Using a Rare Disease Model to Explore the Role of FGF21 in Insulin Resistance

使用罕见疾病模型探索 FGF21 在胰岛素抵抗中的作用

基本信息

批准号：
10216013
负责人：
Stephen Isaac Stone
金额：
$ 16.23万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10216013
关键词：
APOA5 gene Adipocytes Adipose tissue Animals Area Biological Assay Biological Markers Biological Models Biology Biometry Blood Body Composition Body Size Body Weight decreased Brown Fat CRISPR/Cas technology Case Study Cell Line Childhood Clinical Communication Research Complex Data Diabetes Mellitus Disease Disease model Down-Regulation Endocrinology Environment Environmental Risk Factor Epidemic Fatty Liver Fatty acid glycerol esters Fellowship Female Adolescents Fibroblast Growth Factor Fibroblast Growth Factor Receptor 1 Fibrosis Foundations Funding Future GLUT 4 protein Genes Genetic Genomic approach Genomics Glucose Goals Growth Healthcare Hepatic Hormones Human Hyperglycemia Hyperinsulinism IGF1 gene Impairment In Vitro Individual Inflammation Insulin Insulin Resistance Investigation K-Series Research Career Programs Ketone Bodies Knock-in Mouse Knowledge Lead Link Lipodystrophy Lipolysis Literature Liver Liver diseases Measurement Measures Mediating Mentors Metabolic Metabolic syndrome Molecular Molecular Biology Mus Mutant Strains Mice Mutation Non-Insulin-Dependent Diabetes Mellitus Obesity Parents Pathogenicity Patients Pattern Pediatric Hospitals Pediatrics Pharmacology Phenotype Physicians Play Principal Investigator Rare Diseases Recombinant Fibroblast Growth Factor Research Research Ethics Residencies Resistance Role SLC2A1 gene Scientist Serum Siblings Signal Transduction Starvation Syndrome Testing Therapeutic Agents Training Transgenic Mice Transgenic Organisms Triglycerides United States Universities Variant Washington Work adipocyte differentiation apolipoprotein C-II autocrine career development dietary drug testing exome sequencing experience fibroblast growth factor 21 functional genomics genetic approach genome editing glucose uptake improved in vivo induced pluripotent stem cell insulin sensitivity laboratory experience lipid metabolism mouse model mutant non-alcoholic fatty liver disease novel proband public health relevance rare condition receptor response skills transcription factor

项目摘要

Project Summary/Abstract Insulin-mediated pseudoacromegaly (IMPA) is a rare insulin resistance syndrome that is distinct from other insulin resistance syndromes, as it leads to overgrowth and tall stature. Despite the handful of reported cases in the literature, there is no molecular or genetic explanation for this disease. Exome sequencing on an adolescent female with IMPA and her sibling and parents revealed that she carries two rare, potentially deleterious, variants in Fibroblast Growth Factor Receptor 1 (FGFR1) and 𝛽𝛽-Klotho (KLB), which form the receptor complex for Fibroblast Growth Factor 21 (FGF21). We hypothesize that these mutations explain the extreme insulin resistance, tall stature, and lipodystrophy-like phenotype seen in IMPA. To test the function of these FGFR1 and KLB variants in vivo, we used CRISPR/Cas9 genome editing to create a transgenic knock-in mouse line with these variants and corrected the FGFR1 and KLB mutations in an induced pluripotent stem cell (iPSC) line isolated from our proband with IMPA. In this study, we are proposing the following specific aims. In Aim 1, we will determine whether the metabolic and physical phenotype of IMPA mice (with mutations in Fgfr1/Klb) is a consequence of impaired FGF21 signaling. In Aim 2, we will determine if the insulin resistance seen in IMPA is due to impaired adipocyte function. In Aim 3, we will determine the mechanism of fatty liver disease in IMPA mutant mice. In addition to forming a functional genetic approach toward defining a novel insulin resistance syndrome, this four-year proposal outlines a comprehensive strategy for the principal investigator's career development in academic pediatric endocrinology. This strategy logically builds on the principal investigator's previous research experience and clinical training. After obtaining his MD, the principal investigator completed his residency training in general pediatrics, and fellowship in pediatric endocrinology. This proposal now focuses on expanding his scientific skills by attaining additional knowledge and practical research experience in studying a transgenic mouse model of a rare human insulin resistance syndrome. The career development goals will be achieved through a multi-faceted approach involving mentoring by Dr. David Ornitz (fibroblast growth factors and transgenic mouse models) and Dr. Fumihiko Urano (metabolic testing in mice), hands-on laboratory experience, scientific investigation, and training in genetics/genomics, biostatistics, scientific communication, and research ethics. This work will take place in a unique training environment comprised of complementary experiences at Washington University and St. Louis Children's Hospital. Successful completion of this career development award will result in the principal investigator's transition to an independent physician-scientist. Additionally, this work will contribute to a better understanding of the role that FGF21 plays in human insulin resistance, which will provide the foundation for the principal investigator to apply for future R01-level funding.

项目摘要/摘要胰岛素介导的假性肿瘤（IMPA）是一种罕见的胰岛素抵抗综合征，与其他不同胰岛素抵抗综合征会导致过度生长和高雕像。尽管有少数报告的案件文献对这种疾病没有分子或遗传解释。在青少年上进行外显子组测序女性与伊京（Impa）和她的兄弟姐妹和父母一起透露，她携带了两个罕见的，可能被删除的变体在成纤维细胞生长因子受体1（FGFR1）和𝛽𝛽-Klotho（KLB）中成纤维细胞生长因子21（FGF21）。我们假设这些突变解释了极端胰岛素在IMPA中看到的抗性，高统计和类似脂肪营养不良的表型。测试这些FGFR1和 KLB变体在体内，我们使用CRISPR/CAS9基因组编辑来创建带有转基因的型鼠标线这些变体并纠正了诱导多能干细胞（IPSC）系中的FGFR1和KLB突变用impa与我们的概率隔离。在这项研究中，我们提出了以下特定目标。在AIM 1中，我们将确定IMPA小鼠的代谢和物理表型（FGFR1/KLB中的突变）是否是 FGF21信号传导受损的结果。在AIM 2中，我们将确定IMPA中看到的胰岛素抵抗是否是由于脂肪细胞功能受损。在AIM 3中，我们将确定IMPA脂肪肝病的机制突变小鼠。除了形成定义新型胰岛素抵抗综合征的功能遗传方法外，四年提案概述了首席研究者职业发展的全面策略学术儿科内分泌学。从逻辑上讲，该策略是基于主要研究者先前的研究经验和临床培训。获得医学博士后，首席研究员完成了他的长期培训一般儿科和小儿内分泌学研究金。现在，该提议着重于扩大他的科学技能通过获得其他知识和实践研究经验来研究转基因稀有人类胰岛素抵抗综合征的小鼠模型。将实现职业发展目标通过David Ornitz博士的心理涉及的多方面方法（成纤维细胞增长因素和转基因小鼠模型）和Fumihiko Urano博士（小鼠的代谢测试），动手实验室经验，科学研究和遗传学/基因组学，生物统计学，科学交流和研究的培训伦理。这项工作将在独特的培训环境中进行，完成了完整的经验华盛顿大学和圣路易斯儿童医院。成功完成了这一职业发展奖励将导致首席研究人员向独立的身体科学家过渡。另外，这个工作将有助于更好地理解FGF21在人类胰岛素抵抗中所发挥的作用，这将为主要研究人员申请未来R01级资金的基础。