Angelina Londono

Epigenetic modifiers upregulate MHC II and impede ovarian cancer tumor growth

Expression of MHC class II pathway proteins in ovarian cancer correlates with prolonged survival. Murine and human ovarian cancer cells were treated with epigenetic modulators – histone deacetylase inhibitors and a DNA methyltransferase inhibitor. mRNA and protein expression of the MHC II pathway were evaluated by qPCR and flow cytometry. Treatment with entinostat and azacytidine of ID8 cells in vitro increased mRNA levels of Cd74, Ciita, and H2-Aa, H2-Eb1. MHC II and CD74 protein expression were increased after treatment with either agent. A dose dependent response in mRNA and protein expression was seen with entinostat. Combination treatment showed higher MHC II protein expression than with single agent treatment. In patient derived xenografts, CIITA, CD74, and MHC II mRNA transcripts were significantly increased after combination treatment. Expression of MHC II on ovarian tumors in MISIIR-Tag mice was increased with both agents relative to control. Combination treatment significantly reduced ID8 tumor growth in immune-competent mice. Epigenetic treatment increases expression of MHC II on ovarian cancer cells and impedes tumor growth. This approach warrants further study in ovarian cancer patients.

Metabolic risk factors and mechanisms of disease in epithelial ovarian cancer: A review

OBJECTIVE Epithelial ovarian cancer continues to be the deadliest gynecologic malignancy. Patients with both diabetes mellitus and obesity have poorer outcomes, yet research correlating metabolic abnormalities, such as metabolic syndrome, to ovarian cancer risk and outcomes is lacking. This article reviews the literature regarding metabolic derangements and their relationship to epithelial ovarian cancer, with a focus on potential mechanisms behind these associations. METHODS PubMed and Google Scholar were searched for articles in the English language regarding epithelial ovarian cancer, obesity, diabetes mellitus, and metabolic syndrome, with a focus on studies conducted since 1990. RESULTS Obesity, type II diabetes mellitus, and metabolic syndrome have been associated with poor outcomes in epithelial ovarian cancer. More studies investigating the relationship between metabolic syndrome and epithelial ovarian cancer are needed. A variety of pathologic factors may contribute to cancer risk in patients with metabolic derangements, including altered adipokine and cytokine expression, altered immune responses to tumor cells, and changes in pro-tumorigenic signaling pathways. CONCLUSION More research is needed to examine the effects of metabolic syndrome on epithelial ovarian cancer risk and mortality, as well as the underlying pathophysiologies in patients with obesity, diabetes mellitus, and metabolic syndrome that may be targeted for therapeutic intervention.

Molecular Response to Neoadjuvant Chemotherapy in High-Grade Serous Ovarian Carcinoma

While high-grade serous ovarian carcinoma (HGSOC) is the most common histologic subtype of ovarian cancer, significant tumor heterogeneity exists. In addition, chemotherapy induces changes in gene expression and alters the mutational profile. To evaluate the notion that patients with HGSOC could be better classified for optimal treatment based on gene expression, we compared genetic variants [by DNA next-generation sequencing (NGS) using a 50 gene Ion Torrent panel] and gene expression (using the NanoString PanCancer 770 gene Panel) in the tumor from 20 patients with HGSOC before and after neoadjuvant chemotherapy (NACT). NGS was performed on plasma cell free DNA (cfDNA) on a select group of patients (n = 14) to assess the utility of using cfDNA to monitor these changes. A total of 86 genes had significant changes in RNA expression after NACT. Thirty-eight genetic variants (including SNPs) from 6 genes were identified in tumors pre-NACT, while 59 variants from 19 genes were detected in the cfDNA. The number of DNA variants were similar after NACT. Of the 59 variants in the plasma pre-NACT, only 6 persisted, whereas 33 of 38 specific variants in the tumor DNA remained unchanged. Pathway analysis showed the most significant alterations in the cell cycle and DNA damage pathways. Implications: Gene expression profiles at the time of interval debulking provide additional genetic information that could help impact treatment decisions after NACT; although, continued collection and analysis of matched tumor and cfDNA from multiple time points are needed to determine the role of cfDNA in the management of HGSOC. Mol Cancer Res; 16(5); 813–24. ©2018 AACR.

Abstract 4710: The effect of Wnt inhibition combined with paclitaxel on tumor burden and CD8+ T cell infiltration in syngeneic murine models of ovarian cancer

Objective: The Wnt/β-catenin pathway is a major signal transduction pathway involved in ovarian cancer (OVCA) metastasis and resistance to chemotherapy. This pathway downregulates protective immunity mediated by intra-tumoral CD8+T cells in other cancer types. WNT974 is a novel drug that inhibits the enzyme Porcupine, which controls Wnt protein secretion. Our objective was to measure the effect of WNT974 alone and in combination with dose dense paclitaxel (ddPac) on tumor growth and on infiltrating CD8+ T cells in two syngeneic OVCA mouse models. Methods: C57BL/6 mice were injected subcutaneously (SC) (n=20) or intraperitoneally (IP) (n=24) with 7 x 106 ID8 mouse OVCA cells. Mice were treated with vehicle control, ddPac, WNT974, or the combination (combo). C57BL/6 TgMISIIR-Tag-Low transgenic mice were injected with 7 x 106 MOVCAR cells IP (n=8) and treated with vehicle control or combo. WNT974 was given by oral gavage twice a day (5mg/kg for 7 days then decreased to 2.5mg/kg twice a day for up to 4 weeks). Paclitaxel was given IP (5mg/kg) 3 days on and 3 days off for a total of 9 doses. SC tumors were measured with calipers twice per week. In the IP model, mice were sacrificed after 13 days of treatment and tumor weights and ascites volume were determined. Flow cytometry was used to evaluate the presence of CD8+ T cells in IP tumors. Results: In the ID8 SC model, combo therapy reduced tumor size compared to vehicle control (18 vs. 40mm2, p Conclusions: The combination of WNT974 and ddPac reduced tumor size and increased tumor infiltration of CD8+ T cells in syngeneic OVCA mouse models. This suggests that the improved response observed with the ddPac and WNT974 combination is in part due to upregulation of the intra-tumoral immune response. Further investigation of this pathway is warranted as an immune modulator and a potential therapeutic target in ovarian cancer. Citation Format: David W. Doo, Angelina I. Londono, Dylana J. Moore, Selene Meza-Perez, Ashwini A. Katre, Tyler R. McCaw, Haller J. Smith, Carol Y. Lin, Sara J. Cooper, J Michael Straughn, Donald J. Buchsbaum, Lyse A. Norian, Troy D. Randall, Rebecca C. Arend. The effect of Wnt inhibition combined with paclitaxel on tumor burden and CD8+ T cell infiltration in syngeneic murine models of ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4710.

Abstract B29: Analysis of Clinical Cancer Gene Panels by Next Generation Sequencing in Tumor and Circulating Cell-Free DNA Samples in Recurrent Ovarian Cancer Patients

Objective: We investigated the potential role of molecular profiling through next generation sequencing (NGS) to identify actionable gene mutations in tumors of recurrent ovarian cancer patients that could impact clinical care decisions. Methods: Under IRB approval, patients diagnosed with recurrent ovarian cancer from 9/2015 to 3/2016 underwent NGS of archival tumor specimens utilizing a 65 gene panel performed by Washington University9s Genomics and Pathology Services (GPS) or a 315 gene FoundationOne (FO) panel performed by Foundation Medicine. Select patients also underwent NGS of circulating cell free DNA (cfDNA) from blood specimens utilizing a 50 gene panel performed at Circulogene Theranostics. Genomic alterations found from cfDNA in the blood at the time of recurrence were compared to alterations found in archival tumor tissue. The effect of therapies selected on the basis of NGS testing was assessed. Results: Specimens from 95 recurrent ovarian cancer patients have been collected; 54 had the GPS gene panel and 35 had the FO gene panel. Six patient samples were unable to be sequenced due to inadequate tumor DNA. 44 patients were sequenced with the 50 gene cfDNA panel at Circulogene; four patient samples did not have detectable cfDNA and were unable to be sequenced. 30 patients have had both NGS on tumor and cfDNA. A range of 0 to15 mutations were detected by the GPS or FO NGS panel with an average of 3.7 mutations per tumor sample. A range of 0 to 6 mutations were detected on cfDNA panel with an average of 1.7 mutations per patient. The most frequent mutation detected by GPS, FO, and Circulogene was TP53. Of the 30 patients with both tumor and cfDNA sequenced, only 7 patients were found to have a TP53 genomic alteration in both the tumor and cfDNA. In all other patients where both cfDNA and tumor DNA had NGS performed, there was no concordance between the tumor and the cfDNA genomic alterations. Of the 95 patients, three patients were started on targeted therapy based on their results (Olaparib for a somatic BRCA mutation [without a germline mutation], Pazopanib for a FGFR mutation, Everolimus for a CTNNB1 mutation). Targeted therapy is being pursued in 6 additional patients: Trametinib for a patient with a KRAS mutation, Nivolumab for a patient with high microsatellite instability, Everolimus for 3 patients with PIK3CA mutations or a PTEN mutation, and Pertuzumab for a patient with a ERBB3 mutation. A somatic BRCA mutation was found in nine patients, seven of which also have a germline BRCA mutation. Conclusion: Ovarian cancer has a diverse genetic landscape and molecular profiling via NGS offers the opportunity to identify genetic alterations that can be utilized to direct therapy. Approximately 15% of patients with recurrent ovarian cancer have a mutation that can be targeted with a commercially available drug. We have been successful in providing patients with a mutation the NGS-directed therapy. Challenges still exist with the large variation in NGS technology and reporting and further research is needed to better identify actionable oncogenic drivers. Citation Format: Angelina I. Londono, Naveed Farrukh, Mary Kat Smith, Cindy M. Tawfik, Ronald D. Alvarez, Warner K. Huh, Kerri S. Bevis, Charles A. Leath, III, Straughn Michael, Jr., Eddy Shih Hsin Yang, Shuko Harada, Kenneth H. Kim, Rebecca C. Arend. Analysis of Clinical Cancer Gene Panels by Next Generation Sequencing in Tumor and Circulating Cell-Free DNA Samples in Recurrent Ovarian Cancer Patients [abstract]. In: Proceedings of the AACR Precision Medicine Series: Opportunities and Challenges of Exploiting Synthetic Lethality in Cancer; Jan 4-7, 2017; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2017;16(10 Suppl):Abstract nr B29.