VCOM Research Day Program Book 2023

Medical Student Research Biomedical

19 Variant FETUIN-B Expression Mediated Changes on PI3K/Akt/mTOR Activity in Ovarian Adenocarcinoma

Ramu Anandakrishnan; Catherine Bassett; Shourik Dutta; Diego Flores; Trent Kite; Ellis Linder-Elkins; Yaswitha Mikkilineni; Rajvi Patel; Nina Pillai; Veda Prasad; Addison Shenk; David Valenti; Tarana Zaman Corresponding author: dflores@vcom.edu

Edward Via College of Osteopathic Medicine-Virginia Campus

In 2022, an estimated 19,880 people will be diagnosed with ovarian cancer in the United States and approximately 64% of them will die from the disease. Ovarian cancer is the fifth leading cause of death associated with cancer with a 5-year survival rate of 49.4% across all stages. The proliferation of many adenocarcinomas is linked to alterations to the PI3K/ Akt pathway. The PI3K/Akt pathway is involved in cell growth, cell survival, and cell cycle regulation, so mutations of certain genes in this pathway have the propensity to induce or suppress tumorigenesis. One of the genes associated with altered expression of this pathway, and thus, affect the growth of an adenocarcinoma, is FETUB . FETUB encodes Fetuin B glycoprotein—a member of the cysteine protease inhibitor family—which is responsible for diverse functions such as osteogenesis, insulin regulation, and tumor suppressor activity. Prior studies investigating FETUB mutations have shown an association with tumor progression in acute myeloid leukemia and

other carcinomas; however, this gene has not been explored in relation to ovarian adenocarcinomas. We have identified FETUB expression in OVCAR3 samples through immunofluorescence assay. OVCAR3, an epithelial cell line isolated from adenocarcinoma of mammalian ovaries, is widely used to recapitulate ovarian carcinomas in-vitro. Our approach involved the generation of both wild-type (WT) and knockout (KO) FETUB OVCAR3 clones using CRISPR/Cas9 systems, with enrichment of highly efficient KO clones by limiting dilution method. PI3K activity was assayed via immunofluorescence. PI3K activity in FETUB KO clones was compared to that of FETUB WT clones. Significant differences between KO and WT in these assays will provide evidence for a potential role of FETUB in tumorigenesis, leading to prospective investigations to further delineate this role, such as with multidrug resistance. One of the reasons behind the high mortality rate in ovarian cancer is drug

resistance. OVCAR3 is known to have resistance to chemotherapies such as Cisplatin. A future proposal extending from our initial investigation will investigate a dose-dependent effect of cisplatin with respect to both WT and KO OVCAR3 cell line viability.

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