Mp65-12 Foxf1: A Novel Regulator of Metastasis in Urothelial Carcinoma of the Bladder

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You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology III (MP65)1 May 2024MP65-12 FOXF1: A NOVEL REGULATOR OF METASTASIS IN UROTHELIAL CARCINOMA OF THE BLADDER Côme Tholomier, Jan K. Rudzsinki, Alexis Steinmetz, Alberto Martini, Akshay Sood, David J. McConkey, and Colin P. N. Dinney Côme TholomierCôme Tholomier , Jan K. RudzsinkiJan K. Rudzsinki , Alexis SteinmetzAlexis Steinmetz , Alberto MartiniAlberto Martini , Akshay SoodAkshay Sood , David J. McConkeyDavid J. McConkey , and Colin P. N. DinneyColin P. N. Dinney View All Author Informationhttps://doi.org/10.1097/01.JU.0001008756.24343.22.12AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: FOXF1 is a transcription factor that may act as a tumor suppressor gene by interaction with p53. We previously identified FOXF1 to be highly differentially expressed in matched bladder cancer (BCA) primary tumors and nodal metastases. We investigated the mechanistic role of FOXF1 in BCA metastasis. METHODS: Association between FOXF1 expression and survival after cystectomy was assessed in 2 independent clinical cohorts using adjusted Cox models. Quantitative PCR (qPCR) and western blot analysis evaluated FOXF1 expression in parental BCA cell lines. FOXF1 knockout (KO) was achieved with CRISPR-Cas 9. Using the high FOXF1 expressing RT112 cell line, FOXF1 KO was confirmed using western blot and qPCR. The Boyden chamber and scratch assays assessed in-vitro invasion and migration in KO, wild-type (WT), and scramble (SCr) RT112 clones. Orthotopic xenograft models for WT, SCr, and KO were used for in-vivo experiments. End stage tumor weights were compared. DNA qPCR for human DNA assessed BCA metastasis in lymph nodes (LN), liver, and lungs. Kaplan-Meier curves were used for survival analyses. RNA sequencing and ingenuity pathway analysis (IPA) were conducted to understand the mechanistic impact of FOXF1. RESULTS: FOXF1 expression was 3.6-fold lower in LN metastases than paired primary tumors (n=33, p<0.001). High FOXF1 expression was associated with improved cancer specific survival (HR=0.35, p=0.046) and overall survival (HR=0.45, p=0.006) in our clinical cohorts (MDA Discovery n=73 and GSE13507 n=55). In-vitro, compared to WT or SCr, KO clones showed significant increase in invasion (p≤0.05) and migration (p≤0.05). In-vivo, FOXF1 KO mice had the largest primary tumors and lowest survival. Human DNA confirmed metastatic disease with high reliability. While most mice developed lung metastases, more KO mice had LN or liver metastases compared to WT or SCr (33% and 33% vs. 18% and 0%, respectively). IPA notably showed consistent positive enrichment in integrin signaling pathways across multiple FOXF1 KO clones. Key genes related to metastatic potential in human cancers were significantly upregulated (e.g., PIC3C2G, ITGB8, ITGA1) or downregulated (AKAT3, CAPN5, CAPN8). CONCLUSIONS: FOXF1 is differentially expressed between primary and metastatic BCA lesions and high FOXF1 expression led to improved outcomes in clinical cohorts. FOXF1 appears to regulate cellular invasion and migration through modulation of the integrin signaling pathway, and FOXF1 KO clones led to more metastasis in mice. These data confirmed our hypothesis that FOXF1 is a potential tumor suppressor gene for BCA metastasis. Source of Funding: Research is supported in part by A.I. Virtanen Institute for Molecular Sciences (Kuopio, Finland) and MD Anderson CCSG program (P30 016672) © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e1081 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Côme Tholomier More articles by this author Jan K. Rudzsinki More articles by this author Alexis Steinmetz More articles by this author Alberto Martini More articles by this author Akshay Sood More articles by this author David J. McConkey More articles by this author Colin P. N. Dinney More articles by this author Expand All Advertisement PDF downloadLoading ...