MiR-21 Exacerbates Cytokine Induced Beta Cell Dysfunction Via Inhibition of mRNAs Regulating Beta Cell Identity

项目摘要 仅在美国，糖尿病就影响了3000多万人，每年花费惊人的2450亿美元。 糖尿病的一个标志是身体或功能性β细胞质量的损失。最近的数据表明， 细胞对炎症和代谢应激的反应最终影响β细胞功能和存活。异常 microRNA（miRNA）在β细胞中的表达已得到证实。miRNAs也被证明可以作为 β细胞发育和功能的重要调节因子，暗示它们与糖尿病期间的β细胞功能障碍有关 发展本申请人的长期目标是建立一个独立的翻译研究事业 探索与β细胞相关的糖尿病病理生理学，特别关注β细胞miRNA的作用 糖尿病的发展。该应用的中心假设是β细胞microRNA-21（miR-21）在细胞内起关键作用。 在抑制β细胞功能和诱导β细胞特性丧失中的作用。提出了两个具体目标来检验这一点 假说.目的1中的实验将使用剂量依赖性miR-21的体外Tet-on INS 1慢病毒系统 过表达，仔细验证增加的β细胞miR-21的生理作用，并描绘直接mRNA miR-21发挥这些作用的靶点。对去分化/可塑性的影响将使用 miR-21过表达的斑马鱼模型中的谱系追踪。目标2中的实验将测试miR-21的作用 使用可诱导miR-21敲除的遗传小鼠模型， 和过度表达。这些目标的完成将首次确定病理性细胞因子的作用- 诱导β细胞miR-21对β细胞功能和特性的增加。这个F30奖项需要一个为期4年的培训计划 旨在实现4个主要目标：1）培训糖尿病研究中最先进的技术和概念 2)培训使用转基因动物模型进行糖尿病研究3）培训口头和书面陈述 研究成果，包括拨款准备和4）将临床经验与研究培训相结合。 申请人将受益于该中心提供的优秀和协作研究环境， 印第安纳州大学医学院的糖尿病和代谢疾病。她的训练也将受益于 一个指导和咨询委员会，由精心挑选和建立的NIH的多元化团队组成 资助的调查员。拟议的培训将为申请人提供一个良好的培训环境， 她可以发展对β细胞生理学的理解，并在现有的 翻译合作团队

PROJECT SUMMARY In the United States alone, diabetes affects over 30 million people and costs a staggering $245 billion annually. A hallmark of diabetes is the loss of physical or functional β cell mass. Recent data suggest that the intrinsic β cell response to inflammatory and metabolic stress ultimately impacts β cell function and survival. Aberrant microRNA (miRNA) expression has been demonstrated in the β cell. MiRNAs have also been shown to serve as important regulators of β cell development and function, implicating them in β cell dysfunction during diabetes development. The long-term goal of this applicant is to establish an independent translational research career exploring diabetes pathophysiology as it relates to β cells, and specifically focusing on the role of β cell miRNAs in diabetes development. This application's central hypothesis is that β cell microRNA-21 (miR-21) plays a critical role in inhibiting β cell function and inducing loss of β cell identity. Two specific aims are proposed to test this hypothesis. Experiments in Aim 1 will use an in vitro Tet-on INS1 lentiviral system of dose dependent miR-21 overexpression to carefully validate physiologic effects of increased β cell miR-21 and delineate direct mRNA targets by which miR-21 exerts these effects. Effects on dedifferentiation/plasticity will be further evaluated using lineage tracing in a zebrafish model of miR-21 overexpression. Experiments in Aim 2 will test effects of miR-21 on β cell function and identity in a mammalian system, using genetic mouse models of inducible miR-21 knockout and overexpression. Completion of these aims will, for the first time, define the role of pathologic cytokine- induced increases in β cell miR-21 on β cell function and identity. This F30 award entails a 4-year training plan designed to achieve 4 main objectives: 1) train in state-of-the-art techniques and concepts in diabetes research 2) train in the use of transgenic animal models for diabetes research 3) train in oral and written presentation of research findings, including grant preparation and 4) integrate clinical experiences with research training.The applicant will benefit from the outstanding and collaborative research environment provided by the Center for Diabetes and Metabolic Diseases at the Indiana University School of Medicine. Her training will also benefit from a mentoring and advisory committee consisting of a diverse team of carefully selected and established NIH funded investigators. The proposed training will provide the applicant with a fertile training environment in which she can develop an understanding of β cell physiology and expand her molecular biology toolkit within an existing translational collaborative team.