Biophysical Modulation of Cardiac Ion Channels by MicroRNA

MicroRNA 对心脏离子通道的生物物理调节

• 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
• 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

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.

项目摘要 microRNA（miRs）是进化上保守的小的非编码RNA分子，其广泛地参与 调节大多数生物事件;以前的研究集中在典型的mRNA干扰（RNAi） miR机制。在上一个资助期间，我们是第一个推出进化- 在其RNAi机制之外，miR的保守的新生物物理作用。具体来说，我们发现了一个 miR 1的新生物物理作用，miR 1是心脏中最主要的miR， 人类心脏衰竭我们发现miR 1与内向整流钾通道Kir2.1物理结合， 直接抑制IK 1电流，并生物药理学地调节心脏细胞电生理学。重要的是， 我们发现人类miR 1的单核苷酸多态性（hSNP）--hSNP 14 A/G（rs776480338）， 其中第14个核苷酸“A”突变为“G”，是一种仅RNAi的变体，其特异性地消除了生物物理活性。 同时维持miR 1的RNAi功能，验证生物物理调节是独立的 关于RNAi我们的发现表明miR通过两种不同的机制调节心脏稳态： 1)规范的RNAi，调节蛋白质的表达，包括离子通道，和2）新发现的 与蛋白质直接结合并迅速导致功能调节的机制。与这个重要 新的发现，现在必须调查多个心脏离子通道是否被生物调制， miR 1的生物物理学作用在调节心肌细胞凋亡中的特异性生理学影响。 （电）生理学。根据我们发表的研究结果和初步数据，我们假设， 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 并且在心脏的体内平衡中起着关键作用。我们将研究这一点，具体目标如下。第一章 研究miR 1对心脏离子通道的生物物理调节，2）了解miR 1对心脏离子通道的生理调节， miR的生物物理作用对心脏的影响，3）揭示指导miR的生物物理作用的一般机制。 心脏离子通道的生物物理调节。除了广泛的细胞活动调节， 大量的miR（>200种中> 30，000个miR）和离子通道，我们的研究将显著和 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，具有广泛的意义。 我们的发现开创了miR生物学的一个新领域，并将提供一个机制基础和新的 RNA药物开发用于抗肿瘤治疗的途径。