The Experts below are selected from a list of 248043 Experts worldwide ranked by ideXlab platform
Ronny Drapkin - One of the best experts on this subject based on the ideXlab platform.
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abstract gmm 036 comprehensive genomic proteomic and Experimental Characterization of ovarian clear cell carcinoma cell lines for improved drug development
Clinical Cancer Research, 2019Co-Authors: Yasuto Kinose, Dorothy Hallberg, Gordon B Mills, Tan A Ince, Victor E Velculescu, Fiona Simpkins, Ronny DrapkinAbstract:PURPOSE: Ovarian cancer is a heterogeneous disease with multiple histological subtypes and a wide range of genetic aberrations. While high-grade serous carcinoma (HGSC) is the most common type, clear cell carcinoma (CCC) of the ovary is notoriously the most challenging to treat and exhibits very low response rates to standard therapies. However, efforts to understand CCC and develop new therapeutic approaches to this subtype have been limited because it represents a minority of ovarian cancer cases in the United States and Europe. In contrast, CCC accounts for more than 30% of all ovarian cancer in Japan. To improve the survival of patients with ovarian CCC, a deeper understanding of the molecular features of available model systems is needed. Our goal is to characterize a panel of CCC cell lines genomically and functionally and identify those that can serve as tractable model systems for future in vivo drug discovery studies. METHOD: First, we characterize 9 CCC cell lines (ES-2, TOV21G, OVTOKO, OVMANA, OCI-C5x, JHOC-5, JHOC-7, JHOC-9, and OVISE) with whole exome sequence and proteomics approaches (reverse phase protein array; RPPA). Secondly, we performed in vitro assays, including soft-agar colony formation assays and MTT assays using standard chemotherapies. Thirdly, to test in vivo tumorigenic potential, by injecting 5 million cells of luciferized CCC lines in NSG female mice using both the subcutaneous route and the intraperitoneal route. Imaging was performed weekly using the In Vivo Imaging System. RESULTS: Among the 9 CCC lines, seven (TOV21G, OVTOKO, OVMANA, OCI-C5x, JHOC-5, JHOC-9, and OVISE) harbor ARID1A mutation, which is the most prevalent mutation in CCC (50%). The second common mutation in CCC (40%), PIK3CA mutation, and was detected in 4 cells (TOV21G, OVMANA, OCI-C5x, and OVISE). The ES-2 line has TP53 and BRAF mutation and its genomic profile is inconsistent with CCC. A BRCA2 mutation was found in OVTOKO and JHOC-5, finding not previously reported. Principal component analysis of RPPA data showed distinct clusters between the 9 CCC lines and the 6 HGSC lines. Interestingly, we also observed two distinct clusters within the CCC lines. Consistent with our genomic analysis, ES-2 correlated with HGSC lines based on RPPA data. In our in vitro drug studies, OVTOKO, OCI-C5x, and OVISE exhibited a Carboplatin/Paclitaxel resistance phenotype. In xenograft study, 4 cell lines (ES-2, TOV21G, OVTOKO, and OCI-C5x) formed tumor within a month, suggesting they are useful tools for in vivo studies. In contrast, OVMANA, JHOC-7, JHOC-9, and OVISE take more than 100 days to form the tumors. Soft-agar colony formation results correlated with how long it took to develop xenograft tumor in each line. CONCLUSION: Our genomic studies identified aberrations in CCC lines not previously described. The proteomics data revealed two clusters within the CCC lines that might represent functionally distinct groups. Finally, we identified four cell lines (ES-2, TOV21G, OVTOKO, and OCI-C5x) that readily form tumors in mice and could be used for future in vivo drug studies. However, ES-2 appears to cluster more closely with HGSC and may not represent the CCC histotype. Citation Format: Yasuto Kinose, Dorothy Hallberg, Gordon Mills, Tan Ince, Victor Velculescu, Fiona Simpkins, Ronny Drapkin. COMPREHENSIVE GENOMIC, PROTEOMIC, AND Experimental Characterization OF OVARIAN CLEAR CELL CARCINOMA CELL LINES FOR IMPROVED DRUG DEVELOPMENT [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr GMM-036.
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abstract 1065 comprehensive molecular and Experimental Characterization of ovarian clear cell carcinoma cell lines forin vivodrug development
Cancer Research, 2019Co-Authors: Yasuto Kinose, Dorothy Hallberg, Kai Doberstein, Gordon B Mills, Tan A Ince, Victor E Velculescu, Fiona Simpkins, Ronny DrapkinAbstract: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.
Yasuto Kinose - One of the best experts on this subject based on the ideXlab platform.
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abstract gmm 036 comprehensive genomic proteomic and Experimental Characterization of ovarian clear cell carcinoma cell lines for improved drug development
Clinical Cancer Research, 2019Co-Authors: Yasuto Kinose, Dorothy Hallberg, Gordon B Mills, Tan A Ince, Victor E Velculescu, Fiona Simpkins, Ronny DrapkinAbstract:PURPOSE: Ovarian cancer is a heterogeneous disease with multiple histological subtypes and a wide range of genetic aberrations. While high-grade serous carcinoma (HGSC) is the most common type, clear cell carcinoma (CCC) of the ovary is notoriously the most challenging to treat and exhibits very low response rates to standard therapies. However, efforts to understand CCC and develop new therapeutic approaches to this subtype have been limited because it represents a minority of ovarian cancer cases in the United States and Europe. In contrast, CCC accounts for more than 30% of all ovarian cancer in Japan. To improve the survival of patients with ovarian CCC, a deeper understanding of the molecular features of available model systems is needed. Our goal is to characterize a panel of CCC cell lines genomically and functionally and identify those that can serve as tractable model systems for future in vivo drug discovery studies. METHOD: First, we characterize 9 CCC cell lines (ES-2, TOV21G, OVTOKO, OVMANA, OCI-C5x, JHOC-5, JHOC-7, JHOC-9, and OVISE) with whole exome sequence and proteomics approaches (reverse phase protein array; RPPA). Secondly, we performed in vitro assays, including soft-agar colony formation assays and MTT assays using standard chemotherapies. Thirdly, to test in vivo tumorigenic potential, by injecting 5 million cells of luciferized CCC lines in NSG female mice using both the subcutaneous route and the intraperitoneal route. Imaging was performed weekly using the In Vivo Imaging System. RESULTS: Among the 9 CCC lines, seven (TOV21G, OVTOKO, OVMANA, OCI-C5x, JHOC-5, JHOC-9, and OVISE) harbor ARID1A mutation, which is the most prevalent mutation in CCC (50%). The second common mutation in CCC (40%), PIK3CA mutation, and was detected in 4 cells (TOV21G, OVMANA, OCI-C5x, and OVISE). The ES-2 line has TP53 and BRAF mutation and its genomic profile is inconsistent with CCC. A BRCA2 mutation was found in OVTOKO and JHOC-5, finding not previously reported. Principal component analysis of RPPA data showed distinct clusters between the 9 CCC lines and the 6 HGSC lines. Interestingly, we also observed two distinct clusters within the CCC lines. Consistent with our genomic analysis, ES-2 correlated with HGSC lines based on RPPA data. In our in vitro drug studies, OVTOKO, OCI-C5x, and OVISE exhibited a Carboplatin/Paclitaxel resistance phenotype. In xenograft study, 4 cell lines (ES-2, TOV21G, OVTOKO, and OCI-C5x) formed tumor within a month, suggesting they are useful tools for in vivo studies. In contrast, OVMANA, JHOC-7, JHOC-9, and OVISE take more than 100 days to form the tumors. Soft-agar colony formation results correlated with how long it took to develop xenograft tumor in each line. CONCLUSION: Our genomic studies identified aberrations in CCC lines not previously described. The proteomics data revealed two clusters within the CCC lines that might represent functionally distinct groups. Finally, we identified four cell lines (ES-2, TOV21G, OVTOKO, and OCI-C5x) that readily form tumors in mice and could be used for future in vivo drug studies. However, ES-2 appears to cluster more closely with HGSC and may not represent the CCC histotype. Citation Format: Yasuto Kinose, Dorothy Hallberg, Gordon Mills, Tan Ince, Victor Velculescu, Fiona Simpkins, Ronny Drapkin. COMPREHENSIVE GENOMIC, PROTEOMIC, AND Experimental Characterization OF OVARIAN CLEAR CELL CARCINOMA CELL LINES FOR IMPROVED DRUG DEVELOPMENT [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr GMM-036.
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abstract 1065 comprehensive molecular and Experimental Characterization of ovarian clear cell carcinoma cell lines forin vivodrug development
Cancer Research, 2019Co-Authors: Yasuto Kinose, Dorothy Hallberg, Kai Doberstein, Gordon B Mills, Tan A Ince, Victor E Velculescu, Fiona Simpkins, Ronny DrapkinAbstract: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.
Victor E Velculescu - One of the best experts on this subject based on the ideXlab platform.
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abstract gmm 036 comprehensive genomic proteomic and Experimental Characterization of ovarian clear cell carcinoma cell lines for improved drug development
Clinical Cancer Research, 2019Co-Authors: Yasuto Kinose, Dorothy Hallberg, Gordon B Mills, Tan A Ince, Victor E Velculescu, Fiona Simpkins, Ronny DrapkinAbstract:PURPOSE: Ovarian cancer is a heterogeneous disease with multiple histological subtypes and a wide range of genetic aberrations. While high-grade serous carcinoma (HGSC) is the most common type, clear cell carcinoma (CCC) of the ovary is notoriously the most challenging to treat and exhibits very low response rates to standard therapies. However, efforts to understand CCC and develop new therapeutic approaches to this subtype have been limited because it represents a minority of ovarian cancer cases in the United States and Europe. In contrast, CCC accounts for more than 30% of all ovarian cancer in Japan. To improve the survival of patients with ovarian CCC, a deeper understanding of the molecular features of available model systems is needed. Our goal is to characterize a panel of CCC cell lines genomically and functionally and identify those that can serve as tractable model systems for future in vivo drug discovery studies. METHOD: First, we characterize 9 CCC cell lines (ES-2, TOV21G, OVTOKO, OVMANA, OCI-C5x, JHOC-5, JHOC-7, JHOC-9, and OVISE) with whole exome sequence and proteomics approaches (reverse phase protein array; RPPA). Secondly, we performed in vitro assays, including soft-agar colony formation assays and MTT assays using standard chemotherapies. Thirdly, to test in vivo tumorigenic potential, by injecting 5 million cells of luciferized CCC lines in NSG female mice using both the subcutaneous route and the intraperitoneal route. Imaging was performed weekly using the In Vivo Imaging System. RESULTS: Among the 9 CCC lines, seven (TOV21G, OVTOKO, OVMANA, OCI-C5x, JHOC-5, JHOC-9, and OVISE) harbor ARID1A mutation, which is the most prevalent mutation in CCC (50%). The second common mutation in CCC (40%), PIK3CA mutation, and was detected in 4 cells (TOV21G, OVMANA, OCI-C5x, and OVISE). The ES-2 line has TP53 and BRAF mutation and its genomic profile is inconsistent with CCC. A BRCA2 mutation was found in OVTOKO and JHOC-5, finding not previously reported. Principal component analysis of RPPA data showed distinct clusters between the 9 CCC lines and the 6 HGSC lines. Interestingly, we also observed two distinct clusters within the CCC lines. Consistent with our genomic analysis, ES-2 correlated with HGSC lines based on RPPA data. In our in vitro drug studies, OVTOKO, OCI-C5x, and OVISE exhibited a Carboplatin/Paclitaxel resistance phenotype. In xenograft study, 4 cell lines (ES-2, TOV21G, OVTOKO, and OCI-C5x) formed tumor within a month, suggesting they are useful tools for in vivo studies. In contrast, OVMANA, JHOC-7, JHOC-9, and OVISE take more than 100 days to form the tumors. Soft-agar colony formation results correlated with how long it took to develop xenograft tumor in each line. CONCLUSION: Our genomic studies identified aberrations in CCC lines not previously described. The proteomics data revealed two clusters within the CCC lines that might represent functionally distinct groups. Finally, we identified four cell lines (ES-2, TOV21G, OVTOKO, and OCI-C5x) that readily form tumors in mice and could be used for future in vivo drug studies. However, ES-2 appears to cluster more closely with HGSC and may not represent the CCC histotype. Citation Format: Yasuto Kinose, Dorothy Hallberg, Gordon Mills, Tan Ince, Victor Velculescu, Fiona Simpkins, Ronny Drapkin. COMPREHENSIVE GENOMIC, PROTEOMIC, AND Experimental Characterization OF OVARIAN CLEAR CELL CARCINOMA CELL LINES FOR IMPROVED DRUG DEVELOPMENT [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr GMM-036.
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abstract 1065 comprehensive molecular and Experimental Characterization of ovarian clear cell carcinoma cell lines forin vivodrug development
Cancer Research, 2019Co-Authors: Yasuto Kinose, Dorothy Hallberg, Kai Doberstein, Gordon B Mills, Tan A Ince, Victor E Velculescu, Fiona Simpkins, Ronny DrapkinAbstract: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.
Tan A Ince - One of the best experts on this subject based on the ideXlab platform.
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abstract gmm 036 comprehensive genomic proteomic and Experimental Characterization of ovarian clear cell carcinoma cell lines for improved drug development
Clinical Cancer Research, 2019Co-Authors: Yasuto Kinose, Dorothy Hallberg, Gordon B Mills, Tan A Ince, Victor E Velculescu, Fiona Simpkins, Ronny DrapkinAbstract:PURPOSE: Ovarian cancer is a heterogeneous disease with multiple histological subtypes and a wide range of genetic aberrations. While high-grade serous carcinoma (HGSC) is the most common type, clear cell carcinoma (CCC) of the ovary is notoriously the most challenging to treat and exhibits very low response rates to standard therapies. However, efforts to understand CCC and develop new therapeutic approaches to this subtype have been limited because it represents a minority of ovarian cancer cases in the United States and Europe. In contrast, CCC accounts for more than 30% of all ovarian cancer in Japan. To improve the survival of patients with ovarian CCC, a deeper understanding of the molecular features of available model systems is needed. Our goal is to characterize a panel of CCC cell lines genomically and functionally and identify those that can serve as tractable model systems for future in vivo drug discovery studies. METHOD: First, we characterize 9 CCC cell lines (ES-2, TOV21G, OVTOKO, OVMANA, OCI-C5x, JHOC-5, JHOC-7, JHOC-9, and OVISE) with whole exome sequence and proteomics approaches (reverse phase protein array; RPPA). Secondly, we performed in vitro assays, including soft-agar colony formation assays and MTT assays using standard chemotherapies. Thirdly, to test in vivo tumorigenic potential, by injecting 5 million cells of luciferized CCC lines in NSG female mice using both the subcutaneous route and the intraperitoneal route. Imaging was performed weekly using the In Vivo Imaging System. RESULTS: Among the 9 CCC lines, seven (TOV21G, OVTOKO, OVMANA, OCI-C5x, JHOC-5, JHOC-9, and OVISE) harbor ARID1A mutation, which is the most prevalent mutation in CCC (50%). The second common mutation in CCC (40%), PIK3CA mutation, and was detected in 4 cells (TOV21G, OVMANA, OCI-C5x, and OVISE). The ES-2 line has TP53 and BRAF mutation and its genomic profile is inconsistent with CCC. A BRCA2 mutation was found in OVTOKO and JHOC-5, finding not previously reported. Principal component analysis of RPPA data showed distinct clusters between the 9 CCC lines and the 6 HGSC lines. Interestingly, we also observed two distinct clusters within the CCC lines. Consistent with our genomic analysis, ES-2 correlated with HGSC lines based on RPPA data. In our in vitro drug studies, OVTOKO, OCI-C5x, and OVISE exhibited a Carboplatin/Paclitaxel resistance phenotype. In xenograft study, 4 cell lines (ES-2, TOV21G, OVTOKO, and OCI-C5x) formed tumor within a month, suggesting they are useful tools for in vivo studies. In contrast, OVMANA, JHOC-7, JHOC-9, and OVISE take more than 100 days to form the tumors. Soft-agar colony formation results correlated with how long it took to develop xenograft tumor in each line. CONCLUSION: Our genomic studies identified aberrations in CCC lines not previously described. The proteomics data revealed two clusters within the CCC lines that might represent functionally distinct groups. Finally, we identified four cell lines (ES-2, TOV21G, OVTOKO, and OCI-C5x) that readily form tumors in mice and could be used for future in vivo drug studies. However, ES-2 appears to cluster more closely with HGSC and may not represent the CCC histotype. Citation Format: Yasuto Kinose, Dorothy Hallberg, Gordon Mills, Tan Ince, Victor Velculescu, Fiona Simpkins, Ronny Drapkin. COMPREHENSIVE GENOMIC, PROTEOMIC, AND Experimental Characterization OF OVARIAN CLEAR CELL CARCINOMA CELL LINES FOR IMPROVED DRUG DEVELOPMENT [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr GMM-036.
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abstract 1065 comprehensive molecular and Experimental Characterization of ovarian clear cell carcinoma cell lines forin vivodrug development
Cancer Research, 2019Co-Authors: Yasuto Kinose, Dorothy Hallberg, Kai Doberstein, Gordon B Mills, Tan A Ince, Victor E Velculescu, Fiona Simpkins, Ronny DrapkinAbstract: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.
Gordon B Mills - One of the best experts on this subject based on the ideXlab platform.
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abstract gmm 036 comprehensive genomic proteomic and Experimental Characterization of ovarian clear cell carcinoma cell lines for improved drug development
Clinical Cancer Research, 2019Co-Authors: Yasuto Kinose, Dorothy Hallberg, Gordon B Mills, Tan A Ince, Victor E Velculescu, Fiona Simpkins, Ronny DrapkinAbstract:PURPOSE: Ovarian cancer is a heterogeneous disease with multiple histological subtypes and a wide range of genetic aberrations. While high-grade serous carcinoma (HGSC) is the most common type, clear cell carcinoma (CCC) of the ovary is notoriously the most challenging to treat and exhibits very low response rates to standard therapies. However, efforts to understand CCC and develop new therapeutic approaches to this subtype have been limited because it represents a minority of ovarian cancer cases in the United States and Europe. In contrast, CCC accounts for more than 30% of all ovarian cancer in Japan. To improve the survival of patients with ovarian CCC, a deeper understanding of the molecular features of available model systems is needed. Our goal is to characterize a panel of CCC cell lines genomically and functionally and identify those that can serve as tractable model systems for future in vivo drug discovery studies. METHOD: First, we characterize 9 CCC cell lines (ES-2, TOV21G, OVTOKO, OVMANA, OCI-C5x, JHOC-5, JHOC-7, JHOC-9, and OVISE) with whole exome sequence and proteomics approaches (reverse phase protein array; RPPA). Secondly, we performed in vitro assays, including soft-agar colony formation assays and MTT assays using standard chemotherapies. Thirdly, to test in vivo tumorigenic potential, by injecting 5 million cells of luciferized CCC lines in NSG female mice using both the subcutaneous route and the intraperitoneal route. Imaging was performed weekly using the In Vivo Imaging System. RESULTS: Among the 9 CCC lines, seven (TOV21G, OVTOKO, OVMANA, OCI-C5x, JHOC-5, JHOC-9, and OVISE) harbor ARID1A mutation, which is the most prevalent mutation in CCC (50%). The second common mutation in CCC (40%), PIK3CA mutation, and was detected in 4 cells (TOV21G, OVMANA, OCI-C5x, and OVISE). The ES-2 line has TP53 and BRAF mutation and its genomic profile is inconsistent with CCC. A BRCA2 mutation was found in OVTOKO and JHOC-5, finding not previously reported. Principal component analysis of RPPA data showed distinct clusters between the 9 CCC lines and the 6 HGSC lines. Interestingly, we also observed two distinct clusters within the CCC lines. Consistent with our genomic analysis, ES-2 correlated with HGSC lines based on RPPA data. In our in vitro drug studies, OVTOKO, OCI-C5x, and OVISE exhibited a Carboplatin/Paclitaxel resistance phenotype. In xenograft study, 4 cell lines (ES-2, TOV21G, OVTOKO, and OCI-C5x) formed tumor within a month, suggesting they are useful tools for in vivo studies. In contrast, OVMANA, JHOC-7, JHOC-9, and OVISE take more than 100 days to form the tumors. Soft-agar colony formation results correlated with how long it took to develop xenograft tumor in each line. CONCLUSION: Our genomic studies identified aberrations in CCC lines not previously described. The proteomics data revealed two clusters within the CCC lines that might represent functionally distinct groups. Finally, we identified four cell lines (ES-2, TOV21G, OVTOKO, and OCI-C5x) that readily form tumors in mice and could be used for future in vivo drug studies. However, ES-2 appears to cluster more closely with HGSC and may not represent the CCC histotype. Citation Format: Yasuto Kinose, Dorothy Hallberg, Gordon Mills, Tan Ince, Victor Velculescu, Fiona Simpkins, Ronny Drapkin. COMPREHENSIVE GENOMIC, PROTEOMIC, AND Experimental Characterization OF OVARIAN CLEAR CELL CARCINOMA CELL LINES FOR IMPROVED DRUG DEVELOPMENT [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr GMM-036.
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abstract 1065 comprehensive molecular and Experimental Characterization of ovarian clear cell carcinoma cell lines forin vivodrug development
Cancer Research, 2019Co-Authors: Yasuto Kinose, Dorothy Hallberg, Kai Doberstein, Gordon B Mills, Tan A Ince, Victor E Velculescu, Fiona Simpkins, Ronny DrapkinAbstract: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.
