Monogenic forms of juvenile onset diabetes: towards novel insights in β-cell development, function and survival

青少年发病糖尿病的单基因形式：对β细胞发育、功能和生存的新见解

• 批准号: 406674944
• 负责人: Professor Dr. Alexander Kleger
• 金额: --
• 依托单位: Klinik für Innere Medizin I
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/406674944?language=en
• 关键词: Monogenic; forms; juvenile; onset; diabetes

A small subset of juvenile onset insulin-treated diabetes (JOD) is caused by single gene mutations, contrasting with the general multifactorial inheritance of common diabetes. Several genes responsible for monogenic diabetes have been identified to date, and these affect β-cell development, function and survival. The overall aim of this project is to identify and genetically characterize new monogenic entities of JOD and perform in-depth functional studies of these genes. The unique approach of this joint proposal is fostered by two partners having complementary track records in human genetics and functional biology. In the first stage, we will extend ongoing genetic studies of highly selected patients and families enriched in monogenic forms of diabetes to identify causative genes. We will perform extended large-scale re-sequencing of selected genes emerging from our exome sequencing data to genetically validate them, as well as of already known monogenic diabetes genes, in a large collection of 1000 JOD patients and controls. Thereby, we will identify monogenic causative mutations in these genes depending on the population, familial structure and clinical characteristics of patients. This will provide for the first time a reliable estimation of monogenic contributions to JOD in various settings. In preliminary studies, we have already identified several genes with monogenic contributions, as well as new mutations in previously reported genes, validating the power of our genetic strategy. In the second stage, starting simultaneously, we will perform in-depth functional studies of selected groups of these genes, starting with two genes: one gene (X) that we have previously identified as mutated in a new form of monogenic diabetes (homozygous mutated status: neonatal diabetes, heterozygous status: early-onset type 2 diabetes), and one gene (Y) that we recently validated as a monogenic diabetes gene with the identification of 7 new mutations in 8 unrelated JOD patients. First, we will extend functional data that characterize gene X using a human embryonic stem cell-derived be-cell platform. Second, we will study the specific functions of gene Y using a state-of-the-art phenotyping model, patient-specific induced pluripotent stem cells (hiPSCs) and genome-edited human embryonic stem cells (hESCs). The latter will be differentiated into pancreatic β-cells for detailed functional analyses. Third, we will create the newly identified mutations in these genes by genomic editing in hESCs using an iCRISPR platform. Thus, we will develop a whole patient-specific cell-bank for detailed cellular investigations aiming to i) model diabetes in a dish to identify the molecular basis of the disease, ii) address novel therapeutic possibilities and iii) define a complete molecular network essential for endocrine pancreatic development as well as β-cell function and maintenance. This study will establish the basis for true “personalized medicine” in diabetes therapy.

青少年发病的胰岛素治疗糖尿病（JOD）的一小部分是由单基因突变引起的，这与普通糖尿病的一般多因素遗传形成鲜明对比。迄今为止，已鉴定出几种与单基因糖尿病有关的基因，这些基因会影响 β 细胞的发育、功能和存活。该项目的总体目标是鉴定和遗传表征 JOD 新的单基因实体，并对这些基因进行深入的功能研究。这项联合提案的独特方法是由在人类遗传学和功能生物学方面拥有互补记录的两个合作伙伴共同提出的。在第一阶段，我们将扩大对富含单基因糖尿病的精心挑选的患者和家庭正在进行的遗传学研究，以确定致病基因。我们将对我们的外显子组测序数据中出现的选定基因以及 1000 名 JOD 患者和对照的已知单基因糖尿病基因进行大规模的重新测序，以进行基因验证。因此，我们将根据人群、家族结构和患者的临床特征来识别这些基因中的单基因致病突变。 This will provide for the first time a reliable estimation of monogenic contributions to JOD in various settings.在初步研究中，我们已经确定了几个具有单基因贡献的基因，以及先前报道的基因中的新突变，验证了我们的遗传策略的力量。在第二阶段，同时开始，我们将对选定的这些基因组进行深入的功能研究，从两个基因开始：一个基因（X），我们之前已鉴定为在一种新形式的单基因糖尿病中发生突变（纯合突变状态：新生儿糖尿病，杂合状态：早发2型糖尿病），以及一个基因（Y），我们最近验证为单基因糖尿病 糖尿病基因，在 8 名不相关的 JOD 患者中鉴定出 7 个新突变。首先，我们将使用人类胚胎干细胞衍生的 be-cell 平台扩展表征基因 X 的功能数据。其次，我们将使用最先进的表型模型、患者特异性诱导多能干细胞（hiPSC）和基因组编辑的人类胚胎干细胞（hESC）来研究基因 Y 的特定功能。后者将分化为胰腺 β 细胞以进行详细的功能分析。第三，我们将使用 iCRISPR 平台通过 hESC 中的基因组编辑来创建这些基因中新发现的突变。因此，我们将开发一个完整的患者特异性细胞库，用于详细的细胞研究，旨在 i) 在培养皿中建立糖尿病模型，以确定疾病的分子基础，ii) 解决新的治疗可能性，以及 iii) 定义内分泌胰腺发育以及 β 细胞功能和维持所必需的完整分子网络。这项研究将为糖尿病治疗中真正的“个性化医疗”奠定基础。