Discovery of Pancreatic Signatures in Type 2 Diabetes Mellitus

 描述(由申请人提供)：2型糖尿病(T2 DM)是由于胰岛素分泌受损、胰岛素抵抗和肝脏葡萄糖生成增加所致，胰岛素分泌不足可能是是否发展为T2 DM的关键决定因素。然而，我们并不了解人类T2 DM患者的？细胞质量减少和/或a/？细胞功能障碍的原因。这是因为有关T2 DM？或a细胞的模型和假设通常来自对啮齿动物模型的研究，或者尚未在人类样本中得到证实。此外，大多数关于T2 DM人胰腺的研究没有充分结合患者或疾病阶段的临床表型，因此结合了不同阶段的特征。此外，以前研究的T2 DM胰腺标本的收集方式并不完全允许新的分子分析。为了克服我们知识上的这些不足，我们建立了一个新的基础设施，用于获取人类T2 DM胰腺标本，用于新技术的分析，并从相同的胰腺中分离胰岛。此外，我们还组建了一支由具有人类胰岛生物学专业知识的科学家和研究人员组成的跨学科团队，他们从神经科学和癌症领域带来了组织和细胞图谱的新技术。使用这一新的基础设施和技术，我们建议：1)识别胰岛中独特的蛋白质和RNA特征，并利用RNA测序、组织清除和单细胞表型等技术从临床表型为短(即5年)和长(即10年)的T2 DM捐赠者中分离出a和？细胞。2)利用组织成像质谱仪，鉴定T2 DM胰岛中差异表达的脂质、蛋白质和代谢物。3)将分子特征与功能T2 DM胰岛图谱相结合，并测试来自脂质、代谢物和/或mRNA/蛋白质数据集的候选分子对a和？细胞活性和存活率的影响。为了与R24机制的目标保持一致，我们的基于发现的方法将产生新的资源和数据集，为人类T2 DM的a/？细胞功能障碍创建新的范例，并促进基础性发现，这些发现不仅将提高我们对？细胞功能障碍/丢失如何发生的理解，而且可能导致治疗干预。

 DESCRIPTION (provided by applicant): Type 2 diabetes (T2DM) results from impaired insulin secretion, insulin resistance, and increased hepatic glucose production with deficits in insulin secretion likely the key determinant of whether T2DM develops. However, we do not understand the cause of reduced ß cell mass and/or a/ß cell dysfunction in human T2DM. This is because models and hypotheses about T2DM ß or a cells generally arise from studies of rodent models or have not been confirmed in human samples. Furthermore, most studies on the T2DM human pancreas do not adequately incorporate the clinical phenotype of the patient or the disease stage and, consequently, combine profiles from different stages. In addition, previously studied T2DM pancreatic specimens have been collected in ways that do not completely allow newly available molecular analyses. To overcome these deficits in our knowledge, we have established a new infrastructure for procuring human T2DM pancreatic specimens for analysis by new technologies and isolating islets from the same pancreas. In addition, we have assembled an interdisciplinary team of scientists with human islet biology expertise and investigators who bring new technologies for tissue and cell profiling from the neuroscience and cancer arenas. Using this new infrastructure and technologies, we propose to: 1) Identify unique protein and RNA signatures in the pancreatic islets and isolated a and ß cells from clinically phenotyped T2DM donors of short (i.e. <5 years)- and long (i.e. >10 years)-duration with technologies such as RNA-sequencing, tissue-clearing, and single cell phenotyping. 2) Using tissue imaging mass spectrometry, identify differentially expressed lipids, proteins, and metabolites in T2DM islets. 3) Integrate molecular signatures with functional T2DM islet profiles and test the impact of candidate molecules from the lipid, metabolite, and/or mRNA/protein datasets on a and ß cell activity and viability. In keeping with the goals of the R24 mechanism, our discovery-based approaches will generate new resources and datasets, create new paradigms for a/ß cell dysfunction in human T2DM, and foster fundamental discoveries that will not only improve our understanding of how ß cell dysfunction/loss occurs, but potentially lead to therapeutic interventions.