A novel mouse model to distinguish the specific physiological significance of RNAi and biophysical mechanisms of microRNA

区分RNAi特定生理意义和microRNA生物物理机制的新型小鼠模型

• 批准号: 10351415
• 负责人: Jidong Fu
• 金额: $ 23.63万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10351415
• 关键词: 3' Untranslated Regions; Adult; Age; Animal Model; Apoptosis; Arrhythmia; Binding; Biological; Biophysical Process; Biophysics; Canes; Cardiac Myocytes; Cell Proliferation; Cells; Colon; Crossbreeding; Defect; Development; Doxycycline; Duchenne muscular dystrophy; Ectopic Expression; Electrophysiology (science); Event; Evolution; Gene Expression; Genes; Genetic Transcription; Genitourinary system; Heart; Heart Abnormalities; Heart Diseases; Heart failure; Homeostasis; Hour; Human; Human Genome; Investigation; Ion Channel; Knock-in Mouse; Knock-out; Knockout Mice; Long-Term Effects; Lung; Malignant Epithelial Cell; Malignant Neoplasms; Metabolism; MicroRNAs; Modeling; Mus; Muscle; Muscle Cells; Muscular Dystrophies; Myocardial dysfunction; Nucleotides; Organ; Organogenesis; Phenotype; Physiological; Physiology; Play; Primary carcinoma of the liver cells; Proteins; Pump; RNA Interference; Regulation; Role; Single Nucleotide Polymorphism; Skeletal Muscle; Symptoms; Tetanus Helper Peptide; Thyroid Gland; Tissues; Transgenic Mice; Translations; Untranslated RNA; Weaning; biological adaptation to stress; body system; cancer type; cardiogenesis; cell growth; human disease; in vivo; inward rectifier potassium channel; knock-down; mRNA Transcript Degradation; mature animal; mouse genome; mouse model; novel; novel therapeutic intervention; postnatal; protein expression; sarcoma; tool; tumorigenesis

Project Summary MicroRNAs (miRs) are evolutionally conserved small non-coding RNA molecules and control most biological events, including apoptosis, cell proliferation, metabolism, cell fate determination, organogenesis, development, stress responses, and tumorigenesis. Classically, miRs are known to negatively regulate gene expression through RNA interference (RNAi) mechanism. Recently, 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 discovered that miR1directly binds to inward rectifier potassium channel Kir2.1, resulting in direct suppression of the IK1 current and leading to biophysical modulation of cardiomyocyte cellular electrophysiological functions. Our studies suggest that miR1 modulates the development and homeostasis of tissues/organs through two different mechanisms: the immediate effect (seconds to minutes) of newly-discovered biophysical modulation and long-term effect (hours to days) of RNAi. With this important new finding, it now becomes essential to understand how these two distinct miRs mechanisms of action coordinate to regulate the development and homeostasis of our body. However, there is no valid model that can distinguish the specific physiology significance of biophysical modulation versus RNAi mechanism. We found that an arrhythmia-associated hSNP14A/G specifically defects the biophysical action while maintaining miR1’s RNAi function; therefore, we propose to develop a unique transgenic mouse model that can separate the specific contribution coming from the biophysical modulation and dissect the pure contribution of RNAi in maintain the homeostasis of multi organs/systems. We will develop miR1-full-KO/muscle-specific inducible hSNP14A/G-knock-in mice, and we hypothesize that an expression of hSNP14A/G in muscle cells could rescue the postnatal lethality of miR1-full- KO mice. We will investigate if lacking biophysical function of hSNP14A/G induces any abnormal phenotypes (Aim 1), such as arrhythmia, heart failure, and abnormal contractility of skeletal muscle, which will demonstrate the specific role of miR1’s biophysical modulation in regulation of the homeostasis in vivo. We will also turn off the expression of hSNP14A/G by administration of doxycycline and investigate the specific physiological significance of miR1’s RNAi mechanism in the heart (Aim 2). This unique animal model will be very valuable to investigate the critical role of miR1 in multiple organs/systems, including the heart, skeletal muscle, various types of cancers. Understanding the specific contributions of miR’s biophysical modulation and RNAi in vivo will expand the biological significance of miRs and guide us to develop new therapeutic approaches for human diseases through targeting of miRs.

项目概要 MicroRNA (miR) 是进化上保守的小非编码 RNA 分子，控制着大多数生物 事件，包括细胞凋亡、细胞增殖、代谢、细胞命运决定、器官发生、发育、 众所周知，miR 会负向调节基因表达。 最近，我们揭示了 miR1 的一种新的生物物理作用。 是心脏中最重要的 miR，并且在人类心力衰竭中下调。 miR1直接结合内向整流钾通道Kir2.1，导致IK1的直接抑制 当前并导致心肌细胞细胞电生理功能的生物物理调节。 研究表明miR1通过两个途径调节组织/器官的发育和稳态 不同的机制：新发现的生物物理调节的即时效果（几秒到几分钟） 有了这一重要的新发现，现在就必须研究 RNAi 的长期影响（数小时至数天）。 了解这两种不同的 miR 作用机制如何协调来调节发育和 然而，没有有效的模型可以区分特定的生理学。 生物物理调节与 RNAi 机制的重要性我们发现与心律失常相关。 hSNP14A/G 在维持 miR1 的 RNAi 功能的同时，特异性地缺陷了生物物理作用；因此，我们 开发一种独特的转基因小鼠模型提案，可以将来自于的特定贡献分开 生物物理调节并剖析 RNAi 在维持多种体内稳态中的纯粹贡献 我们将开发 miR1-full-KO/肌肉特异性诱导型 hSNP14A/G 敲入小鼠，并且我们 研究发现，肌肉细胞中 hSNP14A/G 的表达可以挽救 miR1-full- 的产后致死性 我们将研究缺乏 hSNP14A/G 的生物物理功能是否会导致任何异常表型。 （目标1），例如心律失常、心力衰竭和骨骼肌收缩力异常，这将证明 我们还将关闭 miR1 的生物物理调节在体内稳态调节中的具体作用。 给予强力霉素后 hSNP14A/G 的表达并研究其具体生理学 miR1 的 RNAi 机制在心脏中的重要性（目标 2）对于这种独特的动物模型将非常有价值。 研究 miR1 在多个器官/系统中的关键作用，包括心脏、骨骼肌、各种 了解 miR 的生物物理调节和体内 RNAi 的具体贡献。 将扩大 miR 的生物学意义并指导我们为人类开发新的治疗方法 通过靶向 miR 来治疗疾病。