Although intravesical BCG immunotherapy has been the gold standard for non-surgical management of non-muscle-invasive bladder cancer (BC), a considerable number of patients exhibit resistance to the adjuvant treatment with unexplained mechanisms. This study aimed to investigate whether and how androgen receptor (AR) signals modulate BCG cytotoxicity in BC. AR knockdown or overexpression in BC lines resulted in induction or reduction, respectively, in intracellular BCG quantity and its cytotoxic activity. Microarray screening identified Rab27b, a small GTPase known to mediate bacterial exocytosis, which was up-regulated in BCG-resistant cells and down-regulated in AR-shRNA cells. Knockdown of Rab27b or its effector SYTL3, or overexpression of Rab27b also induced or reduced, respectively, BCG quantity and cytotoxicity. Additionally, treatment with GW4869, which was previously shown to inhibit Rab27b-dependent secretion, induced them and reduced Rab27b expression in BC cells. Meanwhile, AR expression was up-regulated in BCG-resistant lines, compared with respective controls. In a mouse orthotopic xenograft model, Rab27b/SYTL3 knockdown or GW4869 treatment enhanced the amount of BCG within tumors and its suppressive effect on tumor growth. Moreover, in non-muscle-invasive BC specimens from patients subsequently undergoing BCG therapy, positivity of AR/Rab27b expression was associated with significantly higher risks of tumor recurrence. AR activation thus correlates with resistance to BCG treatment, presumably via up-regulating Rab27b expression. Mechanistically, it is suggested that BCG elimination from urothelial cells is induced by Rab27b/SYTL3-mediated exocytosis. Accordingly, Rab27b inactivation, potentially via anti-androgenic drugs, and/or exocytosis inhibition are anticipated to sensitize the efficacy of BCG therapy, especially in patients with BCG-refractory AR/Rab27b-positive BC.
Molecular cancer therapeutics. 2020 Jul 31 [Epub ahead of print]
Taichi Mizushima, Guiyang Jiang, Takashi Kawahara, Peng Li, Bin Han, Satoshi Inoue, Hiroki Ide, Ikuma Kato, Mehrsa Jalalizadeh, Etsuko Miyagi, Mitsunori Fukuda, Leonardo O Reis, Hiroshi Miyamoto
Department of Pathology & Laboratory Medicine, University of Rochester Medical Center., Pathology, College of Basic Medical Sciences and First Affiliated Hospital, China Medical University., Johns Hopkins University School of Medicine., Molecular Biology, University of Occupational and Environmental Health., University of Rochester Medical Center., Department of Urology, Keio University School of Medicine., Molecular Pathology, Yokohama City University Graduate School of Medicine., Obstetrics and Gynecology, Yokohama City University., Department of Integrative Life Sciences, Tohoku University Graduate School of Life Sciences., Johns Hopkins School of Medicine., Department of Pathology & Laboratory Medicine, University of Rochester Medical Center .