Biophysical Modulation of Cardiac Ion Channels by MicroRNA

MicroRNA 对心脏离子通道的生物物理调节

基本信息

批准号：
10660561
负责人：
Isabelle Deschenes
金额：
$ 65.14万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-01 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10660561
关键词：
Acute Adult Age Anti-Arrhythmia Agents Antibodies Arrhythmia Binding Biological Biological Assay Biology Biophysical Process Biophysics Biotinylation CRISPR/Cas technology Cardiac Cardiac Electrophysiologic Techniques Cardiac Myocytes Consensus Data Development Disease Electrophysiology (science)Environment Event Evolution Foundations Funding Genes Heart Heart Abnormalities Heart failure Homeostasis Homo Human Immunoprecipitation Ion Channel Ion Channel Protein Kir2.1 channel Knock-out Knowledge Mass Spectrum Analysis Mediating Medicine Membrane Potentials Membrane Proteins Messenger RNA MicroRNAs Molecular Mus Mutate Mutation Neonatal Nucleotides Physiological Physiology Play Potassium Protein Binding Domain Proteins Publishing RNA RNA Interference Regulation Rest Role Single Nucleotide Polymorphism Small RNA Solid Testing Transgenic Mice Transgenic Organisms Untranslated RNA Variant Weaning cardiogenesis crosslink heart function improved innovation inward rectifier potassium channel mortality mutant novel overexpression postnatal protein expression tool transcriptome sequencing

项目摘要

Project Summary MicroRNAs (miRs) are evolutionally conserved small non-coding RNA molecules that are broadly involved in regulating most biological events; previous studies have focused on the canonical mRNA interference (RNAi) mechanism of miRs. During the previous funding period, we were the first to unveil an evolutionarily- conserved novel biophysical action for miRs beyond its RNAi mechanism. Specifically, we revealed a novel biophysical action of miR1, which is the most predominant miR in the heart and is downregulated in human heart failure. We found that miR1 physically binds to an inward rectifier potassium channel Kir2.1, directly suppresses the IK1 current and biophysically modulates cardiac cellular electrophysiology. Importantly, we found that a human single nucleotide polymorphism (hSNP) of miR1–– hSNP14A/G (rs776480338), in which the 14th nucleotide “A” is mutated to “G”, is a RNAi-only variant that specifically abolishes the biophysical action while maintaining the RNAi function of miR1, validating that the biophysical modulation is independent of RNAi. Our discoveries suggest that miRs modulate cardiac homeostasis through two different mechanisms: 1) canonical RNAi that regulates the expression of proteins, including ion channels, and 2) newly-discovered mechanism of direct binding with proteins that quickly results in functional modulation. With this important new finding, it is now imperative to investigate if multiple cardiac ion channels are biophysically modulated by miRs and to elucidate the specific physiological impact of miR1’s biophysical action in the regulation of cardiac (electro)physiology. Based on our published findings and preliminary data, we hypothesize that the biophysical modulation of cardiac ion channels by miRs is a general regulatory mechanism that exists broadly and plays a critical role in the homeostasis of the heart. We will study this with the following specific aims. 1) To investigate the biophysical modulation of cardiac ion channels by miR1, 2) To understand the physiological impact of miRs’ biophysical action on the heart, 3) To unveil the general mechanisms guiding miRs’ biophysical modulation of cardiac ion channels. In addition to a broad range of cellular activities regulated by the large number of miRs (>30,000 miRs in >200 species) and ion channels, our study will significantly and innovatively expand the biological significance of miR biology and ion channel biology with broad implications. Our discoveries have pioneered a new field in miR biology and will provide a mechanistic foundation and new avenue of RNA-medicine development for antiarrhythmic therapy.

项目概要 MicroRNA (miR) 是进化上保守的小非编码 RNA 分子，广泛参与调节大多数生物事件；之前的研究主要集中在典型的 mRNA 干扰 (RNAi) 上在之前的资助期间，我们是第一个揭示了 miR 的进化机制的人。具体而言，我们揭示了 miR 超出其 RNAi 机制的保守的新生物物理作用。 miR1 的新生物物理作用，它是心脏中最主要的 miR，并且在心脏中下调我们发现 miR1 与内向整流钾通道 Kir2.1 物理结合。直接抑制 IK1 电流并从生物物理角度调节心脏细胞电生理学。我们发现 miR1–hSNP14A/G (rs776480338) 的人类单核苷酸多态性 (hSNP) 其中第 14 个核苷酸“A”突变为“G”，是一种仅 RNAi 的变体，专门废除了生物物理学作用，同时维持 miR1 的 RNAi 功能，验证生物物理调节是独立的我们的发现表明 miR 通过两种不同的机制调节心脏稳态： 1) 调节蛋白质表达（包括离子通道）的经典 RNAi，以及 2) 新发现的与蛋白质直接结合的机制，可快速导致功能调节。新发现，现在有必要研究多个心脏离子通道是否受到生物物理调节 miRs 并阐明 miR1 的生物物理作用在心脏调节中的具体生理影响（电）生理学。根据我们发表的研究结果和初步数据，我们发现 miR对心脏离子通道的生物物理调节是广泛存在的一般调节机制并在心脏的稳态中发挥着关键作用，我们将通过以下具体目标来研究这一点：1）。研究 miR1 对心脏离子通道的生物物理调节，2) 了解生理学 miRs的生物物理作用对心脏的影响，3）揭示指导miRs的一般机制除了受调节的广泛细胞活动外，心脏离子通道的生物物理调节。大量的 miR（超过 200 个物种中超过 30,000 个 miR）和离子通道，我们的研究将显着且创新性地扩展了 miR 生物学和离子通道生物学的生物学意义，具有广泛的影响。我们的发现开创了 miR 生物学的新领域，并将提供机制基础和新的用于抗心律失常治疗的 RNA 药物开发途径。