Molecular target/Disease orientation Education session

Sarcoma

Lecture Time
09:50 - 10:10
Speakers
  • Jean-Yves Blay, Lyon, CEDEX, France, Centre Léon Bérard
Location
Fleming room, Queen Elizabeth II Centre, London, United Kingdom
Date
08.11.2019
Time
08:50 - 10:30
Molecular pathology and single cell analysis: New technologies coming to practise Workshop

How to analyse data from digital pathology

Lecture Time
14:30 - 15:00
Speakers
  • Trevor Graham, London, United Kingdom, Barts Cancer Institute-Queen Mary University of London
Location
Abbey room, Queen Elizabeth II Centre, London, United Kingdom
Date
07.11.2019
Time
14:00 - 17:00
Lunch and poster viewing Poster display

15P - Mechanistic insight into anti-cancer activity of plumbagin in endocrine resistant and HER2-overexpressed breast cancer cells

Lecture Time
13:15 - 13:15
Speakers
  • Nithidol Sakunrangsit, Pathumwan, Thailand, Chulalongkorn University - Faculty of Medicine
Session Name
Lunch and poster viewing
Location
Britten room, Queen Elizabeth II Centre, London, United Kingdom
Date
08.11.2019
Time
13:15 - 14:00

Abstract

Background

Plumbagin (PLB) is a naphthoquinone compound isolated from the root of Plumbago indica. In our previous study reported that PLB was able to inhibit cell growth in estrogen receptor (ER)-positive and endocrine resistant cells in vitro. The overexpression of human epidermal growth factor receptor 2 (HER2) and nuclear receptor coactivator 3 (NCOA3) was reported in endocrine resistant cells. However, the molecular mechanism underlying of inhibitory effect of PLB remains to be elucidated. We further investigated the mechanisms of PLB on cell apoptosis and HER2 signaling in endocrine resistant and HER2-overexpressing breast cancer.

Methods

The experiments were performed on MCF-7/LCC9 (endocrine resistant) and SKBR3 (HER2-overexpressing) cell lines. Basal levels of HER2 downstream signaling and NCOA3 in cancer cells used this study were determined by quantitative PCR (qPCR) and western blot. Functional and molecular studies of PLB included cell proliferation and apoptosis were evaluated by colony-forming assay, flow cytometry, qPCR and western blot analyses.

Results

PLB significantly inhibited the proliferation of MCF-7/LCC9 and SKBR3 cells in a concentration-dependent manner. PLB dramatically induced apoptosis in both cell lines and reduced apoptosis-related gene expression. NCOA3 mRNA and AKT phosphorylation significantly reduced in MCF-7/LCC9 cells after treatment with PLB. PLB also decreased phosphorylation of SKBR3 cells.

Conclusions

These findings demonstrated the NCOA3 and AKT signaling might be involved in anti-cancer mechanism of PLB in endocrine resistant HER2-overexpressed breast cancer, supporting further investigations on the therapeutic potential of PLB in animal model and/or phase I clinical trials.

Legal entity responsible for the study

The authors.

Funding

Ratchadaphiseksomphot Endowment Fund (RA62/034 to W.K.) and Special Task Force for Activating Research (STAR) Ratchadaphiseksomphot Endownment Fund to Overcoming Cancer Drug Resistance Research Group.

Disclosure

All authors have declared no conflicts of interest.

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Lunch and poster viewing Poster display

34P - The importance of p53 signaling pathway in the evolution of gastric cancer

Lecture Time
13:15 - 13:15
Speakers
  • Simona Gurzu, Targu Mures, Romania, University of Medicine and Pharmacy Targu-Mures
Session Name
Lunch and poster viewing
Location
Britten room, Queen Elizabeth II Centre, London, United Kingdom
Date
08.11.2019
Time
13:15 - 14:00

Abstract

Background

Although p53 pathway was previously explored in human carcinogenesis, the clinical implication of this gene is still unknown. The aim of this paper was to explore the particularities of TP53gene, at molecular and protein level, in gastric cancer.

Methods

In 197 surgical specimens provided from patients with gastric carcinomas, consecutive cases with curative resection, without preoperative therapy, the clinicopathological parameters were correlated with immunohistochemical (IHC) expression of p53, HER-2 and the DNA mismatch repair protein MLH-1. Mutations in exons 2-11 of the TP53 were also screened using Sanger sequencing.

Results

The rate of TP53 mutation was 28.43% (n = 56). Although it was a widely distribution among the exons 2 and 11 (except exon 9 which did not present mutations), in half of the cases mutations were located in the exons 5 (n = 13) and 8 (n = 15). No statistical correlations were seen between TP53 mutation rate and most of the clinicopathological parameters (e.g. age and gender, tumor localization, microscopic aspect, presence of intestinal metaplasia, HER-2 expression, overall survival). Compared to the non-mutated cases, those that showed mutations, independently from the involved exon, displayed more frequent p53 protein in over 50% of tumor cells (p = 0.004). Although no correlation with pT (p = 0.63) and pN stage (p = 0.84) was seen, a direct correlation was found with the Dukes-MAC-like staging system for gastric cancer, which was proposed by our team in 2017. Based on this system, a significant high number of cases diagnosed in stage pT3N1-3 showed TP53 mutation, whereas mutations were rare in pT3N0 cases and pT1-2 stage, independently from lymph node status (p = 0.03). Another significant correlation regarded the MLH-1 protein, which was more frequently (p = 0.01) negative in non-mutated versus mutated cases (19.86% vs 7.14%).

Conclusions

As MLH-1 negativity is considered an indicator of microsatellite instability, the present study conclude that TP53 gene seems to not be involved in gastric cancers microsatellite instability pathway and do not iterract with HER-2 pathway. Occurrence of TP53 gene mutations induce overexpression of p53 protein in tumor cells and increases risk of lymph node metastases, especially in locally advanced cases.

Legal entity responsible for the study

Romanian National Authority for Scientific Research, CNCS – UEFISCDI.

Funding

Romanian National Authority for Scientific Research, CNCS – UEFISCDI, project number 20 PCCF/2018, code: PN-III-P4-ID-PCCF-2016-0006.

Disclosure

All authors have declared no conflicts of interest.

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Lunch and poster viewing Poster display

73P - Overexpression of EGFR, COX2 and p53 in oral squamous cell carcinoma patients of Pakistan and correlation with prognosis

Lecture Time
13:15 - 13:15
Speakers
  • S. M. Adnan Ali, Karachi, Pakistan, The Aga Khan University
Session Name
Lunch and poster viewing
Location
Britten room, Queen Elizabeth II Centre, London, United Kingdom
Date
08.11.2019
Time
13:15 - 14:00

Abstract

Background

Oral squamous cell carcinoma (OSCC), the sixth leading cancer worldwide, ranks as the most common cancer in males, and the third most common in females in Pakistan. It is influenced by risk factors which are widely consumed in our population. The epidermal growth factor receptor (EGFR) is a tyrosine kinase receptor that is mutated and overexpressed in a variety of cancers. Cyclooxygenase 2 (COX2) is a rate-limiting enzyme that is upregulated in many cancers. It is involved in angiogenesis, apoptosis and metastasis of neoplasia. TP53 mutation and overexpression have been correlated with poor survival in many cancers including oral squamous cell carcinoma (OSCC). The aim of our study was to observe the expression of EGFR, COX2 and p53 in OSCC and correlate the expression with patient’s overall survival, and validate these markers for personalized therapy.

Methods

Formalin-fixed paraffin-embedded tissues of OSCC cases (n = 100) were used. Immunohistochemistry for EGFR, COX2 and p53 was performed using Dako monoclonal antibodies and EnVision system.

Results

There was a majority of males (55%) in our data set and the larger portion of patients was above 40 years of age (82%). Risk factor consumption was found in 73% of cases. EGFR overexpression was observed in 70%, COX2 in 55% and p53 in 67% of OSCC patients. Survival analysis revealed EGFR positive patients had markedly low overall survival (p = 0.098). Similarly, COX2 proved a good prognostic factor for patient survival (p = 0.013), while p53 overexpression was insignificant for survival (p = 0.107).

Conclusions

Our findings emphasize the role of EGFR, COX2 and p53 overexpression in OSCC patients. Molecular therapies targeted towards EGFR, COX2 and p53 may be used against Pakistani OSCC patients to improve prognosis.

Legal entity responsible for the study

Aga Khan University, Karachi, Pakistan.

Funding

Higher Education Commission, Pakistan.

Disclosure

All authors have declared no conflicts of interest.

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Industry satellite - Roche: Novel approaches in personalised healthcare Industry satellite symposium

Interactive patient case focused on comprehensive genomic profiling resulting in treatment with combination therapies

Lecture Time
12:19 - 12:28
Speakers
  • Jason Sicklick, La Jolla, United States of America, University of California San Diego - UCSD
Location
Fleming room, Queen Elizabeth II Centre, London, United Kingdom
Date
09.11.2019
Time
12:05 - 12:50
Targeting cancer neoantigens Keynote Lecture

Targeting cancer neoantigens

Lecture Time
13:30 - 14:15
Speakers
  • Özlem Türeci, Mainz, Germany, BioNTech AG
Location
Fleming room, Queen Elizabeth II Centre, London, United Kingdom
Date
09.11.2019
Time
13:30 - 14:15
Lunch and poster viewing Poster display

49P - An exceptional response to immunotherapy doublet in combined hepatocellular carcinoma-cholangiocarcinoma

Lecture Time
13:15 - 13:15
Speakers
  • Esther Tahover, Jerusalem, Israel, Shaare Zedek Medical Center
Session Name
Lunch and poster viewing
Location
Britten room, Queen Elizabeth II Centre, London, United Kingdom
Date
08.11.2019
Time
13:15 - 14:00

Abstract

Background

We present here a case of a 67-year-old male. He presented in May 2018 with severe abdominal pain and weight loss of 25%. Alpha-feto protein (AFP) and Carbohydrate antigen 19-9 (CA19-9) were within normal limits, and CA125 was 5 times upper normal limit (ULN). Computed tomography showed liver masses and enlarged retroperitoneal lymph nodes. Biopsy from a liver mass showed Combined Hepatocellular Carcinoma (HCC)-Cholangiocarcinoma (CC) (CHC). This a rare tumor, with an incidence of less than 10% of primary liver tumors. In whole exome analysis, mutations suggested the bipotent cell origin and stem cell origin. There are no guidelines or randomized trials regarding treatment. In an analysis of 36 patients who were treated with chemotherapy, the progression-free survival was 2.8 months, with an overall response rate of 5.6%. Targeted agents had minimal effect on survival.

Methods

In our case, extensive genomic, transcriptomic and proteomic testing was performed. No genomic alterations were identified, tumor mutational burden was low and microsatellite status was stable. 7 of 9 immune checkpoint genes were overexpressed. A variant in CDK12 was noted, which was shown to be associated with elevated neoantigen burden and may predict benefit from immune checkpoint therapy.

Results

Blood tests of case

Before immunotherapyAfter 11 months of immunotherapyNormal range
Alk Phos4568338-150
GGT5887215-73
Ca125210212-35

The patient began immunotherapy with ipilimumab and nivolumab followed by nivolumab, which he is continuing. The only side effects were hypothyroidism and Addison’s disease which are being treated. His clinical response was dramatic, he regained all the lost weight, and discontinued high dose opiate treatment. ECOG performance status improved from 3 to 0. Repeated PET-CT showed near complete response, ca125 decreased by 90% and liver function tests normalized.

Conclusions

We present here an exceptional case of a rare tumor, where the patient had a clinical, laboratory and radiological response and a significant improvement in quality of life, suggesting that these tumors are sensitive to immunotherapy. No published cases of this tumor have yet been treated with immunotherapy doublet.

Legal entity responsible for the study

The author.

Funding

Has not received any funding.

Disclosure

The author has declared no conflicts of interest.

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Clonal evolution Education session

4O - Tumour cells mirror embryonic developmental programs to acquire invasive and metastatic capabilities

Lecture Time
12:10 - 12:20
Speakers
  • Sadegh Saghafinia, Lausanne, Switzerland, EPFL - Swiss Institute for Experimental Cancer Research (ISREC)
Session Name
Location
Fleming room, Queen Elizabeth II Centre, London, United Kingdom
Date
08.11.2019
Time
11:00 - 12:20

Abstract

Background

Pancreatic Neuroendocrine Tumours (PanNETs) are the second most common form of pancreatic cancer. We previously identified two principal subtypes of PanNET: insulinomas (islet tumors; IT) and metastasis-like primaries (MLP), corresponding to the low- and high-grade classification of human PanNETs. This study describes the mechanisms by which PanNETs progress from IT to more aggressive MLP tumors and eventually metastasize.

Methods

We profiled single cell transcriptomes, bulk mRNA and miRNA transcriptomes and the proteomes of primary and metastasis specimens from the genetically engineered mouse model of PanNETs (Rip1 Tag2).

Results

We found that the IT tumours maintained the expression of mature ß cell markers. In contrast, the MLP tumours expressed pancreatic progenitor markers. By integrating the data on ß-cell differentiation from pancreatic progenitors to mature ß cells, we demonstrated that the tumour progression from IT to MLP follows the reverse embryonic and postnatal developmental path. Furthermore, we identified Hmgb3 and miR-181cd cluster as the MLP master regulators. Over-expression of the miR-181cd cluster in IT-like cancer cell-lines resulted in the acquisition of the MLP gene signature and MLP morphologic phenotypes, in addition to activation Hmgb3 expression. This suggested a central role for Hmgb3 in initiating the MLP program. Furthermore, inhibiting the expression of Hmgb3 in MLP-like cancer cell-lines resulted in a significant growth decrease, demonstrating the importance of Hmgb3 in the maintenance of MLP-like cell state in vitro. Using transcriptomic data from human patients, we evaluated the relevance of the MLP program in human tumours. We established that aggressive human PanNET tumours also follow the same reverse developmental trajectory of dedifferentiation. Notably, patients with high MLP genes expression had a worse prognosis.

Conclusions

These data demonstrate dedifferentiation as a mechanism by which malignant neuroendocrine cancer cells acquire progenitor-like features, enabling them to become more aggressive and metastatic. In addition, miR-181cd cluster and Hmgb3 act as the core regulators in the initiation of dedifferentiation and maintenance of progenitor-like features in tumour cells.

Legal entity responsible for the study

CMSO Group at ISREC, EPFL.

Funding

Swiss National Science Foundation.

Disclosure

All authors have declared no conflicts of interest.

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Clonal evolution Education session

3O - Temporal dissection of altered pathways during the evolution of cancer

Lecture Time
12:00 - 12:10
Speakers
  • Johanne Ahrenfeldt, Aarhus, Denmark, Institute of Clinical Medicine, Aarhus University Hospital Skejby
Session Name
Location
Fleming room, Queen Elizabeth II Centre, London, United Kingdom
Date
08.11.2019
Time
11:00 - 12:20

Abstract

Background

Cancer arise as normal cells acquire insults to the genome through both mutations and gross copy number alterations. Our current understanding of the molecular makeup of cancer is based on a snap-shot, a biopsy taken at a single time point. Large-scale studies have shown an abundance of genomic alterations, but few are shared across samples which makes interpretation of effect difficult, particularly as different processes may be dominant at different time points in the development of cancer. Here, we have performed pathway analysis on timed mutational driver events to investigate if specific pathways are preferentially affected at different time points during the life-history of cancer.

Methods

We analyzed mutation data and copy number information from bladder cancer acquired from The Cancer Genome Atlas, to time the mutations as early and late relative to copy number events affecting the same genomic location (McGranahan et al., 2015). The mutations were annotated to genes and potential driver mutations were annotated (Jamal-Hanjani et al., 2017). Timed driver events across each cohort were combined into early and late, and subjected to pathway analysis using Reactome (Fabregat et al., 2017).

Results

Focusing on bladder cancer, 28% of the timed mutations are early and 72% are late. The most frequent early driver mutations are TP53 (12%), MUC4 (8%), and PIK3CA (5%), while in late they are PABPC1 (21%), KMT2C (16%), HLA-A (9%). Pathway analysis on early drivers showed how events that affects cell cycle and proliferation dominates, while late drivers are particularly enriched in pathways associated with the immune system.

Conclusions

Our results demonstrate a clear time-separated preference for specific events. These indicates that selection in early cancer development may be primarily focused on genomic alterations that increase proliferation. Conversely, late cancer development enriches in events targeting pathways associated with the immune response, consistent with increased immune pressure as the cancer develops. These results show how timing of events may provide novel insights into how cancer develops, and may help determine the evolutionary developmental stage of cancer with potential implications for improved prognostics and differential therapy.

Legal entity responsible for the study

The authors.

Funding

The Lundbeck Foundation.

Disclosure

All authors have declared no conflicts of interest.

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Lunch and poster viewing Poster display

69P - A laboratory assay for improved prediction of drug responses in acute myeloid leukaemia cells

Lecture Time
13:15 - 13:15
Speakers
  • Eiman Aleem, London, United Kingdom, London South Bank University
Session Name
Lunch and poster viewing
Location
Britten room, Queen Elizabeth II Centre, London, United Kingdom
Date
08.11.2019
Time
13:15 - 14:00

Abstract

Background

The five-year-survival-rate for paediatric patients with Acute Myeloid Leukaemia (AML) is from 60-70%. The most common causes of death are disease relapse and chemo-resistance. Typically, drug dose response testing is performed at atmospheric oxygen (21%) in a 2-Dimensional (2-D) format; however, AML cells reside in the hypoxic (1% O2), 3-Dimensional (3-D) bone marrow structure along with other cells. Therefore, the drug response observed in typical culture conditions (2D, 21% O2) do not always translate to the same response in patients. The objectives of the present study were to: (1) develop an in vitro assay reproducing key elements of the bone marrow microenvironment (BMM), and to (2) use this system to evaluate the sensitivity/resistance of AML cell lines towards selected therapeutic agents.

Methods

A functional drug screen was performed first in 2D normoxia and hypoxia to evaluate the response of AML cell lines to 50 compounds. RNA sequencing was performed to determine global changes of gene expression. Overrepresented genes were further matched to potential pathways using Ingenuity Pathway Analysis. Results were validated using real-time RT-PCR. MV-4-11, an AML cell line with a FLT3 mutation was used to study the effects of 3-D structure on drug response. Cells were seeded in 3D and allowed to grow for 72h, compounds were added for an additional 24 h and cell viability was measured using Cell Titer Glo assay.

Results

Twelve cell lines were used in the present study. In the initial screen THP1 cells for example were more sensitive in hypoxia to 8 out of 50 different compounds. In MV-4-11 cells, hypoxia and 3-D structure conferred resistance towards YM155, a BIRC5 targeting compound. Mechanistic studies revealed that MCL-1 stabilization may underlie this response. A functional drug screen using 50 compounds in 3-D showed that 10 out of 50 compounds resulted in significantly reduced viability of MV-4-11 cells compared to vehicle indicating that this assay can be used for high-throughput drug screening (HTS).

Conclusions

Hypoxia and 3D structure alter drug response in AML cells. The proposed 3D assay can be used for HTS. Clinical application of this study may lead to more accurate predictions of therapeutic outcomes.

Legal entity responsible for the study

The authors.

Funding

Children Cancer Network.

Disclosure

All authors have declared no conflicts of interest.

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Single cell analysis / single cell sequencing Education session

Subclonal heterogeneity and behaviour in TNBC

Lecture Time
08:30 - 08:50
Speakers
  • Leif W. Ellisen, Boston, United States of America, Massachusetts General Hospital, Harvard Medical School
Location
Fleming room, Queen Elizabeth II Centre, London, United Kingdom
Date
09.11.2019
Time
08:30 - 10:00