bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract ED03-05: Innovative technology development to advance biospecimen science.
Authors: Dickherber Anthony
Published: August 2014

In 1998, the National Cancer Institute (NCI) established the groundwork for a highly successful program focused on innovative technology development to overcome persistent barriers for cancer researchers and clinicians. Unlike other initiatives of the time, the Innovative Molecular Analysis Technologies (IMAT) Program solicited only the most cutting-edge ideas, thus restricting its application pool to those projects that had the potential to be truly transformative. By doing so, the program filled a void that no other program at the NCI or NIH filled at the time. In 2005, the IMAT program introduced an additional focus on supporting innovative technology development that uniquely advanced the field of biospecimen science. Technologies supported by this mechanism have seen a disproportionately high level of success. The IMAT program continues to represent a unique resource for highly innovative technology development, but runs alongside several active programs at the NCI for supporting cancer-relevant technologies.

The full reauthorization of the IMAT program in fall of 2011 by the NCI for 2012-2013 demonstrates a significant commitment, especially under ever tightening fiscal constraints, to supporting innovative technology development for cancer research and clinical care. The IMAT program utilizes unique funding mechanisms to better support investigators through both the early and advanced stages of technology development over traditional funding mechanisms for exploratory research. Moreover, receipt of an IMAT award offers membership to a truly unique community of investigators that meet annually to share progress and exchange ideas. A variety of IMAT-supported research projects will be highlighted to demonstrate the variety and level of innovation evident in the IMAT portfolio of supported research. Potential projects of interest and identified technology gaps specific to the field of biospecimen science and with relevance to cancer health disparities will also be discussed.

Currently available funding opportunity announcements are active with three receipt dates open through September, 2013. Enthusiasm from the NCI Board of Scientific Advisors and NCI leadership for the IMAT program indicates that these solicitations are likely to be active beyond these dates. Technology developers from all fields of research are encouraged to apply.

Citation Format: Anthony Dickherber. Innovative technology development to advance biospecimen science. [abstract]. In: Proceedings of the Fifth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2012 Oct 27-30; San Diego, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2012;21(10 Suppl):Abstract nr ED03-05.
Abstract A02: Targeted deep sequencing of second biopsy tissues from patients with acquired resistance (AR) to EGFR tyrosine kinase inhibitors harboring EGFR mutations: Preliminary results.
Authors: Ahn Myung-Ju , Kim Youngwook , Kim Kyoung-Mee , Choi Yoon-la , Han Joungho , Jho EunHye , Sun Joung-Mu , Ahn Jin-Sock , Park Keunchil
Published: August 2014

Background: Patients with EGFR activating mutation in non-small cell lung cancer (NSCLC) eventually develop AR to EGFR tyrosine kinase inhibitors (TKI) after durable and high response. Several mechanisms of AR to EGFR TKIs identified include T790M gatekeeper mutation, PIK3CA mutation, C-met amplification, transformation into small cell carcinoma or epithelial-mesenchymal transition (EMT). In the present study, to identify other genetic alterations, we performed targeted deep sequencing for over 700 hotspots in 50 cancer-related genes using next generation sequencing (NGS) technology.

Methods: Genomic DNA was extracted from FFPE tissue or pleural/pericardial fluid or primary culture cells of 57 NSCLC patients who developed AR to EGFR TKIs. Amplicons NGS libraries for 50 oncogenes included in Ion AmpliSeqTM Cancer Hotspot Panel v.2 were generated and sequenced in the Ion PGM Sequencer. Variant caller included in Torrent Suite Software was utilized to identify mutations in the samples, annotation was performed with IonTorrentTM Software.

Results: All 57 patients were treated with EGFR TKIs, gefitinib (n=39), erlotinib (n=17), or afatinib (n=1) as first-line or later line of therapy. 65% were female, 74% never smokers, median age of 58 years (range 25-77) and 98% were adenocarcinoma. 31 patients have exon 19 deletion and 14 with L858R at baseline. The median PFS for EGFR TKIs was 11 months (range 4-63). Targeted deep sequencing of second biopsy identified recurrent coexisting mutations of T790M in 52.6%. Other frequently altered genes include mutation of CTNNB1 (n=9), c-KIT (n=7), c-met (n=7), SMARCB1 (n=5), PDGFRA (n=5), PTEN (n=2), NRAS (n=2), and PIK3CA (n=1). However, in order to investigate whether these recurrent mutations observed in second biopsy represent genes associated with AR or pre-existing genes, targeted deep sequencing of available paired baseline samples will be performed and updated.

Conclusions: We found that NGS technology is feasible in FFPE tissues. Although T790M mutation confers the most common resistant mechanism to EGFR TKI, several detected genetic mutations are novel observations which might provide potential candidates for AR mechanisms. Further validation results will be discussed.

Citation Format: Myung-Ju Ahn, Youngwook Kim, Kyoung-Mee Kim, Yoon-la Choi, Joungho Han, EunHye Jho, Joung-Mu Sun, Jin-Sock Ahn, Keunchil Park. Targeted deep sequencing of second biopsy tissues from patients with acquired resistance (AR) to EGFR tyrosine kinase inhibitors harboring EGFR mutations: Preliminary results. [abstract]. In: Proceedings of the AACR-IASLC Joint Conference on Molecular Origins of Lung Cancer; 2014 Jan 6-9; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2014;20(2Suppl):Abstract nr A02.
Abstract B02: Protein Kinase C iota is required for the maintenance of a tumor-initiating cell phenotype in lung squamous cell carcinoma.
Authors: Justilien Verline , Walsh Michael P. , Ali Syed A. , Thompson E. A. , Fields Alan P.
Published: August 2014

Background: Protein kinase C iota (PKCι) is an oncogene that is frequently targeted for tumor-specific gene amplification in human cancers harboring the 3q26 amplicon including Non-Small Cell Lung Carcinoma (NSCLC). PKCι is required for the maintenance of the transformed phenotype of NSCLC cells in vitro and in vivo. In addition, PKCι is critical for lung tumorigenesis in the LSL-K-rasG12D mouse model of NSCLC. Recently, we demonstrated that genetic disruption of Prkcι/λ; in LSL-K-rasG12D mice blocks K-ras-mediated expansion and morphological transformation of bronchioalveolar stem cells (BASCs), the first identifiable step in K-ras-mediated transformation in this model. These results demonstrate that PKCι plays a requisite role in tumor initiation, and suggest that PKCι may serve a similar role in the tumor-initiating activity of lung cancer stem cells. Our current studies aim to identify and characterize the molecular signaling pathway(s) by which PKCι regulates the biology of human lung squamous cell carcinoma (LSCC) tumor initiating cells (TICs) isolated from LSCC cell lines and primary tumors.

Methods: Cultures enriched in TICs were established from LSCC lines or primary human LSCC tumors that harbor PRKCI amplification. PKCι expression in TIC cultures was inhibited by lentiviral shRNA, and the effect of PKCι knockdown on their stem-like properties was assessed. Whole genome deep sequencing analysis was performed on mRNA from parental, control TIC and PKCι knockdown TIC cultures in order to identify PKCι-regulated targets and signaling pathways in TICs.

Results: LSCC TIC cultures express putative stem cell markers, and exhibit clonal expansion, self-renewal, enhanced anchorage-independent growth and tumorigenic potential in vivo. RNAi-mediated knockdown of PKCι inhibited TIC growth, clonal expansion, transformed growth and tumor initiation. mRNA sequencing revealed that PKCι regulates the expression and function of critical molecules involved in the stem-like phenotype of LSCC TICs. Details of PKCι-mediated signaling in LSCC TICs will be discussed.

Conclusions: PKCι is required for maintenance of a highly tumorigenic stem-like population of cells in human LSCC. Given the prevalence of 3q26 amplification in human cancer, PKCι may regulate the biology of TICs in tumor types other than LSCC. Our results provide a rationale for therapeutic targeting of PKCι in LSCC TICs.

Citation Format: Verline Justilien, Michael P. Walsh, Syed A. Ali, E. A. Thompson, Alan P. Fields. Protein Kinase C iota is required for the maintenance of a tumor-initiating cell phenotype in lung squamous cell carcinoma. [abstract]. In: Proceedings of the AACR-IASLC Joint Conference on Molecular Origins of Lung Cancer; 2014 Jan 6-9; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2014;20(2Suppl):Abstract nr B02.
Abstract IA18: Early but promising data: PD-1 inhibitors in non-small cell lung cancer.
Authors: Garon Edward B. , Wolf Brian R. , Wong Deborah JL , Goldman Jonathan W.
Published: August 2014

Blockade of the programmed death (PD-1) checkpoint has emerged as a promising approach for immunotherapies in the treatment of non-small cell lung cancer (NSCLC). Immune checkpoints, such as PD-1, an inhibitory T-cell immunoreceptor, and the receptor's ligands, PD-L1 and PD-L2, assist in hindering autoimmunity and protecting peripheral tissues to minimize damage (1). However, in cancers, the interaction between PD-1 and its ligands also can result in the promotion of an immunosuppressive tumor microenvironment (1).

Several agents targeting PD-1 are currently undergoing clinical development. Clinical trials of anti-PD-1 antibodies nivolumab (BMS-936558) and MK-3475 have produced encouraging results in non-small cell lung cancer patients with regards to objective response rates (ORR) and progression free survival (PFS) (2,3). In a pretreated advanced NSCLC population, nivolumab produced durable responses, particularly at 3 and 10 mg/kg doses (2). For 38 previously-treated NSCLC patients, MK-3475, administered at 10 mg/kg every three weeks, led to an ORR of 24 percent based on irRC (21 percent based on RECIST 1.1) (3). Median overall survival was 51 weeks with a PFS, which had not been reached at a time when the minimum PFS would be 62 weeks among the responders (3). ORR was 67% in patients with tumors having PD-L1 expression above a cutpoint versus 4% in those below the cutpoint (3). Based on the favorable response rates relative to historical controls, and the durability of responses in clinical trials, the development of PD-1 inhibitors in lung cancer is proceeding rapidly.

Future trials utilizing anti PD-1 antibodies will explore further biomarkers, and combination with other therapies.

Citation Format: Edward B. Garon, Brian R. Wolf, Deborah JL Wong, Jonathan W. Goldman. Early but promising data: PD-1 inhibitors in non-small cell lung cancer. [abstract]. In: Proceedings of the AACR-IASLC Joint Conference on Molecular Origins of Lung Cancer; 2014 Jan 6-9; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2014;20(2Suppl):Abstract nr IA18.
Abstract IA41: A new TRacK in lung cancer: NTRK1 gene fusions as a therapeutic target.
Authors: Doebele Robert C.
Published: August 2014

Non-small cell lung cancer (NSCLC) is a heterogeneous disease that can be organized into groups designated by the dominant oncogene or oncogenic ‘driver’. Classification of NSCLC by oncogenes is critical because oral kinase inhibitors or other targeted therapeutic strategies can often effectively inhibit these oncogenes leading to dramatic tumor shrinkage and thus significant clinical benefit for patients. Several of these oncogenes are the result of chromosomal translocations that generate gene fusions in which the kinase domain of a receptor tyrosine kinase is aberrantly expressed and activated by the addition of sequence from an unrelated gene. Recent examples of such gene fusions in NSCLC include those involving the ALK, ROS1, RET and FGFR1/2/3 genes. We have recently discovered previously uncharacterized gene fusions involving the NTRK1 gene in patients with lung adenocarcinoma whose tumors did not harbor other known oncogenes. These novel oncogenes contain the 3 portion of the neurotrophic tyrosine kinase, receptor, type 1 (NTRK1) gene, which encodes the kinase domain of the TRKA receptor. Cloning and ectopic expression of these novels genes in non-tumor cells demonstrate their oncogenic potential. Treatment of cells expressing NTRK1 gene fusions with TRK inhibitors inhibits autophosphorylation of the chimeric TRKA proteins, important downstream signaling pathways, and cellular proliferation. Gene fusions involving NTRK1 (TRKA), NTRK2 (TRKB), and NTRK3 (and TRKC) have now been identified across multiple tumor types. As clinical next generation sequencing becomes increasingly common, cancer patients with these somatic gene abnormalities will be identified and therefore it is critical to understand their biology, utility as a therapeutic target, and mechanisms of intrinsic and acquired drug resistance in order to advance a precision medicine strategy for the treatment of NSCLC.

Citation Format: Robert C. Doebele. A new TRacK in lung cancer: NTRK1 gene fusions as a therapeutic target. [abstract]. In: Proceedings of the AACR-IASLC Joint Conference on Molecular Origins of Lung Cancer; 2014 Jan 6-9; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2014;20(2Suppl):Abstract nr IA41.
Abstract PR02: Recurrent oncogenic mutations in the small GTPase RIT1 activate PI3K and MEK.
Authors: Berger Alice H. , Duke Fujiko , Imielinski Marcin , Kaplan Nathan , Wala Jeremiah , Shi Geng-Xian , Andres Douglas A. , Meyerson Matthew
Published: August 2014

Recently, we identified somatic mutations in the RAS-family small GTPase gene RIT1 in 2.4% (10/413) of lung adenocarcinoma tumors. Mutation of RIT1 is mutually exclusive with other oncogenic mutations in the receptor tyrosine kinase/RAS pathway, suggesting that RIT1 mutations are rare driver mutations in lung adenocarcinoma. Oncogenic RIT1 mutations cluster in the switch II domain of the protein and are observed not only in lung adenocarcinoma but also in myeloid malignancies, breast carcinoma, cervical carcinoma, and in the germline of Noonan syndrome patients. Here we functionally characterize 26 RIT1 point mutations and in-frame insertions/deletions in murine fibroblasts and human immortalized lung epithelial cells and find that the majority of somatic RIT1 mutations are able to transform cells. Furthermore, we identified a human lung cancer cell line, NCI-H2110, with an endogenous RIT1 p.M90I mutation. Knockdown of RIT1 in NCI-H2110 cells revealed the role of RIT1 in regulation of PI3K and MEK signaling. Consistently, combination PI3K/MEK inhibition could inhibit cellular transformation induced by ectopic expression of mutated RIT1. Furthermore, PI3K inhibition suppressed in vivo growth of NCI-H2110 cells. These data identify RIT1 as an oncogene and nominate combined PI3K and MEK inhibition as a therapeutic strategy in RIT1-mutated cancers.

This abstract is also presented as Poster A04.

Citation Format: Alice H. Berger, Fujiko Duke, Marcin Imielinski, Nathan Kaplan, Jeremiah Wala, Geng-Xian Shi, Douglas A. Andres, Matthew Meyerson. Recurrent oncogenic mutations in the small GTPase RIT1 activate PI3K and MEK. [abstract]. In: Proceedings of the AACR-IASLC Joint Conference on Molecular Origins of Lung Cancer; 2014 Jan 6-9; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2014;20(2Suppl):Abstract nr PR02.
Functional Kinomics Identifies Candidate Therapeutic Targets in Head and Neck Cancer
Authors: Moser Russell , Xu Chang , Kao Michael , Annis James , Lerma Luisa Angelica , Schaupp Christopher M. , Gurley Kay E. , Jang In Sock , Biktasova Asel , Yarbrough Wendell G. , Margolin Adam A. , Grandori Carla , Kemp Christopher J. , Méndez Eduardo
Published: August 2014

Purpose: To identify novel therapeutic drug targets for p53-mutant head and neck squamous cell carcinoma (HNSCC).

Experimental Design: RNAi kinome viability screens were performed on HNSCC cells, including autologous pairs from primary tumor and recurrent/metastatic lesions, and in parallel on murine squamous cell carcinoma (MSCC) cells derived from tumors of inbred mice bearing germline mutations in Trp53, and p53 regulatory genes: Atm, Prkdc, and p19Arf. Cross-species analysis of cell lines stratified by p53 mutational status and metastatic phenotype was used to select 38 kinase targets. Both primary and secondary RNAi validation assays were performed on additional HNSCC cell lines to credential these kinase targets using multiple phenotypic endpoints. Kinase targets were also examined via chemical inhibition using a panel of kinase inhibitors. A preclinical study was conducted on the WEE1 kinase inhibitor, MK-1775.

Results: Our functional kinomics approach identified novel survival kinases in HNSCC involved in G2–M cell-cycle checkpoint, SFK, PI3K, and FAK pathways. RNAi-mediated knockdown and chemical inhibition of the WEE1 kinase with a specific inhibitor, MK-1775, had a significant effect on both viability and apoptosis. Sensitivity to the MK-1775 kinase inhibitor is in part determined by p53 mutational status, and due to unscheduled mitotic entry. MK-1775 displays single-agent activity and potentiates the efficacy of cisplatin in a p53-mutant HNSCC xenograft model.

Conclusions: WEE1 kinase is a potential therapeutic drug target for HNSCC. This study supports the application of a functional kinomics strategy to identify novel therapeutic targets for cancer. Clin Cancer Res; 20(16); 4274–88. ©2014 AACR.
Abstract A32: Fractalkine receptor, CX3CR1, Can regulate the response of ovarian carcinoma cells to radiation therapy
Authors: Xie Jia , Gurler Hilal , Barbolina Maria
Published: August 2014

Ovarian carcinoma is a leading cause of death from gynecologic malignancies. The current treatment, a combination of chemotherapy and radiation, fails to keep patients in remission. Therefore, mechanistical understanding of the disease and modifications of the therapy could offer benefits to overcome current problems and combat the disease. Fractalkine receptor (CX3CR1) belongs to a chemokine family of G protein-coupled receptors, the most frequently and successfully drug-targeted group of proteins. Previous publications show that interaction of chemokines with their receptors plays an important role in homing distant metastasis, as well as promoting migration, adhesion, and proliferation in many types of cancer. X-Ray radiotherapy is a well-developed method of anti-cancer treatment, however, it is seldom used against ovarian carcinoma due to multiple reasons, including dose-related effects. Hence, strategies for reducing the dose of radiation while keeping its efficiency at a therapeutically effective level could provide rational for developing approaches for a more wide-spread use of this method.

Our data indicate that transient downregulation of CX3CR1 in p53-null cells and p53-mutant cells but not in p53 wild-type ovarian carcinoma cell lines can lead to approximately 50% sensitization to x-ray radiation, as determined by the clonogenic assay. Importantly, our in vivo studies demonstrated that a transient downregulation of CX3CR1 in combination with x-ray radiation led to approximately 50% reduction of tumor weights compared to either downregulation of CX3CR1 or x-ray radiation alone. We are currently characterizing the detailed mechanisms behind the CX3CR1-dependent radiosensitization, including those related to changes in cell cycle progression, involvement of cellular reactive oxygen species accumulation and regulation of DNA single and double strand break repair. Taken together, our date indicate that disruption of the CX3CR1 signaling could sensitize ovarian carcinoma to radiation therapy.

Citation Format: Jia Xie, Hilal Gurler, Maria Barbolina. Fractalkine receptor, CX3CR1, Can regulate the response of ovarian carcinoma cells to radiation therapy. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr A32.
Abstract A40: New approaches to target mevalonate biosynthesis
Authors: Casella Cinzia , Brodsky Alexander S.
Published: August 2014

To support rapid growth, tumors need to synthesize lipids and cholesterol, and stopping production of these metabolites can inhibit tumors. Statins are among the most prescribed and well-tolerated drugs in the world to treat cardiovascular disease by inhibiting cholesterol synthesis. Statins are known to be toxic to cancer cells and have attracted renewed attention because of the increased appreciation of the importance of the regulation of metabolism in tumors. Statins have not progressed beyond clinical trials because some patients significantly benefit, while others do not in a range of cancers. Our strategy is to improve our understanding of the mechanisms of tumor responses to statins and thereby enhance statin toxicity to cancer cells. We have found that combining statins with oxysterols synergistically enhances statin toxicity by 10-fold. Oxysterols are oxidized cholesterol metabolites that naturally occur during cholesterol synthesis. We hypothesize that oxysterols inhibit the expression of cholesterol synthesis through a master regulator of metabolism, SREBP-2, making cells more sensitive to statins. We find that oxysterols enhance statin induced apoptosis and suppress the expression of SREBP-2 along with SREBP-2 targets including HMGCR. We have tested a number of oxysterols that work with a range of efficacies in seven representative ovarian cancer cell lines. P53 status is not a major factor in mediating oxysterol activity. Oxysterols inhibit expression of SREBP-2 leading to lower expression of the statin target, HMGCR, along with lower expression of most of the mevalonate and cholesterol biosynthesis pathways. These observations suggests that the cell's response to statins and not initial expression levels may be most critical to determine which patients may benefit from statin therapy. Current studies are testing the combination in mouse models and evaluating the function and regulation of SREBP-2 in ovarian cancer. In summary, we have discovered an approach to suppress cancer cell's response to statins leading to greatly improved statin efficacy highlighting the importance of the mevalonate and cholesterol pathways in mediating growth and survival.

Citation Format: Cinzia Casella, Alexander S. Brodsky. New approaches to target mevalonate biosynthesis. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr A40.
Abstract A53: Biomarkers of chemotherapy resistance in serous epithelial ovarian cancer identified by integrative genomic and transcriptomic analysis
Authors: Koti Madhuri , Gooding Robert J. , Nuin Paulo , Haslehurst Alexandria , Crane Colleen , Weberpals Johanne , Chids Timothy , Bryson Peter , Dharsee Moyez , Evans Kenneth R. , Feilotter Harriet E. , Park Paul C. , Squire Jeremy A.
Published: August 2014

Resistance to platinum-based chemotherapy remains a major impediment in the treatment of serous epithelial ovarian cancer. The objective of this study was to use gene expression and copy number profiling to delineate major deregulated pathways and biomarker networks associated with the development of intrinsic chemotherapy resistance with exposure to standard first-line therapy for ovarian cancer. The study cohort comprised 28 high grade serous ovarian cancer patients divided into two groups based on their varying sensitivity to first-line chemotherapy using progression free survival (PFS) as a surrogate of response. Twelve patient tumors demonstrating relative resistance to platinum based chemotherapy corresponding to shorter PFS (less than 6 months) were compared to 16 tumors from platinum-sensitive patients (PFS more than 18months). Molecular profiling was performed using Affymetrix high-resolution microarray platforms to permit global comparisons of gene expression levels and copy number profiles between tumors from the resistant group with the sensitive group. Microarray data analysis using statistical methods revealed a set of 204 discriminating genes of which expression levels may be influencing differential chemotherapy response between the two groups. Pathway analysis of the differentiating genes showed IGF1 network to be significantly altered between the two groups in addition to PI3K, NFkB, distinguishing the chemotherapy resistant with the sensitive group. Copy number analysis showed differences in the chromosomal regions, 4q31.22, 5q13.2, 9p24.3, 2p23.2, 16q21, 6q14.1, 7p22.3, 12p13 and Xq.

Integrative copy number and gene expression profiling will delineate the drivers of chemotherapy resistance in patients undergoing standard platinum-based treatment of ovarian cancer. Future studies to validate these markers are necessary to apply this knowledge to biomarker-based clinical trials.

Citation Format: Madhuri Koti, Robert J. Gooding, Paulo Nuin, Alexandria Haslehurst, Colleen Crane, Johanne Weberpals, Timothy Chids, Peter Bryson, Moyez Dharsee, Kenneth R. Evans, Harriet E. Feilotter, Paul C. Park, Jeremy A. Squire. Biomarkers of chemotherapy resistance in serous epithelial ovarian cancer identified by integrative genomic and transcriptomic analysis. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr A53.
Abstract A56: Identification of a micro RNA profile to predict response to therapy and improve patient survival in ovarian cancer
Authors: Rigau Marina , Majem Blanca , Altadill Tatiana , Lanau Lucia , Sánchez-Iglesias José-Luis , Castellví Josep , Pérez-Benavente Assumpció , Cabrera Silvia , García Angel , Xercavins Jordi , Alameda Francesc , Matias-Guiu Xavier , Gil-Moreno Antonio , Campo Josep-Maria Del , Llauradó Marta , Reventós Jaume
Published: August 2014

Introduction & Objectives: Ovarian cancer (OC) is the leading cause of death among gynecological malignancies in developed countries. The standard treatment is surgery plus platinum-paclitaxel (CP/TX) chemotherapy. Nevertheless, 70% of these patients are diagnosed in advanced stages, and, among those, 75% will recur, become resistant to therapies and die. We believe that an improvement in OC survival might be related with an early identification of those patients, since long survival (SV) is related to chemosensitivity. Our aim is to identify an expression profile of micro RNAs (miRNAs) associated with SV of patients with advanced OC.

Material & Methods: Formalin-fixed paraffin-embedded (FFPE) primary tumor tissues from late stage (III/IV) OC patients were collected at the time of surgery for the study. All patients were treated with optimal surgery and standard chemotherapy (CP/TX; after surgery). Samples were divided into two groups; (a) patients with “long SV” (SV>10 years after diagnosis) and (b) patients with “short SV”. The identification of miRNAs associated with each profile of SV was performed using TaqMan Array MicroRNA microfluidic cards in 30 patients.

Results: The identification phase has shown that the majority of the miRNAs appeared to be down-regulated in the group of “short SV” versus the “long SV”. A validation phase in a new group of tumor tissue samples is ready to validate the promising miRNA candidates that seem to be associated with the resistant (short SV) and the sensitive (long SV) profiles.

Conclusions: The ability to identify markers associated with “long SV” in patients with advanced OC could allow transforming the impact of the disease through the reduction of chemotherapy treatments and the reduction of unnecessary health care costs. Moreover, it could permit the generation of knowledge applicable to the future development of new therapies for the “short SV” since they are supposed to be resistant to the therapy.

Citation Format: Marina Rigau, Blanca Majem, Tatiana Altadill, Lucia Lanau, José-Luis Sánchez-Iglesias, Josep Castellví, Assumpció Pérez-Benavente, Silvia Cabrera, Angel García, Jordi Xercavins, Francesc Alameda, Xavier Matias-Guiu, Antonio Gil-Moreno, Josep-Maria Del Campo, Marta Llauradó, Jaume Reventós. Identification of a micro RNA profile to predict response to therapy and improve patient survival in ovarian cancer. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr A56.
Abstract B15: Unveiling the mechanism of ovarian cancer dissemination through a comparative study among primary tumors, ascites and metastases
Authors: Lanau Lucia , Rigau Marina , Majem Blanca , Altadill Tatiana , Castellví Josep , Sánchez-Iglesias José-Luis , Pérez-Benavente Assumpció , Cabrera Silvia , García Angel , Xercavins Jordi , Campo Josep-Maria Del , Gil-Moreno Antonio , Ruiz Anna , Reventós Jaume , Llauradó Marta
Published: August 2014

Introduction & Objectives: Epithelial ovarian cancer (OC) is the most lethal gynecological malignancy and the fifth cause of cancer deaths in women in the western world. Largely asymptomatic, OC is frequently detected at late stage (75-80%). Five-year survival rate for women with advanced stage disease is less than 20%, while the cure rate is almost 90% when are diagnosed at early stages. Epithelial OC metastasizes by direct extension from the ovary seeding the cells into the peritoneal wall and to neighbouring organs. Our aim is to study the OC dissemination comparing human paired ovarian primary tumors, ascites and metastases.

Material & Methods: Fresh ovarian primary tumor, ascites and peritoneal metastases from patients suffering advanced serous OC were collected at the surgery room and processed for further analysis. Formalin-fixed paraffin-embedded (FFPE) tissues were collected from the Pathology Department for further immunohistochemical analysis. Discovery phase: We determined the global gene expression profile of 5 fresh-paired samples (primary tumor, ascites and metastases) by microarray analysis. Validation phase: We analysed mRNA and protein levels from 10 unpaired fresh samples by RTqPCR and Western-blot, respectively. We further evaluated protein expression by immunohistochemistry from 10-paired tumor and metastasis FFPE-samples.

Results: GREM1 was shown to be upregulated in metastases versus ascites and in metastases versus primary tumors. FABP4 and INHBA were significantly overexpressed in metastases when compared to primary tumors. INHBA and FABP4 were validated at RNA and protein level, whereas GREM1 only at RNA level.

Conclusions: The present study highlights the role of previously unknown candidates in OC dissemination that might be used as tumor biomarkers, to clinically monitor the progression of the disease, or as target therapies, to block the OC dissemination.

Citation Format: Lucia Lanau, Marina Rigau, Blanca Majem, Tatiana Altadill, Josep Castellví, José-Luis Sánchez-Iglesias, Assumpció Pérez-Benavente, Silvia Cabrera, Angel García, Jordi Xercavins, Josep-Maria Del Campo, Antonio Gil-Moreno, Anna Ruiz, Jaume Reventós, Marta Llauradó. Unveiling the mechanism of ovarian cancer dissemination through a comparative study among primary tumors, ascites and metastases. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr B15.
Abstract B16: Resources for research: Identification and validation of ovarian cancer cases from the Synthetic Derivative
Authors: Beeghly-Fadiel Alicia , Lovett Whitney , Delahanty Ryan J. , Khabele Dineo , Zhang Wei
Published: August 2014

As a rare and highly lethal cancer, research on ovarian cancer is prone to sample size limitations. To build a research resource, we set out to identify women with ovarian cancer from a large database of de-identified electronic medical records (EMR) of subjects who received medical care from the Vanderbilt University Medical Center. Known as the Synthetic Derivative (SD), this database was initiated in 2006, and now includes more than 20 years of EMR for approximately 1.7 million subjects. We developed a computational algorithm to identify subjects whose EMR included a minimum of three occurrences of an International Classification of Disease (ICD) code of 183 (Malignant neoplasm of ovary and other uterine adnexa) or with local Tumor Registry data that included a primary cancer site of ovary (C569) or fallopian tube (C570). A total of 1,755 women were identified; review of EMR and data abstraction is currently underway. Automated data downloads were conducted for Tumor Registry data, demographics and vital status, CA125 levels and dates, and surgical types and dates. Of the 1,755 women, the majority were Caucasian (83.7%) and deceased (61.7%). Tumor Registry data was available for 1,500 women; epithelial ovarian or fallopian tube cancers were confirmed for 1,141 women. In agreement with national numbers, the 5 year survival rate of these women was only 42.7%. Stored DNA is currently available for 424 women; genotyping data from a variety of platforms is available for 132 women. After additional EMR review and data abstraction, we anticipate being able to verify approximately 1,415 cases of epithelial ovarian cancer. This approach can be utilized at other institutions in the Electronic Medical Records and Genomics (eMERGE) Network; our goal is to build a large resource of validated ovarian cancer cases to support both etiologic and prognostic investigations of this insidious disease.

Citation Format: Alicia Beeghly-Fadiel, Whitney Lovett, Ryan J. Delahanty, Dineo Khabele, Wei Zhang. Resources for research: Identification and validation of ovarian cancer cases from the Synthetic Derivative. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr B16.
Abstract B24: Using a unique genetically modified ovarian cancer cell line model to identify the targets for siRNA directed therapies
Authors: Tonin Patricia N. , Greenwood Celia MT , Provencher Diane MT , Mes-Masson Anne-Marie
Published: August 2014

We reported that though >90% of high-grade serous ovarian carcinomas (HGSC) harbor somatic TP53 mutations, cases with missense mutations have significantly longer progression free and overall survival than cases with “null” mutations, and that HGSCs defined by TP53 mutation type exhibit unique differences in their genomic landscapes. Large-scale molecular genetic analyses by The Cancer Atlas Group (TCGA) have identified numerous genes/molecular pathways that could be targeted for therapy in HGSC. A large number of candidates were reported, whereby the results were affected by heterogeneity of samples. We have analyzed the transcriptomes from a unique HGSC cell line model with altered in vivo tumorigenic and in vitro growth characteristics to understand the biology of HGSC as we have shown in previous analyses that the pathways affected intersected those found altered in tumors. We posit that investigating phenotypically defined cancer cell line models could facilitate the identification of targets for siRNA-based therapeutics (as an example). Towards this goal, we have characterized the transcriptomes (n~700 RNAs) derived from our parental HGSC cell line model and derived genetically modified cell lines. The cell line was derived from long term passage of ovarian ascites, harbors a somatic missense TP53 mutation and exhibits suppression of tumorigenicity in murine models. The genetically modified derivative cell lines have lost tumorigenic potential and exhibit altered growth characteristics. We compared their transcriptomes, derived a list a genes correlated with these alterations and then compared them to transcriptomes from public data sets: ovarian surface epithelial cells (n=10) and HGSC (n=56); and (2) TCGA samples (n~300). A defined list of known and new genes was identified by these comparative analyses pointing to targets that could be used to affect tumorigenic potential. The genetically modified model provides a new avenue of research that has the potential to elucidate pathways important in HGSC, especially those involved in tumorigenicity, as well as define targets for the development of RNA-based targeted therapeutics.

Citation Format: Patricia N. Tonin, Celia MT Greenwood, Diane MT Provencher, Anne-Marie Mes-Masson. Using a unique genetically modified ovarian cancer cell line model to identify the targets for siRNA directed therapies. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr B24.
Abstract B61: Proteomic profiling of ovarian cancer and mesothelial cells reveals novel mediators implicated in the metastatic progression of ovarian cancer to the peritoneum
Authors: Musrap Natasha , Karagiannis George S. , Diamandis Eleftherios P.
Published: August 2014

Over the past decade, numerous studies have highlighted the impact of the cancer microenvironment on tumor progression. It is now well understood that tumor cell behaviour is largely influenced by various stromal cell types and extracellular matrix components within their surrounding environments. A key hallmark of epithelial ovarian cancer (EOC) progression is the attachment of cancerous cells to the mesothelium and the formation of invasive peritoneal implants. Given that metastatic dissemination to the peritoneum is associated with poor prognosis, delineating the factors involved in cancer-peritoneal cell interaction, and the mechanisms they employ, is vital to improving patient survival as it may lead to the discovery of novel therapeutic targets. As such, we aimed to elucidate proteins that participate in this interaction by comparing the secreted proteome (referred to as the secretome) of an in vitro co-culture model containing ovarian cancer (OVCAR-5) and mesothelial cells (LP-9) to that of their monoculture secretomes. Using two-dimensional liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), we identified 1584 proteins that were secreted by co-cultures, and 1644 and 1420 proteins by OVCAR-5 and LP-9 cells, respectively. After comparative analysis and application of our filtering criteria, we identified 53 proteins that were differentially secreted during cancer and mesothelial interaction, and of these, mucin 5AC (MUC5AC) displayed significantly elevated secretion in the co-culture secretomes. Relative mRNA expression of candidates was assessed using quantitative PCR, which also revealed an increase in MUC5AC gene expression in three different co-culture models (OVCAR-5 and LP-9; BG-1 and LP-9; OV-90 and LP-9) (p<0.05). Immunocytochemistry analysis also revealed increased expression of MUC5AC in ovarian cancer and peritoneal co-cultures. Taken together, these findings reveal novel proteins that may potentially regulate the progression of ovarian cancer to the peritoneum, particularly MUC5AC, and elucidation of its role in EOC warrants further investigation.

Citation Format: Natasha Musrap, George S. Karagiannis, Eleftherios P. Diamandis. Proteomic profiling of ovarian cancer and mesothelial cells reveals novel mediators implicated in the metastatic progression of ovarian cancer to the peritoneum. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr B61.
Abstract B73: Human omental-derived adipose stem cells: Modulator of ovarian cancer proliferation, migration and chemoresistance
Authors: Nowicka Aleksandra , Solley Travis , Marini Frank C. , Sharp Hadley J. , Broaddus Russell R. , Mikhail Kolonin G. , Mok Samuel C. , Woodward Wendy A. , Lu Karen H. , Klopp Ann H.
Published: August 2014

Objectives: Adipose tissue contains a population of multipotent mesenchymal stromal cells (ASC) which exhibit tumor tropism, similar to bone marrow derived mesenchymal stem cells (MSC). Excess visceral adipose tissue increases the risk of ovarian cancer. The omentum is a prominent site for ovarian cancer metastasis. We hypothesize that the omentum serves as a source of ASC which promote ovarian cancer progression.

Materials and Methods: ASC were isolated from the omentum (O-ASCs) of three patients with ovarian cancer. O-ASC1 was isolated from a patient with synchronous adenocarcinoma of endometrial and ovarian cancer without peritoneal metastasis; and O-ASC4 and O-ASC5 were isolated from patients with peritoneally disseminated serous ovarian cancer. Gene expression array profiling was performed with using Nimblegen arrays (Roche NimbleGen, Inc., Madison, WI). The impact of stromal cells on proliferation and chemoresistane and radio-protection of ovarian cancer cells was tested with co-culture assays using luciferase-labeled human ovarian cancer cell lines. Transwell migration assays were performed with conditioned media from O-ASC and control cell lines. To evaluate of O-ASCs tumor tropism in-vivo experiment was performed.

Results: Human O-ASCs were detected engrafted within the stroma of human ovarian cancer xengrafts. O-ASC significantly promoted in-vitro proliferation and migration of ovarian cancer cell lines OVCA 429, OVCA 433, A2780. Co-culture of ovarian cancer cells with O-ASCs increased resistance to chemotherapeutic drugs and radiation. Gene expression array analysis revealed significant differences in expression profiles for group of 415 genes in sub-populations of O-ASCs.

Conclusions: Adipose stem cells derived from human omentum promoted the proliferation, migration, chemoresistance and radioresistance of ovarian cancers. Clinical isolates demonstrate heterogenous effects in- vitro. Future studies will determine if tumor promoting effects of O-ASC can be predicted on the basis of clinical or disease related characteristics.

Citation Format: Aleksandra Nowicka, Travis Solley, Frank C. Marini, Hadley J. Sharp, Russell R. Broaddus, Kolonin G. Mikhail, Samuel C. Mok, Wendy A. Woodward, Karen H. Lu, Ann H. Klopp. Human omental-derived adipose stem cells: Modulator of ovarian cancer proliferation, migration and chemoresistance. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr B73.
Abstract IA13: The Fanconi anemia-BRCA pathway and chemosensitivity of cancer cells
Authors: Taniguchi Toshiyasu
Published: August 2014

The Fanconi anemia-BRCA pathway has emerged as an important pathway in cancer biology. Fanconi anemia (FA) is a rare genetic disease characterized by chromosomal instability, cancer-susceptibility and cellular sensitivity to interstrand DNA crosslink (ICL)-inducing agents. Sixteen FA genes have been identified (FANCA, FANCB, FANCC, FANCD1/BRCA2, FANCD2, FANCE, FANCF, FANCG, FANCI, FANCJ/BRIP1, FANCL, FANCM, FANCN/PALB2, FANCO/RAD51, FANCP/SLX4 and FANCQ/XPF). The FA proteins and breast/ovarian cancer susceptibility proteins, BRCA1 and BRCA2, cooperate in a common pathway required for cellular resistance to ICL-inducing agents. This pathway is called the FA-BRCA pathway or FA pathway. The main function of the pathway is to coordinate multiple DNA repair mechanisms (including homologous recombination) during ICL repair.

Inactivation of this pathway in cancer cells can lead to sensitivity to anti-cancer ICL-inducing agents, such as cisplatin, while reactivation of this pathway is implicated in acquired resistance to ICL-inducing agents. Since a part of the FA-BRCA pathway regulates homologous recombination, integrity of the pathway is also important for cellular resistance to PARP inhibitor. Therefore, inhibition of the FA-BRCA pathway is an attractive therapeutic strategy to overcome drug resistance of tumor cells.

In this talk, I will introduce what the FA-BRCA pathway is and how this pathway is inactivated in human cancer. I will also mention our finding that secondary BRCA1/2 mutations, which restore normal function of BRCA1/2 proteins, are involved in acquired drug resistance of BRCA1/2-mutated cancer.

Citation Format: Toshiyasu Taniguchi. The Fanconi anemia-BRCA pathway and chemosensitivity of cancer cells. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr IA13.
Abstract IA15: Targeting acquired platinum resistance in ovarian cancer
Authors: Gabra Hani
Published: August 2014

Platinum is an important cornerstone of chemotherapy for ovarian cancer. Platinum resistance is a multifactorial process, and may be intrinsic to the presenting tumor (refractory disease) or more commonly acquired after initial platinum sensitivity (acquired resistant disease). Although induction of acquired platinum resistance through adaptation of the primary tumor is thought to be one expression of these multifactorial mechanisms, there is evidence that selection of pre-existing resistance generated through clonal heterogeneity prior to clinical presentation may be a significant process that results in selection of acquired platinum resistant clinical relapse. This hypothesis has led to the identification by expression microarray difference analysis of upregulated targets which through targeting may reverse acquired platinum resistance. These targets include HDAC4, STAT1, FOLR2, and PI3K components, particularly AKT. In acquired platinum resistance, but not platinum sensitive ovarian cancer, there is nuclear relocalization of AKT with DNA damage mediated nuclear phosphorylation-activation of AKT resulting in survival signaling. Functional validation of these targets in-vitro and preclinically in-vivo has led to development of a translational proof of concept platform in the clinic with sequential biopsy within trials to test the hypothesis of targeting acquired resistance derived through clonal diversity at the outset of disease. Through this science, subsequent clinical trials of AKT inhibitors in platinum resistant ovarian cancer have demonstrated activity, underscoring the validity of this approach. The clinical development of such targeting of acquired resistance through international consortia within the EUTROC group will be discussed. It is hoped that this approach will improve the prognosis of ovarian cancer by early intervention to delete resistant disease before it becomes a clinical problem.

Citation Format: Hani Gabra. Targeting acquired platinum resistance in ovarian cancer. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr IA15.