Discovery of Pancreatic Signatures in Type 2 Diabetes Mellitus

 描述（由申请方提供）：2型糖尿病（T2 DM）由胰岛素分泌受损、胰岛素抵抗和肝葡萄糖生成增加引起，胰岛素分泌缺陷可能是是否发生T2 DM的关键决定因素。然而，我们不了解人T2 DM中β细胞质量减少和/或α/β细胞功能障碍的原因。这是因为关于T2 DM β细胞或α细胞的模型和假设通常来自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纳入患者的临床表型或疾病分期，因此，联合收割机组合了不同分期的特征。此外，先前研究的T2 DM胰腺标本的采集方式不完全允许进行新的分子分析。为了克服我们知识中的这些缺陷，我们建立了一个新的基础设施，用于采购人类T2 DM胰腺标本，通过新技术进行分析，并从同一胰腺中分离胰岛。此外，我们还组建了一个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分析的新技术。使用这种新的基础设施和技术，我们提出：1）使用诸如RNA测序、组织清除和单细胞表型分析的技术，从短期（即<5年）和长期（即>10年）的临床表型T2 DM供体中鉴定胰岛和分离的α和β细胞中的独特蛋白质和RNA特征。2)使用组织成像质谱法，鉴定T2 DM胰岛中差异表达的脂质、蛋白质和代谢物。3)将分子特征与功能性T2 DM胰岛特征整合，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α和β细胞活性和活力的影响。为了与R24机制的目标保持一致，我们基于发现的方法将产生新的资源和数据集，为人类T2 DM中的α/β细胞功能障碍创造新的范例，并促进基础发现，这不仅将提高我们对β细胞功能障碍/损失如何发生的理解，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.