Poster Display session Poster Display session

3P - Dissecting the spatial heterogeneity of single CTCs reveals immune evasion through MAX regulated CCL5 overexpression in hepatocellular carcinoma (ID 208)

Presentation Number
3P
Lecture Time
12:30 - 12:30
Speakers
  • Y. Sun (Shanghai, China)
Session Name
Poster Display session
Location
Room B, Geneva Palexpo, Geneva, Switzerland
Date
14.12.2018
Time
12:30 - 13:00
Authors
  • Y. Sun (Shanghai, China)
  • X. Yang (Shanghai, China)
  • J. Fan (Shanghai, China)

Abstract

Background

The prognosis of hepatocellular carcinoma (HCC) is closely associated with recurrence and metastasis which has been proposed to be initiated by circulating tumor cells (CTCs). Yet, the transcriptomic plasticity and immune evasion mechanism of CTCs during circulation are not well defined.

Methods

Blood was drawn from 4 different vascular sites, including hepatic vein (HV), peripheral artery (PA), peripheral vein (PV) and portal vein (PoV) of 10 localized HCC patients. Single CTCs were isolated by negative enrichment and robotic micromanipulator, followed by single-cell RNA sequencing (scRNA-seq). After filtering, 113 CTCs with qualified data were subjected to bioinformatics analysis. The scRNA-seq results were further validated in three independent HCC cohorts.

Results

Our scRNA-seq data revealed remarkable intra- and inter-vascular heterogeneity among CTCs from four vascular sites. We determined CTC transcriptional dynamics during transportation through consecutive vascular sites and revealed their adaptation mechanisms under biomechanical stress. We further classified CTCs from different vascular sites into two subsets, namely dormant and activated CTCs. Dormant CTCs were associated with a non-cycling state and upregulation of EMT/angiogenic signatures and showed stronger prognostic ability for early relapse than activated CTCs. Furthermore, we discovered an immune escape mechanism by which CTCs recruited regulatory T cells (Tregs) via expression of CCL5, consequently promoting the formation of an immunosuppressive microenvironment favorable for their survival in bloodstream and distant colonization.We proved that MAX, activated through the p38 pathway, was the key transcriptional factor regulating CCL5 overexpression, which was validated by ChIP, luciferase reporter gene and in vitro/vivo knockdown assays. And we further determined that Tregs-derived TGF-β1 can heighten MAX expression, thus amplifying the CCL5 expression.

Conclusions

Collectively, our results reveal a previously unappreciated spatial heterogeneity of CTCs and a CTC immune-escape mechanism, which may aid in designing new anti-metastasis therapeutic strategies in HCC.

Legal entity responsible for the study

Jia Fan.

Funding

The State Key Program of National Natural Science of China.

Disclosure

All authors have declared no conflicts of interest.

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Poster Display session Poster Display session

4P - Prospective study of circulating tumor cells in long survivors of immunotherapy (ID 334)

Presentation Number
4P
Lecture Time
12:30 - 12:30
Speakers
  • M. A. Brenes Fernández (Madrid, Spain)
Session Name
Poster Display session
Location
Room B, Geneva Palexpo, Geneva, Switzerland
Date
14.12.2018
Time
12:30 - 13:00
Authors
  • M. A. Brenes Fernández (Madrid, Spain)
  • A. Garcia Grande (Madrid, Spain)
  • F. Franco (Madrid, Spain)
  • M. Coronado (Madrid, Spain)
  • V. Calvo (Madrid, Spain)
  • L. Gutierrez Sanz (Madrid, Spain)
  • J. C. Sanchez (Madrid, Spain)
  • M. Torrente Regidor (Madrid, Spain)
  • B. Núñez (Madrid, Spain)
  • R. Gómez Bravo (Madrid, Spain)
  • M. Provencio Pulla (Majadahonda, Madrid, Spain)

Abstract

Background

Up to date, there isn’t current method or parameter that allows identifying long survivors in treatment with immunotherapy (IT) in a simple and accessible way. We made a prospective study of the usefulness of quantifying circulating tumor cells (CTCs) and CTCs/PDL1+ in patients treated with immunotherapy.

Methods

Patients, diagnosed with non-small cell lung cancer and in second-line treatment with IT were analyzed prospectively. CTCs from peripheral blood samples were isolated by double density gradient and immunomagnetic separation with AutoMACS equipment (M.Biotec). Quantification of CTCs was performed by Cytometry and Confocal Microscopy. The combination of both methodologies allows greater sensitivity and specificity. Samples were acquired in a MACSQuant cytometer (M.Biotec) and TCS SP5 Confocal Microscope (Leica).Data analysis was performed using MACSQuantify and LASF Lite. Determination of CTCs was made at the beginning of the treatment and every 3 months and up to 12 months during IT, together with radiological evaluation in each extraction. We selected those patients who had no progression of the disease for at least 12 months.

Results

Determination of CTCs alone did not allow us to obtain any pattern of recurrence or response. However, we were able to identify 7 patients in the group of long survivors (> 12 months of treatment with IT) using the marker PDL1. The study of their CTCs showed that none of them had circulating CTCs+/PD-L1+. Two of these patients were PDL1+ in tissue sample but negative CTC/PDL1 in blood. We also observed the presence of non-tumor WBC (white blood cells)/PDL1+, but their meaning is not clear in patients with relapse. To emphasize, another patient PDL1 + in tissue, CTC/PDL1+ in serum and low number of WBC, showed positive imaging (PET) of the progression of the disease and its CTC number increased in later determinations.

Conclusions

The absence of circulating CTCs/PDL1 + can predict a sustained response to long-term IT. Isolated CTCs without quantifying their associated expression of PDL1 are not associated with a particular pattern nor appear to be useful in identifying long survivors. WBCs that express PDL1 were associated with the appearance of relapse. Larger studies are needed to validate our results.

Legal entity responsible for the study

Hospital Universitario Puerta de Hierro Majadahonda.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

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Poster Display session Poster Display session

5P - Baseline CRP predicts severe immune related adverse events (ID 294)

Presentation Number
5P
Lecture Time
12:30 - 12:30
Speakers
  • R. Verheijden (Utrecht, Netherlands)
Session Name
Poster Display session
Location
Room B, Geneva Palexpo, Geneva, Switzerland
Date
14.12.2018
Time
12:30 - 13:00
Authors
  • R. Verheijden (Utrecht, Netherlands)
  • K. Suijkerbuijk (Utrecht, Netherlands)

Abstract

Background

The PD-1 inhibitors nivolumab and pembrolizumab are currently widely used as treatment of advanced melanoma. While being tolerated very well by some patients, they cause severe and even fatal immune related adverse events (irAEs) in others. Factors associated with increased risk of toxicity have barely been identified. With the perspective of long-term survival in stage IV disease, and increasing numbers of patients receiving adjuvant immunotherapy in stage III melanoma, being able to predict severe (≥grade 3) toxicity becomes more and more important. In this study, we aimed to assess factors associated with severe irAEs in clinical data and routine peripheral blood tests.

Methods

Patients with advanced (stage III-IV) melanoma, treated with nivolumab or pembrolizumab with at least 3 months follow-up were included. Demographical parameters together with lactate dehydrogenases (LDH), C-reactive protein (CRP), leukocytes, eosinophils, neutrophils and lymphocytes at baseline and at last measurement before toxicity were retrieved retrospectively. IrAEs were graded according to CTCAE 4.03. Non parametric tests were used to assess the difference between groups.

Results

60 patients treated with anti-PD1 monotherapy were included, of whom 7 developed ≥grade 3 irAEs. Median CRP at baseline was significantly higher in patients that experienced severe irAEs (25mg/L) compared to patients who did not (3 mg/L; p = 0,038). Furthermore, in patients with severe irAEs, median eosinophil level at last measurement before toxicity (0,27x109/L) was significantly increased compared to baseline (0,15x109/L; p = 0,047).

Conclusions

Our data indicate that baseline CRP is a potential predictor of severe anti-PD1 induced toxicity. If confirmed in a larger study, CRP could play a role in clinical decision making at the start of treatment. Furthermore, our observation that increase of eosinophils precedes ≥grade 3 toxicity underpins its possible role in the pathogenesis of irAEs and could help identify and treat irAEs at an early stage.

Legal entity responsible for the study

University Medical Center Utrecht.

Funding

Has not received any funding.

Disclosure

K. Suijkerbuijk: Consulting/advisory relationship: Bristol-Myers Squibb, MSD; Honoraria (paid to institution): Novartis, Roche. All other authors have declared no conflicts of interest.

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Poster Display session Poster Display session

6P - Predictive score using clinical and blood biomarkers in advanced non-small cell lung cancer (aNSCLC) patients treated with immunotherapy (ID 459)

Presentation Number
6P
Lecture Time
12:30 - 12:30
Speakers
  • A. Prelaj (Milan, Italy)
Session Name
Poster Display session
Location
Room B, Geneva Palexpo, Geneva, Switzerland
Date
14.12.2018
Time
12:30 - 13:00
Authors
  • A. Prelaj (Milan, Italy)
  • S. Rebuzzi (Genova, Italy)
  • P. Pizzutilo (Bari, Italy)
  • M. Montrone (Bari, Italy)
  • F. Pesola (Bari, Italy)
  • V. Longo (Bari, Italy)
  • V. Lapadula (Bari, Italy)
  • F. Cassano (Bari, Italy)
  • P. Petrillo (Bari, Italy)
  • D. Bafunno (Bari, Italy)
  • N. Varesano (Bari, Italy)
  • V. Lamorgese (Bari, Italy)
  • A. Mastrandrea (Bari, Italy)
  • D. Ricci (Bari, Italy)
  • A. Catino (Bari, Italy)
  • G. Domenico (Bari, Italy)

Abstract

Background

Despite the overall survival (OS) benefit, only 18-20% of aNSCLC patients respond to immune-checkpoint inhibitors (ICI) as second-line therapy with a median progression-free survival (mPFS) of 2-4 months. The identification of predictive and prognostic biomarkers to select patients most likely to respond to ICI is greatly needed in guiding clinical practice.

Methods

We conducted a retrospective monocentric analysis of 154 aNSCLC patients receiving single-agent Nivolumab or Pembrolizumab as second-line (68%) and >3rd line (32%). We collected complete blood cell count at baseline and evaluated LDH, absolute neutrophil count (ANC), lymphocyte count (ALC), monocyte count (AMC) and eosinophil count (AEC) and their ratio such as neutrophil-lymphocyte ratio (NLR), derived-NLR (dNLR) and lymphocyte-monocyte ratio (LMR). Univariate and multivariate analyses were performed to identified indipendent predictors factors for immunotherapy (using Kaplan–Meier and Cox Progression analyses).

Results

The multivariate analysis on clinical factors showed the negative predictive role of ECOG PS 2 and liver metastasisand the positive predicitive role of smoking status. The multivariate analysis for PFS showed the negative predictive role of higher ANC (>6000/mL) and LDH (>400 mg/dl) and positive predictive role of higher ALC (>2200/mL). Also, according to stepwise regression analyses, NLR>4 playsa negative predictive and prognostic role at baseline. Finally, five predictive clinical and blood biomarkers at baseline (smoking status, ECOG PS, liver metastases, LDH and NLR), were used to create a predictive score for immunotherapy. Three predictive groups were defined as high, intermediate and low with a mPFS of 10.2 vs 4.9 vs 1.7 months respectively (HR 4.18 95% IC 2.64–6.62, p < 0.001).

Predictive FactorAssessmentPoint
ECOG PS0-1 20 1
Smoking (pack-years)> 43 < 430 1
Liver metastasesNo Yes0 1
LDH (mg/dl)< 400 > 4000 1
NLR< 4 > 40 1
Predictive groups (Points): 1 = 0 2 = 1-2 3 = 3-5PFS (months): 10.2 4.9 1.7HR 4.18 95% IC (2.64 – 6.62) p < 0.001

Conclusions

In advanced NSCLC patients treated with second-line immunotherapy, the identification of five and predictive clinical and blood biomarkers at baseline, combined in a predictive score, may help identify patients most likely to benefit from immunotherapy.

Legal entity responsible for the study

Clinical Cancer Center Giovanni Paolo II, Bari, Italy.

Funding

Has not received any funding.

Disclosure

G. Domenico: Advisory board: Bristol-Myers Squibb. All other authors have declared no conflicts of interest.

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Poster Display session Poster Display session

7P - Monocyte-to-lymphocye ratio (MLR) and LDH level in metastatic colorectal cancer (mCRC) patients (pts) (ID 430)

Presentation Number
7P
Lecture Time
12:30 - 12:30
Speakers
  • D. Basile (Aviano, Italy)
Session Name
Poster Display session
Location
Room B, Geneva Palexpo, Geneva, Switzerland
Date
14.12.2018
Time
12:30 - 13:00
Authors
  • D. Basile (Aviano, Italy)
  • L. Gerratana (Udine, Italy)
  • C. Corvaja (Udine, Italy)
  • G. Pelizzari (Udine, Italy)
  • S. Garattini (Udine, Italy)
  • C. Lisanti (Udine, Italy)
  • M. Bartoletti (Udine, Italy)
  • L. Bortot (Udine, Italy)
  • V. Fanotto (Udine, Italy)
  • E. Ongaro (Udine, Italy)
  • F. Cortiula (Udine, Italy)
  • A. Parnofiello (Udine, Italy)
  • M. G. Vitale (Udine, Italy)
  • L. Da Ros (Aviano, Italy)
  • P. Di Nardo (Aviano, Italy)
  • E. Torrisi (Aviano, Italy)
  • M. Guardascione (Aviano, Pordenone, Italy)
  • G. Miolo (Aviano, Italy)
  • A. Buonadonna (Aviano, Italy)
  • F. Puglisi (Aviano, Italy)

Abstract

Background

Tumor microenvironment (TME) is a pitch for multiple players, where the crosstalk between tumor and immune cells determines the fate of tumor progression. In fact, immune system may either destroy or paradoxically promote cancer growth, by recruiting immunosuppressive and inflammatory cells. MLR and LDH levels could be dynamic biomarkers that provide indirect information about TME and are associated with poor prognosis in many tumors. Therefore, we evaluated their prognostic impact in mCRC pts.

Methods

We conducted a retrospective cohort study evaluating consecutive data of 165 mCRC pts treated in 2004-2017 at the Unit of Medical Oncology and Oncology Prevention, CRO of Aviano. The prognostic impact of MLR and LDH levels on overall survival (OS) was analyzed through uni- and multivariate Cox regression analysis.

Results

At median follow-up of 61.87 months, median OS was 30.74 months. Overall, 100 pts (62%) were aged <65, 63 pts (39%) had a left tumor, 9 (6%) had a BRAF mutation and 45 (28%) had a KRAS mutation, underestimated for the missing data. High levels of LDH (>480 U/L) and MLR (>0.44) were discovered in 35 (21%) and 40 pts (24%), respectively. By univariate analysis, resection (HR 0.25, P < 0.001, 95% CI 0.14-0.45), metastasectomy (HR 0.46, P < 0.001, 95%CI 0.30-0.70) and sidness (left tumors: HR 0.58, P = 0.035, 95%CI 0.36-0.96) were associated with better OS. Conversely, older age (HR 1.63, P = 0.014, 95%CI 1.10-2.42), KRAS mutation (HR 1.77, P = 0.017, 95%CI 1.19-2.85), LDH or MLR high (HR 2.92, P < 0.001, 95% CI 1.77-4.79) or both (HR 4.02, P < 0.001, 95%CI 1.93-8.37) were associated with worse OS. By multivariate analysis, metastasectomy (HR 0.53, P = 0.04, 95%CI 0.29-0.97), KRAS mutation (2.10, P = 0.014, 95% CI 1.16-3.79), LDH or MLR high (HR 3.05, P < 0.001, 95%CI 1.68-5.55) or both (HR 2.65, P = 0.039, 95%CI 1.05-6.68) confirmed their impact on OS. Interestingly, high MLR was associated with right and rectum tumors P = 0.007).

Conclusions

We showed that high baseline LDH, MLR or both are poor prognostic factors in mCRC pts adding further evidence of the interlink between immune system, inflammation and cancer. However, further prospective and translational studies are needed to better deepen this topic.

Legal entity responsible for the study

CRO, IRCCS, National Cancer Institute of Aviano.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.

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Poster Display session Poster Display session

8P - Anti-PD1 therapy increases peripheral blood NKT cells and chemokines in metastatic melanoma patients (ID 337)

Presentation Number
8P
Lecture Time
12:30 - 12:30
Speakers
  • H. Hakanen (Helsinki, Finland)
Session Name
Poster Display session
Location
Room B, Geneva Palexpo, Geneva, Switzerland
Date
14.12.2018
Time
12:30 - 13:00
Authors
  • H. Hakanen (Helsinki, Finland)
  • M. Hernberg (Helsinki, Finland)
  • S. Mäkelä (Helsinki, Finland)
  • B. Yadav (Helsinki, Finland)
  • O. Brück (Helsinki, Finland)
  • S. Juteau (Helsinki, Finland)
  • L. Kohtamäki (Helsinki, Finland)
  • M. Ilander (Helsinki, Finland)
  • S. Mustjoki (Helsinki, Finland)
  • A. Kreutzman (Helsinki, Finland)

Abstract

Background

Anti-PD1 therapy has proven to be effective in various cancer types, but the overall impact of the treatment on the immune system is not yet fully understood. Therefore, we aimed in this study to discover the effects of anti-PD1 therapy on the immune system, especially on NK and NKT cells, which are less studied, but known to be involved in antitumor immune events.

Methods

Peripheral blood from immuno-oncology naive metastatic melanoma patients (n = 19) were obtained before the first infusion of anti-PD1, then 1 and 3 months later. From each time-point, complete blood counts (CBC) were obtained, and comprehensive immunophenotyping of NK, NKT, and T cells was performed. Also, 92 serum cytokines were measured using the Olink inflammation panel.

Results

Patients were categorized into two cohorts: responders (R, n = 6, PFS=17.0 months) and progressive disease (PD, n = 9, PFS=5.0 months). 4 patients were excluded due to challenging clinical evaluation of response. CBC indicated a significant decrease in the mean frequency of lymphocytes in PD but not in the R cohort. The responders had also higher frequency of lymphocytes at 1- and 3-month time points and lower frequency of neutrophils before initiation and after 1 and 3 months of treatment. The CBC absolute counts revealed that the responders had less neutrophils and monocytes after 3 months of treatment when compared to PD. Immunophenotyping revealed no changes in the frequency of T and NK cells during treatment, however the frequency of NKTbrightcells increased significantly. Moreover, R had more NKTdimcells before and after 3 months of therapy. Also, cytotoxic CD56dimNK cells expressed increased CD25 and CD45RO after 1 month of therapy. The cytokine assay indicated that anti-PD1 significantly increased the levels of CXC-family cytokines in the serum; CXCL9, CXCL11, CXCL10. Also, IL-12B and TNFRSF9 levels were increased. Further comparison of the two cohorts showed that CXCL9 was only increased in R cohort.

Conclusions

Anti-PD1 therapy increases the levels of NKT cells, inflammation related chemokines in blood and the expression of markers linked to enhanced cytotoxicity of NK cells. Therefore, further studies to investigate the role of NK and NKT cells in anti-PD1 induced responses are warranted.

Legal entity responsible for the study

Anna Kreutzman.

Funding

Academy of Finland, Cancer Society of Finland, Sigrid Juselius Foundation.

Disclosure

M. Hernberg, S. Mäkelä, L. Kohtamäki: Trial funding: MSD, BMS, Amgen. S. Mustjoki: Honoraria, research funding: Novartis, Bristol-Myers Squibb, Ariad, Pfizer. All other authors have declared no conflicts of interest.

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Poster Display session Poster Display session

9P - Identification of serum microRNAs predicting the response to nivolumab in patients with advanced gastric cancer (ID 182)

Presentation Number
9P
Lecture Time
12:30 - 12:30
Speakers
  • T. Miyamoto (Chuo-ku, Japan)
Session Name
Poster Display session
Location
Room B, Geneva Palexpo, Geneva, Switzerland
Date
14.12.2018
Time
12:30 - 13:00
Authors
  • T. Miyamoto (Chuo-ku, Japan)
  • K. Kato (Tokyo, Japan)
  • J. Matsuzaki (Tokyo, Japan)
  • S. Takizawa (Tokyo, Japan)
  • K. Sudo (Tokyo, Japan)
  • H. Shoji (Tokyo, Japan)
  • S. Iwasa (Tokyo, Japan)
  • Y. Honma (Tokyo, Japan)
  • A. Takashima (Tokyo, Japan)
  • N. Okita (Tokyo, Japan)
  • H. Sakamoto (Tokyo, Japan)
  • N. Boku (Kawasaki, Japan)
  • O. Takahiro (Tokyo, Japan)

Abstract

Background

Recent studies have reported that serum microRNAs (miRNAs) are potentially useful cancer biomarkers. In patients with gastric cancer (GC), the efficacy of nivolumab (Nivo), a PD-1 inhibitor, was shown in a phase III study, but predictive biomarkers of Nivo for gastric cancer have not been found. We investigated whether serum miRNAs could be predictive markers of the efficacy of Nivo in GC.

Methods

The subjects of this study were 20 patients who were enrolled in the phase III study (ONO-4538-12) and received Nivo 3 mg/kg IV Q2W at our institution. Expressions of 2565 miRNAs in the serum samples before and during treatment were analyzed using “3D-Gene” Human miRNA Oligo Chip (Toray Industries, Inc.). We explored miRNAs that were significantly associated with the treatment response using receiver operating characteristic analysis and Cox regression analysis.

Results

Median progression-free survival (PFS) was 2.3 month, and partial response was achieved in four of the 20 patients who received Nivo. Serum samples before (n = 20) and 4 weeks after (n = 17) the treatment of Nivo were available. Two different miRNAs, one before and the other after treatment, were identifiedwhich were related to response to Nivo. AUC of miR-A identified before treatment was 0.88, whereas that of miR-B after the first treatment was 0.85. The overall response rate (ORR) was 44% (4/9) in miR-A-high patients and 0% (0/11) in miR-A-low patients. The ORR was 50% (4/8) in miR-B-high patients and 0% (0/9) in miR-B-low patients. Median PFS was 14.3 m (95%CI: N/A-35.1) in miR-A-high patients and 1.6 m (95%CI: 0.9-2.3) in miR-A–low patients (HR = 0.19, log rank: p = 0.01), and those were 5.6 m (95%CI: N/A-35.1) and 1.6 m (95%CI: 0.9-2.3) in miR-B-high patients (HR = 0.21, log rank: p = 0.01), respectively.

Conclusions

The two miRNAs in GC pts may be a predictive marker for identifying pts who derived greater benefit from Nivo therapy.

Legal entity responsible for the study

Ken Kato.

Funding

Has not received any funding.

Disclosure

S. Takizawa: Personal fees: Toray Industries, Inc., outside the submitted work. O. Takahiro: Grants: Kewpie Corporation, Takeda, BioMimetics Sympathies, Rohto Pharmaceutical Co., Ltd., Inter Stem, Japan Atherosclerosis Research Foundation, Ono Pharmaceutical Co., Ltd., Daiichi-Sankyo Company. All other authors have declared no conflicts of interest.

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Poster Display session Poster Display session

10P - Clinical and laboratory predictors of immune checkpoint inhibitor efficacy in non-small cell lung cancer (ID 245)

Presentation Number
10P
Lecture Time
12:30 - 12:30
Speakers
  • P. Christopoulos (Heidelberg, Germany)
Session Name
Poster Display session
Location
Room B, Geneva Palexpo, Geneva, Switzerland
Date
14.12.2018
Time
12:30 - 13:00
Authors
  • P. Christopoulos (Heidelberg, Germany)
  • J. Kohlhäufl (Heidelberg, Germany)
  • F. Bozorgmehr (Heidelberg, Germany)
  • J. Kuon (Heidelberg, Germany)
  • M. Schneider (Heidelberg, Germany)
  • O. Neumann (Heidelberg, Germany)
  • S. Liersch (Heidelberg, Germany)
  • C. Heussel (Heidelberg, Germany)
  • H. Winter (Heidelberg, Germany)
  • F. Herth (Heidelberg, Germany)
  • S. Rieken (Heidelberg, Germany)
  • T. Muley (Heidelberg, Germany)
  • M. Meister (Heidelberg, Germany)
  • H. Bischoff (Heidelberg, Germany)
  • F. Lasitschka (Heidelberg, Germany)
  • A. Stenzinger (Heidelberg, Germany)
  • M. Thomas (Heidelberg, Germany)

Abstract

Background

Treatment with immune checkpoint inhibitors (ICI) prolongs overall survival (OS) and confers long-term disease control in 15-20% of non-small cell lung cancer (NSCLC) patients. Patient selection currently depends on the levels of PD-L1 expression, but correlation with outcome is weak.

Methods

We retrospectively analyzed the clinical course of ICI-treated stage IV NSCLC patients at our institution.

Results

A total of 453 patients were identified with a median age of 64 years having received nivolumab (57%), pembrolizumab (35%), PD-L1 inhibitors (7%) or PD1 blockade in combination with chemotherapy or CTLA4 inhibitors (1%). Progression-free survival (PFS) under ICI was significantly longer for patients receiving ICI in the first (21%, 7 months in median) compared to second (47%, 4 months in median) and later treatment lines (2 months in median, p < 0.001), for current and ex-smokers (91% of cases, p = 0.034), in case of adenocarcinoma (66%) compared to squamous cell carcinoma (28%) and other histologies (9%, p < 0.001), while age, sex and ECOG status at initial diagnosis had no influence. The total number of metastatic sites and the presence of liver metastases at start of IO treatment were associated with shorter ICI responses (p = 0.018 and p = 0.005, respectively), while metastases to other organs, especially brain, did not play a role. Blood markers, like the Lymphocyte-to-Neutrophile-Ratio (LNR) as well as CRP and LDH as indicators of inflammation and tumor load, had the highest discriminatory value (≥ 2x longer median PFS for cases with higher LNR or lower CRP or LDH, p < 0.0001 for each), while the occurrence of immune-related adverse events (irAE) conferred longer PFS (p < 0.001) as well as overall survival (OS) from the start of IO treatment (p < 0.01). PFS under ICI was similar for cases with PD-L1 <1% vs. 1-49% (14% and 34%, respectively), while cases with PD-L1 expression >50% had an ICI-PFS twice as long (p = 0.011).

Conclusions

Several clinical and blood parameters appear to correlate with ICI benefit better than tissue PD-L1 expression levels in NSCLC patients and could be used along with molecular markers to improve predictive tools for lung cancer immunotherapy.

Legal entity responsible for the study

Thoraxklinik Heidelberg.

Funding

Has not received any funding.

Disclosure

F. Bozorgmehr: Research funding: BMS; Travel grants: BMS, MSD. J. Kuon: Research funding: AstraZeneca, Celgene. C. Heussel: Consultation, lecture and other fees: Novartis, Siemens, Chiesi, Intermune, MEDA Pharma, Bracco, Pfizer, MSD, Roche, Lilly, AstraZeneca, Schering-Plough, Essex, Gilead; Ownership of GSK stocks. F. Herth: Advisory board fees and honoraria: Lilly, Roche, AstraZeneca, Novartis, Boehringer, Chiesi, Teva, Pulmonx BTG, Olympus; Research funding: Lilly, Roche, AstraZeneca, Novartis, Boehringer, Chiesi, Teva. T. Muley: Research funding, patents: Roche. A. Stenzinger: Advisory board honoraria: BMS, AstraZeneca, ThermoFisher, Novartis; Speaker’s honoraria: BMS, Illumina, AstraZeneca, Novartis, ThermoFisher, MSD, Roche; Research funding: Chugai. M. Thomas: Advisory board honoraria: Novartis, Lilly, BMS, MSD, Roche, Celgene, Takeda, AbbVie, Boehringer; Speaker’s honoraria: Lilly, MSD, Takeda; Research funding: AstraZeneca, BMS, Celgene, Novartis, Roche.  All other authors have declared no conflicts of interest.

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11P - A SNP germinal signature for predicting checkpoint inhibitor treatment outcome (ID 435)

Presentation Number
11P
Lecture Time
12:30 - 12:30
Speakers
  • G. Milano (Nice, France)
Session Name
Poster Display session
Location
Room B, Geneva Palexpo, Geneva, Switzerland
Date
14.12.2018
Time
12:30 - 13:00
Authors
  • G. Milano (Nice, France)
  • S. Refae (Nice, France)
  • J. Gal (Nice, France)
  • N. Ebran (Nice, France)
  • J. Otto (Nice, France)
  • S. Shell (San Diego, United States of America)
  • R. Everts (San Diego, United States of America)
  • E. Chamorey (Nice, France)
  • E. Saada-Bouzid (Nice, France)

Abstract

Background

Cumulated clinical experience with checkpoint inhibitors (CPIs) points to a strong need for the identification of predictive biomarkers. Surprisingly, the potential role of the host has not been advocated so far. We developed a custom designed panel of single nucleotide polymorphisms (SNPs) from genes potentially implicated in the response to CPIs.

Methods

We studied 94 patients treated in Centre Antoine Lacassagne (Nice, France) with CPI (anti PD-1/PD-L1). High-throughput genotyping of germinal DNA was performed by MassARRAY ImmunoCarta (AGENA Bioscience®) using a custom-panel of 173 SNPs across 90 selected genes (minor allelic frequency ≥5% in the Caucasian population). All tested SNPs were in Hardy-Weinberg equilibrium, and linkage disequilibrium analyses were performed (r2>0.8). A Ridge-penalized logistic regression with 5-fold cross validation was used for the SNP identification to predict objective response (complete or partial response) to treatment.

Results

Median age of patients was 68 (range: 32-85), 67% were male, 51% had non-small cell lung carcinoma (NSCLC), 14% had head and neck squamous cell carcinoma (HNSCC), 15% had renal cell carcinoma (RCC), 13% had melanoma and 7% had another cancer type. Median follow-up was 16.3 months (95%CI: 12.5-18.3). The following SNPs’ decreasing intrinsic weight according to response were selected by the multivariate modeling approach (CCR2: rs1799864, FAS: rs2234767, CD3G: rs3753058, CTLA4: rs5742909, CCL2: rs13900, TNXB: rs12153855, Il1RN: rs419598, PD1: rs11568821, IL17A: rs2275913, IL12B: rs3212227, TLR3: rs7668666, CXCR3: rs2280964, IL10: rs1800871, IL6: rs2069837, TRAF3: rs7145509, VEGFR3: rs307821). In the training set, the accuracy was 0.87 (95%CI: 0.76-0.95; p < 0.001) associated with a sensitivity and a specificity of 0.90 and 0.83, respectively, with a ROC curve AUC at 0.93 (95%CI: 0.87-0.99). In the validation set, the accuracy decreased to 0.71 (95%CI: 0.52-0.85; p < 0.02) associated with a sensitivity and a specificity of 0.82 and 0.60, respectively, and with a ROC curve AUC of 0.85 (95%CI: 0.72-0.98).

Conclusions

These preliminary results point to the feasibility of a signature based on host characteristics for predicting response to CPI.

Legal entity responsible for the study

Centre Antoine Lacassagne, Nice, France.

Funding

Has not received any funding.

Disclosure

G. Milano: Paid scientific consultant: Agena Bioscience. S. Shell, R. Everts: Employee of Agena Bioscience. All other authors have declared no conflicts of interest.

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Poster Display session Poster Display session

12P - MSI status plus immunoscore to select metastatic colorectal cancer patients for immunotherapies (ID 401)

Presentation Number
12P
Lecture Time
12:30 - 12:30
Speakers
  • J. Galon (Paris, France)
Session Name
Poster Display session
Location
Room B, Geneva Palexpo, Geneva, Switzerland
Date
14.12.2018
Time
12:30 - 13:00
Authors
  • J. Galon (Paris, France)
  • B. Mlecnik (Paris, France)
  • F. Hermitte (Marseille, France)
  • F. Marliot (Paris, France)
  • C. Bifulco (Portland, OR, United States of America)
  • A. Lugli (Bern, Switzerland)
  • I. D. Nagtegaal (Nijmegen, Netherlands)
  • A. Hartmann (Erlangen, Germany)
  • M. Van den Eynde (Brussels, Belgium)
  • M. Roehrl (New-York, AL, United States of America)
  • P. Ohashi (Toronto, ON, Canada)
  • E. Zavadova (Prague, 2, Czech Republic)
  • T. Torigoe (Sapporo, Japan)
  • P. Patel (Ahmedabad, India)
  • Y. Wang (Xian, China)
  • Y. Kawakami (Tokyo, Japan)
  • F. M. Marincola (Redwood City, CA, United States of America)
  • P. A. Ascierto (Napoli, Italy)
  • B. Fox (Portland, United States of America)
  • F. Pagès (Paris, France)

Abstract

Background

Only a fraction of patients with metastatic disease respond to immune checkpoint inhibitors (ICI). Responders are more likely to be defective in mismatch repair genes (MSI+). MSI+ patients are over-represented in the group of tumors with high densities of CD8 T-cells. Patients with High T-cell infiltration have a higher expression of PD-1 and PDL1, and are more likely to respond to ICI. These observations suggest that the response to ICI is strongly dependent on the presence of an established in situ adaptive immune reaction (Mlecnik Immunity 2016). MSI + colon cancer stage IV patients are eligible to ICI. However this only concerns a minority of patients. Taking into account the evaluation of the in situ immune reaction in the primary tumor together with MSI status could clarify and extend the range of patients eligible to ICI.

Methods

Immunoscore is a robust and validated IVD test measuring the host immune reaction (CD3+ and CD8+ cells) at the tumor site. Immunoscore classifies patients’ tumors into High, Intermediate or Low. We investigated the Immunoscore (IS) distribution of primary tumors of UICC-TNM stage I-III patients who experienced metachronous metastase(s) from the Immunoscore international validation cohort (Pagès The Lancet 2018). MSI status was assessed with the molecular new Bethesda panel and by IHC.

Results

We recorded 1579 UICC-TNM stage I/II/III patients with available MSI status and IS. 318 patients (20%) experienced a relapse. Among those patients, 36 patients (11%) were MSI+ whereas the vast majority of them (89%; 282 patients) were MSS. 43 patients (13%) were IS High, 136 patients (43%) were IS Intermediate and 139 patients (44%) were IS Low. Importantly, 33 patients (12%) were classified IS High in MSS tumors. Combining MSI+ status with IS high to select patients for ICI extends from 11% to 22% the patients eligible to such immunotherapy.

Conclusions

The proportion of metastatic colon cancer patients eligible for ICI could be refined and extended by the characterization of their primary tumor immune infiltrate based on the Immunoscore status, as suggested by Le et al (Cancer Immunol Res 2017). Interventional trials are now needed to validate the predictive value of Immunoscore.

Legal entity responsible for the study

Society for Immunotherapy of Cancer SITC.

Funding

French National Institute of Health and Medical Research, LabEx Immuno-oncology, Transcan ERAnet Immunoscore European project, Association pour la Recherche contre le Cancer, CARPEM, AP-HP, Institut National du Cancer, Italian Association for Cancer Research, national grants, Society for Immunotherapy of Cancer.

Disclosure

J. Galon: HalioDx Co-founder /SAB; Consult/ Grts: PE, IObiotech, MedImmune, Janssen, BMS, AstraZeneca, Novartis, Definiens, Merck Serono, IObiotech, Nanostring, Illum., Northwest, Actelion, Amgen, Kite Pharma, Roche, GSK, Compugen, Mologen. F. Hermitte: Employee and co-founder: HalioDx. M. Roehrl: Consulting/Advisory roles: Member, Scientific advisory board: Proscia, Baltimore, MD - Member, Scientific advisory board, Trans-Hit, Montreal, QC - Advisor, Gerson Lehrman Group F.M. Marincola: Employee: Abbvie. B. Fox, F. Pagès: Immunoscore Patent (INSERM) co-inventor. All other authors have declared no conflicts of interest.

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Poster Display session Poster Display session

13P - Correlative analysis of gene expression changes and PD-L1 immunoexpression in non-small cell lung cancer (ID 225)

Presentation Number
13P
Lecture Time
12:30 - 12:30
Speakers
  • I. M. Guerreiro (Porto, Portugal)
Session Name
Poster Display session
Location
Room B, Geneva Palexpo, Geneva, Switzerland
Date
14.12.2018
Time
12:30 - 13:00
Authors
  • I. M. Guerreiro (Porto, Portugal)
  • D. Barros-Silva (Porto, Portugal)
  • P. Lopes (Porto, Portugal)
  • A. Cunha (Porto, Portugal)
  • J. Lobo (Porto, Portugal)
  • A. Rodrigues (Porto, Portugal)
  • M. Soares (Porto, Portugal)
  • L. Antunes (Porto, Portugal)
  • R. Henrique (Porto, Portugal)
  • C. Jerónimo (Porto, Portugal)

Abstract

Background

Lung cancer (LC) cells frequently express programmed death-ligand 1 (PD-L1). Although this expression grossly correlates with likelihood of response to checkpoint inhibitors, prediction of response is rather imperfect and, thus, more accurate predictive biomarkers are mandatory. The aim of this study was to investigate the association of immune checkpoint PD-L1 and DNA methylation status of DNA repair genes (RAD51B and XXRC3) as well as vimentin (VIM) expression in non-small cell lung cancer (NSCLC), correlating with patients' outcome.

Methods

A cohort of NSCLC patients diagnosed between August 2014 and June 2017 were enrolled after informed consent. Expression of PD-L1 was determined by IHQ. Evaluation of the methylation status of DNA repair genes (RAD51B and XRCC3) and VIM expression was performed by quantitative methylation-specific PCR (qMSP) using SybrGreen methodology. Predictors of PD-L1 expression were determined using logistic regression multivariable models. Impact on overall survival was determined using Cox analysis.

Results

A total of 75 patients with NSCLC were assessed for PD-L1 immunoexpression, 60 (80%) were male and the median age was 64 years old (range: 29-88). Fifty patients (66.7%) presented adenocarcinoma, 24 (32%) squamous cell carcinoma and one NSCLC NOS. Thirty-nine (52%) cases depicted positivity for PD-L1. RAD51B promoter methylation levels and VIM expression were significantly higher in PD-L1 positive cases compared to negative group (p = 0.01). This significant association was maintained in multivariable analyses: per each unit increase in RAD51B promoter methylation level and VIM expression, the OR for PD-L1 expression was 20.4 (CI95%: 1.5-275.2) and 1.23 (CI95%: 1-1.4), respectively. Association between XXRC3 promoter methylation and PD-L1 expression was not found. None of the analyzed markers associated with patients’ overall survival.

Conclusions

Herein, higher RAD51B methylation levels and VIM expression are independently associated with PD-L1 immunoexpression. Further studies with an extended cohort and follow-up period are warranted to validate these results.

Legal entity responsible for the study

Instituto Português de Oncologia do Porto.

Funding

Instituto Português de Oncologia do Porto.

Disclosure

All authors have declared no conflicts of interest.

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Poster Display session Poster Display session

14P - Relation between IDH1 status, histologic grade, immune-cell infiltration and expression of immune-related genes in patients with gliomas (ID 386)

Presentation Number
14P
Lecture Time
12:30 - 12:30
Speakers
  • S. Cabezas-Camarero (Madrid, Spain)
Session Name
Poster Display session
Location
Room B, Geneva Palexpo, Geneva, Switzerland
Date
14.12.2018
Time
12:30 - 13:00
Authors
  • S. Cabezas-Camarero (Madrid, Spain)
  • R. Pérez-Alfayate (Madrid, Spain)
  • I. Casado Fariñas (Madrid, Spain)
  • M. Sáiz-Pardo Sanz (Madrid, Spain)
  • I. Subhi-Issa (Madrid, Spain)
  • P. Pérez-Segura (Madrid, Spain)
  • V. García-Barberán (Madrid, Spain)

Abstract

Background

Very few data exist regarding the immune profile of gliomas. Our aim was to study different immunological parameters according to IDH1 mutational status and histologic grade.

Methods

Patients with grade II to IV gliomas were prospectively enrolled. Immunohistochemistry (IHC) was used to quantify CD4+ and CD8+ T cells and to study IDH1 and PDL1 (> 5%) expression on tumor cells. These parameters were correlated to mRNA expression of 20 immune-related genes (CSF1R, CTLA4, CXCL9, CXCL10, CXCL13, FOXP3, GZMA, HLADRA, IDO1, INFG, PD1, PDL1, PDL2, PRF1, STAT1, STAT3, TIGIT, TIM3, VISTA, TLR7) according to IDH1 status and histologic grade.

Results

Between February 2017 and July 2018, 20 patients (pts) were enrolled, 14 glioblastomas (GB: 9 IDHWT, 5 IDH1MUT) and 6 lower-grade gliomas (LGG: 2 IDHWT, 4 IDHMUT). Among 18 pts: PDL1 IHC was positive (PDL1IHC+) in 10 GB (3 IDH1MUT) and 1 LGG (IDH1MUT); CD4+ T cell infiltration (intratumoral (IT)/perivascular(PV)) in 5 GB (2 IDH1MUT); CD8+ T cell infiltration in 12 GB (8 IT+PV, 3 IT, 1 PV) and 5 LGG (1 IT, 4 IT+PV). PDL1IHC+ more frequent in GB + LGGIDH1wt vs LGGIDH1mut (P = 0.06). PDL1IHC+ tumors shoed lower expression of STAT1 (P = 0.042), TIM3 (P = 0.02) and TLR7 (P = 0.06). Tumors with > 20 CD8+ T cells/HPF showed higher expression of PD1 (P = 0.03), PRF1 (P = 0.03) and STAT1 (P = 0.03).

Conclusions

Histologic grade and IDH1 status identify glioma groups with different immunophenotypes. This results should be confirmed in a larger sample.

Legal entity responsible for the study

Santiago Cabezas.

Funding

GEINO (Grupo Español de Investigación en Neuro-Oncología).

Disclosure

All authors have declared no conflicts of interest.

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