Role of microRNAs in insulin production and secretion in pancreatic beta-cells

描述(由申请人提供)： 这位候选人之前一直在使用RNAi技术研究胰岛素调节的葡萄糖在3T3-L1脂肪细胞中的转运机制，后来专注于RNAi的机制和癌细胞和小鼠组织中microRNAs的图谱。为了继续她在糖尿病领域的训练，她最近加入了Sabire Ozcan博士的实验室。厄兹坎博士的研究主要集中在胰岛β细胞中葡萄糖调节的胰岛素基因转录。应聘者的直接职业目标是更好地熟悉胰岛素基因表达调节和葡萄糖调节胰岛β细胞功能的实验程序和应用。这项培训将使她能够实现她的长期目标，即作为一名独立研究者，研究miRNAs在调节β细胞功能中的作用，并阐明miRNAs在II型糖尿病发展中的作用。 最近的研究表明，一种新的细胞小RNA群体，称为microRNAs(MiRNAs)，可能在2型糖尿病的发生发展中发挥重要作用。这些miRNAs的表达可能在各种环境和生理条件下受到高度调控；这些小RNAs的失调可能导致2型糖尿病的发生。例如，miR-375已被发现在胰岛β细胞中特异表达，并在胰岛素分泌中发挥重要作用。大多数miRNAs在β细胞中的功能有待研究。P.I.的假设是，miRNAs并不是单独发挥作用的，而是由多个miRNAs共同作用，以调节β细胞的发育以及胰岛素的产生和分泌，以响应葡萄糖。本项目的具体目标是：(1)利用新近发展的Rational miRNA微阵列技术，研究胰腺β细胞中miRNA的表达谱，以确定新的细胞特异性和葡萄糖调节的miRNAs；(2)通过转染合成的miRNAs和2-O-甲基RNA抗原体，研究它们在MIN6细胞中的功能，以研究其过度表达的效果和与其下调相关的作用。这些活动的结果可能会导致发现新的miRNAs或miRNA网络在2型糖尿病发展中的新作用，以及治疗或预防这种疾病的潜在治疗方法。

DESCRIPTION (provided by applicant): The candidate has been previously studying the mechanisms of insulin-regulated glucose transport in 3T3-L1 adipocytes using RNAi technology and later focused on the mechanisms of RNAi and profiling of microRNAs in cancer cells and mouse tissues. To continue her training in the field of diabetes, she has recently joined Dr. Sabire Ozcan's laboratory. Dr. Ozcan's research focuses mainly on glucose-regulated insulin gene transcription in pancreatic beta-cells. The immediate career goal of the candidate is to better acquaint herself with experimental procedures and applications in the regulation of insulin gene expression and glucose regulation of pancreatic beta-cell function. This training will allow her to pursue her long-term goal, which is to study the role of miRNAs in regulating beta-cell function and to elucidate the role of miRNAs in the development of type-ll diabetes, as an independent investigator. Recent studies indicate that a novel population of cellular small RNAs, termed microRNAs (miRNAs), may play an important role in the development of type 2 diabetes. The expression of these miRNAs may be highly regulated under various environmental and physiological conditions; dysregulation of these small RNAs may lead to the development of type 2 diabetes. For example, miR-375 has been discovered to be specifically expressed in pancreatic beta cells and be important in insulin secretion. The functions of most miRNAs in beta cells await to be investigated. The hypothesis of the P.I. is that miRNAs are not functioning alone, but rather, multiple miRNAs act in cohort to regulate beta cell development and insulin production and secretion in response to glucose. The specific aims of this project are: (1) To profile miRNA expression in pancreatic beta cells, in order to identify novel cell-specific and glucose-regulated miRNAs with a recently developed rational miRNA microarray technology; (2) To study the function of the identified glucose-regulated and beta cell-specific miRNAs in MIN6 cells by transfecting with synthetic miRNAs to study overexpression effects and with 2-O-methyl RNA antagomirs to investigate the effects associated with their down-regulation. The results obtained from the proposed activities may lead to the discovery of novel miRNAs or new roles of miRNA network in the development of type 2 diabetes and potential therapeutic approaches to treat or prevent this disease.