Dissecting the mechanisms behind biphasic beta cell disease caused by mutations in HNF1alpha

剖析 HNF1α 突变引起的双相 β 细胞疾病背后的机制

• 批准号: 10488267
• 负责人: Karla Fitzgerald Leavens
• 金额: $ 14.89万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-30 至 2025-06-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10488267
• 关键词: Acute; Advisory Committees; Age; Amino Acids; Applications Grants; Area; Automobile Driving; B-Lymphocytes; Basic Science; Beta Cell; Biological Models; Candidate Disease Gene; Cell Maturation; Cell model; Cells; Clinical; Communities; Complex; Coupled; Data; Defect; Development Plans; Diabetes Mellitus; Disease; Down-Regulation; Educational workshop; Environment; Exhibits; Foundations; Functional disorder; Future; Gene Mutation; Generations; Genes; Glucose; Glycolysis; Goals; Human; Hyperinsulinism; Individual; Insulin; Insulin deficiency; Insulin-Dependent Diabetes Mellitus; Knowledge; Learning; Link; Measures; Mediator of activation protein; Mendelian disorder; Mentors; Metabolic; Metabolism; Methods; Modeling; Mus; Mutation; NADH; NADP; Non-Insulin-Dependent Diabetes Mellitus; Nutrient; Pathogenesis; Pathway interactions; Patients; Pediatric Hospitals; Pennsylvania; Persistent Hyperinsulinemia Hypoglycemia of Infancy; Pharmaceutical Preparations; Philadelphia; Process; Production; Publications; Publishing; RNA Interference; Regulation; Research; Role; Signal Pathway; Structure of beta Cell of islet; Techniques; Testing; Training; Translational Research; Universities; Work; Xenograft procedure; base; career development; directed differentiation; disease-causing mutation; energy balance; experimental study; genome editing; glucose metabolism; human stem cells; insulin secretion; knowledge base; loss of function; loss of function mutation; medical schools; mutant; novel; response; skills; stem cells; transcription factor; transplant model

Project Summary In order for the b-cell to precisely secrete insulin in response to glucose, glucose metabolism to ATP is tightly coupled to insulin secretion through the ATP-sensitive K (KATP) channel. Any disruption along this pathway leads to b-cell dysfunction and ensuing diabetes or, more rarely, congenital hyperinsulinism. While these diseases mostly arise from multiple factors, some cases are caused by single gene mutations, termed monogenic diseases. Interestingly, the same loss-of-function heterozygous mutations in the transcription factor HNF1a can result in both hyperinsulinism and diabetes, though presenting at different ages and through unknown mechanisms. In the proposed project, we will investigate the underlying causes of both HNF1a- related diabetes and hyperinsulinism. Our prior published work and additional preliminary data show that HNF1A-deficient human stem cell-derived β-cells exhibit increased basal and decreased glucose-stimulated insulin secretion, recapitulating the clinical disease. HNF1A-deficient β-cell models exhibited significantly decreased glycolysis, suggesting decreased ATP production in response to glucose. Expression analyses revealed broad defects in HNF1A-deficient β-cells in genes regulating cellular metabolism and decreased expression of KATP channel genes. Based on these previous findings, we hypothesize that HNF1a regulates both KATP channel expression and ATP production in response to glucose. Loss of HNF1a leads to uncoupling of glucose metabolism and insulin secretion at these two points in the insulin secretion cascade, impacting basal and stimulated insulin secretion in opposing directions and resulting in hyperinsulinism and diabetes, respectively. Using a novel and manipulatable human b-cell model system combined with a variety of analytical techniques, we will rigorously test our hypotheses and advance our knowledge of b-cell dysfunction in complex disease states. My long-term goal is to have a basic and translational research lab that studies the diseases resulting from pancreatic β-cell dysfunction. My proposed K08 project will help me learn new technical methods, develop a new intellectual foundation and generate preliminary data that will be instrumental in helping me start my own independent lab. I chose my co-mentors as they have complimentary research approaches and varied areas of expertise. I have a career development plan which relies on my extensive mentoring relationships, including regular interaction with my co-mentors and my advisory committee, which contains individuals within the local and national research community with varying areas of expertise. I have developed a plan of training which takes full advantage of the collaborative research environment at the Children’s Hospital of Philadelphia and University of Pennsylvania School of Medicine, including participation in trainings, seminars, and workshops to advance my knowledge base and professional skills.

项目摘要 为了使b细胞响应于葡萄糖而精确地分泌胰岛素，葡萄糖代谢为ATP是必需的。 通过ATP敏感性K（KATP）通道与胰岛素分泌紧密结合。任何沿着这条路的中断 途径导致B细胞功能障碍和随后的糖尿病，或更罕见的先天性高胰岛素血症。而 这些疾病大多数是由多种因素引起的，有些病例是由单个基因突变引起的， 单基因疾病有趣的是，转录因子中相同的功能丧失杂合突变 HNF 1a可导致高胰岛素血症和糖尿病，尽管表现在不同的年龄和通过 未知的机制。在拟议的项目中，我们将调查HNF 1a- 相关的糖尿病和高胰岛素血症。我们先前发表的工作和额外的初步数据表明， HNF 1A缺陷的人干细胞衍生的β细胞表现出增加的基础和减少的葡萄糖刺激 胰岛素分泌，概括了临床疾病。HNF 1A缺陷的β细胞模型表现出显著的 糖酵解减少，表明响应于葡萄糖的ATP产生减少。表达分析 发现HNF 1A缺陷β细胞在调节细胞代谢的基因中存在广泛缺陷， KATP通道基因的表达。基于这些先前的发现，我们假设HNF 1a调节 KATP通道的表达和ATP的产生。HNF 1a的缺失导致解偶联 葡萄糖代谢和胰岛素分泌在这两个点在胰岛素分泌级联，影响 在相反方向上的基础和刺激胰岛素分泌并导致高胰岛素血症和糖尿病， 分别使用一种新颖的和可操纵的人类b细胞模型系统，结合各种分析， 技术，我们将严格测试我们的假设，并推进我们的知识的b细胞功能障碍，在复杂的 疾病状态。 我的长期目标是建立一个基础和转化研究实验室，研究由此产生的疾病， 胰腺β细胞功能障碍我提出的K 08项目将帮助我学习新的技术方法， 开发新的知识基础，并生成有助于帮助我的初步数据 开始我自己的独立实验室我选择了我的共同导师，因为他们有互补的研究方法， 不同的专业领域。我有一个职业发展计划，这取决于我的广泛指导 关系，包括与我的共同导师和咨询委员会的定期互动，其中包括 地方和国家研究界中具有不同专业领域的个人。I have developed 一项培训计划，充分利用儿童研究所的合作研究环境， 费城医院和宾夕法尼亚大学医学院，包括参加培训， 研讨会和讲习班，以提高我的知识基础和专业技能。