(searched for: Generation and Characterization of A Carboplatin Resistant Ovarian Cancer Model)
Cells, Volume 9; doi:10.3390/cells9112408
Ovarian clear cell carcinoma (OCCC) is a rare subtype of gynecological cancer for which well-characterized and authenticated model systems are scarce. We provide an extensive characterization of ‘105C’, a cell line generated from an adenocarcinoma of the clear cell histotype using targeted next-generation sequencing, cytogenetic microarrays, along with analyses of AKT/mTOR signaling. We report that that the 105C cell line is a bona fide OCCC cell line, carrying PIK3CA, PTEN, and ARID1A gene mutations, consistent with OCCC, yet maintain a stable genome as reflected by low copy number variation. Unlike KOC-7c, TOV-21G, and RMG-V OCCC lines also mutated for the above genes, the 105C cells do not carry mutations in mismatch repair genes. Importantly, we show that 105C cells exhibit greater resistance to mTOR inhibition and carboplatin treatment compared to 9 other OCCC cell lines in 3D spheroid cultures. This resistance may be attributed to 105C cells remaining dormant in suspension culture which surprisingly, contrasts with several other OCCC lines which continue to proliferate in long-term suspension culture. 105C cells survive xenotransplantation but do not proliferate and metastasize. Collectively, we show that the 105C OCCC cell line exhibits unique properties useful for the pre-clinical investigation of OCCC pathobiology.
Published: 1 July 2020
Poster Presentations - Proffered Abstracts, Volume 26; doi:10.1158/1557-3265.ovca19-b78
Conference: Abstracts: AACR Special Conference on Advances in Ovarian Cancer Research; September 13-16, 2019; Atlanta, GA
The publisher has not yet granted permission to display this abstract.
Published: 1 January 2020
Journal of Cancer Science and Clinical Therapeutics, Volume 4, pp 144-153; doi:10.26502/jcsct.5079060
Acquisition of drug resistance is one of the major hurdles in the treatment of several cancers including ovarian carcinoma. There exist very few models that aid in understanding the mechanism of drug resistance in cancer cells. Carboplatin is administered as a first line chemotherapeutic drug in the treatment of ovarian cancer. Majority of ovarian cancer patients respond well to chemotherapy in the initial stages, but 60-70% report recurrence post therapy. The underlying molecular mechanisms contributing to resistance to therapy remain elusive. Establishing a clinically relevant in vitro tumor model that could closely mimic the drug resistance pattern in the patient would provide an opportunity to understand drug resistance. The present study focuses on generating a clinically relevant drug resistant model of ovarian cancer.
Published: 15 December 2019
PurposeThere remains an unmet need for preclinical models to enable personalized therapy for ovarian cancer (OC) patients. Recently, patient-derived organoid (PDO) cultures of patients with OC have been established that faithfully represent the histopathological features and genomic landscape of the patient tumor. In this study, we evaluate the capacity of OC PDOs to predict clinical drug response and functional consequences of tumor heterogeneity.Experimental design36 genomically characterized PDOs from 23 patients with known clinical histories were exposed to chemotherapeutics and targeted drugs.ResultsOC PDOs maintained genomic features of the original tumor lesion and recapitulated patient response to neoadjuvant carboplatin and paclitaxel combination treatment, according to distinct clinical outcomes (histopathological, biochemical and radiological). PDOs displayed inter-as well as intrapatient drug response heterogeneity, which could in part be explained by genetic aberrations. All PDOs were resistant to PARP-inhibitors, in accordance with homologous recombination pathway fidelity and genome-wide mutation context. KRAS, BRAF and NRAS mutation status predicted response to BRAF-inhibitor vemurafenib and pan-HER-inhibitor afatinib, and explained differential response among four PDOs derived from distinct tumor locations of an individual patient. Importantly, PDO drug screening identified sensitivity to at least one drug for the majority of patients (88%).ConclusionsOC PDOs are a valuable preclinical model system that can provide insights in drug response for individual patients with OC, complementary to genetic testing. Generating PDOs of multiple tumor locations can improve clinical decision making and increase our knowledge on genetic and drug response heterogeneity.
Tumor Biology, Volume 79, pp 1065-1065; doi:10.1158/1538-7445.sabcs18-1065
Purpose: Ovarian cancer is heterogeneous with multiple histological subtypes and a wide range of genetic aberrations. While high-grade serous ovarian carcinoma (HGSOC) is the most common, clear cell ovarian carcinoma (CCOC) is the most challenging to treat and exhibits low response rates to standard therapies. However, efforts to understand CCOC and develop new therapies have been limited because it represents a minority of ovarian cancers in the U.S. and Europe. In contrast, CCOC accounts for approximately 30% of all ovarian cancer in Japan. To improve the survival of patients with CCOC, a deeper understanding of the molecular features that define available model systems is needed. Our goal is to comprehensively characterize a panel of CCOC lines using next generation sequencing and functional in vitro and in vivo experiments to define the lines that are most faithful to CCOC and are tractable for subsequent in vivo drug discovery. Method: We obtained 9 CCOC cells from ATCC, Riken Cell Bank, and University of Miami (ES-2, TOV21G, OVTOKO, OVMANA, OCI-C5x, JHOC-5, JHOC-7, JHOC-9, and OVISE). Genomic DNA, RNA, and protein were isolated and subjected to whole exome DNA-seq, RNA-seq, and reverse phase protein array (RPPA), respectively. We performed in vitro MTT assays to test the sensitivity of these lines to chemotherapies. Tumorigenicity was evaluated by injecting 5 million cells of luciferized CCOC lines into NSG female mice using both the subcutaneous route and the intraperitoneal route. Imaging was performed weekly using the In Vivo Imaging System. Results: ARID1A mutation, the most prevalent mutation in CCOC, is present in 8 of the 9 cell lines (TOV21G, OVTOKO, OVMANA, OCI-C5x, JHOC-5, JHOC-7, JHOC-9, and OVISE), whereas the second common mutation in CCOC (PIK3CA) is detected in 5 lines (TOV21G, OVMANA, OCI-C5x, JHOC-7, and OVISE). The ES-2 cell line has both TP53 and BRAF mutations and its genomic profile is not typical of CCOC. Principal component analysis of RPPA showed distinct groups between the 9 CCOC lines and the 6 HGSOC lines. Interestingly, we also observed two distinct clusters within the CCOC lines. Consistent with our genomic analysis, the ES-2 cell line correlated more closely with the HGSOC lines based on RPPA data. In our in vitro drug studies, OVTOKO and OCI-C5x exhibited resistance to Carboplatin/Paclitaxel. In xenograft study, 4 cell lines (ES-2, TOV21G, OVTOKO, and OCI-C5x) formed measurable tumor within a month. In contrast, OVMANA, JHOC-7, JHOC-9, and OVISE took over 100 days to form tumors. Conclusion: Our data suggests that there may exist two functionally distinct groups within CCOC that warrants further study. In vitro and in vivo studies identified 4 cell lines that represent tractable models for rigorous therapeutic studies: ES-2, TOV21G, OVTOKO, and OCI-C5x. However, ES-2 appears to cluster more closely with HGSOC and may not represent the CCOC histotype. Citation Format: Yasuto Kinose, Dorothy Hallberg, Kai Doberstein, Gordon Mills, Tan Ince, Victor Velculescu, Fiona Simpkins, Ronny Drapkin. Comprehensive molecular and experimental characterization of ovarian clear cell carcinoma cell lines for in vivo drug development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1065.
Published: 1 August 2018
Abstracts: AACR Special Conference: Addressing Critical Questions in Ovarian Cancer Research and Treatment; October 1-4, 2017; Pittsburgh, PA, Volume 24; doi:10.1158/1557-3265.ovca17-a54
Conference: Abstracts: AACR Special Conference: Addressing Critical Questions in Ovarian Cancer Research and Treatment; October 1-4, 2017; Pittsburgh, PA
The publisher has not yet granted permission to display this abstract.