Cell cycle ontrol of B-cell mass

描述(由申请人提供)： 了解调节胰岛β细胞发育的机制对于开发新的方法来增强1型糖尿病患者的胰岛细胞的再生和活性至关重要。我们最近的研究表明，两个细胞周期基因P57Kip2和Cyclin D2在胚胎和出生后的胰岛β细胞分化和增殖中起关键作用。我们在这些R01协作性提案中的目标是使用新的“功能获得”方法进一步探索p57kip2和细胞周期蛋白D2在出生后a细胞新生和增殖中的作用。我们的具体目标是：(1)确定出生后α-细胞新生是否源于胰管上皮细胞以及p57Kip2在介导出生后β细胞分化中的作用；这些研究将结合一种新的模型，即胰管上皮细胞来自GFP转基因小鼠的移植骨髓，和慢病毒载体表达p57Kip2；(2)确定细胞周期蛋白D2的固有表达是否足以诱导出生后β细胞复制；这些研究将使用体外(病毒载体介导的基因在胰岛培养中的表达)和体内(细胞周期蛋白D2在小鼠的可诱导表达)模型。这些合作的R01应用程序汇集了来自胰腺发育生物学(Anil Bhushan博士)、造血干细胞生物学(Gay Crooks博士)和使用病毒载体的基因转移和表达(Donald Kohn博士)领域的三名研究人员。这组不同的研究人员提供的专业知识的结合为与1型糖尿病相关的创新和协同研究创造了一个独特的机会。

DESCRIPTION (provided by applicant): Understanding the mechanisms that regulate pancreatic beta-cell development is critical to the development of novel approaches to enhance regeneration and viability of beta-cells in patients with Type1 Diabetes. Our recent studies suggest key roles for two cell cycle genes, p57kip2 and cyclin D2, in differentiation and proliferation of pancreatic beta-cells during embryonic and postnatal life. Our goal in these collaborative R01 proposals is to explore further the roles of p57 kip2 and cyclin D2 in postnatal a-cell neogenesis and proliferation using novel "gain of function" approaches. Our Specific Aims are (1) To determine if postnatal a-cell neogenesis originates from the epithelial cells of the ducts and the role of p57kip2 expression in mediating post-natal beta-cell differentiation; these studies will use the combination of a novel model in which epithelial cells of the pancreatic ducts are derived from the transplanted bone marrow of GFP transgenic mice, and lentiviral vectors to express p57kip2; (2) To determine whether constitutive expression of cyclin D2 is sufficient to induce post natal beta-cell replication; these studies will use both in vitro (viral vector mediated gene expression in islet cultures) and in vivo (inducible expression of cyclin D2 in transgenic mice) models. These collaborative R01 applications bring together three investigators from the fields of developmental biology of the pancreas (Dr. Anil Bhushan), hematopoietic stem cell biology (Dr. Gay Crooks) and gene transfer and expression using viral vectors (Dr. Donald Kohn). The combination of expertise provided by this diverse group of investigators generates a unique opportunity for innovative and synergistic research relevant to Type 1 Diabetes.