PD42-02 THE EFFECTS OF RECOMBINANT BACILLUS CALMETTE-GUERIN RESISTANT TO ANTIMICROBIAL PEPTIDES ON ORTHOTOPIC BLADDER CANCER MOUSE MODEL

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology II (PD42)1 Sep 2021PD42-02 THE EFFECTS OF RECOMBINANT BACILLUS CALMETTE-GUERIN RESISTANT TO ANTIMICROBIAL PEPTIDES ON ORTHOTOPIC BLADDER CANCER MOUSE MODEL Jae Hun Shim, Joonhee Gook, Se Young Choi, Byung Hoon Chi, Jin Wook Kim, Tae-Hyoung Kim, Soon Chul Myung, and In Ho Chang Jae Hun ShimJae Hun Shim More articles by this author , Joonhee GookJoonhee Gook More articles by this author , Se Young ChoiSe Young Choi More articles by this author , Byung Hoon ChiByung Hoon Chi More articles by this author , Jin Wook KimJin Wook Kim More articles by this author , Tae-Hyoung KimTae-Hyoung Kim More articles by this author , Soon Chul MyungSoon Chul Myung More articles by this author , and In Ho ChangIn Ho Chang More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002056.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Although Mycobacterium bovis Bacillus Calmette-Guerin (BCG) is the most widely used bladder cancer immunotherapy, innate immune responses involving antimicrobial peptides (AMPs) cause BCG failure. Here, we developed genetically modified recombinant BCG (rBCG) strains which escape AMPs and evaluate the efficacy and effects of rBCG. METHODS: We constructed rBCG strains expressing Streptococcal inhibitor of complement (Sic), which confers resistance to human α-defensin-1 and cathelicidin, and d-alanyl carrier protein ligase (dltA), which confers resistance to cationic AMPs. Sic and dltA were separately cloned into the pMV306 plasmid and introduced into BCG via electroporation. The efficacy of the Sic and dltA gene electroporation into BCG was evaluated by quantitative real-time polymerase chane reaction (qRT-PCR). The internalization rates and anti-cancer effects of the rBCG strains containing Sic (rBCG-Sic) and dltA (rBCG-dltA) was evaluated by the orthotopic bladder cancer mouse model. RESULTS: The cycle quantification (Cq) values of rBCG-Sic (y=-4.8823x+13.645, R2=0.9996) and rBCG-dltA (y=-5.438x+11.641, R2=0.9995) were inverse correlations to the amount of Sic and dltA genes dose dependently. The mean introduction proportions of Sic and dltA genes into BCG by electroporation were 22.2%, 27.5% and showed constant efficacy. In the orthotopic bladder cancer mouse model, the relative internalization number of rBCG-Sic, and rBCG-dltA into bladder cell in mouse bladder were higher than that of BCG and the tumor volume at rBCG-Sic were lower than at BCG and rBCG-dltA at 11, 14 m and 18 days. CONCLUSIONS: Our results showed that constructed rBCG-Sic and rBCG-dltA by electroporation and the rBCG-Sic and rBCG-dltA can effectively escape BCG-stimulated AMPs, and significantly improved immunotherapeutic tools to treat bladder cancer in orthotopic bladder cancer mouse model. Source of Funding: None © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e724-e724 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jae Hun Shim More articles by this author Joonhee Gook More articles by this author Se Young Choi More articles by this author Byung Hoon Chi More articles by this author Jin Wook Kim More articles by this author Tae-Hyoung Kim More articles by this author Soon Chul Myung More articles by this author In Ho Chang More articles by this author Expand All Advertisement Loading ...