Targeting of MyD88 Homodimerization by Novel Synthetic Inhibitor TJ-M2010-5 in Preventing Colitis-Associated Colorectal Cancer

JNCI: Journal of the National Cancer Institute, Apr 2016

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Targeting of MyD88 Homodimerization by Novel Synthetic Inhibitor TJ-M2010-5 in Preventing Colitis-Associated Colorectal Cancer

JNCI J Natl Cancer Inst ( Targeting of M yD88 Ho m odim erization by Novel Synthetic Inhibitor TJ-M 2010-5 in Preventing Colitis-Associated Colorectal Cancer Lin Xie 0 1 2 3 4 Feng-Chao Jiang 0 1 2 3 4 Li-Min Zhang 0 1 2 3 4 Wen-Tao He 0 1 2 3 4 Jian-Hua Liu 0 1 2 3 4 Ming-Qiang Li 0 1 2 3 4 Xue Zhang 0 1 2 3 4 Shuai Xing 0 1 2 3 4 Hui Guo 0 1 2 3 4 Ping Zhou 0 1 2 3 4 Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (WTH). 0 PZ); Academy of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology , Wuhan , China ( FCJ); Department of endocrinology , Tongji 1 of Organ Transplantation, Ministry of Health, and Key Laboratory of Organ Transplantation, Ministry of Education , Wuhan, China (LX, LMZ, JHL, MQL, XZ, SX, HG 2 Affiliations of authors: Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology , Key Laboratory 3 The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions , please 4 Wuhan 430030 , China for antitumor therapy. Background: The TLR/MyD88 signaling pathway is an important driver of inflammation and cancer and is a possible target Methods: We generated a MyD88 inhibitor (TJ-M2010-5), which was designed to bind to the TIR domain of MyD88 to interfere with its homodimerization, and the TLR/MyD88 signal pathway. We utilized a mouse model of azoxymethane/ dextran sodium sulfate (AOM/DSS)-induced colitis-associated cancer (CAC) in combination with TJ-M2010-5 administration to investigate the anti-inflammation-related cancer effect of MyD88 inhibitor in vivo. Data were analyzed with one-way and repeated measures analysis of variance. Differences in survival between groups were compared using the log rank test. All statistical tests were two-sided. Results: TJ-M2010-5 inhibited MyD88 homodimerization in transfected HEK293 cells in a concentration-dependent manner and suppressed MyD88 signaling in LPS-responsive RAW 264.7 cells in vitro. In a 10-week CAC mouse model (n = 30 per group), TJ-M2010-5 treatment statistically significantly reduced AOM/DSS-induced colitis and completely prevented CAC development with less related body mass loss, resulted in 0% mortality of treated mice (compared with 53% mortality of control mice), decreased cell proliferation, and increased apoptosis in colon tissue. TJ-M2010-5 treatment also inhibited Novel - M TJ-M production of inflammatory cytokines and chemokines (TNαF,- IL-6,G-CSF, MIP-1β, TGF-β1, IL-11, IL-17A, IL-22 and IL-23) and infiltration of immune cells (macrophages, dendritic cells, neutropihls an d+4CTDcells) in colon tissues of mice. Conclusions: Our findings suggest that TLR/MyD88 signaling may be a therapeutic target for CAC intervention and MyD88 inhibitors may be a promising therapeutic modality for treating patients with colitis or CAC. Inflammatory microenvironment around intestinal epithelial molecule myeloid differentiation factor 88 (MyD88) signaling cells in inflammatory bowel disease plays a role in carcinog-en pathway ( 3,4 ). MyD88 is one of the important adapter molecules esis and inflammation-related epithelial cell injury and repair, of TLRs for the activation of downstream NκFB- ( 5 ) and mitogenwhich contribute to the development of colitis-associated c-an activated protein kinase (MAPK)6() pathways. Aberrant activ-a cer (CAC) ( 1,2 ). Microbial infection or tissue injury can trigger tion of TLR/MyD88 signaling is crucial for inflammation and inflammation by activating the toll-like receptor (TLR)/adaptoris associated with the development of inflammation-related tumorigenesis (18). death domain (DD) and a C-terminal TIR domain separated by a short linker region1( 9 ). MyD88 can be recruited to TLR complexes as a dimer, which is stabilized by homophilic intera-c cancers ( 7,8 ) in the skin ( 9,10 ), ovary (11), liver ( 12,13 ), pancreas ( 14 ), and colon ( 15–17 ). TLR/MyD88 signaling has been thought to vehicle. HEK293 cells were collected 48 hours after transfection and Tumor Formation collected for immunoprecipitation (IP) using protein A/G a-ga The entire colon of each mouse was opened longitudinally, and rose beads (Santa Cruz Biotech). Then, immunocomplexes ca-p the number of tumors in each individual colon was counted. tured by protein beads were eluted in SDS-PAGE sample buffer With a sliding caliper, the average diameter of each tumor n-od for western blot (WB) analysis. ule was measured. Paraffin-embedded colon tissue sections (4  µm) were stained by hematoxylin and eosin (H&E), and the Kit (BD Pharmingen) according to the manufacturer’s suggested pathological severity of colitis was evaluated by a six-grade c-las protocols. Apoptosis was detected using an In Situ Cell Death sification system S(upplementary Method,savailable online). Detection Fluoresce in Kit (Beyotime Biotech) with DAPI. Immunohistochemistry, Bromodeoxyuridine, and TUNEL (...truncated)


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Xie, Lin, Jiang, Feng-Chao, Zhang, Li-Min, He, Wen-Tao, Liu, Jian-Hua, Li, Ming-Qiang, Zhang, Xue, Xing, Shuai, Guo, Hui, Zhou, Ping. Targeting of MyD88 Homodimerization by Novel Synthetic Inhibitor TJ-M2010-5 in Preventing Colitis-Associated Colorectal Cancer, JNCI: Journal of the National Cancer Institute, 2016, Volume 108, Issue 4, DOI: 10.1093/jnci/djv364