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.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 平滑肌肥大发生在多种内脏肌肉对病理生理条件的反应。我们建立了一种小肠部分梗阻(PO)模型，在该模型中，SM肥大是口腔性的，而不是流产到PO的部位，以研究肥大和/或增殖过程中平滑肌细胞(SMC)的重构。MicroRNAs(MiRNAs)调节SMC的重要细胞功能，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显著不同。我们还发现了一组独特的miRNAs，它们在小鼠和人类的小肠中高度表达，其中miR-143和miR-145尤其丰富。这些miRNAs也在排序的SMC中大量表达，并且在PO模型中都下调。这导致了一种假设，即SM肥大部分由SMC依赖的miRNAs调控。为了确定miRNAs在小肠肥大发生发展中的作用，将涉及以下三个目标：目的1：鉴定在小肠SMC中表达的miRNAs以及在SMC肥大发生过程中发生的变化；目的2：鉴定在小肠SMC中表达的信使RNAs(MRNAs)及其在肥大发展过程中发生的变化；目的3：鉴定特定调控肥大相关基因的miRNAs。几种分子、遗传和生物信息学方法将被用来在肥大的SMC中发展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小鼠产生的PO模型中研究SMC肥大，该模型允许高度特异的SMC分选和454测序(焦测序)技术。对于目标2，我们将使用从分离的SMC中分离的mRNAs的基因芯片分析来识别肥大依赖基因。对于目标3，我们将首先使用生物信息学将miRNAs与靶基因联系起来，然后使用尖端的分子生物学技术验证这些关系，从而识别调控肥大基因的miRNAs。我们将主要关注miR-143和miR-145及其靶基因，以了解它们如何调控SMC在增殖和/或肥大过程中的发育。了解SMCs基因表达的变化为理解SM病理生理条件下的表型调控提供了一个令人兴奋的新机会。识别与肥大相关的遗传标记将有助于miRNA药物的开发，这些药物有可能使导致肥厚的mRNA靶标正常化，从而逆转这些疾病中发生的一些不必要的病理变化。