HNF1A in human endocrine cell development and function

HNF1A 在人类内分泌细胞发育和功能中的作用

• 批准号: 10646185
• 负责人: PAUL J GADUE
• 金额: $ 51.89万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-20 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10646185
• 关键词: Affect; Age; Alpha Cell; Animals; Autoimmune Diseases; Beta Cell; Biological Assay; Biological Models; CRISPR/Cas technology; Cell Line; Cell Separation; Cell physiology; Cells; Cellular Stress; Cellular biology; Child; Classification; Code; Complement; Complex; Cre driver; Data Set; Defect; Development; Diabetes Mellitus; Diagnosis; Disease; Down-Regulation; Endocrine; Gene Expression Profiling; Genes; Genetic; Genome engineering; Genomics; Glycolysis; Goals; Heterozygote; High Prevalence; Human; Hypoxia; In Vitro; Individual; Inherited; Insulin; Insulin-Dependent Diabetes Mellitus; Islets of Langerhans; Link; LoxP-flanked allele; Metabolic; Metabolic Diseases; Mitochondria; Modeling; Mouse Strains; Mus; Mutate; Mutation; Non-Insulin-Dependent Diabetes Mellitus; Patients; Penetrance; Phenotype; Pluripotent Stem Cells; Proto-Oncogene Proteins c-akt; Rare Diseases; Regulation; Rodent; Rodent Model; Role; Signal Transduction; Untranslated RNA; Variant; Xenograft Model; cohort; diabetic; diabetic patient; differentiation protocol; endocrine pancreas development; exhaustion; experimental study; genome wide association study; genome-wide; human disease; human embryonic stem cell line; human model; human pluripotent stem cell; human stem cells; in vitro Model; in vivo; insulin secretion; maturity onset diabetes of the young; metabolic phenotype; mouse genome; mouse model; mutant; novel; overexpression; pancreas development; response; stem; stem cell model; stem cells; stressor; transcription factor; type I and type II diabetes; virtual

Diabetes is a complex disorder with three main subtypes, type I, type II and monogenic. Type I is an autoimmune disorder characterized by loss of the insulin secreting beta cell. Type II is metabolic disorder characterized by poor insulin responsiveness and eventual beta cell exhaustion and loss. Monogenic forms are caused by mutations in a number of genes that are generally important in beta cell development and/or function. This proposal will focus on the study on HNF1A, a transcription factor that when mutated leads to the most common cause on monogenic diabetes. In addition, HNF1A has been linked to both type I and type II diabetes in GWAS studies. Mouse models of HNF1A deficiency are available, but unfortunately do not appropriately mimic the human disease. Therefore, use of a human model system is important. We will use the human pluripotent stem cell model to study the role of HNF1A in both endocrine cell development and function, utilizing CRISPR-Cas9 based genome engineering to generate isogenic mutant lines. Robust in vitro differentiation protocols exist and will be used to generate beta cells in the human stem cell model. In preliminary studies, we have uncovered human specific HNF1A targets that have been validated in a dataset from an individual with monogenic diabetes due to a HNF1A mutation. We have also discovered phenotypes in metabolic functions in HNF1A mutant stem cell derived beta cells, offering a potential link between the association of HNF1A with type II and possibly type I diabetes. Lastly, one of the human specific targets of HNF1A is a long non-coding RNA that when deleted in stem cells, mimics some of the metabolic phenotypes seen in the HNF1A mutants. These studies will help define the developmental and functional requirements of HNF1A in pancreatic endocrine cells and have the potential to directly impact the treatment of not only rare monogenic forms of diabetes but also type I and type II diabetes.

糖尿病是一种复杂的疾病，主要有三种亚型：I型、II型和单基因型。I型是一种自身免疫性疾病，其特征是分泌胰岛素的β细胞缺失。II型是一种代谢紊乱，以胰岛素反应性差和最终的β细胞耗竭和损失为特征。单基因形式是由一些基因的突变引起的，这些基因通常在细胞发育和/或功能中很重要。本提案将重点研究HNF1A，这是一种转录因子，当突变时导致单基因糖尿病最常见的原因。此外，在GWAS研究中，HNF1A与I型和II型糖尿病都有关联。HNF1A缺乏的小鼠模型是可用的，但不幸的是不能适当地模拟人类疾病。因此，使用人类模型系统很重要。我们将利用人多能干细胞模型研究HNF1A在内分泌细胞发育和功能中的作用，利用基于CRISPR-Cas9的基因组工程产生等基因突变系。稳健的体外分化方案存在，并将用于在人类干细胞模型中产生β细胞。在初步研究中，我们发现了人类特异性的HNF1A靶点，这些靶点已在来自HNF1A突变引起的单基因糖尿病患者的数据集中得到验证。我们还发现了HNF1A突变干细胞衍生β细胞代谢功能的表型，提供了HNF1A与II型和可能的I型糖尿病之间的潜在联系。最后，HNF1A的人类特异性靶点之一是一个长链非编码RNA，当在干细胞中删除时，模仿HNF1A突变体中看到的一些代谢表型。这些研究将有助于确定HNF1A在胰腺内分泌细胞中的发育和功能需求，并有可能直接影响罕见的单基因糖尿病以及I型和II型糖尿病的治疗。

项目成果

Sexually dimorphic roles for the type 2 diabetes-associated C2cd4b gene in murine glucose homeostasis.

• DOI: 10.1007/s00125-020-05350-x
• 发表时间: 2021-04
• 期刊: Diabetologia
• 影响因子: 8.2
• 作者: Mousavy Gharavy SN;Owen BM;Millership SJ;Chabosseau P;Pizza G;Martinez-Sanchez A;Tasoez E;Georgiadou E;Hu M;Fine NHF;Jacobson DA;Dickerson MT;Idevall-Hagren O;Montoya A;Kramer H;Mehta Z;Withers DJ;Ninov N;Gadue PJ;Cardenas-Diaz FL;Cruciani-Guglielmacci C;Magnan C;Ibberson M;Leclerc I;Voz M;Rutter GA
• 通讯作者: Rutter GA

Genome Engineering Human ESCs or iPSCs with Cytosine and Adenine Base Editors.

使用胞嘧啶和腺嘌呤碱基编辑器对人类 ESC 或 iPSC 进行基因组工程。

• DOI: 10.1007/7651_2022_461
• 发表时间: 2022
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Pavani,Giulia;Klein,JoshuaG;French,DeborahL;Gadue,Paul
• 通讯作者: Gadue,Paul

Generation of a double insulin and somatostatin reporter line, SCSe001-A-3, for the advancement of stem cell-derived pancreatic islets.

• DOI: 10.1016/j.scr.2020.102112
• 发表时间: 2020-12-08
• 期刊: Stem cell research
• 影响因子: 1.2
• 作者: Leavens KF;Liao CM;Gagne AL;Kishore S;Cardenas-Diaz FL;French DL;Gadue P
• 通讯作者: Gadue P

Genome Editing Human Pluripotent Stem Cells to Model β-Cell Disease and Unmask Novel Genetic Modifiers.

• DOI: 10.3389/fendo.2021.682625
• 发表时间: 2021
• 期刊: Frontiers in endocrinology
• 影响因子: 5.2
• 作者: George MN;Leavens KF;Gadue P
• 通讯作者: Gadue P

Chromatin 3D interaction analysis of the STARD10 locus unveils FCHSD2 as a regulator of insulin secretion.

Stard10基因座的染色质3D相互作用分析揭示了FCHSD2作为胰岛素分泌的调节剂。

• DOI: 10.1016/j.celrep.2021.108703
• 发表时间: 2021-02-02
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Hu M;Cebola I;Carrat G;Jiang S;Nawaz S;Khamis A;Canouil M;Froguel P;Schulte A;Solimena M;Ibberson M;Marchetti P;Cardenas-Diaz FL;Gadue PJ;Hastoy B;Almeida-Souza L;McMahon H;Rutter GA
• 通讯作者: Rutter GA