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miRNA Regulation of the Mitochondrial Genome

线粒体基因组的 miRNA 调控

基本信息

批准号：
9130443
负责人：
John M Hollander
金额：
$ 40.97万
依托单位：
WEST VIRGINIA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-08 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9130443
关键词：
Address Animals Automobile Driving Bioenergetics Cardiac Cardiac Myocytes Cell Line Chronic Code Complex Data Development Diabetes Mellitus Diabetic mouse Experimental Models Functional disorder Health Health Care Costs Heart Heart Atrium Heart failure Human Immunoprecipitation In Vitro Incidence Intervention Knowledge Life Messenger RNA MicroRNAs Mission Mitochondria Mitochondrial RNA Modeling Molecular Motor Non-Insulin-Dependent Diabetes Mellitus Outcome Pathogenesis Patients Polyribonucleotide Nucleotidyltransferase Preventive Intervention Protein Import Proteins Protons Public Health Regulation Regulatory Pathway Research Testing Therapeutic Therapeutic Intervention Translating Translations Treatment Efficacy Untranslated RNA Work burden of illness crosslink db/db mouse deep sequencing diabetic diabetic patient in vivo inhibitor/antagonist innovation insight interest mitochondrial dysfunction mitochondrial genome mouse model non-diabetic outcome forecast overexpression preclinical evaluation prophylactic protein expression ratiometric research study response treatment strategy

项目摘要

DESCRIPTION (provided by applicant): Type 2 diabetes mellitus incidence has increased dramatically. Among the life threatening complications is heart failure, which is preceded by bioenergetic dysfunction. Using mouse (db/db) and human (patient) type 2 diabetic models; we observed pronounced mitochondrial dysfunction culminating in a decreased ability to generate ATP for cardiac contraction. MicroRNAs (miRs) are non-coding RNAs that regulate translation. Using cross-linking immunoprecipitation and deep sequencing, we made the exciting observation, in both db/db and type 2 diabetic patients that miRs translocate into and out of cardiac mitochondria. Of particular interest was an increased miR- 378 presences in a functional regulatory context with mitochondrial genome-encoded ATP6 mRNA which codes for a subunit of the F0 proton motor that is part of the ATP synthase complex. Decreased ATP synthase functionality promotes bioenergetic deficit in the heart, promoting heart failure. Nevertheless, it is currently unclear whether miR-378 blockade can reduce mitochondrial dysfunction associated with the type 2 diabetic heart by direct interaction with the mitochondrial genome. Further, the mechanisms responsible for the dynamic flux of miRs into the mitochondrion are undefined. One potential mechanism involves the participation of the mitochondrial RNA import protein polynucleotide phosphorylase (PNPase) which we have observed to be increased in mitochondria from db/db mice and type 2 diabetic patients. The studies being proposed address these gaps in knowledge and integrate in vitro cellular approaches with animal and human experimental models in an effort to translate the findings to the type 2 diabetic patient. The central hypothesis of this application is that inhibition of miR-378 disrupts its ability to down-regulate ATP6 in the mitochondrion, preserving ATP synthase function as well as ATP levels, and limiting cardiac contractile dysfunction in the type 2 diabetic hearts. Further, miR-378 flux into the mitochondrion can be modulated by PNPase manipulation. The objectives of this application are: (1) determine the impact in vivo of a prophylactically delivered miR-378 inhibitor in a type 2 diabetic mouse model for restoring mitochondrial ATP6 protein expression and ATP generating capacity in the heart; (2) evaluate the therapeutic efficacy of a miR-378 inhibitor delivered to isolated human cardiomyocytes from type 2 diabetic patients; and (3) assess the contribution of PNPase to the mechanisms driving miR-378 flux into the mitochondrion. To test this hypothesis, an innovative approach has been proposed which employs antagomir intervention to manipulate mitochondrial genome-encoded proteins in an effort to mitigate diabetes- associated contractile dysfunction. The combination of work proposed is significant because it will provide insight into the mechanisms regulating miR distribution in the mitochondrion while providing translational insight into the therapeutic potential of miR-378 inhibition as a treatment strategy. Our approach merges mechanistic examination of a previously unexplored regulatory pathway contributing to mitochondrial dysfunction in the diabetic heart with preclinical evaluation of key molecular constituents participating in the axis.

描述（由申请人提供）：2型糖尿病发病率急剧增加。危及生命的并发症之一是心力衰竭，其之前是生物能功能障碍。使用小鼠（db/db）和人类（患者）2型糖尿病模型;我们观察到明显的线粒体功能障碍，最终导致产生心脏收缩ATP的能力降低。microRNA（miRs）是调节翻译的非编码RNA。使用交联免疫沉淀和深度测序，我们在db/db和2型糖尿病患者中进行了令人兴奋的观察，miR易位进入和离开心脏线粒体。特别令人感兴趣的是，在线粒体基因组编码的ATP 6 mRNA的功能调控背景下，miR- 378的存在增加，ATP 6 mRNA编码作为ATP合酶复合物一部分的F0质子马达亚基。ATP合酶功能降低促进心脏中的生物能缺陷，从而促进心力衰竭。但它目前尚不清楚miR-378阻断是否可以通过与线粒体基因组的直接相互作用来减少与2型糖尿病心脏相关的线粒体功能障碍。此外，负责miR动态通量进入微扰子的机制尚未确定。一个潜在的机制涉及线粒体RNA输入蛋白多核苷酸磷酸化酶（PNIPs）的参与，我们已经观察到它在db/db小鼠和2型糖尿病患者的线粒体中增加。正在提出的研究解决了这些知识上的差距，并将体外细胞方法与动物和人类实验模型相结合，以努力将研究结果转化为2型糖尿病患者。本申请的中心假设是miR-378的抑制破坏了其在2型糖尿病心脏中下调ATP 6的能力，从而保留ATP合酶功能以及ATP水平，并限制心脏收缩功能障碍。此外，miR-378流入到PNDs中可以通过PNDs操作来调节。本申请的目的是：（1）确定经药理学递送的miR-378抑制剂的体内影响在2型糖尿病小鼠模型中用于恢复心脏中线粒体ATP 6蛋白表达和ATP产生能力;（2）评估递送至来自2型糖尿病患者的分离的人心肌细胞的miR-378抑制剂的治疗功效;和（3）评估PNTR对驱动miR-378通量进入心肌细胞的机制的贡献。为了检验这一假设，已经提出了一种创新方法，其采用Eschomir干预来操纵线粒体基因组编码的蛋白质，以减轻糖尿病相关的收缩功能障碍。所提出的工作组合是重要的，因为它将提供对调节miR在转录子中分布的机制的洞察，同时提供对miR-378抑制作为治疗策略的治疗潜力的翻译洞察。我们的方法合并了一个以前未探索的调节途径，有助于线粒体功能障碍的糖尿病心脏与参与轴的关键分子成分的临床前评价机制检查。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Targeting Diabetic Cardiomyopathy: LncRNA Kcnq1ot1 Rescues Mitochondrial ATP Synthase via Sponging of MiR-378a-5p.

靶向糖尿病心肌病：LncRNA Kcnq1ot1 通过 MiR-378a-5p 海绵作用拯救线粒体 ATP 合酶。

DOI：
发表时间：
2022
期刊：
FASEB journal : official publication of the Federation of American Societies for Experimental Biology
影响因子：
0
作者：
Durr,AndryaJ;Hathaway,QuincyA;Kunovac,Amina;Taylor,AndrewD;Rizwan,Saira;Cook,ChrisC;Hollander,JohnM
通讯作者：
Hollander,JohnM

Mitochondrial miRNAs in diabetes: just the tip of the iceberg.