靶向小鼠肝 miR-221/222 治疗非酒精性脂肪性肝炎。
Targeting hepatic miR-221/222 for therapeutic intervention of nonalcoholic steatohepatitis in mice.
机构信息
Shanghai Clinical Center for Endocrine and Metabolic Diseases, Shanghai Key Laboratory for Endocrine Tumors, Rui-Jin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai 200025, China.
Shanghai Clinical Center for Endocrine and Metabolic Diseases, Shanghai Key Laboratory for Endocrine Tumors, Rui-Jin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai 200025, China; Laboratory of Endocrinology and Metabolism, Institute of Health Sciences, Shanghai Institutes for Biological Sciences (SIBS), Chinese Academy of Sciences (CAS), Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai 200025, China.
出版信息
EBioMedicine. 2018 Nov;37:307-321. doi: 10.1016/j.ebiom.2018.09.051. Epub 2018 Oct 10.
BACKGROUND
Effective targeting therapies for common chronic liver disease nonalcoholic steatohepatitis (NASH) are in urgent need. MicroRNA-targeted therapeutics would be potentially an effective treatment strategy of hepatic diseases. Here we investigated the functional role of miR-221/222 and the therapeutic effects of antimiRs-221/222 in NASH mouse models.
METHODS
We generated the miR-221/222 mice on a C57BL/6 J background and the hepatic miR-221/222 knockout (miR-221/222-LKO) mice. The mice were challenged with the methionine and choline deficient diet (MCDD) or chronic carbon tetrachloride (CCl) treatment to generate experimental steatohepatitis models. Adenovirus-mediated re-expression of miR-221/222 was performed on the MCDD-fed miR-221/222-LKO mice. The MCDD and control diet-fed mice were treated with locked nucleic acid (LNA)-based antimiRs of miR-221/222 to evaluate the therapeutic effects. Histological analysis, RNA-seq, quantitative PCR and Western blot of liver tissues were carried out to study the hepatic lipid accumulation, inflammation and collagen deposition in mouse models.
FINDINGS
Hepatic deletion of miR-221/222 resulted in significant reduction of liver fibrosis, lipid deposition and inflammatory infiltration in the MCDD-fed and CCl4-treated mouse models. The hepatic steatosis and fibrosis were dramatically aggravated by miR-221/222 re-expression in MCDD-fed miR-221/222-LKO mice. AntimiRs of miR-221/222 could effectively reduce the MCDD-mediated hepatic steatosis and fibrosis. Systematically mechanistic study revealed that hepatic miR-221/222 controlled the expression of target gene Timp3 and promoted the progression of NASH.
INTERPRETATION
Our findings demonstrate that miR-221/222 are crucial for the regulation of lipid metabolism, inflammation and fibrosis in the liver. LNA-antimiRs targeted miR-221/222 could reduce steatohepatitis with prominent antifibrotic effect in NASH mice. FUND: This work is supported by the Natural Science Foundation of China (81530020, 81390352 to Dr. Ning and 81522032 to Dr. Cao and 81670793 to Dr. Jiang); National Key Research and Development Program (No. 2016YFC0905001 and 2017YFC0909703 to Dr. Cao); the Shanghai Rising-Star Program (15QA1402900 to Dr. Cao); Shanghai Municipal Education Commission-Gaofeng Clinical Medicine Grant (20171905 to Dr. Jiang).
背景
目前急需针对常见慢性肝病非酒精性脂肪性肝炎（NASH）的有效靶向治疗方法。miRNA 靶向治疗可能是肝脏疾病的一种有效治疗策略。在这里，我们研究了 miR-221/222 的功能作用以及抗 miR-221/222 在 NASH 小鼠模型中的治疗效果。
方法
我们在 C57BL/6J 背景下生成了 miR-221/222 小鼠和肝 miR-221/222 敲除（miR-221/222-LKO）小鼠。用蛋氨酸和胆碱缺乏饮食（MCDD）或慢性四氯化碳（CCl）处理使这些小鼠产生实验性脂肪性肝炎模型。在 MCDD 喂养的 miR-221/222-LKO 小鼠上进行 miR-221/222 的腺病毒介导再表达。用锁定核酸（LNA）为基础的 miR-221/222 抗 miR 对 MCDD 喂养的对照组和 MCDD 喂养的小鼠进行治疗，以评估治疗效果。对肝组织进行组织学分析、RNA-seq、定量 PCR 和 Western blot，以研究小鼠模型中的肝脂质蓄积、炎症和胶原沉积。
结果
MCDD 喂养和 CCl4 处理的小鼠模型中，肝 miR-221/222 的缺失导致肝纤维化、脂质蓄积和炎症浸润明显减少。在 MCDD 喂养的 miR-221/222-LKO 小鼠中，miR-221/222 的再表达导致肝脂肪变性和纤维化显著加重。miR-221/222 的抗 miR 可有效降低 MCDD 介导的肝脂肪变性和纤维化。系统的机制研究表明，肝 miR-221/222 控制靶基因 Timp3 的表达，并促进 NASH 的进展。
结论
我们的研究结果表明，miR-221/222 对肝脏脂质代谢、炎症和纤维化的调节至关重要。针对 miR-221/222 的 LNA 抗 miR 可减少 NASH 小鼠的脂肪性肝炎，具有显著的抗纤维化作用。
资助
本研究得到了中国自然科学基金（81530020、81390352 给宁博士、8122032 给曹博士和 81670793 给蒋博士）、国家重点研发计划（No. 2016YFC0905001 和 2017YFC0909703 给曹博士）、上海市启明星计划（15QA1402900 给曹博士）和上海市教委高峰学科建设计划（20171905 给蒋博士）的支持。
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