SMOOTH MUSCLE HYPERTROPHY REGULATED BY MICRORNAS AND THEIR TARGET GENES

微核及其靶基因调控平滑肌肥大

• 批准号: 8360519
• 负责人: Seungil Ro
• 金额: $ 20.86万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8360519
• 关键词: Address; Apoptosis; Bioinformatics; Cell Separation; Cell physiology; Centers of Research Excellence; Development; Disease; Funding; Gene Expression; Gene Targeting; Genes; Genetic Markers; Grant; Human; Hyperplasia; Hypertrophy; Link; Messenger RNA; MicroRNAs; Microarray Analysis; Modeling; Molecular Biology Techniques; Molecular Genetics; Mus; Muscle; National Center for Research Resources; Obstruction; Oral; Pharmaceutical Preparations; Phenotype; Principal Investigator; Regulation; Research; Research Infrastructure; Resources; Role; Site; Small Intestines; Smooth Muscle; Smooth Muscle Myocytes; Sorting - Cell Movement; Source; Techniques; United States National Institutes of Health; Visceral; cost; muscle hypertrophy; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Smooth muscle (SM) hypertrophy occurs in a variety of visceral muscles in response to pathophysiological conditions. We have developed a small intestine partial obstruction (PO) model in which the SM hypertrophy is oral but not aboral to the site of PO, to study the remodeling that occurs in smooth muscle cells (SMCs) during hypertrophy and/or hyperplasia. microRNAs (miRNAs) regulate vital cellular functions in SMCs such as differentiation, proliferation, and apoptosis. Our preliminary microarray analysis reveals that gene expression in the PO model is dramatically different from that of control muscles. We have also identified a unique set of miRNAs that are highly expressed in the small intestine of mice and humans with miR-143 and miR-145 being particularly abundant. These miRNAs are also abundantly expressed in sorted SMCs and both are down-regulated in the PO model. This has led to the hypothesis that SM hypertrophy is regulated, in part, by SMC-dependent miRNAs. To determine the role of miRNAs in the development of small intestine hypertrophy, the following three aims will be addressed: Aim 1: Identify the miRNAs expressed in small intestine SMCs and the changes which occur with the development of SMC hypertrophy; Aim 2: Identify the messenger RNAs (mRNAs) expressed in small intestine SMCs and the changes which occur with the development of hypertrophy; Aim 3: Identify the miRNAs specifically regulating the genes associated with hypertrophy. Several molecular, genetic, and bioinformatic approaches will be used to develop a functional link between miRNA and target gene expression in the hypertrophic SMCs. For Aim 1, SMC hypertrophy will be studied in our PO model generated in the SM-specific Cre/eGFP mice which permit highly specific SMC sorting and 454 sequencing (pyrosequencing) techniques. For Aim 2, we will identify hypertrophy-dependent genes using GeneChip analyses of the mRNAs isolated from sorted SMCs. For Aim 3, we will identify the miRNAs regulating hypertrophy genes by first relating miRNAs to target genes using bioinformatics and then validating these relationships using cutting edge molecular biology techniques. We will primarily focus on miR-143 and miR-145 and their target genes to see how they regulate the development of SMCs during hyperplasia and/or hypertrophy. Understanding changes in gene expression in SMCs provides an exciting new opportunity for understanding the regulation of phenotype in SM pathophysiological conditions. Identifying the genetic markers associated with hypertrophy will aid in the development of miRNA drugs that have the potential to normalize mRNA targets that are responsible for hypertrophy and thus reverse some of the unwanted pathological changes that occur in these disorders.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 平滑肌（SM）肥大发生在多种内脏肌中，以响应病理生理条件。我们开发了一种小肠部分梗阻 (PO) 模型，其中 SM 肥大是口腔性的，但不是 PO 部位的肠外性，以研究平滑肌细胞 (SMC) 在肥大和/或增生期间发生的重塑。 microRNA (miRNA) 调节 SMC 中的重要细胞功能，例如分化、增殖和凋亡。我们的初步微阵列分析表明，PO 模型中的基因表达与对照肌肉的基因表达显着不同。我们还鉴定了一组独特的 miRNA，它们在小鼠和人类的小肠中高表达，其中 miR-143 和 miR-145 特别丰富。这些 miRNA 在分选的 SMC 中也大量表达，并且在 PO 模型中均下调。这导致了这样的假设：SM 肥大部分受到 SMC 依赖性 miRNA 的调节。为了确定 miRNA 在小肠肥大发展中的作用，将解决以下三个目标： 目标 1：识别小肠 SMC 中表达的 miRNA 以及随着 SMC 肥大的发展而发生的变化；目标2：识别小肠SMC中表达的信使RNA（mRNA）以及随着肥大的发展而发生的变化；目标 3：鉴定特异性调节与肥大相关基因的 miRNA。几种分子、遗传和生物信息方法将用于开发肥大 SMC 中 miRNA 和靶基因表达之间的功能联系。对于目标 1，我们将在 SM 特异性 Cre/eGFP 小鼠中生成的 PO 模型中研究 SMC 肥大，该模型允许高度特异性的 SMC 分选和 454 测序（焦磷酸测序）技术。对于目标 2，我们将使用从分选的 SMC 中分离的 mRNA 的 GeneChip 分析来识别肥大依赖性基因。对于目标 3，我们将首先使用生物信息学将 miRNA 与靶基因相关联，然后使用尖端分子生物学技术验证这些关系，从而识别调节肥大基因的 miRNA。我们将主要关注 miR-143 和 miR-145 及其靶基因，以了解它们如何在增生和/或肥大期间调节 SMC 的发育。了解 SMC 基因表达的变化为了解 SM 病理生理条件下表型的调节提供了令人兴奋的新机会。识别与肥大相关的遗传标记将有助于开发 miRNA 药物，这些药物有可能使导致肥大的 mRNA 靶点正常化，从而逆转这些疾病中发生的一些不需要的病理变化。