Generation of low passage high grade serous ovarian cancer cell lines from primary tumors
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ABSTRACT: High grade serous ovarian cancers (HGSC) are deadly malignancies that relapse despite carboplatin chemotherapy. Many commercially ovarian cancer cell lines are not good models for HGSC. Here we demonstrate that 3 low passage cell lines derived from HGSC have similar transcriptomes to their parental bulk tumors. These cell lines recapitulated tumor characteristics of the primary cancer and had responded to therapy in the same manner as primary HGSC cells, demonstrating they are accurate models for HGSCs. mRNA profiles of low passage high grade serous tumor cell lines and their parental tumors, generated by next generation sequencing, were compared.
Project description:High grade serous ovarian cancers (HGSC) are deadly malignancies that relapse despite carboplatin chemotherapy. Here we show that 16 independent primary HGSCs contain a CA125 negative population enriched for carboplatin resistant cancer initiating cells. Transcriptome analysis reveals up-regulation of homologous recombination DNA repair and anti-apoptotic signals in this population. While treatment with carboplatin enriches for CA125 negative cells, co-treatment with carboplatin and birinapant eliminates these cells in HGSCs expressing high levels of the inhibitor of apoptosis protein cIAP in the CA125 negative population. Birinapant sensitizes CA125 negative cells to carboplatin by mediating degradation of cIAP causing cleavage of caspase-8 and restoration of apoptosis. This co-therapy significantly improved disease free survival in vivo compared to either therapy alone in tumor-bearing mice. These findings suggest that therapeutic strategies that target CA125 negative cells may be useful in the treatment of HGSC. mRNA profiles of CA125 positive and negative populations, generated by next generation sequencing of populations FACS isolated from 10 independent dissociated primary human high grade serous ovarian cancers, were compared.
Project description:High grade serous ovarian cancers (HGSC) are deadly malignancies that relapse despite carboplatin chemotherapy. Many commercially ovarian cancer cell lines are not good models for HGSC. Here we demonstrate that 3 low passage cell lines derived from HGSC have similar transcriptomes to their parental bulk tumors. These cell lines recapitulated tumor characteristics of the primary cancer and had responded to therapy in the same manner as primary HGSC cells, demonstrating they are accurate models for HGSCs.
Project description:High-grade serous ovarian cancer (HGSC) is a challenging disease, especially for patients with immunologically 'cold' tumors devoid of tumor-infiltrating lymphocytes (TIL). HGSC exhibits among the highest levels of MYCN transcriptional signature in human cancer, which is strongly linked to diminished features of anti-tumor immunity. We used microarrays to identify genes potentially dysregulated by the oncogene N-MYC in HGSC cell lines.
Project description:Identification and validation of potential prognostic biomarkers in older ovarian cancer patients with high-grade serous adenocarcinoma (HGSC)
Project description:Low grade serous ovarian cancer (LGSC) is a rare subtype of ovarian cancer, characterized by a slow growth rate, resistance to current treatment regimens, multiple recurrences and poor survival. LGSC arise from serous borderline tumor (SBT), however the mechanism of transformation is poorly understood. To better understand the biology of serous ovarian tumors, we performed whole proteome profiling of LGSC, SBT and the more common high grade serous (HGSC) ovarian tumors. Proteins associated with the tumor microenvironment were differentially expressed between LGSC and SBT or HGSC. In particular, fibroblast activation protein (FAP), a protein expressed in cancer associated fibroblasts, is abundantly expressed in LGSC. Furthermore, Tregs and M2 macrophages are more abundant in the stroma of LGSC compared to SBT. Together these data suggest that the tumor microenvironment provides a supportive environment for LGSC tumorigenesis and progression, and that targeting the tumor microenvironment of LGSC may be a viable therapeutic strategy.
Project description:Here we show that ovarian progesterone is a crucial endogenous factor inducing metastatic ovarian cancer in a mouse model of high-grade serous carcinoma (HGSC). To examine the molecular signaling pathways underlying progesterone (P4)-induced HGSC development, we performed gene-expression profiling using RNA sequencing.
Project description:Purpose: One of the goals of this study is to compare the transcriptome profiles of tumors from mice inoculated with specific engineered fallopian tube cells of high-grade serous tubo-ovarian cancer (HGSC) models (this study) using scRNA sequencing Methods: Tumors from mice inoculated with specific engineered fallopian tube cells of high-grade serous tubo-ovarian cancer (HGSC) models were compared using scRNA sequencing. Conclusions: We conclude that scRNA-seq based transcriptome characterization would expedite genetic network analyses and permit the dissection of complex biological functions.
Project description:Cisplatin- and carboplatin-resistant high-grade serous ovarian cancer (HGSC) cells were generated and new lines were created from single cell clones. All cells were derived from a common ancestral line, Ovcar4. RNA sequencing was performed to examine transcriptional changes related to acquisition of platinum resistance.
Project description:Here, we performed proteomics on whole cell lysate of three high-grade serous ovarian cancer patient derived cancer associated fibroblast (CAF) lines, four commercially available high-grade serous ovarian epithelial cancer cell lines (Kuramochi, PEO4, OVCAR8, ES2) and two immortalized normal Fallopian tube secretory epithelial cell lines (FT237, FT194). N-glycoproteomics using hydrazide-based enrichment was performed on 8/9 models. Integration of data from in vitro models with publicly available primary tissue data identified multiple novel CAF proteins that are associated with poor clinical outcomes in HGSC.
Project description:Purpose: One of the goals of this study is to compare the transcriptome profiles of tumors from mice inoculated with specific engineered fallopian tube cells of high-grade serous tubo-ovarian cancer (HGSC) models (this study) using next-generation sequencing (NGS)-derived RNA-seq. Methods: Tumors from mice inoculated with specific engineered fallopian tube cells of high-grade serous tubo-ovarian cancer (HGSC) models were compared using RNA sequencing. Conclusions: We conclude that RNA-seq based transcriptome characterization would expedite genetic network analyses and permit the dissection of complex biological functions.