MicroRNA-143 Targets ATG2B to Inhibit Autophagy and Increase Inflammatory Responses in Crohn's Disease
MicroRNA-143 Targets ATG2B to Inhibit Autophagy and Increase Inflammatory Responses in Crohn's Disease
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DOI：
10.1093/ibd/izx075
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发表时间：
2018-04-01
影响因子：
4.9
通讯作者：
He, Xiao-sheng
中科院分区：
文献类型：
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作者：
Lin, Xu-tao;Zheng, Xiao-bin;He, Xiao-sheng
背景：功能失调的自噬被认为是许多慢性炎症性疾病，包括克罗恩病（CD）的一个促成因素。方法：采用多聚腺苷酸化位点选择性测序（SAPAS）方法，对3例CD患者手术标本（包括炎性组织和相应的非炎性组织）的总mRNA序列3'端进行比较。使用免疫印迹和定量逆转录聚合酶链反应比较了炎症组织和非炎症组织中自噬相关2B（ATG 2B）、LC 3和miR-143的水平。使用荧光素酶测定来验证miR-143与ATG 2B之间的相互作用。通过LC 3的免疫印迹分析和透射电子显微镜测量自噬。结果：ATG 2B的串联重复序列3 '-UTR在炎症组织中比相应的非炎症组织中更长，并且含有miR-143靶位点。在炎症组织中，miR-143表达升高，而ATG 2B和LC 3-II表达下调。通过3 '-UTR双荧光素酶报告基因测定验证miR-143和ATG 2B之间的直接相互作用。结论：miR-143可能通过调控ATG 2B和自噬而诱导肠道炎症反应，提示miR-143可能在CD的发生发展中起重要作用。因此，miR-143可能是CD患者基因治疗的一个有前途的新靶点。
Background: Dysfunctional autophagy is recognized as a contributing factor in many chronic inflammatory diseases, including Crohn's disease (CD). Genetic analyses have found that microRNA (miRNA) levels are altered in the intestinal tissues of CD patients.Methods: The Sequencing Alternative Poly-Adenylation Sites (SAPAS) method was used to compare the 3' end of the total mRNA sequence of 3 surgical specimens of CD patients (including inflamed tissues and corresponding noninflamed tissues in each case). The levels of autophagy-related 2B (ATG2B), LC3, and miR-143 were compared between inflamed tissues and noninflamed tissues using immunoblot and quantitative reverse transcription polymerase chain reaction. Luciferase assays were used to verify the interactions between miR-143 and ATG2B. Autophagy was measured by immunoblot analyses of LC3 and transmission electron microscopy. Inflammatory cytokines and I kappa B alpha were analyzed to evaluate the effect of miR-143 on inflammatory response.Results: The tandem repeat 3'-UTR of ATG2B was longer in inflamed tissues than in corresponding noninflamed tissues and contained an miR-143 target site. miR-143 expression was elevated, whereas ATG2B and LC3-II were downregulated in inflamed tissues. The direct interaction between miR-143 and ATG2B was verified by a 3'-UTR dual-luciferase reporter assay. Constitutive expression of miR-143 or depletion of ATG2B in cultured intestinal epithelial cells inhibited autophagy, reduced I kappa B alpha levels, and increased inflammatory responses.Conclusions: miR-143 may induce bowel inflammation by regulating ATG2B and autophagy, suggesting that miR-143 might play a critical role in the development of CD. Therefore, miR-143 could be a promising novel target for gene therapy in CD patients.