods

TNBC (ER&PgR<10%) patients treated with neoadjuvant A-T +/-Cb failing to achieve pCR were included. Stromal TILs were evaluated on baseline biopsy and matched samples of RD. The non-parametric Wilcoxon test was applied to compare TIL levels from baseline biopsy to RD.

Results

79 patients with matched samples of baseline TILs and RD-TILs were included: 62% treated with A-T, 38% with A-TCb. Mean and median baseline TILs were 8.8% and 5%; mean and median RD-TILs were 14.1% and 8%. The distribution of TIL categories in matched samples of baseline biopsy and RD is shown in the table. RD-TILs were significantly higher than baseline TILs in the overall population (p=0.004) and in both A-T (p=0.027) and A-TCb subgroups (p=0.043). A significant increase of TIL levels after NACT was also seen in patients with low baseline TILs, in the overall population (p<001) and in both treatment subgroups (A-T, p=0.002, A-TCb, p=001). Table: 26P

Conclusions

We reported in a population of TNBC patients failing to achieve pCR after standard NACT, that A-T +/- Cb is capable of enhancing tumor immune infiltration from pre-treatment tumor to RD. Importantly, we observed that TIL infiltration was enhanced after A-T +/- Cb also in patients with “immune-cold” tumors at baseline.

Legal entity responsible for the study

The authors.

Funding

Univ.

Disclosure

F. Miglietta: Financial Interests, Personal, Invited Speaker, outside the submitted work: Roche. G. Griguolo: Financial Interests, Personal, Invited Speaker: Novartis; Financial Interests, Personal, Invited Speaker: Eli Lilly; Other, Travel Support: Novartis; Other, Travel Support: Amgen; Other, Travel Support: Pfizer; Other, Travel Support: Daiichi Sankyo. M. Fassan: Financial Interests, Personal, Advisory Board, outside the submitted work: Astellas Pharma; Financial Interests, Personal, Advisory Board, outside the submitted work: QED Therapeutics; Financial Interests, Personal, Advisory Board, outside the submitted work: Diaceutics; Financial Interests, Personal, Advisory Board, outside the submitted work: Tesaro; Financial Interests, Personal, Advisory Board, outside the submitted work: Roche; Financial Interests, Personal, Advisory Board, outside the submitted work: Eli Lilly; Financial Interests, Personal, Advisory Board, outside the submitted work: Novartis. C.A. Giorgi: Financial Interests, Personal, Invited Speaker, outside the submitted work: Novartis. V. Guarneri: Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Invited Speaker: Eli Lilly; Financial Interests, Personal, Advisory Board: Eli Lilly; Financial Interests, Personal, Advisory Board: Novartis; Financial Interests, Personal, Invited Speaker: Novartis; Financial Interests, Personal, Advisory Board: MSD; Financial Interests, Personal, Advisory Board: Gilead; Financial Interests, Institutional, Invited Speaker: Eli Lilly; Financial Interests, Institutional, Invited Speaker: Roche; Financial Interests, Institutional, Invited Speaker: BMS; Financial Interests, Institutional, Invited Speaker: Novartis; Financial Interests, Institutional, Invited Speaker: AstraZeneca; Financial Interests, Institutional, Invited Speaker: MSD; Financial Interests, Institutional, Invited Speaker: Synton Biopharmaceuticals; Financial Interests, Institutional, Invited Speaker: Merck. M.V. Dieci: Financial Interests, Personal, Invited Speaker: Eli Lilly; Financial Interests, Personal, Advisory Board: Novartis; Financial Interests, Personal, Advisory Board: Eli Lilly; Financial Interests, Personal, Invited Speaker: Pfizer; Financial Interests, Personal, Advisory Board: Seagen; Financial Interests, Personal, Advisory Board: Exact Science; Financial Interests, Personal, Other, Consultancy: Pfizer; Financial Interests, Institutional, Research Grant: Veneto Institute of Oncology IOV-IRCCS; Financial Interests, Institutional, Research Grant: Italian Ministry of health; Financial Interests, Institutional, Research Grant: University of Padova. All other authors have declared no conflicts of interest.

Background

Tumor-infiltrating lymphocytes (TILs) and their subsets contribute to breast cancer prognosis. We investigated the prognostic impact of CD8+ TILs in patients with early intermediate/high-risk breast cancer treated with adjuvant anthracycline-based chemotherapy.

Methods

All patients received dose-dense sequential chemotherapy with E-T-CMF regimen. We examined their tumors for total, stromal (s) and intratumoral (i) CD8 lymphocyte density (counts/mm2) on tissue-microarray cores by immunohistochemistry. Morphological sTIL density was also evaluated. Impact of CD8+ TILs counts on DFS and OS and its correlation with breast cancer (IHC4) subtypes and standard clinicopathological parameters were investigated along with efficacy and safety data.

Results

990 patients were enrolled from which 627 had available CD8 data. The median number of sCD8 and iCD8 counts/mm² was 113.7 and 3.7, respectively. 156 (24.9%) had high expression of sCD8, iCD8 and total CD8 and were strongly correlated with higher Ki67, TILs density, ER/PgR negativity and higher histological grade. At a median follow-up of 132.5 months, the median DFS and OS have not been reached yet. 5-year DFS and OS rates were 86.1% and 91.4% respectively. Patients with high intratumoral and total CD8 had longer DFS and OS as compared to those with low counts/mm² (DFS: HR=0.58, p=0.011 and HR=0.65, p=0.034 and OS: HR=0.63, p=0.043 and HR=0.58, p=0.020, respectively). Upon adjustment for clinicopathological parameters, iCD8 and total CD8 retained their favorable prognostic significance for DFS and OS, whereas high sCD8 was significantly associated with prolonged DFS. Menopausal status, tumor size and nodal status retained their prognostic significance in all examined multivariate models. TILs density and CD8 status did not interact with BC subtypes.

Conclusions

CD8+ TILs and especially their intratumoral subset, represent a potential favorable prognostic factor and impact survival rates in early breast cancer. Further research is needed to establish their role in breast cancer prognosis.

Clinical trial identification

ACTRN12615000161527.

Legal entity responsible for the study

Hellenic Cooperative Oncology Group.

Funding

Hellenic Cooperative Oncology Group.

Disclosure

All authors have declared no conflicts of interest.

Background

Deregulated lipogenesis is essential for tumor cell survival. Regardless of circulating lipid levels, a wide range of tumors presents accelerated de novo fatty acid synthesis. Oncogenic lipogenesis is manifested by enhanced activity and coordinated expression of multiple lipogenic enzymes including ATP-citrate lyase (ACLY) in tumor cells. Therefore, the present study was aimed at understanding the influence of ACLY on breast tumorigenesis and its role in inducing immune escape.

Methods

We analyzed The Cancer Genome Atlas (TCGA) and Clinical Proteomic Tumor Analysis Consortium (CPTAC) datasets using an R statistical environment to understand the correlation between lipogenic enzymes and immune regulatory genes. Expression levels of ACLY and PDL-1/AKT/mTOR signaling proteins in different breast cancer cells were quantified using immunoblotting. The siRNA-mediated ACLY knockdown was conducted and the subsequent effect on cell cycle and cell death were quantified using flow cytometry. The expression levels of inflammatory markers are quantified using enzyme-linked immunosorbent assay (ELISA). The expression of immune-regulatory genes in ACLY knockdown cells were quantified using real-time PCR.

Results

Our TCGA and CPTAC analysis revealed that lipogenic enzymes- ACLY, FASN, ACACA were deregulated in breast cancer and were correlated with immune regulatory genes including PD-L1. The siRNA-mediated knockdown demonstrated that ACLY modulated PDL-1 via AKT/mTOR signaling. ACLY knockdown also resulted in significant cell cycle arrest and cell death which was also reflected in the expression of Bcl-2, survivin, and caspase. The expression of TNFa and IL-8 were found to be elevated while IL-4 was found to be unaltered in the conditioned medium of ACLY knckdown cells. Furthermore, the ACLY knockdown in breast cancer cells led to aberrant expression of immune regulatory genes including CD27, CD40, CD47 and TNFSF4 indicating deregulated expression of ACLY in breast tumor cells mediates immune escape.

Conclusions

This is the first study that shows ACLY influences PD-L1 leading to aberrant expression of immune regulatory genes and tumor cell survival in breast cancer.

Legal entity responsible for the study

The authors.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

Background

Mucoepidermoid carcinoma (MEC) of the breast is an extremely rare form of salivary gland-type tumor morphologically characterized by four types of cells (basaloid, intermediate, epidermoid, and mucinous) arranged in solid and cystic patterns. Despite its triple-negative phenotype, breast MECs are reported to have a relatively good clinical behavior. However, no data on their biology and therefore no guidelines for their clinical management are currently available. Here, we sought to characterize the molecular landscape of breast MECs.

Methods

Thirteen cases were included and histologically reviewed. Tumor infiltrating lymphocytes (TILs), along with the immunohistochemical expression of programmed death ligand 1 (PD-L1, clone 22C3), EGFR, and amphiregulin (AREG) were assessed. The presence of MAML2 and EWSR1 rearrangements was investigated by fluorescent in situ hybridization. Eight cases with available enough tissue material were subjected to a next-generation sequencing (NGS) panel targeting 161 cancer-related genes.

Results

Most cases were of low histological grade (n=10, 77%) with low proliferation (Ki67<30%). TILs were found in 2 (17%) cases, while PD-L1 combined positive score ranged from 0 to 20 (median 12.5). All cases showed EGFR overexpression and 2 of them were AREG-positive, among which one patient developed metastasis. No MAML2 or EWSR1 rearrangements were detected. Pathogenic mutations in PIK3CA were highly recurrent (n=4/8; 50%), though only one case harbored TP53 mutations. Additional somatic mutations affecting cancer-related genes found in MECs included CDK2, NF1/2, AKT1, SMARCB1, MYC, KRAS, CDK4, NOTCH1, and FGFR3/4.

Conclusions

This is the first study providing molecular data on breast MECs and the broadest collection of cases presented so far. Here we show that breast MECs lack the hallmark TP53 mutation found in high-grade forms of TNBC but also the MAML2 or EWSR1 rearrangements found in salivary MECs. The low level of TILs and PD-L1 suggest an immunoediting deficiency and that MECs may be unlikely to respond to immunotherapy combination strategies. Finally, the EGFR-AREG axis activation, coupled with the complex patterns of mutations in PI3K/AKT/mTOR and cell cycle regulation pathways warrant caution in considering MECs as low-grade TNBCs.

Legal entity responsible for the study

The authors.

Funding

Italian ministry of health (Ricerca Corrente Funds).

Disclosure

All authors have declared no conflicts of interest.

Background

The oxysterols 27-hydroxycholesterol (27-HC) and 25-HC were identified as endogenous estrogen receptor (ER) modulators and are hypothesized to play a role in breast cancer progression. We assessed associations between circulating 27-HC, 25-HC, and breast cancer prognosis.

Methods

A total of 2,282 breast cancer patients with blood collection post-diagnosis were included from the Mamma Carcinoma Risk Factor Investigation (MARIE) study. 438 total deaths, 237 breast cancer (BC)-specific deaths and 322 recurrences occurred over median follow-up time of 11 years. 86% of participants had ER+/progesterone receptor (PR)+ disease. 27-HC and 25-HC levels were quantified by liquid chromatography–mass spectrometry, and estradiol levels were measured with an ELISA. We calculated hazard ratios (HR) and 95% confidence intervals [CI] for quartiles of oxysterol levels and associations with overall mortality, BC-specific mortality and risk of recurrence using multivariable Cox proportional hazards models.

Results

We observed no significant associations between 27-HC (median concentration=200.8 nanomolar (nM)) and breast cancer prognosis. There was no heterogeneity by ER/PR-status, but significant heterogeneity was observed in subgroups defined by median estradiol levels (22.1 ng/ml). 27-HC was associated with higher risk of overall death (HR_Q4vs.Q1= 1.51 [CI 1.01-2.26]) in the low estradiol group, while a non-significant inverse trend was observed in the high estradiol group. Higher 27-HC levels were associated with higher risk of death among participants not using endocrine therapy. Higher 25-HC levels were associated with lower risk of recurrence (HR _Q4vs.Q1=0.71 [CI 0.51-0.97]). Among women with low estradiol, higher 25-HC was associated with higher risk of overall death (HR= 1.49 [CI 1.01-2.20]), whereas an inverse association was observed for BC-specific mortality (HR=0.55 [CI 0.31-0.97]) and recurrence risk (HR= 0.60 [CI 0.37-0.98]) among women with high estradiol levels. Heterogeneity was observed by tamoxifen use.

Conclusions

This study provides novel data on circulating 27-HC, 25-HC and breast cancer prognosis. Associations differ by estradiol levels and endocrine therapy. Further studies on circulating and tissue-based oxysterols are warranted.

Legal entity responsible for the study

The authors.

Funding

U.S. Army Medical Research and Development Command, Fort Detrick, Maryland 21702-5012.

Disclosure

All authors have declared no conflicts of interest.

Background

Angiogenesis is associated with poor survival. However, previous clinical studies assessing anti-angiogenesis methods demonstrated no clear overall survival benefits due to the lack of appropriate biomarkers. Therefore, detecting biomarkers that identify patients who might benefit from targeted anti-angiogenesis agents may help determine which patients would benefit from these therapies. This study aimed to identify biomarkers that reflect tumor angiogenesis and serve as prognostic factors.

Methods

Patients with stage I-III breast cancer who completed the planned treatment were assessed. Data were retrospectively collected from the Wonju Severance Christian Hospital database of Yonsei University and the Korean National Cancer Center database.

Results

A total of 534 patients were enrolled. Patients were divided into two groups based on the cut-off value, 31.4% of the proportional ratio between the maximum Hounsfield unit (HU) of the tumor and maximum HU of the aortic arch (maximum tumor-aorta ratio, TAR). The Kaplan-Meier curve and log-rank test revealed that the high TAR group exhibited significantly worse distant recurrence-free survival rates (p=0.001). The Cox proportional hazard model indicated that age, ER negativity, and high TAR were significant risk factors for distant relapse.

Conclusions

TAR measured by computed tomography before treatment is a potential prognostic factor for overall and distant relapse-free survival in patients with breast cancer. TAR is a potential biomarker for patients who will benefit from anti-angiogenesis agents.

Legal entity responsible for the study

The authors.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

Background

Ki-67 a cellular proliferation marker is used in breast cancer (BC) to guide therapy decision and predict prognosis in some clinical cases. It could signify treatment responsiveness/ resistance before, during and after neoadjuvant therapy. However, despite having potential applicability Ki-67 has still not been completely integrated as a standard point of care tool in clinical decision making or routine pathological reporting. Neutrophil to lymphocyte ratio (NLR) has emerged as a potential prognostic factor in various cancers, including breast cancer. Neutrophil: Lymphocyte roughly translates as tumor: immunity and this ratio in fact becomes more significant with polarization state of neutrophils and recognition of pro-tumor N2 neutrophils.

Methods

73 histopathologically diagnosed cases of BC were included in this study, their pre-treatment NLR was noted and Ki-67 expression graded. NLR was calculated from 2 ml peripheral blood as a ratio of absolute neutrophil count to absolute lymphocyte count and was categorized as low < 3 or high ≥ 3. Ki-67 immunohistochemistry was visually graded as low ≤ 5%, intermediate 6-29% and high ≥ 30%. Various clinical and histopathological characteristics of these patients were also noted. Statistical tests were performed using SPSS version 25.0, p value of < 0.05 was taken as significant.

Results

The Ki-67 immunoexpression showed a statistically significant correlation (p < 0.05) with tumor grade (modified SBR grading), tumor size, lymph node status, and clinical stage. Correlation of NLR with tumor SBR grade turned out to be non-significant (p > 0.05), however, NLR showed a statistically significant correlation with tumor size, lymph node status, and clinical stage (p< 0.05). Amongst themselves, the peripheral NLR and tumor Ki-67 immunoexpression showed a statistically significant correlation (p< 0.05).

Conclusions

The tumor proliferative potential and its subsequent therapeutic and prognostic implications as deducted from the Ki-67 index could very well be signified by peripheral NLR. Baseline NLR and its dynamics in relation to breast tumor biology could serve as a minimally invasive personalized marker for early diagnosis, guiding therapy, prognosis and follow-up in clinical settings.

Legal entity responsible for the study

The authors.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

Background

Patients with Breast Cancer (BC) often display elevated levels of circulating neutrophils (Ns). Evidence from mouse models of Early Breast Cancer (EBC) shows that Ns can either have a tumour promoting or inhibitory role. CD62L (L-selectin) is a cell adhesion molecule found on the surface of Ns which helps them bind to the endothelium and migrate into tissues and is also rapidly shed upon N activation. There is limited knowledge whether different circulatory N subsets are present in patients with BC and if this is influenced by BC subtype. In order to address these questions, we phenotyped Ns from BC patients.

Methods

We investigated changes in CD62L expression via flow cytometry in Ns from patients and healthy volunteers (HVs). Since kinases govern many aspects of intracellular signalling, we also assessed the kinase activity (n = 340 kinases) within Ns using a Pamgene kinase assay. We recruited 44 patients with EBC (14 Node +, 30 node -; 10 T1 and 26 T2 and 8 T3) and paired HVs (age matched) to compare the phenotype of Ns isolated from blood. We recruited patients 15 with oestrogen receptor-positive (ER+) and HER2- BC, 7 patients with ER + HER2 – BC, 8 patients with ER- HER2- BC and 6 patients with triple-negative BC between June 2018 and June 2021. No patient had overt metastatic disease and all were screened for exclusion factors.

Results

ER+ patients had lower level of CD62L^low N subsets compared to HVs (P< 0.0019). There was a significant increase in CD62L^low N subsets (2-fold higher) for ER- patients compared to HVs (P< 0.0331). There was a general upregulation in neutrophil kinase activity for patients with ER+ HER2 – BC compared to paired HVs. The presence of HER2+ resulted in a general downregulation in the tyrosine kinase neutrophil activity for BC patients (regardless of ER status) compared to paired HVs. The functional implications of these differences in kinase activity are being actively investigated.

Conclusions

We have detected changes in CD62L expression and kinase activity within circulatory Ns in patients with EBC and the nature of these changes appear to be intrinsically linked to ER subtype. These findings may have important implications for use as part of an early diagnostic multi-omic platform.

Legal entity responsible for the study

Imperial College London.

Funding

Medical Research Council.

Disclosure

All authors have declared no conflicts of interest.

Background

PD-L1 expression has shown to be predictive of response to immunotherapy in patients with metastatic triple-negative breast cancer (TNBC). It is unclear if PD-L1 expression by currently available validated assays has predictive capacity for response to chemotherapy in patients with non-metastatic TNBC.

Methods

We conducted a systematic review and meta-analysis of clinical studies to assess the PD-L1 expression as a predictor of response to chemotherapy in non-metastatic TNBC using validated assays. The primary endpoint was pathological complete response (pCR) rate to neoadjuvant chemotherapy. Secondary endpoints included the prevalence of PD-L1 expression and its impact on disease-free survival (DFS) and overall survival (OS). Moreover, RNA sequence data from the TCGA breast cancer cohort was used to define the relationship between PDCD1 and response to chemotherapy and prognosis.

Results

Nineteen studies were eligible for the meta-analysis with a total of 2319 patients with non-metastatic TNBC disease. The PD-L1-positive cohort had a significantly higher probability of achieving pCR with neoadjuvant chemotherapy (pooled OR =1.95; 95% CI: 1.39-2.73, p <0.0001). In studies which reported long-term outcomes, PD-L1 positivity was associated with better DFS and OS compared to PD-L1-negative patients (pooled HR= 0.51; 95% CI: 0.35-0.74, p< 0.0001 and pooled HR= 0.51; 95% CI: 0.27-0.94, p=0.031, respectively). RNA sequence data suggested that PD-L1 expression is a surrogate marker for the upregulation of key immune-related genes that mediate response to chemotherapy in TNBC.

Conclusions

This meta-analysis clearly shows that patients with PD-L1-negative TNBC respond less to neoadjuvant chemotherapy and are associated with poorer survival compared to patients with PD-L1-positive tumors. The newly distinct quadruple-negative breast cancer (QNBC) subtype could be considered the BC subtype with the poorest outcome.

Legal entity responsible for the study

The authors.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

Background

Several Breast Cancer Multigene Signatures (BCMS) are available to profile early breast cancer (eBC) but knowledge regarding their use in clinical practice is scarce. PROCURE Project preliminary results were presented at ESMO Breast Virtual Congress 2021 (Abstract 549). Final results regarding the panelist opinion on the utility of BCMS in treatment decision making for different eBC patient profiles are now available.

Methods

The Delphi questionnaire developed by the Scientific Committee was administered twice to 133 experts across 11 European countries. The questionnaire included 5 sections: 1) Panelists’ profile and experience with BCMS, 2) Current clinical practice in eBC and use of BCMS, 3) Panelists’ opinion on the utility of the BCMS in eBC according to patient profiles, 4) Agreement with a set of recommendations on the use of BCMS in clinical practice and 5) Identification of unmet needs and future applications of BCMS. 70% agreement was used to determine consensus on a topic.

Results

Panelists agreed on the clinical utility of BC molecular intrinsic subtypes for prognosis or residual risk of recurrence with standard of care in eBC HR+ (76%) and to identify a group of patients that can safely avoid chemotherapy (75%). Additionally, panelist agreed on the importance of BCMS prognostic results when deciding chemotherapy in the adjuvant setting in eBC patients with node negative (88%) or with 1 to 3 positive lymph nodes (75%). Regarding BCMS use in different patient profiles, panelists agreed on their utility in post-menopausal eBC patients (90%) and on the absence of utility in the metastatic setting (74%), in eBC patients with an HER2 overexpressed profile (82%) and in eBC triple negative (TN) (74%). Also, just 27% of the panelist considered that BCMS can be useful in the neoadjuvant setting.

Conclusions

The low percentage of panelists performing BCMS in the neoadjuvant setting, the misconception regarding their predictive value on chemotherapy benefits and the fact that some experts consider BCMS useful in TN, HER2+ eBC or in the metastatic setting show that there is a need of education on how to interpret BCMS results.

Legal entity responsible for the study

Veracyte Inc.

Funding

Veracyte Inc.

Disclosure

G. Curigliano: Financial Interests, Personal, Invited Speaker: Roche; Financial Interests, Personal, Advisory Board: Ellipsis; Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Invited Speaker: AstraZeneca; Financial Interests, Personal, Advisory Board: AstraZeneca; Financial Interests, Personal, Invited Speaker: Daiichi Sankyo; Financial Interests, Personal, Advisory Board: Daiichi Sankyo; Financial Interests, Personal, Invited Speaker: Novartis; Financial Interests, Personal, Advisory Board: Lilly; Financial Interests, Personal, Invited Speaker: Pfizer; Financial Interests, Personal, Advisory Board: Pfizer; Financial Interests, Personal, Invited Speaker: Pfizer; Financial Interests, Personal, Advisory Board: Veracyte; Financial Interests, Personal, Advisory Board: BMS; Financial Interests, Personal, Advisory Board: Merck; Financial Interests, Personal, Advisory Board: Exact Sciences; Financial Interests, Institutional, Research Grant, Investigator Initiated Trial: Merck; Financial Interests, Institutional, Funding, Phase I studies: BMS; Financial Interests, Institutional, Funding, Phase I studies: Novartis; Financial Interests, Institutional, Funding, Phase I studies: AstraZeneca; Financial Interests, Institutional, Funding, Phase I studies: Daiichi Sankyo; Financial Interests, Institutional, Funding, Phase I studies: Roche; Financial Interests, Institutional, Funding, Phase I studies: Blueprint Medicine; Financial Interests, Institutional, Funding, Phase I studies: Kymab; Financial Interests, Institutional, Funding, Phase I studies: Astellas; Financial Interests, Institutional, Funding, Phase I studies: Sanofi; Financial Interests, Institutional, Funding, Phase I studies: Philogen; Non-Financial Interests, Officer, Italian National Health Council as Advisor for Ministry of Health: Consiglio Superiore di Sanità; Non-Financial Interests, Advisory Role, Member of the Scientific Council. Patient advocacy association: Europa Donna; Non-Financial Interests, Advisory Role, Cancer Research Foundation: Fondazione Beretta; Non-Financial Interests, Invited Speaker, No compensation for this role. This a public national company for cancer prevention: Lega Italiana Lotta ai Tumori. F. Cardoso: Financial Interests, Personal, Other, Consultancy: Amgen; Financial Interests, Personal, Other, Consultancy: Astellas/Medivation; Financial Interests, Personal, Other, Consultancy: AstraZeneca; Financial Interests, Personal, Other, Consultancy: Celgene; Financial Interests, Personal, Other, Consultancy: Daiichi Sankyo; Financial Interests, Personal, Other, Consultancy: Eisai; Financial Interests, Personal, Other, Consultancy: GE Oncology; Financial Interests, Personal, Other, Consultancy: Genentech; Financial Interests, Personal, Other, Consultancy: GlaxoSmithKline; Financial Interests, Personal, Other, Consultancy: Macrogenics; Financial Interests, Personal, Other, Consultancy: Medscape; Financial Interests, Personal, Other, Consultancy: Merck-Sharp; Financial Interests, Personal, Other, Consultancy: Merus BV; Financial Interests, Personal, Other, Consultancy: Mylan; Financial Interests, Personal, Other, Consultancy: Mundipharma; Financial Interests, Personal, Other, Consultancy: Novartis; Financial Interests, Personal, Other, Consultancy: Pfizer; Financial Interests, Personal, Other, Consultancy: Pierre Fabre; Financial Interests, Personal, Other, Consultancy: prIME Oncology; Financial Interests, Personal, Other, Consultancy: Roche; Financial Interests, Personal, Other, Consultancy: Sanofi; Financial Interests, Personal, Other, Consultancy: Samsung Bioepis; Financial Interests, Personal, Other, Consultancy: Seagen; Financial Interests, Personal, Other, Consultancy: Teva; Financial Interests, Institutional, Invited Speaker: Amgen; Financial Interests, Institutional, Invited Speaker: AstraZeneca; Financial Interests, Institutional, Invited Speaker: Boehringer-Ingelheim; Financial Interests, Institutional, Invited Speaker: Bristol Myers Squibb; Financial Interests, Institutional, Invited Speaker: Bayer; Financial Interests, Institutional, Invited Speaker: Daiichi; Financial Interests, Institutional, Invited Speaker: Eisai; Financial Interests, Institutional, Invited Speaker: Fresenius GmbH; Financial Interests, Institutional, Invited Speaker: Genentech; Financial Interests, Institutional, Invited Speaker: GlaxoSmithKline; Financial Interests, Institutional, Invited Speaker: Ipsen; Financial Interests, Institutional, Invited Speaker: Incyte; Financial Interests, Institutional, Invited Speaker: Nektar Therapeutics; Financial Interests, Institutional, Invited Speaker: Nerviano; Financial Interests, Institutional, Invited Speaker: Novartis; Financial Interests, Institutional, Invited Speaker: Macrogenics; Financial Interests, Institutional, Invited Speaker: Medigene; Financial Interests, Institutional, Invited Speaker: MedImmune; Financial Interests, Institutional, Invited Speaker: Merck; Financial Interests, Institutional, Invited Speaker: Millenium; Financial Interests, Institutional, Invited Speaker: Pfizer; Financial Interests, Institutional, Invited Speaker: Pierre Fabre; Financial Interests, Institutional, Invited Speaker: Roche; Financial Interests, Institutional, Invited Speaker: Sanofi-Aventis; Financial Interests, Institutional, Invited Speaker: Sonus; Financial Interests, Institutional, Invited Speaker: Taiho Oncology; Financial Interests, Institutional, Invited Speaker: Tesaro; Financial Interests, Institutional, Invited Speaker: Tigris; Financial Interests, Institutional, Invited Speaker: Wilex; Financial Interests, Institutional, Invited Speaker: Wyeth; Non-Financial Interests, Leadership Role, President: ABC Global Alliance and ABC Consensus Conference and Guidelines; Non-Financial Interests, Member: ESMO; Non-Financial Interests, Member: ESO; Non-Financial Interests, Member: EORTC; Non-Financial Interests, Member: BCG; Non-Financial Interests, Member: IBCSG; Non-Financial Interests, Member: SOLTI; Non-Financial Interests, Member: ASCO; Non-Financial Interests, Member: AACR; Non-Financial Interests, Member: EACR; Non-Financial Interests, Member: SIS; Non-Financial Interests, Member: ASPIC. M.I. Gnant: Financial Interests, Personal, Invited Speaker: Amgen; Financial Interests, Personal, Advisory Board: Daiichi Sankyo; Financial Interests, Personal, Invited Speaker: AstraZeneca; Financial Interests, Personal, Advisory Board: Eli Lilly; Financial Interests, Personal, Other, Consulting on scientific matters: LifeBrain; Financial Interests, Personal, Expert Testimony: Veracyte; Financial Interests, Personal, Invited Speaker: Novartis; Financial Interests, Personal, Invited Speaker: Pierre Fabre; Financial Interests, Personal, Full or part-time Employment: ABCSG GmbH; Financial Interests, Invited Speaker: Pfizer; Other, Spouse is employed by: Sandoz. N. Harbeck: Financial Interests, Invited Speaker: Amgen; Financial Interests, Invited Speaker: AstraZeneca; Financial Interests, Invited Speaker: Daiichi Sankyo; Financial Interests, Invited Speaker: Exact Sciences; Financial Interests, Invited Speaker: Gilead; Financial Interests, Invited Speaker: Lilly; Financial Interests, Invited Speaker: MSD; Financial Interests, Invited Speaker: Novartis; Financial Interests, Invited Speaker Pierre Fabre; Financial Interests, Invited Speaker: Pfizer; Financial Interests, Invited Speaker: Roches; Financial Interests, Invited Speaker: Seagen; Financial Interests, Advisory Board: AstraZeneca; Financial Interests, Advisory Board: Daiichi Sankyo; Financial Interests, Advisory Board: Gilead; Financial Interests, Advisory Board: Lilly; Financial Interests, Advisory Board: MSD; Financial Interests, Advisory Board: Novartis; Financial Interests, Advisory Board: Pfizer; Financial Interests, Advisory Board: Roche; Financial Interests, Advisory Board: Sandoz; Financial Interests, Advisory Board: Seagen. J. King: Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Advisory Board: Pfizer; Financial Interests, Personal, Advisory Board: AstraZeneca; Financial Interests, Personal, Advisory Board: Gilead; Financial Interests, Personal, Advisory Board: Lilly; Financial Interests, Personal, Speaker’s Bureau: Pfizer; Financial Interests, Personal, Speaker’s Bureau: Roche; Financial Interests, Personal, Speaker’s Bureau: Seagen; Financial Interests, Personal, Speaker’s Bureau: Novartis. A. Laenkholm: Financial Interests, Advisory Board: Merck; Financial Interests, Advisory Board: Agilent; Financial Interests, Funding: Novartis; Financial Interests, Funding: Roche. F. Penault-Llorca: Financial Interests, Funding: Veracyte; Financial Interests, Funding: Myriad; Financial Interests, Funding: Illumina; Financial Interests, Advisory Board: Veracyte; Financial Interests, Advisory Board: Exact Science; Financial Interests, Advisory Board: Myriad; Financial Interests, Advisory Board: Agendia; Financial Interests, Advisory Board: Illumina; Financial Interests, Other: Myriad; Financial Interests, Other: Veracyte. A. Prat: Financial Interests, Personal, Invited Speaker: Roche; Financial Interests, Institutional, Other, Contracted research:: Roche; Financial Interests, Personal, Invited Speaker, Lecture fees: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Novartis; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Novartis; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Amgen; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Amgen; Financial Interests, Personal, Invited Speaker, Lecture fees: BMS; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: BMS; Financial Interests, Personal, Invited Speaker, Lecture fees: NanoString; Financial Interests, Institutional, Other, Contracted research: NanoString; Financial Interests, Institutional, Invited Speaker, Lecture fees: NanoString; Financial Interests, Institutional, Other, Contracted research: Novartis; Financial Interests, Institutional, Other, Contracted research: Roche; Financial Interests, Institutional, Other, Contracted research: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Contracted research; Financial Interests, Personal, Invited Speaker, Lecture fees: Daiichi Sankyo; Financial Interests, Institutional, Invited Speaker, Clinical trials: Daiichi Sankyo; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Puma; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Oncolytics; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: MSD; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Guardan Health; Financial Interests, Personal, Expert Testimony, Advisory role/consultancy: Peptomyc; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Lilly; Financial Interests, Institutional, Other, Clinical trials: Lilly; Financial Interests, Institutional, Other, Contracted research: Boehringer; Financial Interests, Institutional, Other, Contracted research: Sysmex Europa GmbH; Financial Interests, Institutional, Other, Contracted research: Medica Scientia inno. Research; Financial Interests, Institutional, Other, Contracted research: Celgene; Financial Interests, Institutional, Other, Contracted research: Astellas; Financial Interests, Institutional, Other, Clinical trials: Roche; Financial Interests, Institutional, Other, Clinical trials: Amgen; Financial Interests, Personal, Invited Speaker, Leadership role: Reveal Genomics, SL.; Financial Interests, Institutional, Other, Clinical trials: Boehringer; Non-Financial Interests, Institutional, Other, Leadership roles: Patronage committee: SOLTI Foundation; Non-Financial Interests, Institutional, Other, Leadership roles: Patronage committee: Actitud Frente al Cáncer Foundation.

Background

Platforms that enable multiplexed analysis of biomarkers from limited amounts of poor-quality material provide value to studies of archival tissue material. The NanoString nCounter® technology allows nonamplified measurements of up to 800 targets within one sample of archived formalin fixed paraffin embedded (FFPE) material. We aimed to compare gene expression in young vs old breast cancer (BC) patients, using archived FFPE material to gain insight into BC biology of the young.

Methods

The NanoString BC360 panel was explored by NanoString nCounter® to investigate gene expression in an in-house BC FFPE archive cohort (diagnosed 1996-2003; age <50 years at diagnosis; n= 129). Data were analysed in nSolver and ROSALIND. Protein-protein interaction (PPI) networks were explored. The Cancer Genome Atlas global gene expression BC data (n=520) was analyzed for validation.

Results

We identified 58 differentially expressed genes (DEGs) in BC samples from young (aged <40) and older patients. In PPI networks, representing the DEGs, we identified highly interconnected regions (sub-clusters), pointing to DEGs indicating increased immune infiltration and inflammatory mechanisms, as well as oncogenic signaling favoring tumor promotion, in BC of the young. Validation in TCGA BC data demonstrated gene expression patterns reflecting overall increased T-cell activation and higher levels of T regulatory immune cells and PD-1 in BC of the young (P<0.04).

Conclusions

The current study indicates increased immune cell activation supporting an immune evasive environment in young BC. Also, higher oncogenic signaling in this patient subset supports increased tumor-promoting processes.

Legal entity responsible for the study

The authors.

Funding

Helse Vest and University of Bergen.

Disclosure

All authors have declared no conflicts of interest.

Background

Triple-negative breast cancer (TNBC) is the most aggressive breast cancer sub-type accounting for nearly 15% of all invasive breast cancer (BC) cases worldwide. Lack of a well-established membrane receptor limits the therapeutic options for this disease. Therefore, identification of a novel receptor can make a paradigm shift in the treatment of TNBC. Recent studies have shown that SLC22A17, a cell surface receptor that is involved in iron transport plays an important role in the pathogenesis of various cancers. Therefore, the current study was aimed at evaluating the role of this protein and its potential as a therapeutic target in TNBC.

Methods

Bioinformatics tools were used to investigate the role of this protein in TNBC data sets. Immunohistochemistry was performed to analyze the expression levels of SLC22A17 in different BC subtypes. In addition, expression of SLC22A17 and signaling proteins including Akt/mTOR, JAK/STAT in TNBC cells were examined by immunoblotting. Moreover, the siRNA-mediated knockdown of SLC22A17 followed by proliferation, clonogenic, wound healing, immunoblotting, and annexin V apoptotic assays were performed to unravel the role of SLC22A17 in TNBCs.

Results

The present study showed that SLC22A17 was significantly overexpressed in TNBC tissues and cells and inversely correlated with the survival of patients. In addition, the expression of this protein was correlated with different stages of TNBC. Moreover, knockdown of SLC22A17 resulted in reduced proliferation, migration, increased apoptosis, and autophagy by inhibiting Akt/mTOR and STAT3 signaling pathways in MDA-MB-231 and MDA-MB-468 cells. Further, the silencing of this protein inhibited epithelial-mesenchymal transition of TNBC cells. Furthermore, the treatment of proinflammatory cytokine TNF-α was shown to induce the expression of this protein in TNBC cell lines and regulated different hallmarks, while silencing of this protein suppressed these processes.

Conclusions

Overall the current study showed that SLC22A17 plays a major role in TNBC by altering Akt/mTOR/STAT3 pathway and can be used as a novel therapeutic target for this disease.

Legal entity responsible for the study

The authors.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

Background

Tumour infiltrating lymphocytes (TILs) in triple-negative and HER2-positive breast cancer has been associated in higher responses to treatment and potentially a prognostic marker. Hormone-positive, HER2-negative (ER+/HER2-) breast cancer is considered a less immunogenic tumour and the role of TILs in this cohort is less clear, partly due to the lack of data. We aimed to assess overall number of CD45-positive tumour-infiltrating lymphocytes in early-stage, ER+/HER2- breast cancer and its relationship with proliferation index, grade, menopausal status and the 21-gene recurrence score.

Methods

Archival patient tissue samples of early-stage, node negatie patients with ER+/HER2- breast cancer were used with associated clinical data and 21-gene recurrence score. Staining was completed via the automated Leica Bond III system with anti-CD45 antibody (DAKA clone M0701) on full face sections. QuPath, an open-source digital pathology software was used to detect and calculate the percentage of overall CD45+ cells (CD45%) within the tumour area. CD45% was further correlated with proliferation index, grade, and 21-gene recurrence score.

Results

A total of 409 samples were included in the final analysis. The mean CD45% for the entire cohort was 5.96, with samples ranging from 0.13 to 78.86. Statistically higher levels of CD45-positive lymphocytes were associated with higher grade (p value = 0.00011) and high 21-gene recurrence score (p value= 0.0015). The correlation with Ki67 proliferation index was less clear, with higher CD45% in the subset with Ki67 >20% (p value= 0.0014) however linear correlation not being evident. No statistically significant difference was seen in regards to menopausal status. A trend toward worse disease free survival was seen in those with a higher CD45% in the whole cohort however did not reach statistical significance (p value = 0.11).

Conclusions

Higher TILs in early stage ER+/HER2- breast cancer seem to be associated with higher grade and higher 21-gene-recurrence score. The impact on recurrence and response to treatment remains In this cohort remains unclear. Further research into spatial distribution and immune phenotypes of TILs is needed.

Legal entity responsible for the study

RCSI.

Funding

RCSI.

Disclosure

W.M. Gallagher: Financial Interests, Personal, Stocks/Shares: OncoMark; Financial Interests, Personal, Stocks/Shares: OncoAssure; Financial Interests, Personal, Leadership Role: OncoMark; Financial Interests, Personal, Leadership Role: OncoAssure. All other authors have declared no conflicts of interest.

Background

Cancer can be defined as cellular mutation and uncontrolled cell division. Many genes are involved in during the process of cell mutation with multifactorial etiology. The genomic biomarkers are the key to investigate and recognize the process behind mutational changes in the genomes. Development of breast cancer develops due to uncontrolled growth of alveolar cell in mammary gland. Genomic mutation investigations helped us to understand breast cancer genetics and early detection of disease progression. Current study was design to analyze the HOXB13 and SRC gene involvement in disease progression and metastization of breast cancer in females. SRC plays role in regulation of phosphorylate RNA binding proteins and pre mRNA processing and HOXB13 gene that belongs to transcription factors protein family that produces and attaches protein to DNA.

Methods

Breast cancer samples were arranged from PINUM, DHQ and Allied hospital. The biopsy samples were preserved in 10% formalin for histopathology and in RNA Later to extract the mRNA. RNA was isolated in TRIZOL. To check gene expression analysis qRT-PCR was performed.

Results

The results revealed that Beta catenin signaling cascade and micro-RNA’s crosstalk associated with SRC and HOXB13 gene expression in female breast cancer. Results revealed that SRC and HOXB13 gene was significantly (p<0.05) highly expressed in female breast cancer associated with microRNA (microRNA-302, microRNA-238 and microRNA-145) expression level of was also analyzed and results revealed that microRNA was significantly (p<0.05) higher expressed in female breast cancer. Histopathological examination showed multi-layering, hyperplasia, complete distortion of ductal and glandular epithelial of the breast gland in the female breast cancer patients.

Conclusions

Overall results concluded that microRNA (microRNA-302, microRNA-238 and microRNA-145) also key role in the caner development and progression. The expression of SRC and HOXB13 gene is associated with microRNA. Corresponding author contact detail: aizakamalkhan92@gmail.com; Co-corresponding author contact detail: m.naeem.faisal@uaf.edu.pk.

Legal entity responsible for the study

The authors. Institute of Physiology and Pharmacology, University of Agriculture Faisalabad, Pakistan.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

Background

The search for effective therapeutic targets for the highly aggressive triple-negative breast cancer (TNBC) subtype is critical. Connexins are channel-forming transmembrane proteins with both gap junction-dependent and independent functions.

Methods

Immuno-histochemistry/fluorescence, western blot, qPCR, flow cytometry, parachute assay, proliferation and migration assays.

Results

Large-scale immunohistochemical staining of connexin43 (Cx43) showed TNBC as the breast cancer subtype with the lowest levels. Cx43 restoration in TNBC primary and metastatic cell lines partially restituted gap junction intercellular communication (GJIC), partially reverted EMT, increased SASP (senescence-associated secretory phenotype) production, reduced 2D and 3D-spheroid proliferation, diminished 2D and 3D-spheroid migration and colony formation and resensitized TNBC cells to Anoikis. These results were validated in physiologically relevant 3D heterospheroid models, comprising TNBC cells and patient-derived ACTA2+FAP+ITGB1+ breast cancer-associated fibroblasts (CAFs). Cx43 restoration in TNBC cells significantly reduced heterospheroid proliferation and migration. These primary CAFs were characterized by an abrupt reduction of Cx43 mRNA and protein, as well as GJIC impairment, when compared to their paired normal fibroblasts (NAFs). Interestingly, upon coculture of CAFs/NAFs with TNBC cell lines, heterocellular GJIC only happened when Cx43 was restored in TNBC, unidirectionally from CAFs/NAFs to Cx43-restituted tumour cells. Furthermore, in cocultures of patient-derived, immortalized and EGFR-CAR-modified natural killer (NK) cells with the primary TNBC cell line, Cx43 restoration in tumour cells significantly boosted the antitumour NK immune response, presumably in a gap junction independent manner.

Conclusions

These results support Cx43 tumour-suppresive role in primary and metastatic TNBC cells, both in 2D and in physiologically relevant 3D homo- and hetero-spheroid models including breast CAFs. Moreover, this work also presents a novel approach to enhance NK cytotoxicity against TNBC upon Cx43 restitution.

Legal entity responsible for the study

The authors.

Funding

Ministerio de Universidades, European Molecular Biology Organization, Xunta de Galicia.

Disclosure

All authors have declared no conflicts of interest.

Background

Breast cancer patients who achieve pathological complete response (pCR) after neoadjuvant therapy are shown to have a significantly improved disease free survival. However, there is currently no way to determine which patients are most likely to respond to neoadjuvant therapy.

Growing evidence implicates tumour infiltrating lymphocytes as contributors to HER2+ breast cancer (BC) outcome. We hypothesize that a deeper investigation of the immune landscape of HER2+ patient tumours may improve patient stratification and identify an immune profile associated with pCR.

Methods

3plex immunohistochemistry (IHC) panels targeting immune cell subpopulations and tertiary lymphoid structures (TLS) were applied to HER2+ BC tumours of patients of the neoadjuvant TCHL clinical trial (NCT01485926), who received HER2-targetting therapy. Artificial intelligence models for quantitative image analysis of CD3+, CD20+, DC-Lamp+, CD4+, and CD8+ cells were trained and validated. Models were applied for analysis of multi-labelled TCHL tumour sections and patient-specific densities of each cell type in tumour regions were determined to facilitate investigation of pre-treatment differences in target immune subpopulations in responders versus non-responders.

Results

Statistical analysis was performed on 28 patient samples (pre-treatment N=19, post-treatment N=9). This demonstrated a trend for higher densities of each immune subpopulation in pre-treatment biopsies of pCR vs. non-pCR patients. Assessment of association of pre-treatment immune infiltrate with clinicopathological features demonstrated trends for higher levels of T-cell infiltration in later stage and hormone receptor-negative patients.

Conclusions

Multiplex IHC is an optimal technique for the concurrent assessment of multiple target markers in patient tissue, which can be quantitatively analysed in a high throughput manner using AI models. The findings of this preliminary study warrant further research into immune subpopulations and their impact on pCR in HER2+ BC.

Clinical trial identification

NCT01485926.

Legal entity responsible for the study

Camille Hurley.

Funding

Science Foundation Ireland, European Union, European Association for Cancer Research, European Federation of Immunological Societies.

Disclosure

All authors have declared no conflicts of interest.

Background

Biomarkers predicting response to bevacizumab (BEV) containing therapy in breast cancer are of clinical need. In a retrospective single-institution analysis we have previously shown the association of the endothelin-1 genetic polymorphism rs5370-TT with shorter overall survival (OS) and a trend towards worse progression-free survival (PFS) compared with the alternate genotypes combined. This negative predictive value was seen in metastatic breast cancer patients treated with chemotherapy plus BEV but not in patients treated with chemotherapy alone (Gampenrieder SP et al. Pharmacogenomics J. 2017. 17(4):344-350). Here, we present the validation of these findings within the prospective phase III trial TANIA (Vrdoljak E et al. Ann Oncol. 2016. 27(11):2046-52) randomizing 494 patients with HER2-negative metastatic breast cancer to chemotherapy plus BEV or chemotherapy alone for two consecutive treatment lines (2^nd- and 3^rd-line).

Methods

DNA samples from 193 patients consenting to optional translational research within the TANIA trial were analyzed retrospectively. One-hundred patients were treated with chemotherapy plus BEV and 93 patients with chemotherapy alone. DNA was isolated from archival primary or metastatic FFPE tumor samples collected before study entry. The majority of samples (88%) were from the primary tumor. SNP analysis in EDN1 was performed using TaqMan® SNP Genotyping.

Results

In the BEV-containing arm, patients with the genotype rs5370 TT did not have a significantly different 2^nd-line PFS (HR 0.99, 95%CI 0.48-2.06; P=0.99) or OS (HR 0.91, 95%CI 0.41-2.05; P=0.83), respectively, compared with the alternate genotypes combined. In addition, there were no differences in 3^rd-line PFS and in the combination of 2^nd- and 3^rd-line PFS between rs5370 TT and rs5370 GG+GT. In the control arm no association of the EDN1-genotype with outcome was seen as well, suggesting neither a predictive nor a prognostic value of the EDN1-SNP in metastatic breast cancer.

Conclusions

The SNP rs5370 in endothelin-1 was not confirmed as a predictive biomarker for BEV efficacy and should therefore not influence treatment decisions in metastatic breast cancer patients.

Legal entity responsible for the study

Richard Greil.

Funding

Roche.

Disclosure

S.P. Gampenrieder: Financial Interests, Personal, Invited Speaker: Roche; Financial Interests, Institutional, Research Grant: Roche; Financial Interests, Personal, Other, Travel grant: Roche; Financial Interests, Personal, Advisory Role: Roche. C. Hufnagl: Financial Interests, Personal, Other, Travel Grant: Roche. G. Rinnerthaler: Financial Interests, Personal, Invited Speaker: Roche; Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Institutional, Research Grant: Roche; Financial Interests, Personal, Other, Travel grant: Roche. T. Westphal: Financial Interests, Personal, Other, Travel grant: Roche. F. Huemer: Financial Interests, Personal, Other, Travel grant: Roche. C. Monzo-Fuentes: Financial Interests, Personal, Other, Travel grant: Roche. P. Morre: Financial Interests, Personal, Other, Travel grant: Roche. C. Hauser-Kronberger: Financial Interests, Personal, Other, Travel Grant: Roche. R. Greil: Financial Interests, Institutional, Research Grant: Roche; Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Other, Travel grant: Roche; Financial Interests, Personal, Invited Speaker: Roche. All other authors have declared no conflicts of interest.

Background

The exact impact of chemotherapy on the immune system of older patients with breast cancer is not well known. This longitudinal study was performed investigating the evolution of the blood immune profile during and after chemotherapy in this population.

Methods

The study included 39 patients receiving adjuvant chemotherapy (chemotherapy group, CTG, median age, 73 years) and 32 patients receiving only hormone therapy (control group, CG, median age, 76 years). A 10-gene panel associated with immunosenescence was measured in peripheral blood mononuclear cells (PBMC) before (T1), at 3 months (T2) and at 12 months (T3) after initiation of adjuvant therapy. Nutrition status was assessed by using a mini nutritional assessment scale (≥12 normal; <12 abnormal). Longitudinal mixed model analyses were performed for trajectory evolution, with or without adjusting for age, tumor stage, breast cancer phenotype, and/or corresponding baseline gene levels.

Results

Six genes relating to T cell activation (CD28, CD27, CD86, LCK, GRAP, LRRN3 ), and two genes relating to oxidative stress (PRDX6, HMOX1) exhibited a significant group-by-time effect, even after adjusting covariates (p≤ 0.01). In CTG, the T cell activation genes substantially declined from T1 to T2 and bounced back to a level higher than baseline at T3 (p<0.03), which was not observed in CG (p>0.26). Patients with malnutrition detected at T1 experienced more pronounced perturbation regarding CD27, LCK, CD69, VAMP5, and LRRN3 (p<0.05).

Conclusions

Chemotherapy leads to transient perturbation of immune-related gene expression and potentially stimulates immunity in the long term. Well-nourished patients experience less impact of chemotherapy on immune-related gene expression profiles.

Clinical trial identification

This is a sub-study of NCT00849758.

Legal entity responsible for the study

The authors.

Funding

Fonds voor Wetenschappelijk Onderzoek – Vlaanderen.

Disclosure

All authors have declared no conflicts of interest.

Background

Breast cancer (BC) is a heterogeneous disease. The expression levels of receptors (ER/PR/HER) define four phenotypically and clinically distinct BC subtypes. Population-based studies in resource-rich countries have demonstrated that more aggressive BC subtypes such as triple negative (ER-/PR-/HER-) and HER2+ are more commonly observed in younger patients. However, the incidence of BC subtypes in resource-restricted countries is less clear. The purpose of this study is to examine the distribution of the BC subtypes in young (≤35 years) versus elderly (>65 years) patients across Georgian cancer centers.

Methods

Clinical and histopathology (ER, PR and HER2 status) data was retrospectively collected for 1007 female patients diagnosed with BC in the 2013 to 2015 time period. Demographic data included age at diagnosis, ER/PR and HER2 status. Incidence rates of subtypes (triple-negative [TNBC]; HR+/HER2 -; HR+/HER2+; HR- /HER2+) were calculated to evaluation association with age.

Results

In this study, the median age of presentation was 55.8 (Range, 20 to 90 years). The incidence of BC was 20.4%, 55.4%, and 24.2% for diagnosis 45 years and under, 46 to 65 years, and over 65 years, respectively; ___% were diagnosed in women 35 years and younger. The most common BC subtype was HR+/HER2- across all age groups (67,1%); the least common was HR-/HER2+ (5.4 %). We identified TNBC in 23.5% of this sample. While no statistically significant differences in BC subtype distributions were observed across age groups, younger women had a much higher incidence of TNBC compared to elderly women (11.4 vs 5.4%, p=0.7).

Conclusions

The overall distribution of BC subtypes among young adults did not vary from that observed in other countries. However, we identified a higher incidence of TNBC, which is approximately twice higher compared to US population (23.5% vs 12.2% respectively), and as also reported, there were numerically more cases of TNBC in younger women. [JNCI J Natl Cancer Inst (2014) 106(5): dju055 doi:10.1016/jnci/dju055] Additional analysis is required to explore the subtype distributions and differences in survival in Georgia.

Legal entity responsible for the study

The authors.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

Background

Breast cancer is a highly heterogeneous disease. Prognosis and treatment are usually decided based on the expression of 3 molecular markers; one being the human epidermal growth factor receptor 2 (HER2). Classification of HER2 tumors generally follows a binary system, either HER2 positive or HER2 negative, the latter encompasses the triple-negative breast cancer (TNBC) types. However, HER2 heterogeneity to a new nomenclature: HER2-positive, HER2-low, and HER2-negative. Here we present a state-of-the art technique to facilitate the identification of HER2-low patients by capturing the spatial distribution of HER2 expression across the entire tumor. This identifies TNBC patients that express HER2-low and for whom there are no targeted therapies and who could benefit from anti-HER2 drugs.

Methods

We identified the range of HER2 expression present from microarrays run on biopsies from HER2-positive (n=48) and TNBC (n=84) patients from a subset of the ISPY1 & 2 trials and characterized the overlap in both HER2 expression and expression of HER2 signaling-responsive genes. We next integrated single cell RNA-seq data with patient-specific tumor morphology using a biophysical modeling platform, SimBioSys TumorScope, and statistical matching algorithms to characterize HER2 heterogeneity within individual patient tumors. We then visualized HER2 heterogeneity within tumors using 3D maps of HER2 expression.

Results

We identified a subset of TNBC patients (17/84) that both expressed HER2 at low levels and a previously published HER2-responsive gene expression signature. Additionally, we characterized the 3D intra-tumoral heterogeneity of HER2 expression predicted to occur within these tumors.

Conclusions

Biophysical modeling can integrate multiple large datasets together and capture tumor heterogeneity at high resolution and large scale. These advances enable a more comprehensive understanding of tumor biology and heterogeneity, and ultimately improve clinical decision making across solid tumors. In our study, this translates to the identification of a subset of TNBC patients that express HER2-low which could benefit from HER2-targeted therapy, like T-DXd which, unlike trastuzumab or T-DM1, has shown promising results in HER2-low patients.

Legal entity responsible for the study

SimBioSys, Inc.

Funding

SimBioSys, Inc.

Disclosure

D. Cook, J. Cole: Financial Interests, Personal and Institutional, Stocks/Shares, Employed by and has stock options with: SimBioSys, Inc. All other authors have declared no conflicts of interest.

Background

Glucocorticoid receptor (GR) shows structural and functional similarities with progesterone receptor (PR) and share common ligands and binding sites, but studies on crosstalk of GR with PR isoforms are lacking. We explored co-expression of GR and PR isoforms in breast cancer (BC) and its influence on prognosis.

Methods

We examined presence of PR and GR expression by immunohistochemistry (IHC) in a cohort of estrogen receptor-positive (ER+) BC patients with long term follow-up. Tumor cells showing more than 1% nuclear stain were considered positive. Tumors were grouped based on combined presence/absence of GR and PR expression. PRA protein expression was estimated by IHC (clone-hPRa7) and PRAB/PRB transcript ratio was calculated by qPCR using sequence specific primers. Tumors were categorized into high and low groups based on median cut off. Clinical characteristics, Ki-67 and overall survival (OS) was examined between tumor groups based on GR and PR expression and high and low PR isoform expression.

Results

Of the total 186 ER+ tumors, 89.25% (n=166) tumors were PR+ and 77.12% (91/118) were GR+ by IHC. Of these 68.65% (81/118) co-expressed GR and PR (GR+PR+) and 2.54% (3/118) had neither. Protein expression quantified by clone-hPRa7 correlated more with full length PR protein indicating it was not specific to PRA isoform, hence only PRB and PRAB/PRB transcript ratio were considered for further analysis. When OS was examined by co-expression of GR and PR, GR+PR+ tumors had significantly better prognosis compared to GR+PR- (p<0.0001) and GR-PR+ (p=0.008) tumors. GR+PR- tumors had higher grade, Ki-67 and PRAB/PRB ratio indicating higher expression of PRA isoform (p<0.05). Further, examination of co-expression of GR with PR isoforms showed GR+PRB high tumors had better OS than GR+PRB low (p=0.028), GR-PRB high (p=0.013) and GR-PRB low (p=0.001) tumors. On the contrary, GR+PRA low tumors had better prognosis suggestive of differential crosstalk of GR with PR isoforms.

Conclusions

Though PR and GR expression is associated with better prognosis, our results indicate GR expression with specific PR isoform influences prognosis within ER+ tumors. Understanding this crosstalk may help in developing better treatment approaches.

Legal entity responsible for the study

The authors.

Funding

DBT/Wellcome Trust India Alliance.

Disclosure

All authors have declared no conflicts of interest.

Background

Tumor immune microenvironment is currently recognized as an important participant in carcinogenesis, influencing the behavior of tumor cells and treatment efficacy. It is suggested that defects in BRCA1/2 genes contribute to a high mutation load and high immunogenicity, which modulates immune microenvironment. At the same time, it was shown that BRCA1/2-associated breast cancer tumors do not belong to the category of immunoactive ones. In addition, there is evidence of the additional evaluation of TP53 mutation in these tumors and disruption of the cell death process, which can also be a factor of resistance to therapy.

Methods

40 patients with breast cancer with BRCA1 mutations 185delAG, 4153delA, 5382insC, 3819DELGTAA, 3875delGTCT, 300T>G, 2080delA and BRCA2 (6174delT)) determined by real-time polymerase chain reaction (PCR) were included. Immunohistochemical study was performed using monoclonal antibodies to Ki67, p53, CD8, CD4, CD68, CD163, BCL2.

Results

In the presented study patients with BRCA1-associated breast cancer, had high low tumor CD4/CD8 ratio in 50% of cases, which characterizes an immunosuppressive microenvironment. Mutation BRCA1 5382insC is associated with high level of CD4+ T-lymphocytes (p˂0.05), G3 is associated with a low CD4/CD8 ratio (p=0.056) and a low levels of CD68+M1, CD163+M2-macrophages (p=0,022, p=0.002); T1 correlates with high levels of CD8+ TILs (p=0.038) and high levels of CD163 (p=0.033). High Ki-67 is associated with a lack of BCL2 expression (p=0.04). Negative expression of BCL2 occurred in 52.5% of cases. A high level of p53 expression is described as the main type of expression in these tumors, suggesting a combination of TB53 and BRCA1 mutations and a violation of the mechanism of cell death in these patients.

Conclusions

The immunosuppressive type of microenvironment prevails in the tumors of breast cancer patients with hereditary mutations in the BRCA1 gene, where a violation of the mechanism of cell death in BRCA1-deficient breast cancer was revealed. The main directions of future therapy of these tumors may be modification of the immune microenvironment and activation of the mechanisms of cell death.

Legal entity responsible for the study

Stukan Anastasia.

Funding

Has not received any funding.

Disclosure

The author has declared no conflicts of interest.

Background

Seroma formation (SF) is one of the most frequent complications after breast cancer (BC) surgery. It occurs most frequently in patients after mastectomy (ME). The aim of this trial was to study (auto)immunological processes as an effect for SF. Identification and characterization of immune cell populations as well as specific immune reactions found in seroma and peripheral blood in patients after ME were therefore the main points of interest. Sera of healthy controls (without seroma and breast cancer) were chosen for comparison.

Methods

In a subgroup analysis within a monocentric pilot phase of 16 BC patients who developed seroma after a ME flow cytometry analyses were performed to identify the general immune cell content in seroma fluid. The latter is measured in comparison to peripheral blood of the same patient and in contrast to healthy controls. In addition, subgroups of specific T-cell immune responses were specified.

Results

The population of B-cells, granulocytes and cytotoxic (CD3+/CD8+) T-cells were significantly reduced within the seroma compared to the blood of the patients and the blood of healthy controls. In contrast specific T helper-cells (Th2 and Th17) were significantly elevated in the seroma as well as in patient blood compared to blood results of healthy controls. Th22 showed significant higher values in seroma compared to blood results of healthy controls and of patient blood. Tregs were significantly reduced in blood and seroma fluid of patients compared to healthy controls.

Conclusions

We were able to detect a specific immune response in BC patients with SF. A general upregulation of the T-helper cell population was identified, whereas other immune cells decreased. These findings indicate a high inflammatory process with diminished protective regulatory responses through Tregs. Further investigations are needed to clarify the role of specific T-helper cell immune responses for a SF after BC surgery.

Legal entity responsible for the study

The authors.

Funding

intramural grants (university hospital Augsburg, Germnay).

Disclosure

N. Ditsch: Financial Interests, Personal, Advisory Board: MSD; Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Advisory Board: Novartis; Financial Interests, Personal, Advisory Board: Pfizer; Financial Interests, Personal, Advisory Board: Daiichi Sankyo; Financial Interests, Personal, Invited Speaker: AstraZeneca; Financial Interests, Personal, Invited Speaker: TEVA. J. Sagasser: Financial Interests, Personal, Invited Speaker: Roche; Financial Interests, Personal, Invited Speaker: Novartis; Financial Interests, Personal, Invited Speaker: AstraZeneca; Financial Interests, Personal, Invited Speaker: BMS; Financial Interests, Personal, Invited Speaker: Pfizer. T. Kühn: Financial Interests, Institutional, Funding: Novartis; Financial Interests, Personal, Invited Speaker: Celegene; Financial Interests, Personal, Invited Speaker: Roche; Financial Interests, Personal, Invited Speaker: Pfizer. C. Dannecker: Financial Interests, Personal, Invited Speaker: Roche; Financial Interests, Personal, Invited Speaker: Novartis; Financial Interests, Personal, Invited Speaker: Pfizer; Financial Interests, Personal, Invited Speaker: MSD; Financial Interests, Personal, Invited Speaker: AstraZeneca. All other authors have declared no conflicts of interest.

Background

HER2-low (IHQ score 1+ or 2+ with negative in situ hybridization) has emerged as a new potential biomarker in metastatic breast cancer. However, its prognostic value in early-stage carcinomas needs to be revisited. We aimed to evaluate the association of HER2-low carcinomas with PAM50 risk groups combined with standard clinicopathological variables in early breast cancer.

Methods

We carried out a retrospective analysis of 332 early-stage breast cancer patients from Hospital Universitario 12 de Octubre (Madrid, Spain) with PAM50 signature performed between 2015-2021. Clinical variables were collected from medical records. We estimated adjusted Odds Ratio and 95% confidence interval for high-risk PAM50 subgroup comparing HER2-low vs HER2-zero carcinomas by multivariable logistic regression. P values below 0.05 were deemed statistically significant.

Results

In our sample, 192 (57%) patients were classified as HER2-low carcinomas (Table). The mean age was 57 years and 70% were postmenopausal. The adjusted OR for high-risk PAM50 subgroup when comparing HER2-low vs HER2-zero carcinomas was 1.31 (95% CI: 0.75-2.30, p=0.33). We found significant associations for ki67 (≥20% vs <20%: OR= 4.03, 95%CI: 2.15-7.56, p<0.001), T stage (T2/T3 vs T1: OR= 3.44, 95%CI: 1.96-6.04, p<0.001), progesterone receptor (PR≥20% vs <20%: OR=0.44, 95%CI: 0.23-0.83, p=0.01), nodal stage (N+ vs N0: OR=3.8, 95%CI 1.89-7.62, p<0.001), and histological grade (grade 2 vs grade 1: OR=2.41, 95%CI: 1.01-5.73, p=0.04; grade 3 vs grade 1: OR=5.40, 95%CI: 1.98-14.60, p=0.001). We found no association with nodal micrometastasis (Nmic vs N0: OR=0.61, 95%CI: 0.24-1.47, p=0.27), and lobular histology (lobular vs ductal: OR=0.54, 95%CI: 0.25-1.15, p=0.11). Table: 49P

Conclusions

In this early-stage breast cancer cohort, HER2-low was not associated with the high-risk PAM50 group compared to HER2-zero carcinomas. Ki67≥20%, T2/T3 stage, histological grade 2/3, N+ and PR<20% were significantly associated to high-risk PAM50 group.

Legal entity responsible for the study

The authors.

Funding

Has not received any funding.

Disclosure

R. Sanchez Bayona: Financial Interests, Personal, Invited Speaker: GSK, AstraZeneca, Novartis, ClovisOncology, Lilly; Financial Interests, Personal, Advisory Board: GSK. P. Tolosa: Financial Interests, Personal, Invited Speaker: Lilly, Novartis, Pfizer, AstraZeneca. M. Alba Bianchi: Financial Interests, Personal, Invited Speaker: Novartis, AstraZeneca. A. Madariaga Urrutia: Financial Interests, Personal, Invited Speaker: AstraZeneca; Financial Interests, Personal, Invited Speaker: Clovis; Financial Interests, Personal, Training: GSK. L. Manso: Financial Interests, Personal, Invited Speaker: GSK, AstraZeneca, Novartis, ClovisOncology, Lilly. E.M. Ciruelos: Financial Interests, Personal, Other, Speakers Bureau. Educational activities: Roche; Financial Interests, Personal, Invited Speaker, Symposia and Educational activities: Roche; Financial Interests, Personal, Advisory Board, Non-permanent advisor: Roche; Financial Interests, Personal, Other, Speakers Bureau. Educational activities: Lilly; Financial Interests, Personal, Invited Speaker, Symposia and Education: Lilly; Financial Interests, Personal, Advisory Board, Non-permanent advisor: Lilly; Financial Interests, Personal, Advisory Board, Non-permanent advisor: Novartis; Financial Interests, Personal, Invited Speaker, Educational activities: Pfizer; Financial Interests, Personal, Advisory Board, Non-permanent advisor: Pfizer; Financial Interests, Personal, Advisory Board, Non-permanent advisor: AstraZeneca; Financial Interests, Personal, Advisory Board, Non-permanent advisor: Daiichi Sankyo; Financial Interests, Personal, Advisory Board, Non-permanent advisor: MSD; Financial Interests, Institutional, Funding, PI for Patricia 2 trial (sponsor: SOLTI Group): Pfizer; Financial Interests, Institutional, Funding, PI for Prometeo 2 trial (sponsor: SOLTI Group): Pfizer; Financial Interests, Institutional, Funding, PI for TATEN trial (sponsor: SOLTI Group): MSD; Financial Interests, Institutional, Funding, PI for NEREA trial (sponsor: SOLTI Group): Puma; Financial Interests, Institutional, Funding, PI for ATREZZO trial (sponsor: SOLTI Group): Roche; Non-Financial Interests, Invited Speaker, Non-profit organization dedicated to breast cancer research: SOLTI Cooperative Group; Non-Financial Interests, Advisory Role, Scientific Evaluator at ISCIII (Spanish Government Academic Research Platform): Instituto de Salud Carlos III. All other authors have declared no conflicts of interest.

Background

Bidirectional relationships have been described between brain metabolic activity at 18F-fuorodeoxiglucose(FDG)-PET in breast cancer (BC). Migraine (Mi) and tension-type headache (TTH) seem to be oddly distributed among BC subtypes and their outcomes. The aim of the study is to investigate 18-FDG-PET brain metabolic profile of early BC patients according to subtypes and personal history of headache.

Methods

Consecutive patients with early BC who received neoadjuvant chemotherapy at IRCCS Humanitas Research Hospital were identified. Demographic, tumor and headache features were collected from detailed interview and instrumental investigations. Ki67 was considered high if above 20%. All patients underwent 18F-FDG-PET scan for the evaluation of brain metabolism. Semiquantitative image analysis was performed using Cortex ID software (GE), comparing patient’s scan with a control group by the means of Z-score. The difference between patients’ subgroups were assessed by using One-way ANOVA model.

Results

We recruited 23 BC patients. Median age was 52.5±10.8 years. Ten patients were diagnosed as triple-negative BC, 6 as HER2-positive and 7 as Luminal B/HER2+. Nearly 70% (16/23) of patients had a high ki67. Ten patients had TTH, 9 migraine of whom two with aura, and four patients had no headache. Eleven patients had active headache during the study. Patients with high ki67 had a significant increase of brain metabolism in the left hemisphere(p=0.007), in parietal and general associative areas bilaterally. HER-2 expression was correlated with a reduced metabolism in right temporal lobe (p=0.04). No other tumor characteristics affected PET caption. TTH patients had a reduced activity of right medial frontal lobe compared to those with migraine (p=0.01). FDG uptake was higher in posterior cingulate and reduced in visual networks in patients with active headache (p=0.03).

Conclusions

Although exploratory, this study suggests that BC subtypes could present different metabolic pattern of cerebral activity, particularly in dominant hemisphere and associative areas. Headache - that was highly prevalent in our sample - seems not to influence this pattern since it presents significant activation in other networks, mainly the cingulate and the visual cortex.

Legal entity responsible for the study

The authors.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

Background

Cancer heterogeneity is associated with drug resistance, risk of disease recurrence and metastasis. Pathology allows for the assessment of heterogeneity in the tumor. However, this assessment is limited to an overall assessment of the tumor microenvironment (TME), through measurement of mitotic levels and cancer invasion. We present PhenoScope, a platform for multi-scale analysis and visualization of tumor heterogeneity that integrates cancer data across scales to extract cross modality trends that drive cancer invasion.

Methods

Here we present a Phenoscope analysis of breast tumor pathology slides at three scales. We developed and validated 3 convolutional neural networks (CNN). We combined the outputs of these networks with 2D simulations of the metabolic behavior and growth of cells within the TME. Then, we developed 2 CNNs and implemented an existing CNN that 1) identify cells undergoing mitosis, 2) segment individual cells and classifies their type, and 3) segment five tissues from pathology slides. Additionally, we used transcriptional data to generate patient specific metabolic models which were used with CNN outputs to predict spatial interactions within the TME.

Results

Our results showed: 1) mitosis detection CNN: accuracy of 0.762 (precision=83%, recall=76%) in the test set, 2) classification CNN: Dice Similarity Coefficient (DSC) of 0.821 for segmenting cells and an F1 score for classifying cells ranging from 0.559 (F1i) to 0.756 (F1d); and 3) segmentation CNN: accuracy of 0.781 with DSC of 0.66 - 0.86 depending on tissue. The segmentations and patient-specific metabolic models were analyzed by a proprietary simulation framework to predict the spatial gradients of nutrients, and spatial organization of growth and metabolism. Our simulations identified different metabolic regions in breast tumors dependent on lactate production or consumption, and alanine uptake. These regions were correlated with other indicators such as mitoses and immune cell infiltration.

Conclusions

Overall, we demonstrate a proof-of-principle approach of integrating data across scales, that allows for a more comprehensive understanding of the TME behavior in breast cancer.

Legal entity responsible for the study

SimBioSys, Inc.

Funding

SimBioSys, Inc.

Disclosure

J. Peterson: Financial Interests, Personal and Institutional, Stocks/Shares, Joseph Peterson is employed by and has stock options: SimBioSys, Inc. All other authors have declared no conflicts of interest.

Background

Multigene expression assays for molecular subtypes and biomarkers can aid clinical management of early invasive breast cancer. Based on RNA-sequencing we aimed to develop single-sample predictor (SSP) models for conventional clinical markers, molecular intrinsic subtype and risk of recurrence (ROR).

Methods

A uniformly accrued breast cancer cohort of 7743 patients with RNA-sequencing data from fresh tissue was divided into a training set and a reserved test set. We trained SSPs for PAM50 molecular subtypes and ROR assigned by nearest-centroid (NC) and SSPs for conventional clinical markers from histopathology data. Additionally, SSP classifications were compared with Prosigna® in two external cohorts. Prognostic value was assessed using distant recurrence-free interval.

Results

In the test set, agreement between SSP and NC classifications for PAM50 (five subtypes) and Subtype (four subtypes) was high (85%, Kappa=0.78) and very high (90%, Kappa=0.84) respectively. Accuracy for ROR risk category was high (84%, Kappa=0.75, weighted Kappa=0.90). The prognostic value for SSP and NC was assessed as equivalent. Agreement for SSP and histopathology was very high or high for receptor status, while moderate and poor for Ki67 status and Nottingham histological grade, respectively. SSP concordance with Prosigna® was high for subtype and moderate and high for ROR risk category. In pooled analysis, concordance between SSP and Prosigna® for emulated treatment recommendation for chemotherapy (yes vs. no) was high (85%, Kappa=0.66). In postmenopausal ER+/HER2-/N0 patients SSP application suggested changed treatment recommendations for up to 17% of patients, with nearly balanced escalation and de-escalation of chemotherapy.

Conclusions

SSP models for histopathological variables, PAM50, and ROR classifications can be derived from RNA-sequencing that closely matches clinical tests. Agreement and outcome analyses suggest that NC and SSP models are interchangeable on a group-level and nearly so on a patient level. Retrospective evaluation in postmenopausal ER+/HER2-/N0 patients suggested that molecular testing could lead to a changed therapy recommendation for almost one-fifth of patients.

Legal entity responsible for the study

Division of Oncology, Department of Clinical Sciences Lund, Lund University, Lund, Sweden.

Funding

Swedish Cancer Society, Swedish Research Council, Mrs Berta Kamprad Foundation, Lund-Lausanne L2-Bridge/Biltema Foundation, Mats Paulsson Foundation, Swedish governmental funding ALF grant, Bröstcancerförbundet, Norwegian Cancer Society, South-Eastern Norway Regional Health Authority, Oslo University Hospital.

Disclosure

All authors have declared no conflicts of interest.

Background

The landscape of systemic therapies available for treating BrM has recently expanded to include new HER2-targeted tyrosine kinase inhibitors and antibody-drug conjugates for patients with HER2-positive metastatic breast cancer (MBC). Given that HER2-targeted therapies are now being investigated among patients whose disease does not meet established cut points for HER2 expression, we examined the proportion of patients with “HER2-low” in breast cancer brain metastases BrM.

Methods

A retrospective cohort study of consecutive patients who underwent surgery for breast cancer BrM at Sunnybrook Health Sciences Centre between 2008 and 2018 were identified through the Anatomic Pathology departmental database. Estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor (HER2) status was determined using 2018 ASCO/CAP guidelines. HER2 status was determined based on the immunohistochemistry (IHC) expression and FISH analysis. HER2-0 was defined as IHC 0; HER2-low was defined as IHC 1+ or 2+ and FISH-negative status.

Results

The median patient age at the time of BrM diagnosis was 54.5 (range 23.2-93.2). The majority of patients were symptomatic from BrM at presentation (74%) and 178 patients (23.7%) had leptomeningeal disease. Out of 752 evaluable patients, HER2 status was available in 340 of 752 cases; 181 (53.2%) has HER2-positive, 65 (19.11%) had HER2-low and 94 (27.6%) had HER2-0 disease. Among patients with TNBC, 22 out of 58 cases (40%) were re-classified as being HER2-low. Low HER2-expression was more frequent in the HR+ subgroup compared to the triple-negative subgroup (61.5% vs. 33.8%). There was no significant difference in OS and brain specific PFS between HER2-low and HER2-0.

Conclusions

A high proportion of patients with metastatic TNBC and HR+/HER2-negative breast cancer who have brain metastases have “HER2-low” disease. Given that there are very few CNS-penetrating systemic therapy options for patients with metastatic TNBC, who have a particularly high risk of developing BrM, trials evaluating the efficacy of CNS-active HER2-directed systemic therapies in this patient population are warranted.

Legal entity responsible for the study

Sunnybrook Research Institute.

Funding

Has not received any funding.

Disclosure

K. Jerzak: Financial Interests, Personal, Other, Speaker/Advisor Board/Consultant: Amgen, Apo Biologix, Eli Lilly, Eisai, Genomic Health, Pfizer, Roche, Novartis, Purdue Pharma: Financial Interests, Personal, Other, Research Funding: Eli Lilly, AstraZeneca; Financial Interests, Personal, Other, Advisor/Consultant for: AbbVie, Xpan Medical, Synaptive, Omniscient. All other authors have declared no conflicts of interest.

Background

Although the clinical utility of gene expression signatures for breast cancer is established, access to these prognostic tools is limited to specific patient profiles. The purpose of our study was to identify a set of gene transcripts associated with the likelihood of recurrence in HR+/HER2- breast cancer patients with the highly efficient TempO-Seq platform.

Methods

By targeted expression analysis of FFPE lysate samples from the NHS Greater Glasgow and Clyde biorepository a machine learning approach idendified a subset of transcripts that were prognostic for recurrence. A Cox proportional hazard model for 10-year recurrence was subsequently trained using these transcripts with HR+ samples that lacked overexpression of the HER2 receptor (n = 211) and tested in samples independent of training (n = 50).

Results

The transcript panel analysed on the TempO-Seq platform provided prognostically significant information for breast cancer recurrence in our sample set (LRχ2 = 78.88, p < 0.001; HR = 2.41). These data indicated that patients could be grouped into low- and high-risk cohorts, with significantly different Kaplan-Meier curves for recurrence free survival (p < 0.001) and breast cancer-associated mortality (p = 0.013). Additionally, the prognostic information provided by the TempO-Seq-derived signature was independent of the UK-PREDICT tool (V2), suggesting complementary applications for the technology.

Conclusions

Our novel prognostic signature and testing method based on TempO-Seq profiling was developed using whole transcriptome analysis of a heterogeneously treated cohort in an adjuvant setting. This approach overcomes many of the technical limitations (e.g., does not require extraction) and cost barriers typically encountered with traditional molecular approaches such as qPCR, RNA-Seq, or molecular barcoding. The high throughput test can rapidly generate results from large numbers of patient samples for efficient turnaround times. More widely, the study demonstrates that TempO-Seq technology can be used for accelerated diagnostic panel development and clinical research. The described approach to estimate the risk of breast cancer recurrence can be further validated in prospectively designed trials.

Legal entity responsible for the study

BioClavis Ltd. Teaching and Learning Centre Queen Elizabeth University Hospital, 1345 Govan Rd, Glasgow G51 4TF, United Kingdom.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

Background

Twenty breast cancer patients from Magdalena V. de Martínez Hospital, Buenos Aires, selected by their higher levels of progesterone receptor isoform A (PRA) than isoform B (PRB) were treated for 14 days with 200 mg mifepristone (MFP) p.o. before surgery (MIPRA; NCT02651844). A decrease in the proliferation index, greater than 30% was observed in 70% of the MFP-treated patients [primary endpoint (e.p.)]. RNA-Seq studies performed in 8 paired samples supported these findings and suggested that MFP increased tissue remodeling and immune-related pathways (secondary e.p.). We report herein the morphological changes and the regulation of different hormone receptor-related proteins associated with MFP treatment (secondary e.p.).

Methods

Core needle biopsies (CNB) and post-treatment samples (S) were formalin-fixed and embedded in paraffin for routine morphological and immunohistochemical (IHC) studies. Morphological features were quantitated in 21 paired samples as absent, low, moderate, or high. Tumor-infiltrating lymphocytes (TILs) were quantified following approved guidelines. Due to limited CNB material, IHC assays were performed in selected pair of samples.

Results

The most relevant changes registered were increases in a) loose stroma and/or tissue remodeling (61.9%); b) TILs (80.9%; p<0.001) and c) isolated apoptosis (52.3%). Areas of differentiation and/or the presence of secretory vacuoles were also observed in 33.3% of cases analyzed. Consistent with morphological findings, increases in calregulin (3/4) and active caspase 3 (3/5) expression were observed by IHC. A decrease (p<0.05) in PR, pSer294PR, estrogen receptor alpha (ER), pSer118ER and MYC staining intensity was observed in >50% of the pairs evaluated. No decrease in pSer167ER expression was observed.

Conclusions

The morphological features observed are consistent with Ki67 and RNA-Seq data, and support the therapeutic effects of MFP in patients with higher levels of PRA than PRB. The morphological changes and protein regulations were variable in different patients. The differential regulation in Ki67-responsive and non-responsive patients will be discussed.

Clinical trial identification

MIPRA; NCT02651844.

Legal entity responsible for the study

Hospital Magdalena V de Martínez, General Pacheco, Buenos Aires.

Funding

National Agency for the Promotion of Research, Technological Development and Innovation.

Disclosure

C.C. Lanari, P. Rojas: Financial Interests, Other: Patent (WO2013086379A2) regarding the use of antiprogestins in human breast cancer. All other authors have have declared no conflicts of interest.

Background

High B cell density and low B-cell receptor evenness are predictive and prognostic in triple-negative breast cancers (TNBC). Within the lymph nodes (LNs), germinal centres (GCs) are the sites of B cell maturation and development. Recently we have shown that the formation of GCs in cancer-free (CF) LNs is associated with longer disease-free progression, particularly in TNBC with low tumour infiltrating lymphocytes (TILs). Here, we aimed to investigate the role of GC formation and characterise the B cell populations in human LNs and orthotopic in vivo models of TNBC.

Methods

Multiplex immunofluorescence was performed on sections of involved and CF LNs from 16 patients with low TILs breast cancer. Panels were optimised to detect B cells, memory B cells and plasma cells. For in vivo studies, 7–11-week-old BALB/c mice were administered 50μl of either the 4T1,4T1.2, 67NR or E0771 cell lines into the 4^th mammary fat pad. Mice were sacrificed on days 7, 14, or 21, and the tumour draining(td)-LNs, non-draining (Ntd)-LNs and tumours were harvested. Single-cell LN suspensions were stained with CD19, B220, CD38, CD95, IgM, IgG1 and CD138 for flow cytometry. GC B cells were defined as CD19+B220+CD38^loCD95^hi.

Results

The ratio of plasma to memory B cells was increased within the patient’s primary tumours and in involved LNs, particularly in cases that developed distant metastasis, and negatively correlated with the number of GCs. All TNBC mouse models showed significantly more GC B cells in the td-LNs than in the ntd-LNs 7 days post-inoculation. At days 14 and 21, significantly more GC B cells were obtained in the td-LNs than the nd-LNs in the 4T1.2 (Mann Whitney U test, p < 0.05), with the 4T1 and 67NR models displaying a similar trend. The GC B cell response was radically diminished in the 4T1.2 by day 21 (Mann Whitney U test, p < 0.05), also trending in the other in vivo settings. Unique to the 4T1.2 model, there was an expansion of IgG1+ plasma cells in the td-LN at day 7 and maintained until day 21.

Conclusions

Orthotopic implantation of tumour cell lines in vivo can recapitulate the B cell environment in human tissue and be used to study B cell anti-tumour immunity in TNBC. These preclinical studies will help to elucidate further the role of B cells in TNBCs.

Legal entity responsible for the study

Anita Grigoriadis.

Funding

Medical Research Council (MRC) Cancer Research UK (CRUK) King's College London (KCL).

Disclosure

All authors have declared no conflicts of interest.

Background

The OlympiAD trial evaluated the PARP inhibitor (PARPi) olaparib versus a non-platinum standard chemotherapy in HER2-negative metastatic breast cancer (MBC) patients with a germline BRCA1/2 (gBRCA) mutation. Olaparib resulted in improved progression-free survival (PFS) and doubled the response rate vs chemotherapy. Nevertheless, the response rate to PARPi is »60% in the gBRCA1/2 population with MBC (current approval), suggesting a limited positive predictive value of gBRCA1/2 status. Moreover, patients with other relevant Homologous Recombination Repair defects (HRD) such as PALB2 or RAD51C/D mutation carriers, or HRD epigenetic silencing, are not captured with a gBRCA analysis. We have previoulsy shown that the functional HRD biomarker RAD51, tested in FFPE tumor samples using an optimized immunofluorescence-based assay, is associated with platinum response in early TNBC and PARPi response in preclinical BC models. We hypothesize that the RAD51 test would help to expand the clinical benefit of PARPi by predicting response to olaparib in MBC with germline/somatic BRCA1/2, PALB2 or RAD51C/D mutation and beyond.

Trial design

RADIOLA is an open-label, single-arm, multicentre phase II study evaluating treatment with olaparib in male or female ≥18 years patients with HER2-negative MBC with ≤ two prior chemotherapy lines in two cohorts: cohort 1 (N=41) with known germline/somatic BRCA1/2, PALB2 or RAD51C/D mutation; cohort 2 (N=25) with functional HRD, namely RAD51-low score (≤10%), in wild-type/unknown mutation status at study entry. All patients will receive olaparib 300mg po BID until progression or unacceptable toxicity. Primary objective will assess, in terms of overall response rate (ORR), the capacity of the RAD51 score to predict olaparib efficacy in cohort 1. Secondary objectives include PFS, clinical benefit rate, duration of response, safety in both cohorts and ORR in cohort 2. Recruitment Recruitment (11 sites) started in March 2022.

Legal entity responsible for the study

SOLTI Cancer Research Group.

Funding

AstraZeneca.

Disclosure

J. Balmana: Financial Interests, Personal, Advisory Board: AstraZeneca; Financial Interests, Personal, Advisory Board: Pfizer; Financial Interests, Institutional, Invited Speaker: Pfizer; Financial Interests, Institutional, Invited Speaker: AstraZeneca; Financial Interests, Institutional, Invited Speaker: MedSir. T. Pascual: Financial Interests, Personal, Invited Speaker: Pfizer; Financial Interests, Personal, Invited Speaker: AstraZeneca; Financial Interests, Personal, Invited Speaker: Lilly. I. Blancas López-Barajas: Non-Financial Interests, Institutional, Research Grant: AstraZeneca; Non-Financial Interests, Institutional, Research Grant: Lilly; Non-Financial Interests, Institutional, Research Grant: Roche; Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Advisory Board: Lilly; Financial Interests, Personal, Advisory Board: AstraZeneca; Financial Interests, Personal, Advisory Board: Novartis; Financial Interests, Personal, Advisory Board: Pierre Fabre; Financial Interests, Personal, Advisory Board: Kiowa-Kirin; Financial Interests, Personal, Advisory Board: Veracyte; Financial Interests, Personal, Advisory Board: Bristol Myers Squibb; Financial Interests, Personal, Speaker’s Bureau: Roche; Financial Interests, Personal, Speaker’s Bureau: Lilly; Financial Interests, Personal, Speaker’s Bureau: Novartis; Financial Interests, Personal, Speaker’s Bureau: AstraZeneca; Financial Interests, Personal, Speaker’s Bureau: Pfizer; Financial Interests, Personal, Speaker’s Bureau: Eisai; Financial Interests, Personal, Speaker’s Bureau: Pierre Fabre; Financial Interests, Personal, Speaker’s Bureau: Celgene; Financial Interests, Personal, Sponsor/Funding: Roche; Financial Interests, Personal, Sponsor/Funding: Pierre Fabre; Financial Interests, Personal, Sponsor/Funding: Novartis; Financial Interests, Personal, Sponsor/Funding: Pfizer; Financial Interests, Personal, Sponsor/Funding: Lilly; Financial Interests, Personal, Advisory Role: Roche; Financial Interests, Personal, Advisory Role: Novartis; Financial Interests, Personal, Advisory Role: Lilly; Financial Interests, Personal, Advisory Role: Pfizer; Financial Interests, Personal, Advisory Role: Pierre Fabre. A. Prat: Financial Interests, Personal, Invited Speaker: Roche; Financial Interests, Institutional, Other, Contracted research:: Roche; Financial Interests, Personal, Invited Speaker, Lecture fees: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Novartis; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Novartis; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Amgen; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Amgen; Financial Interests, Personal, Invited Speaker, Lecture fees: BMS; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: BMS; Financial Interests, Personal, Invited Speaker, Lecture fees: NanoString; Financial Interests, Institutional, Other, Contracted research: NanoString; Financial Interests, Institutional, Invited Speaker, Lecture fees: NanoString; Financial Interests, Institutional, Other, Contracted research: Novartis; Financial Interests, Institutional, Other, Contracted research: Roche; Financial Interests, Institutional, Other, Contracted research: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture fees: Contracted research; Financial Interests, Personal, Invited Speaker, Lecture fees: Daiichi Sankyo; Financial Interests, Institutional, Invited Speaker, Clinical trials: Daiichi Sankyo; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Puma; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Oncolytics; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: MSD; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Guardan Health; Financial Interests, Personal, Expert Testimony, Advisory role/consultancy: Peptomyc; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Lilly; Financial Interests, Institutional, Other, Clinical trials: Lilly; Financial Interests, Institutional, Other, Contracted research: Boehringer; Financial Interests, Institutional, Other, Contracted research: Sysmex Europa GmbH; Financial Interests, Institutional, Other, Contracted research: Medica Scientia inno. Research; Financial Interests, Institutional, Other, Contracted research: Celgene; Financial Interests, Institutional, Other, Contracted research: Astellas; Financial Interests, Institutional, Other, Clinical trials: Roche; Financial Interests, Institutional, Other, Clinical trials: Amgen; Financial Interests, Personal, Invited Speaker, Leadership role: Reveal Genomics, SL.; Financial Interests, Institutional, Other, Clinical trials: Boehringer; Non-Financial Interests, Institutional, Other, Leadership roles: Patronage committee: SOLTI Foundation; Non-Financial Interests, Institutional, Other, Leadership roles: Patronage committee: Actitud Frente al Cáncer Foundation. V. Serra Elizalde: Financial Interests, Institutional, Invited Speaker: AstraZeneca; Financial Interests, Personal, Other, WO2019122411A1: Methods based on the detection of rad51 foci in tumor cells: TBD; Financial Interests, Institutional, Research Grant, Testing various novel targeted agents in patient-derived tumour models: AstraZeneca. All other authors have declared no conflicts of interest.

Background

Chemotherapy (CT) followed by endocrine therapy (ET) is the current standard of care for high-risk hormone receptor (HR)-positive/HER2-negative early breast cancer (EBC). The monarchE, with 27-months follow-up, showed that the addition of abemaciclib for 2 years in combination with ET improves the 3-years invasive relapse rate (IRR) and distant relapse rate (DRR) for patients (pts) with high-risk EBC. Obtaining data on the long-term risk of relapse is needed to put into context the magnitude of abemaciclib benefit.

Methods

We performed a retrospective analysis of 1742 HR-positive/HER2-negative EBC pts fulfilling the high-risk criteria of the monarchE in order to estimate their long-term outcomes. They were selected from El Álamo IV registry with pts diagnosed between 2002 and 2005 and 3 adjuvant GEICAM studies (9906, 2003-10 and 2006-10,) carried out from 1999 to 2010. Pts were divided in 2 cohorts (C), similarly to monarchE: C1) ≥ 4 positive axillary lymph nodes (PALN) or 1–3 PALN with tumor size (T) ≥ 5cm and/or histologic grade (G) 3and C2) 1–3 PALN (with T < 5cm and G < 3) and Ki-67 ≥ 20%. Pts had to have undergone surgery with curative intent and had received adjuvant CT (anthracyclines +/- taxanes) and ET.

Results

El Álamo IV registered 6374 luminal EBC pts from which 16.2% were high-risk and included in the analysis (n=1030; 981 C1, 49 C2). In addition 326 pts from 9906 (311 C1, 15 C2), 225 pts from 2003-10 (C1) and 161 pts from 2006-10 (C1) were included. With a median follow-up of 10.2 years, the IRR and DRR at 5 and 10 years were 22% and 34.8% and 19.7% and 31.0%, respectively. Per-year IRR, DRR, and death rate (DR) are described in the table. IRR and DRR increased from year 2, till year 3 followed by a second peak at years 7-8. DR increased steadily from year 1 (7.8% and 24.8% at 5 and 10 years). Table: 62P

Conclusions

The highest yearly risk of recurrence of high-risk EBC, defined as in monarchE, is at 3 years after diagnosis but remains high in the long-term follow-up.

Legal entity responsible for the study

GEICAM Spanish Breast Cancer Group.

Funding

LILLY.

Disclosure

M. Martin Jimenez: Financial Interests, Personal, Advisory Board: ASTRAZENECA; Financial Interests, Personal, Invited Speaker: PFIZER; Financial Interests, Personal, Invited Speaker: ASTRAZENECA; Financial Interests, Personal, Advisory Board: LILLY; Financial Interests, Personal, Invited Speaker: LILLY; Financial Interests, Personal, Invited Speaker: NOVARTIS; Financial Interests, Personal, Advisory Board: ROCHE/GENENTECH; Financial Interests, Personal, Invited Speaker: ROCHE/GENENTECH; Financial Interests, Personal, Advisory Board: Daiichi Sankyo; Financial Interests, Institutional, Research Grant: NOVARTIS; Financial Interests, Institutional, Research Grant: ROCHE; Financial Interests, Institutional, Research Grant: Puma; Non-Financial Interests, Invited Speaker: TRIO; Non-Financial Interests, Leadership Role: GEICAM. B. Bermejo De Las Heras: Financial Interests, Institutional, Advisory Role: MSD, Roche, Pierre Fabre, Novartis, AstraZeneca, Seagen; Financial Interests, Institutional, Speaker’s Bureau: Pfizer, Roche, MSD, Palex, Eisai, Daiichi, AstraZeneca, Seagen. A. Guerrero: Financial Interests, Institutional, Research Grant: Pfizer; Financial Interests, Institutional, Advisory Board: Pfizer, Palex, Novartis, AstraZeneca, Pierre Fabre; Financial Interests, Personal, Other, Travel grant: Pfizer, Novartis; Financial Interests, Institutional, Speaker’s Bureau: Novartis, Pfizer, Lilly, AstraZeneca. A. Santaballa Bertran: Financial Interests, Institutional, Advisory Role: Pfizer. S. Lopez-Tarruella Cobo: Financial Interests, Institutional, Advisory Role: AstraZeneca, Novartis, Roche, Pfizer, Pierre Fabre, Lilly, Seagen, Daiichi Sankyo Europe GmbH, Gilead Sciences,MSD, GalaxoSmithkline, Veracyte; Financial Interests, Personal, Speaker’s Bureau: Lilly. All other authors have declared no conflicts of interest.

Background

Abemaciclib is the first and only approved CDK4 & 6 inhibitor in the adjuvant setting for certain types of HR+, HER2-, node-positive, high-risk early breast cancer (EBC). Patient (Pts) with HR+, HER2- tumors who are premenopausal (preM) may have a different tumor biology and response to endocrine therapy (ET) than postmenopausal (postM) pts, warranting analysis of the prespecified subgroup within monarchE.

Methods

Randomized (1:1) pts received adjuvant ET +/- abemaciclib for 2 years (yrs) plus at least 3 yrs of ET as clinically indicated. Menopausal status (preM vs. postM) at diagnosis was a predefined stratification factor. Standard ET (tamoxifen or an aromatase inhibitor), with or without additional GnRH, was determined by physician’s choice. Baseline disease characteristics and ET choice were described at ESMO, 2021. Efficacy and safety results of abemaciclib + ET vs. ET alone including invasive disease-free survival (IDFS) and distant relapse-free survival (DRFS) were assessed by menopausal status with a data cut-off date on 1 April 2021, at a median follow-up of 27 months.

Results

Among randomized pts, 2451 (43.5%) were preM and 3181 (56.4%) postM. Treatment benefit was consistent in both menopausal status subgroups, with numerically greater effect size in preM subgroup compared to postM subgroup (Table). For preM pts, abemaciclib + ET resulted in a 42.7% and 40.8% reduction in risk of developing an IDFS and DRFS event, respectively. The absolute improvement at 3 yrs was 5.7% for IDFS and 4.4% for DRFS rates. The safety profile for preM pts was consistent with the safety population. Table: 63P

Efficacy data by menopausal status

Conclusions

Abemaciclib + ET demonstrated a clinically meaningful treatment benefit in IDFS and DRFS vs. ET alone regardless of menopausal status, with a numerically greater benefit in the preM population. Safety data in preM pts is consistent with the overall safety profile of abemaciclib.

Clinical trial identification

NCT03155997.

Editorial acknowledgement

Medical writing support was provided by Trish Huynh (Eli Lilly and Company, IN, USA).

Legal entity responsible for the study

Eli Lilly and Company.

Funding

Eli Lilly and Company.

Disclosure

S. Paluch-Shimon: Financial Interests, Institutional, Advisory Board, Advisory board invited speaker honoraria: Roche; Financial Interests, Institutional, Other, Advisory board invited speaker honoraria: Pfizer; Financial Interests, Institutional, Other, Advisory board invited speaker honoraria: Novartis; Financial Interests, Institutional, Other, Advisory board invited speaker honoraria: AstraZeneca; Financial Interests, Institutional, Advisory Board, Advisory board invited speaker honoraria: Exact sciences; Financial Interests, Institutional, Advisory Board, Advisory board invited speaker honoraria: Eli Lilly; Financial Interests, Institutional, Other, Advisory board, speaker's bureau, consultancy: Medison; Financial Interests, Personal and Institutional, Research Grant, Research grant for an RFP independent research put out by SPCC and Pfizer: SPCC (Shared Progress in Cancer Care). P. Neven: Financial Interests, Institutional, Advisory Board: Eli Lilly; Financial Interests, Institutional, Advisory Board: AstraZeneca; Financial Interests, Institutional, Advisory Board: Novartis; Financial Interests, Institutional, Advisory Board: Pfizer; Financial Interests, Institutional, Advisory Board: Roche; Financial Interests, Institutional, Advisory Board: Roularta Belgium; Financial Interests, Personal, Other, Lecture fees: Eli Lilly; Financial Interests, Personal, Other, Lecture fees: Novartis; Financial Interests, Personal, Other, Lecture fees: Pfizer; Financial Interests, Personal, Other, Lecture fees: AstraZeneca. J. Huober: Financial Interests, Research Grant: Celgene; Financial Interests, Research Grant: Novartis; Financial Interests, Research Grant: Hexal; Financial Interests, Research Grant: Eli Lilly and Company; Financial Interests, Other, Honoraria: Eli Lilly and Company; Financial Interests, Other, Honoraria: Novartis; Financial Interests, Other, Honoraria: Roche; Financial Interests, Other, Honoraria: Pfizer; Financial Interests, Other, Honoraria: AstraZeneca; Financial Interests, Other, Honoraria: MSD; Financial Interests, Other, Honoraria: Celgene; Financial Interests, Other, Honoraria: Eisai; Financial Interests, Other, Honoraria: AbbVie; Financial Interests, Other, Honoraria: Seagen; Financial Interests, Other, Honoraria: Gilead; Financial Interests, Advisory Role: Eli Lilly and Company; Financial Interests, Advisory Role: Novartis; Financial Interests, Advisory Role: Roche; Financial Interests, Advisory Role: Pfizer; Financial Interests, Advisory Role: Hexal; Financial Interests, Advisory Role: AstraZeneca; Financial Interests, Advisory Role: MSD; Financial Interests, Advisory Role: Celgene; Financial Interests, Advisory Role: AbbVie; Financial Interests, Advisory Role: Seagen; Financial Interests, Advisory Role: Gilead; Financial Interests, Other, Travel expenses: Roche; Financial Interests, Other, Travel expenses: Pfizer; Financial Interests, Other, Travel expenses: Novartis; Financial Interests, Other, Travel expenses: Celgene; Financial Interests, Other, Travel expenses: Daiichi. Z. Jiang: Financial Interests, Advisory Board: Eli Lilly and Company; Financial Interests, Advisory Board: Roche; Financial Interests, Advisory Board: AbbVie; Financial Interests, Advisory Board: AstraZeneca; Financial Interests, Advisory Board: Pfizer; Financial Interests, Advisory Board: Bayer; Financial Interests, Advisory Board: Novartis; Financial Interests, Advisory Board: Amgen; Financial Interests, Advisory Board: Glaxo Smith Kline; Financial Interests, Advisory Board: Boehringer Ingelheim. M.P. Goetz: Other, Institutional, Other, Consulting: Eagle Pharmaceuticals; Other, Institutional, Other, Consulting: Eli Lilly and Company; Other, Institutional, Other, Consulting: Biovica; Other, Institutional, Other, Consulting: Novartis; Other, Institutional, Other, Consulting: Sermonix; Financial Interests, Institutional, Research Grant: Pfizer; Non-Financial Interests, Institutional, Research Grant: Eli Lilly and Company; Other, Institutional, Other, Consulting: Pfizer; Other, Institutional, Other, Consulting: Biotheranostics; Financial Interests, Institutional, Research Grant: Sermonix; Other, Institutional, Other, Consulting: AstraZeneca; Other, Institutional, Other, Consulting: Blueprint Medicines; Other, Personal, Other, Consulting: Research to Practice; Other, Personal, Other, Consulting: Clinical Education Alliance. C. Shimizu: Financial Interests, Research Grant: Eli Lilly and Company; Financial Interests, Personal, Invited Speaker, Lecture Fee: Daiichi Sankyo; Financial Interests, Personal, Other, Honoraria: Pfizer; Financial Interests, Personal, Invited Speaker, Lecture Fee: Chugai; Financial Interests, Personal, Invited Speaker, Lecture Fee: Taiho; Financial Interests, Personal, Other, Honoraria: AstraZeneca. C. Huang: Financial Interests, Personal, Advisory Board: Eli Lilly and Company; Financial Interests, Personal, Speaker’s Bureau: Amgen; Financial Interests, Personal, Research Grant, Advisory board, Speaker bureau, travel expense: AstraZeneca; Financial Interests, Personal, Advisory Board: EirGenix; Financial Interests, Research Grant: MSD; Financial Interests, Personal, Invited Speaker, Advisory board, speaker bureau: Novartis; Financial Interests, Research Grant: Daiichi Sankyo; Financial Interests, Research Grant: OBI Pharma; Financial Interests, Personal, Research Grant, advisory board, speaker bureau, travel expense: Pfizer; Financial Interests, Personal, Research Grant, advisory board, speaker bureau, travel expense: Roche. R.J. Wei, S.C. Nabinger, T. Forrester: Financial Interests, Personal, Full or part-time Employment: Eli Lilly and Company; Financial Interests, Personal, Stocks/Shares: Eli Lilly and Company. N. Harbeck: Financial Interests, Personal, Other, Lectures, Consulting: Pfizer; Financial Interests, Personal, Other, Lectures, Consulting: AstraZeneca; Financial Interests, Personal, Other, Lectures, Consulting: Eli Lilly and Company; Financial Interests, Personal, Other, Lectures, Consulting: Novartis. All other authors have declared no conflicts of interest.

Background

With survival increases, many breast cancer patients concomitantly use adjuvant hormone therapy and cardiovascular therapy. However, whether discontinuation of these two therapies is associated, and tends to occur at the same time has not been evaluated.

Methods

By linking eight Swedish health registers, we identified 1,811 discontinuers of adjuvant hormone therapy who concomitantly used cardiovascular therapy (cardiovascular drugs, statins, or aspirins) and randomly matched each discontinuer with a continuer on diagnosis year and age at diagnosis (± 3 years). Poisson regression was used to estimate incidence rate ratios (IRRs) of discontinuing cardiovascular therapy from one year before until one year after discontinuation of adjuvant hormone therapy, comparing discontinuers with continuers of adjuvant hormone therapy. We further investigated the association of discontinuing adjuvant hormone therapy with breast cancer-specific and cardiovascular disease-specific mortality using Cox models.

Results

At the time when patients discontinued their adjuvant hormone therapy, 10.1%, 16.2% and 16.4% of breast cancer patients concomitantly discontinued cardiovascular drugs, statins, and aspirins, respectively. Overall, discontinuers of adjuvant hormone therapy had higher risks of discontinuing cardiovascular therapy than continuers, with IRRs of 1.85 (95%CI, 1.42-2.40) and 2.30 (95%CI, 1.74-3.05) within 3 months before and after discontinuation, respectively. Furthermore, discontinuation of adjuvant hormone therapy was statistically significantly associated with death from both breast cancer (HR, 1.43; 95 CI%, 1.02-2.00) and cardiovascular diseases (HR, 1.81; 95 CI%, 1.16-2.81).

Conclusions

Discontinuation of adjuvant hormone therapy and cardiovascular therapy tended to occur concomitantly in breast cancer patients. Multiple disease focus needs to be applied to prevent therapy discontinuation in polypharmacy users, which would subsequently improve overall survival.

Legal entity responsible for the study

The authors.

Funding

Swedish Research Council; Swedish Cancer Society; FORTE; China Scholarship Council; Zhejiang University “Hundred Talents Program”.

Disclosure

All authors have declared no conflicts of interest.

Background

Triple-negative breast cancer (TNBC) is an aggressive type that lack Estrogen, progesterone and HER2 receptors.Targeted treatments aren't available for TNBC like other types of breast cancer but Several studies proved the benefit of adjuvant chemotherapy. This study aims to evaluate the survival outcome of adjuvant chemoradiotherapy compared to different treatment regimens in the management of node-negative T1a,T1b,T1c triple-negative breast cancer (TNBC).

Methods

we used Surveillance, Epidemiology, and End Results (SEER) database to extract the data of 9038 patients with T1abc node-negative triple-negative breast cancer diagnosed from 2010 to 2018, they were treated with different approaches: adjuvant chemoradiotherapy, adjuvant chemotherapy, adjuvant radiotherapy and surgery without adjuvant treatment.

Results

The 5-year disease free survival DFS is better in patients treated with adjuvant chemoradiotherapy compared to adjuvant chemotherapy, adjuvant radiotherapy and surgery without adjuvant treatment respectively(92.4%,91.4%,84.5%,77,9%, P-value >0.00) and the overall 5-year survival outcome was higher for adjuvant chemoradiotherapy and adjuvant chemotherapy(92.3%,91.2%) compared to adjuvant radiotherapy and surgery(84.2%,77.6% with P-value >0.00). Adjuvant chemoradiotherapy shows better survival outcome in T1a, T1b (98.4%,95.3% ) compared to adjuvant chemotherapy (95.7%,91.4%), adjuvant radiotherapy (93.1%,86.8%) and surgery without adjuvant treatment(85.8%,82.5%) while both adjuvant chemoradiotherapy and adjuvant chemotherapy has similar outcome in T1c (90.9%).

Conclusions

Adjuvant chemoradiotherapy has better 5-year Disease Free Survival (DFS), better overall 5-year survival outcome and better survival outcome in T1a,T1b compared to other treatment modalities. The recent guidelines shows the survival benefit of adjuvant chemotherapy on T1c only but this study highlighted the benefit of adjuvant chemoradiotherapy on treatment of T1a,T1b node-negative triple-negative breast cancer. Further evaluation of each patient is necessary to weigh the benefits versus side effects of this treatment regimen.

Legal entity responsible for the study

The authors.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

Background

Bisphosphonates (BP) do not have regulatory approval for use as adjuvant therapy in early breast cancer due to the failure of pivotal trials such as AZURE and NSABP B-34. However, an EBCTBG meta-analysis of randomized data showed clear clinical benefit from adjuvant BP use in postmenopausal (PM) women and both ESMO and ASCO guidelines recommend their use in this population. Retrospective analyses of the MAF biomarker in the AZURE and NSABP B-34 trials demonstrated a significant benefit of adjuvant BP on relapse and death that was restricted to the 80% of tumors that were MAF non-amplified (MAF(-)).

Methods

We studied the relative differences in relapse and death with and without adjuvant BP in PM patients (pts) included in the EBCTBG analyses and all MAF(-)pts in the AZURE + B-34 trials. MAF gene amplification was determined by FISH analysis on the primary tumor using the MAF Test (Inbiomotion, Spain).

Results

For the PM cohorts in the EBCTBG meta-analysis, the rates of relapse and death in untreated control pts (n=5668) were 15.8% and 8.7% respectively compared with 13.4% and 7.5% for those pts (n=6099) BP treated. The relative reductions in risk of recurrence and death were 15.2% and 13.8% respectively For MAF(-)pts in the AZURE and B-34 trials combined the rate of relapse and death events were 17.1% and 8.9% respectively for control pts (n= 1126) and 13.0% and 6.5% for BP treated (n=1170) pts, translating into relative risk reduction for relapse and death events of 23.8% and 26.3% respectively favoring BP treated pts These results contrast markedly with the results for MAF(+)pts; Here recurrence rates were 26.5% without and 30.4% with adjuvant BP while deaths occurred in 13.7% of control (n= 249) and 15.9% of BP treated (n= 265) pts, indicating that BP are ineffective in this sub-group and are associated with a relative increase of 14.6% in relapses and 16.1% in deaths.

Conclusions

The use of the MAF biomarker measured on the primary breast tumor identifies a target population (MAF(-)) for treatment with adjuvant BPs that is independent of menopausal status and provides a beneficial treatment opportunity for a higher proportion of breast cancer pts than are currently recommended by clinical guidelines.

Legal entity responsible for the study

The authors.

Funding

Inbiomotion.

Disclosure

R.E. Coleman: Financial Interests, Personal, Stocks/Shares: Inbiomotion; Financial Interests, Personal, Advisory Role: Amgen; Financial Interests, Personal, Advisory Role: AstraZeneca; Financial Interests, Personal, Advisory Role: Boehringer Ingelheim; Financial Interests, Personal, Advisory Role: Menarini; Financial Interests, Personal, Advisory Role: Sanofi. R.R. Gomis: Financial Interests, Personal, Stocks/Shares: Inbiomotion. All other authors have declared no conflicts of interest.

Background

The approval of abemaciclib (abema) for the adjuvant treatment of patients with high-risk, lymph node (LN)+, hormone-receptor (HR)+ breast cancer (BC) has reshaped treatment algorithms for this disease. However, the real-world features of BC patients eligible for abema remain unknown.

Methods

We reviewed data of all consecutive patients with HR+ BC who underwent surgery at Dana-Farber Brigham Cancer Center from 1/2016 to 3/2021. According to the latest NCCN guidelines, patients were eligible for abema if they had either: (i) 1-3+ LN and tumors either ≥5 cm or grade 3, or (ii) ≥4+ LN. Clinicopathologic features of this population were compared with HR+ BC patients not otherwise meeting eligibility criteria for abema.

Results

Of 4,496 HR+ patients in the database, 499 (11.1%) were eligible for adjuvant abema, whereas 3,997 did not meet eligibility criteria. Compared with non-eligible patients, those eligible for adjuvant abema were significantly younger (median age 53 vs 61, p<0.001), more often pre-menopausal (52% vs 30.3%, p<0.001) or BRCA2-mutant (10.9% vs 2.9%, p=0.002), more often had lobular tumors (20.8% vs. 13.8%, p<0.001) and high-risk (≥26) OncotypeDX recurrence scores (30.7% vs 14.4%, p<0.001). As expected, there were significant differences in receipt of treatment, with abema-eligible patients more often receiving (neo)adjuvant chemotherapy (77.8% vs 11.6%, p<0.001), neoadjuvant endocrine treatment (11.2% vs 3.7%, p<0.001), total mastectomy (64.7% vs 24.3%, p<0.001) and axillary LN dissection (76.0% vs 6.2%, p<0.001). Among eligible patients receiving neoadjuvant chemotherapy (n=202), none experienced a pathological complete response, 3 experienced a residual cancer burden (RCB) score of I, and the remaining patients had extensive residual disease at surgery (RCB II-III). After a median follow up of 10 months, 2-year distant disease-free survival of eligible pts was 90.9% and 2-year overall survival was 96.4%.

Conclusions

Patients eligible for adjuvant abema are younger, more often BRCA2-mutant and more often harbor lobular tumors compared to non-eligible patients. Response to chemotherapy in this cohort appears poor. Our real-world data support recently approved adjuvant treatment strategies to include abema and olaparib.

Legal entity responsible for the study

The authors.

Funding

Has not received any funding.

Disclosure

P. Tarantino: Financial Interests, Personal, Financial Interests, Fee for consulting/advisory role: AstraZeneca. E.A. Mittendorf: Financial Interests, Personal, Invited Speaker: Bristol Myers Squib; Financial Interests, Personal, Invited Speaker: Exact Sciences; Financial Interests, Personal, Invited Speaker: Genentech/Roche; Financial Interests, Personal, Invited Speaker: Lilly; Financial Interests, Personal, Invited Speaker: Merck. T.A. King: Financial Interests, Personal, Financial Interests: Exact Sciences; Financial Interests, Personal, Financial Interests: PrecisCA cancer information service. G. Curigliano: Financial Interests, Personal, Invited Speaker: Roche; Financial Interests, Personal, Advisory Board: Ellipsis; Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Invited Speaker: AstraZeneca; Financial Interests, Personal, Advisory Board: AstraZeneca; Financial Interests, Personal, Invited Speaker: Daiichi Sankyo; Financial Interests, Personal, Advisory Board: Daiichi Sankyo; Financial Interests, Personal, Invited Speaker: Novartis; Financial Interests, Personal, Advisory Board: Lilly; Financial Interests, Personal, Invited Speaker: Pfizer; Financial Interests, Personal, Advisory Board: Pfizer; Financial Interests, Personal, Invited Speaker: Pfizer; Financial Interests, Personal, Advisory Board: Veracyte; Financial Interests, Personal, Advisory Board: BMS; Financial Interests, Personal, Advisory Board: Merck; Financial Interests, Personal, Advisory Board: Exact Sciences; Financial Interests, Institutional, Research Grant, Investigator Initiated Trial: Merck; Financial Interests, Institutional, Funding, Phase I studies: BMS; Financial Interests, Institutional, Funding, Phase I studies: Novartis; Financial Interests, Institutional, Funding, Phase I studies: AstraZeneca; Financial Interests, Institutional, Funding, Phase I studies: Daiichi Sankyo; Financial Interests, Institutional, Funding, Phase I studies: Roche; Financial Interests, Institutional, Funding, Phase I studies: Blueprint Medicine; Financial Interests, Institutional, Funding, Phase I studies: Kymab; Financial Interests, Institutional, Funding, Phase I studies: Astellas; Financial Interests, Institutional, Funding, Phase I studies: Sanofi; Financial Interests, Institutional, Funding, Phase I studies: Philogen; Non-Financial Interests, Officer, Italian National Health Council as Advisor for Ministry of Health: Consiglio Superiore di Sanità; Non-Financial Interests, Advisory Role, Member of the Scientific Council. Patient advocacy association: Europa Donna; Non-Financial Interests, Advisory Role, Cancer Research Foundation: Fondazione Beretta; Non-Financial Interests, Invited Speaker, No compensation for this role. This a public national company for cancer prevention: Lega Italiana Lotta ai Tumori. S.M. Tolaney: Financial Interests, Personal, Advisory Board, Ad Board Participant/Consultant: AstraZeneca; Financial Interests, Personal, Advisory Board, Ad Board Participant/Consultant: Eli Lilly; Financial Interests, Personal, Advisory Board, Ad board participant/Consultant: Pfizer; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Novartis; Financial Interests, Personal, Advisory Board, Ad Board participant/consultant: Merck; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Gilead; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Genentech/Roche; Financial Interests, Personal, Other, Consultant: Nektar; Financial Interests, Personal, Other, Consultant: NanoString; Financial Interests, Personal, Advisory Board, Ad board participant: Bristol Myers Squibb; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Eisai; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Sanofi; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Seagen; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Daiichi Sankyo; Financial Interests, Personal, Advisory Board, Ad board participant/consultant/DSMC: Odonate; Financial Interests, Personal, Other, Steering Committee Member/Consultant: CytomX; Financial Interests, Personal, Other, Consultant: Athenex; Financial Interests, Personal, Advisory Board, Ad board participant: OncoPep; Financial Interests, Personal, Advisory Board, Ad board participant: OncoSec; Financial Interests, Personal, Advisory Board, Ad board participant: Certara; Financial Interests, Personal, Advisory Board, Ad board participant: Mersana Therapeutics; Financial Interests, Personal, Advisory Board, Ad board participant: Ellipses Pharma; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: 4D Pharma; Financial Interests, Personal, Advisory Board, Ad board participant/consultant: Puma; Financial Interests, Personal, Invited Speaker, Invited speaker for pharma supported educational activity: Chugai Pharmaceuticals; Financial Interests, Personal, Advisory Board, Advisory board participant: G1 Therapeutics; Financial Interests, Personal, Other, Consultant: Blueprint Medicines; Financial Interests, Personal, Advisory Board, Advisory Board Participation: Zymeworks; Financial Interests, Personal, Advisory Board, Advisory Board participation: Zentalis; Financial Interests, Personal, Advisory Board, Advisory board participation: Reveal Genomics; Financial Interests, Personal, Advisory Board, Advisory Board participation: OncXerna; Financial Interests, Institutional, Funding: AstraZeneca; Financial Interests, Institutional, Funding: Eli Lilly; Financial Interests, Institutional, Funding: Pfizer; Financial Interests, Institutional, Funding: Sanofi; Financial Interests, Institutional, Funding: Seagen; Financial Interests, Institutional, Funding: Odonate; Financial Interests, Personal and Institutional, Invited Speaker: CytomX; Financial Interests, Institutional, Funding: Cyclacel; Financial Interests, Institutional, Funding: Exelixis; Financial Interests, Institutional, Funding: Gilead; Financial Interests, Institutional, Funding: Bristol Myers Squibb; Financial Interests, Institutional, Funding: Eisai; Financial Interests, Institutional, Funding: Merck; Financial Interests, Institutional, Funding: Novartis; Financial Interests, Institutional, Funding: Nektar; Financial Interests, Institutional, Funding: Genentech/Roche. All other authors have declared no conflicts of interest.

Background

The clinically used breast cancer markers are known to predict short-term survival, but whether they predict long-term survival is unclear. We therefore aimed to investigate the long-term benefit from tamoxifen (TAM) by clinically used markers in ER-positive/HER2-negative breast cancer patients.

Methods

Secondary analysis of ER-positive/HER2-negative patients (n=1242) in the Stockholm tamoxifen trials (1976-1996) randomized to TAM (40 mg) vs no TAM. Complete 20-year follow-up was from Swedish registers. Distant recurrence-free interval (DRFI) by clinically used markers was assessed by Kaplan-Meier, multivariable Cox proportional hazards, and time-varying analysis.

Results

Kaplan-Meier analysis showed significantly (log-rank P<0.05) improved long-term DRFI from TAM vs no TAM for larger tumor size (T1c P<0.001; T2-3 P=0.012), grade 2 (P<0.001), lymph node-negative (P<0.001), PR-positive (P<0.001) and Ki-67-low tumors (P<0.001). Multivariable analysis (Table) showed significant long-term TAM benefit for patients with larger tumor size (T1c HR=0.56; 95% CI, 0.42-0.75; T2 HR=0.67; 95% CI, 0.49-0.92), grade 2 (HR=0.55; 95% CI, 0.42-0.71), lymph node-negative (HR=0.45, 95% CI, 0.33-0.62), PR-positive (HR=0.59, 95% CI, 0.47-0.75) and Ki-67-low tumors (HR=0.55, 95% CI, 0.43-0.71). Time-varying analysis showed that patients with tumor size T1c, grade 2, lymph node-negative, PR-positive, and Ki-67-low tumors significantly benefitted from TAM for 20 years (HR=0.56, 95% CI: 0.35-0.92; HR=0.49, 95% CI: 0.32-0.76; HR=0.50, 95% CI: 0.30-0.84; HR=0.54, 95% CI: 0.36-0.80; HR =0.51, 95% CI: 0.34-0.75 at year 20, respectively). Patients with tumor size T2-3 had significant TAM benefit for 10 years (HR=0.62, 95% CI: 0.40-0.96 at year 10). Table: 68P

Conclusions

Our findings suggest a significant long-term tamoxifen treatment benefit among patients with larger tumors, lymph node-negative tumors, PR-positive tumors, and Ki-67 low tumors.

Clinical trial identification

The trial center for the STO-trials was the Regional Cancer Center Stockholm-Gotland, in Stockholm Sweden. However, the start of the Stockholm Tamoxifen trials in 1976 was well before trial registration started in Sweden, therefore information on trial number is not available.

Legal entity responsible for the study

The authors.

Funding

Swedish Research Council, Swedish Research Council for Health, Working life and Welfare, The Gösta Milton Donation Fund and Swedish Cancer Society.

Disclosure

All authors have declared no conflicts of interest.

Background

Preclinical data suggest a protective role of statins in patients (pts) with breast cancer. Clinical data are controversial with limited data on the benefit for pts with HER2-positive early breast cancer (HER2+ EBC). We investigated the association of statin use with survival outcomes in pts with HER2+ EBC enrolled in APHINITY (BIG 4-11/NCT01358877).

Methods

APHINITY is a prospective, randomized, double-blind, phase III trial testing the addition of pertuzumab to trastuzumab and chemotherapy in pts with HER2+ EBC. All pts evaluated in APHINITY were included in the present analysis (N=4804). Pts who received statins at baseline, or started statins within one year from randomization were considered as statin users. Pts’ characteristics were compared according to statin use by chi-square test. Survival analysis for invasive-disease-free survival (IDFS), distant relapse-free interval (DRFI), and overall survival (OS) were performed using the Kaplan Meier method and log-rank test. A multivariate analysis using a Cox proportional hazard model was used to generate hazard ratios (HR), adjusting for tumor size, nodal status, hormone receptor status, age, menopausal status, body mass index (BMI) ≥ 30 kg/m², treatment arm and statin use.

Results

Median follow-up was 6 years. Statin users (N=423) were more frequently ≥65 years old, postmenopausal, obese (BMI ≥ 30 kg/m²) and were treated more often with breast conserving surgery compared to non-statin users (N=4381) (all p<.001). In multivariate analysis, no significant impact of statin use on any of the survival outcomes was observed (IDFS, HR 1.17, 95% confidence interval [CI] 0.86-1.59; DRFI, HR 1.22, 95% CI 0.83-1.79; OS, HR 1.18, 95% CI 0.80-1.72). No significant interaction between statin use, obesity, treatment arm, hormone receptor status and survival could be detected (p>.05).

Conclusions

In this analysis, statin use was not associated with improved survival outcomes in the relatively young EBC population enrolled in APHINITY. We acknowledge the retrospective and exploratory nature of the present analysis. Further research should explore the potential clinical benefit of statins in a higher risk (e.g. older age) EBC population.

Clinical trial identification

BIG 4-11/ BO 25 126/ TOC 4939g.

Legal entity responsible for the study

F. Hoffmann-La Roche Ltd. and Genentech, Inc.

Funding

Has not received any funding.

Disclosure

C. Maurer: Financial Interests, Personal, Other, Travel grant: Mundipharma; Financial Interests, Personal, Other, Travel grant: Amgen; Financial Interests, Personal, Other, Travel grant: Servier Deutschland GmbH; Financial Interests, Personal, Other, Travel grant: AbbVie; Financial Interests, Personal, Advisory Board: AbbVie; Financial Interests, Personal, Advisory Board: Pfizer; Financial Interests, Personal, Advisory Board: Celgene/BMS. E. Agostinetto: Financial Interests, Personal, Invited Speaker: Eli Lilly; Financial Interests, Personal, Other, Travel grant: Novartis; Financial Interests, Personal, Other, Travel grant: Roche; Financial Interests, Personal, Other, Travel grant: Eli Lilly; Financial Interests, Personal, Other, Travel grant: Genetic. M. Lambertini: Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Advisory Board: AstraZeneca; Financial Interests, Personal, Advisory Board: Lilly; Financial Interests, Personal, Advisory Board: Novartis; Financial Interests, Personal, Invited Speaker: Pfizer; Financial Interests, Personal, Invited Speaker: Takeda; Financial Interests, Personal, Invited Speaker: Sandoz; Financial Interests, Personal, Invited Speaker: Ipsen; Financial Interests, Personal, Invited Speaker: Libbs; Financial Interests, Personal, Invited Speaker: Knight; Financial Interests, Personal, Advisory Board: Exact Sciences; Financial Interests, Personal, Advisory Board: MSD; Financial Interests, Personal, Advisory Board: Seagen; Financial Interests, Personal, Advisory Board: Gilead. S. Martel: Financial Interests, Personal, Advisory Board: Novartis; Financial Interests, Personal, Advisory Board: Pfizer; Financial Interests, Personal, Advisory Board: Knight Therapeutics; Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Advisory Board: Taiho; Financial Interests, Personal, Advisory Board: Exact Science; Financial Interests, Personal, Advisory Board: Apobiologix. N.F. Ponde: Financial Interests, Personal, Full or part-time Employment: IQVIA Biotech. M. Brandão: Financial Interests, Personal, Invited Speaker: Roche/GNE; Financial Interests, Personal, Invited Speaker: Janssen; Financial Interests, Personal, Other, Travel grant: Roche/GNE; Financial Interests, Personal, Other, Travel grant: Sanofi; Financial Interests, Institutional, Research Grant: Roche/GNE; Financial Interests, Institutional, Research Grant: AstraZeneca; Financial Interests, Institutional, Research Grant: GSK/Novartis; Financial Interests, Institutional, Research Grant: Servier. F. Poggio: Financial Interests, Personal, Invited Speaker: Eli Lilly; Financial Interests, Personal, Invited Speaker: Novartis. A.R. Ferreira: Financial Interests, Personal, Advisory Board: Daiichi Sankyo; Financial Interests, Personal, Advisory Board: Gilead; Financial Interests, Personal, Advisory Board: Merck Sharp & Dohme; Financial Interests, Personal, Advisory Board: Novartis; Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Other, Travel grant: Roche. R. Schiff: Financial Interests, Institutional, Funding: AstraZeneca; Financial Interests, Institutional, Funding: GlaxoSmithKline; Financial Interests, Institutional, Funding: Puma; Financial Interests, Institutional, Funding: Biotechnology Inc.; Financial Interests, Institutional, Funding: Gilead Sciences; Financial Interests, Personal, Advisory Board: Eli Lilly; Financial Interests, Personal, Advisory Board: Macrogenics; Financial Interests, Personal, Royalties: UpToDate; Other, Co-inventor in the Patent application # PCT/US21/70543 (Methods for breast cancer treatment and prediction of therapeutic response): Other. C. De Angelis: Financial Interests, Personal, Speaker’s Bureau: Roche; Financial Interests, Personal, Speaker’s Bureau: Eli Lilly; Financial Interests, Personal, Speaker’s Bureau: GSK; Financial Interests, Personal, Speaker’s Bureau: Novartis; Financial Interests, Personal, Speaker’s Bureau: Pfizer; Financial Interests, Personal, Speaker’s Bureau: AstraZeneca; Financial Interests, Institutional, Research Grant: Novartis; Other, Co-inventor in the Patent application # PCT/US21/70543 (Methods for breast cancer treatment and prediction of therapeutic response): Other. R.D. Gelber: Financial Interests, Institutional, Funding: Roche; Financial Interests, Institutional, Funding: Novartis; Financial Interests, Institutional, Funding: AstraZeneca; Financial Interests, Institutional, Funding: Pfizer; Financial Interests, Institutional, Funding: Merck. S. Dent: Financial Interests, Institutional, Funding: Novartis; Financial Interests, Personal, Other, Honoraria: Novartis. C. Thomssen: Financial Interests, Personal, Advisory Board: Amgen; Financial Interests, Personal, Advisory Board: AstraZeneca; Financial Interests, Personal, Advisory Board: Celgene; Financial Interests, Personal, Advisory Board: Daiichi Sankyo; Financial Interests, Personal, Advisory Board: Eisai; Financial Interests, Personal, Advisory Board: Lilly; Financial Interests, Personal, Advisory Board: MSD; Financial Interests, Personal, Advisory Board: Mylan; Financial Interests, Personal, Advisory Board: NanoString; Financial Interests, Personal, Advisory Board: Novartis; Financial Interests, Personal, Advisory Board: Pfizer; Financial Interests, Personal, Advisory Board: Pierre Fabre; Financial Interests, Personal, Advisory Board: Puma; Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Advisory Board: Vifor; Financial Interests, Institutional, Funding, Discount prizes: Amercian Diagnostica; Financial Interests, Institutional, Funding, Discount prizes: Affymetrix; Financial Interests, Institutional, Funding, Discount prizes: NanoString. M. Piccart: Financial Interests, Personal, Invited Speaker: AstraZeneca; Financial Interests, Personal, Other, Consultant: Camel-IDS; Financial Interests, Personal, Advisory Board: Debiopharm; Financial Interests, Personal, Advisory Board: Immunomedics; Financial Interests, Personal, Invited Speaker: Lilly; Financial Interests, Personal, Advisory Board: Menarini; Financial Interests, Personal, Invited Speaker: MSD; Financial Interests, Personal, Invited Speaker: Novartis; Financial Interests, Personal, Advisory Board: Odonate; Financial Interests, Personal, Invited Speaker: Pfizer; Financial Interests, Personal, Advisory Board, Consultant and invited speaker: Roche-Genentech; Financial Interests, Personal, Advisory Board: Seattle Genetics; Financial Interests, Personal, Advisory Board: Immutep; Financial Interests, Personal, Advisory Board: Seagen; Financial Interests, Personal, Invited Speaker, Scientific Board: Oncolytics; Financial Interests, Institutional, Research Grant: AstraZeneca; Financial Interests, Institutional, Research Grant: Immunomedics; Financial Interests, Institutional, Research Grant: Lilly; Financial Interests, Institutional, Funding: Menarini; Financial Interests, Institutional, Funding: MSD; Financial Interests, Institutional, Funding: Novartis; Financial Interests, Institutional, Funding: Pfizer; Financial Interests, Institutional, Funding: Radius; Financial Interests, Institutional, Funding: Roche-Genentech; Financial Interests, Institutional, Funding: Servier; Financial Interests, Institutional, Funding: Synthon. E. de Azambuja: Financial Interests, Personal, Advisory Board: Roche/GNE; Financial Interests, Personal, Advisory Board: Novartis; Financial Interests, Personal, Advisory Board: Seattle Genetics; Financial Interests, Personal, Advisory Board: Zodiac; Financial Interests, Personal, Advisory Board: Libbs; Financial Interests, Personal, Advisory Board: Pierre Fabre; Financial Interests, Personal, Other, Travel grant: Roche/GNE; Financial Interests, Personal, Other, Travel grant: GSK/Novartis; Financial Interests, Institutional, Research Grant: Roche/GNE; Financial Interests, Institutional, Research Grant: AstraZeneca; Financial Interests, Institutional, Research Grant: GSK/Novartis; Financial Interests, Institutional, Research Grant: Servier. All other authors have declared no conflicts of interest.

Background

The emerging treatment paradigm in eBC, with new systemic therapies varying in efficacy, safety and conditions of use, creates a complex patient journey involving many inflection points with respect to treatment sequence, duration and response to initial therapy. We quantified patient preferences for attributes of different treatment pathways in eBC in Germany, Italy and Japan.

Methods

Patients diagnosed with HER2-negative stage I–IIIa breast cancer after 2014 who had undergone surgery and chemotherapy completed an online survey that included a DCE to assess attribute preferences of treatments for eBC. In 12 DCE tasks, patients indicated their preference between 2 hypothetical treatment profiles that varied in 8 attributes. Preference weights for each attribute level and relative attribute importance (RI) were estimated using hierarchical Bayesian modelling and calculated as a percentage based on the difference from the most to least favourable attribute level.

Results

Overall, 452 patients (Germany 151, Italy 151, Japan 150) participated; median (range) age was 48 (19–78) years; 80% were employed and most patients were satisfied (64%) or very satisfied (18%) with the prior therapy they received. Reducing risk of a serious side effect from 77% to 6% was most important (RI=27%), followed by increasing cancer-free survival at 3 years from 67% to 87% (RI=19%), decreasing treatment duration from 18 to 3 months (RI=16%) and reducing risk of nausea from 67% to 2% (RI=14%). Choice of a flexible vs fixed treatment plan was as important as reducing risk of neuropathy from 21% to 1% or fatigue from 41% to 3% (RI=7% for all). Choice of an oral vs intravenous regimen was least important (RI=5%). Some treatment attribute preferences differed by country; notably, efficacy was more important to patients in Japan.

Conclusions

Overall, this international cohort of patients regard the risk of serious side effects, treatment efficacy and treatment duration as highly important. Additionally, these patients would prefer a flexible treatment plan, with treatment escalation/de-escalation in line with response to initial therapy, over a fixed plan. This information may enhance physician–patient interactions in eBC.

Editorial acknowledgement

Editorial assistance was provided by Aaron Borg, PhD of PharmaGenesis Cambridge, Cambridge, UK, with funding from AstraZeneca and Merck Sharp & Dohme Corp., a subsidiary of Merck & Co., Inc.

Legal entity responsible for the study

AstraZeneca.

Funding

This study was funded by AstraZeneca and is part of an alliance between AstraZeneca and Merck Sharp & Dohme Corp., a subsidiary of Merck & Co., Inc., Kenilworth, NJ, USA.

Disclosure

A. Gennari: Financial Interests, Personal, Invited Speaker: Eisai; Financial Interests, Personal, Advisory Board: Eisai; Financial Interests, Personal, Advisory Board: Novartis; Financial Interests, Personal, Advisory Board: AstraZeneca; Financial Interests, Personal, Advisory Board: Roche; Financial Interests, Personal, Expert Testimony: Gentili; Financial Interests, Personal, Advisory Board: Daiichi Sankyo; Financial Interests, Personal, Advisory Board: Pfizer; Financial Interests, Personal, Advisory Board: Lilly; Financial Interests, Personal, Invited Speaker: Lilly. C. Jackisch: Financial Interests, Personal, Other, Honoraria, Consultancy, travel and accommodation expenses: AstraZeneca; Financial Interests, Personal, Other, Consultancy, travel and accommodation expenses: Lilly; Financial Interests, Personal, Leadership Role, Travel and accommodation expenses: Novartis; Financial Interests, Personal, Invited Speaker, Consultancy, travel and accommodation expenses: Roche. S. McCutcheon: Financial Interests, Personal, Full or part-time Employment: AstraZeneca; Financial Interests, Personal, Stocks/Shares: AstraZeneca. E. Flood: Financial Interests, Personal, Full or part-time Employment: AstraZeneca; Financial Interests, Personal, Stocks/Shares: AstraZeneca. B. Murali: Financial Interests, Personal, Full or part-time Employment: Cerner Enviza; Financial Interests, Personal, Other, Provides consultancy services to AstraZeneca as an employee of Cerner Enviza: AstraZeneca. X. Guillaume: Financial Interests, Personal, Full or part-time Employment: Cerner Enviza; Financial Interests, Personal, Other, Provides consultancy services to AstraZeneca as an employee of Cerner Enviza: AstraZeneca. O. Will: Financial Interests, Personal, Full or part-time Employment: Cerner Enviza; Financial Interests, Personal, Other, Provides consultancy services to AstraZeneca as an employee of Cerner Enviza: AstraZeneca. C. Shimizu: Financial Interests, Institutional, Research Grant: Chugai; Financial Interests, Personal, Other, Honoraria: Chugai; Financial Interests, Personal, Other, Honoraria: Eisai; Financial Interests, Personal, Other, Honoraria: Pfizer; Financial Interests, Institutional, Research Grant: AstraZeneca; Financial Interests, Institutional, Research Grant: Eli Lilly; Financial Interests, Institutional, Research Grant: Taiho. S. Mokiou: Financial Interests, Personal, Full or part-time Employment: AstraZeneca; Financial Interests, Personal, Stocks/Shares: AstraZeneca.

Background

Breast cancer remains a common diagnosis. Aromatase inhibitors (AI) are recommended for their mortality benefit in postmenopausal women with hormone sensitive disease. AI bone loss is a recognised adverse event with resultant increase in fracture risk. In 2017, a consensus statement of international bone and cancer societies published an algorithm based on risk factors and bone mineral densities (BMD) threshold for bone active therapeutic intervention.

Methods

Over 7 years, 1001 women were prescribed AI at a university teaching hospital. The new guidelines were adopted in 2017. 361 women had commenced AI prior to the adoption of guidelines and were offered bone active treatment based on NOS 2009 guidelines. 640 women were in the post implementation group and offered treatment based on the 2017 consensus guidelines. Women with osteoporosis were all offered treatment, the difference in guideline is pertinent to osteopenia and we analysed that group.

Results

Mean age was 64 years. 929 (93%) women were Caucasian, 723 (72%) had invasive ductal carcinoma and 863 (86%) were postmenopausal. At diagnosis, 428 (43%) had node-positive disease and 35 (4%) metastases. 91 (9%) had fractures prior to their cancer. First group (n=361): baseline DEXA mean left neck of femur (LNOF) BMD of 0.888 g/cm² (range 0.552-1.222). 143 (40%) women were normal, 174 (48%) osteopenic and 44 (12%) osteoporotic. Osteopenia- 44 women (25%) were treated and 33 had a repeat DEXA after 4 years (mean). Treatment group- LNOF mean BMD remained unchanged from 0.814 g/cm2 to 0.812 g/cm2 at the repeat (p= 0.94). 22 (13%) women had a fracture. Second group (n=640): baseline DEXA mean LNOF BMD of 0.888 g/cm² (range 0.512-1.390). 216 (33%) women were normal, 322 (50%) osteopenic and 107 (17%) osteoporotic. Osteopenia- 127 women (39%) were treated and 56 had a repeat DEXA after 3 years (mean). Treatment group- LNOF mean BMD remained relatively unchanged from 0.822 g/cm2 to 0.829 g/cm2 at the repeat (p= 0.6169). 8 (2.5%) women had a fracture.