microRNAs targeting Kit inhibit the development and maintenance of ICC

microRNAs 靶向试剂盒抑制 ICC 的发展和维持

• 批准号: 8284316
• 负责人: Seungil Ro
• 金额: $ 17.63万
• 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: 2011-06-20 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8284316
• 关键词: Address; Animal Model; Apoptosis; Applications Grants; Bioinformatics; Cell Proliferation; Constipation; Data; Defect; Development; Diabetes Mellitus; Diabetic mouse; Diarrhea; Endothelial Cells; Erythropoiesis; Fecal Incontinence; Fluorescence-Activated Cell Sorting; Functional disorder; Gastrointestinal Diseases; Gastrointestinal Motility; Gastrointestinal tract structure; Genetic; Green Fluorescent Proteins; Growth; Human; Interstitial Cell of Cajal; Lead; Link; Maintenance; Mediating; Messenger RNA; MicroRNAs; Modeling; Molecular; Motor Neurons; Mus; Mutant Strains Mice; Outcome; Patients; Pharmaceutical Preparations; Phenotype; Play; Principal Investigator; Proto-Oncogene Protein c-kit; Receptor Protein-Tyrosine Kinases; Regulation; Role; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Sorting - Cell Movement; Stomach; Surface; Testing; Therapeutic; Translations; angiogenesis; cell motility; diabetic; gastrointestinal; gastrointestinal symptom; genetic regulatory protein; motility disorder; motor disorder; nodal myocyte; novel; programs

DESCRIPTION (provided by applicant): Interstitial cells of Cajal (ICC) are pacemaker cells that generate spontaneous electrical slow waves, and mediate inputs from motor neurons in the gastrointestinal (GI) tract. Loss or defects in ICC networks are directly linked to GI motility disorders. Recent studies have demonstrated that ICC plays an important role in the development of diabetic gastroenteropathy (DGEP). Patients with DGEP show loss or defects in ICC networks that may lead to GI motility disorders. ICC express the receptor tyrosine kinase, KIT, which is the receptor for stem cell factor. KIT signaling is required for the normal development and maintenance of ICC in the GI tract. Therefore, it is possible that loss of ICC in DGEP may be a result of reduced expression of KIT. Recent studies show that expression of Kit is regulated by microRNAs (miRNAs). Thus, our central hypothesis is that miRNAs targeting Kit inhibit the development and maintenance of ICC in DGEP. To address the hypothesis, we propose the following specific aims: 1) Characterization of microRNAome in ICC, 2) Identification of changes in microRNAs during the development of DGEP, and 3) Suppression of the growth and maintenance of ICC by miRNAs targeting Kit. To achieve the specific aims, we will utilize two animal models Kit+/copGFP and type 2 diabetic Kit+/copGFP;Lepob/ob mutant mice that we have recently generated . Both mice express a bright green fluorescent protein copGFP in a Kit-specific manner, which offers a powerful new model to study the function and genetic regulation of ICC phenotypes. Completion of the specific aims of this project will provide information about the miRNAs expressed specifically by ICC and how this body of regulatory molecules is altered in a major GI motor disorder. Our preliminary data suggest that many of the miRNAs in ICC target Kit. This study will describe a novel molecular mechanism involving miRNAs that inhibit KIT expression. This is an exciting opportunity to understand how KIT expression in ICC is regulated and how ICC are lost during the development of DGEP. Identification of the miRNAs inhibiting KIT expression may aid in the development of a therapeutic antisense miRNA drug that might be useful in blocking or reversing damaged ICC networks in DGEP and other GI motility and functional disorders in which loss of ICC occurs.

描述（由申请人提供）：卡哈尔间质细胞（ICC）是起搏细胞，可产生自发电慢波，并介导胃肠道（GI）中运动神经元的输入。 ICC 网络的丢失或缺陷与胃肠道运动障碍直接相关。最近的研究表明ICC在糖尿病胃肠病（DGEP）的发展中发挥着重要作用。 DGEP 患者的 ICC 网络出现缺失或缺陷，可能导致胃肠道运动障碍。 ICC 表达受体酪氨酸激酶 KIT，它是干细胞因子的受体。 KIT 信号传导对于胃肠道中 ICC 的正常发育和维持是必需的。因此，DGEP 中 ICC 的丢失可能是 KIT 表达减少的结果。最近的研究表明 Kit 的表达受到 microRNA (miRNA) 的调节。因此，我们的中心假设是 miRNA 靶向 Kit 抑制 DGEP 中 ICC 的发育和维持。为了解决这一假设，我们提出以下具体目标：1) ICC 中 microRNAome 的表征，2) 鉴定 DGEP 发育过程中 microRNA 的变化，3) 通过 miRNA 靶向试剂盒抑制 ICC 的生长和维持。为了实现特定目标，我们将利用我们最近生成的两种动物模型Kit+/copGFP和2型糖尿病Kit+/copGFP;Lepob/ob突变小鼠。两只小鼠都以 Kit 特异的方式表达亮绿色荧光蛋白 copGFP，这为研究 ICC 表型的功能和遗传调控提供了一个强大的新模型。完成该项目的具体目标将提供有关 ICC 特异性表达的 miRNA 的信息，以及该调节分子主体如何在主要胃肠道运动障碍中发生改变的信息。我们的初步数据表明 ICC 目标试剂盒中的许多 miRNA。这项研究将描述一种涉及抑制 KIT 表达的 miRNA 的新分子机制。这是一个令人兴奋的机会，可以了解 ICC 中的 KIT 表达是如何调节的以及 ICC 在 DGEP 的开发过程中是如何丢失的。抑制 KIT 表达的 miRNA 的鉴定可能有助于开发治疗性反义 miRNA 药物，该药物可能有助于阻断​​或逆转 DGEP 中受损的 ICC 网络以及发生 ICC 丢失的其他胃肠道运动和功能性疾病。