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

项目总结 仅在美国，糖尿病就影响了3000多万人，每年造成的损失高达2450亿美元。 糖尿病的一个特征是失去生理或功能上的β细胞团。最近的数据表明，内在的β 细胞对炎症和代谢应激的反应最终影响β细胞的功能和存活。反常的 微RNA(MiRNA)在β细胞中有表达。MiRNAs也被证明可以作为 β细胞发育和功能的重要调节因子与糖尿病时β细胞功能障碍的关系 发展。该申请者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 在糖尿病的发展中。这一应用的中心假设是β细胞microRNA-21(miR-21)在 在抑制β细胞功能和诱导β细胞特性丧失方面的作用。提出了两个具体的目标来测试这一点 假设。AIM 1中的实验将使用剂量依赖的miR-21的体外Tet-on INS1慢病毒系统 过度表达以仔细验证增加的β细胞MIR-21的生理效应并勾画出DIRECT基因 MIR-21发挥这些作用的靶点。对脱分化/可塑性的影响将进一步评估使用 MiR-21过表达斑马鱼模型的谱系追踪。Aim 2的实验将测试miR-21的效果 利用可诱导miR-21基因敲除的小鼠遗传模型，研究哺乳动物系统中β细胞的功能和身份 和过度表达。这些目标的完成将第一次定义病理细胞因子的作用- 诱导β细胞MIR-21对β细胞功能和特性的影响。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)培训糖尿病研究的最新技术和概念 2)在糖尿病研究中使用转基因动物模型的培训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。 申请者将受益于该中心提供的卓越和协作的研究环境 印第安纳大学医学院的糖尿病和新陈代谢疾病。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的不同团队组成 资助调查人员。拟议的培训将为申请者提供一个肥沃的培训环境， 她可以加深对β细胞生理学的理解，并在现有的 翻译协作团队。

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.