Abstracts from the New England Section of the AUA 2021

© The Canadian Journal of Urology TM : International Supplement, October 2021 Concurrent Poster Session II Extracellular miRNA as a Marker of Invasive Urothelial Cancer Esther L. Finney, MD , Thomas Kalantzakos, BS, Travis Sullivan, MS, Kimberly Rieger-Christ, PhD Lahey Hospital and Medical Center, Burlington, MA, USA Introduction: Determination of muscle invasion is critical in the staging, prognosis, and treatment of urothelial carcinoma (UC). MicroRNA (miRNA) have been shown to be dysregulated in both non- and muscle-invasive UC and are thought to serve an important role in tumor progression. Extracellular vesicle (EV) miRNA can be easily isolated from body fluids, such as urine. They are a novel target for development of diagnostic biomarkers that are non-invasive and overcome the constraints of tissue biopsies. We sought to identify differential expression of EV miRNA between noninvasive and invasive UC and investigate the effect of identified miRNA on tumor pathogenesis in UC cell lines. Materials & Methods: EV miRNA was isolated from the urine of patients with biopsy-proven non-invasive (n= 14) or invasive bladder UC (n=15). Urine was collected pre-operatively and specimens were grouped according to invasion at the time of transurethral resection. TwelvemiRNAwere assayed via PCR and those with increased expression in invasive specimens were further examined. miRNA from four UC cell lines (J82, UMUC, RT112, and CUBIII) and EV miRNA from cell-free culture media from those lines were isolated and PCR performed for in-vitro validation. Two UC cell lines, UMUC (invasive) and CUBIII (non-invasive), were then transfectedwith pre-miRNA of the previously identified miRNA. Cell proliferation was evaluated at 24 and 48 hrs in the transfected cell lines byMTT assays. Potential miRNAtumor suppressor protein targets were identified based on predicted interactions, and their levels were determined via Western blots. Results: Three miRNA (miR-222-3p, miR-320b, and miR-423-5p) had increased expression in the urine EVs of patients with invasive UC (p<0.05). These miRNA were also upregulated in the EVs of cultured cells with an invasive phenotype. Transfection of miR-320b andmiR-222-3pwas successful into both UC cell lines and increased proliferation in both the noninvasive and invasive cell lines (p<0.05). Conclusions: Urine EV miR-320b and miR-222-3p are upregulated in invasive UC and may have a role in tumor progression. They are promising biomarkers that could improve diagnostic precision as well as serve as potential therapeutic targets. Obesity-associated Proinflammatory Cytokines Regulate Prostate Cancer Cell Proliferation & Migration Xiaobo Wu, MD, Christina Sharkey, MS, Zoongwei Wang, PhD, Aria F. Olumi, MD Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA Introduction: Previous studies revealed that obesity is significantly associated with a higher incidence of prostate cancer (PCa). Meanwhile, obesity- associated inflammation plays an important role in tumorigenesis. Here we wished to identify the downstream signaling pathways of obesity-associated proinflammatory cytokines IL-6 and TNF- α effect on prostate cancer cell proliferation and migration. Materials & Methods: Adipocytes and THP-1 macrophages were differentiated and treated with myristic acid (MA) to induce inflammatory cytokines, and conditioned media (CM) were collected to treat prostate epithelial cancer, androgen-dependent LNCaP and androgen-independent PC-3 cell lines. Cells were also treated with interleukin-6 (IL-6) 10ng/ml, tumor necrosis factor-alpha (TNF- α ) 10ng/ml, oncometabolite R-2- hydroxyglutarate 50µM (R-2-HG) and Dutasteride 10µM (SRD5A1 inhibitor). Prostate cancer cell migration and proliferation were assessed by scratch assay (0h, 24h, 48h) and MTT assay (0h, 24h, 48h, 72h) and analyzed by 2-way ANOVA. Enzymes regulating steroid hormone metabolism SRD5A1, AKR1C1, AKR1C3, HSD3B2, CYP17A1 and UGT2B15 were determined by immunocytochemistry and qPCR. The protein level of SRD5A1was measured by western blot. Results: The proliferation of LNCaP and PC-3 cells was significantly stimulated by adding of CM for 24h, 48h, and 72h. The migration of PC-3 cells was also promoted by CM. Addition of testosterone did not change the effect of CM. Administration of IL-6, TNF- α and R-2-HG significantly increased the proliferation and migration of LNCaP and PC-3 cells. IL-6 also increased the expression of SRD5A1. The expression of AKR1C1 andHSD3B2 was downregulated, and UGT2B15 was upregulated by TNF- α . SRD5A1 was upregulated either by R-2-HG alone or combining with IL-6 and TNF- α , but downregulated by Dutasteride alone. Conclusions: Our study suggests that obesity-associated proinflammatory cytokines, IL-6 and TNF- α , affect prostate cancer cells’ proliferation and migration in an androgen-independent manner. Synergistic targeting oncometabolite R-2-HG and proinflammatory cytokines may improve the management of androgen-independent prostate cancer. P28 P27 50