A HUMAN IPSC-BASED ORGANOID PLATFORM FOR STUDYING MATERNAL HYPERGLYCEMIA-INDUCED CONGENITAL HEART DEFECTS

基于人体 IPSC 的类器官平台，用于研究母亲高血糖引起的先天性心脏缺陷

• 批准号: 10752276
• 负责人: Javier Contreras
• 金额: $ 4.13万
• 依托单位: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
• 依托单位国家: 美国
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-02-01 至 2029-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10752276
• 关键词: 3-Dimensional; Address; Adherent Culture; Adult; Affect; Animals; Apoptosis; Biological; Biological Assay; Blood Glucose; Cardiac; Cardiac Myocytes; Cardiovascular system; Cell Culture Techniques; Cell Differentiation process; Cell Lineage; Cell Maturation; Cells; Cellular Metabolic Process; Congenital Abnormality; Congenital Heart Defects; Culture Media; Development; Diabetic mother; Differentiated Gene; Disease; Dose; Embryonic Development; Endothelium; Environment; Environmental Risk Factor; Fluorescence; Gene Expression; Gene Expression Profiling; Generations; Genes; Genetic Risk; Gestational Diabetes; Glucose; Goals; Heart; Heart Abnormalities; Heterogeneity; Human; Hyperglycemia; Immunofluorescence Immunologic; Impairment; Incidence; Infant Mortality; Knowledge; Live Birth; Measures; Mesoderm; Mesoderm Cell; Metabolic; Metabolism; Mitochondria; Modeling; Mus; Myocardial; Organ; Organoids; Oxidative Stress; Pathogenesis; Phenotype; Physiological; Physiology; Population; Pregnancy; Pregnant Women; Production; Reactive Oxygen Species; Reporting; Respiration; Risk; Sampling; Signal Transduction; Specific qualifier value; Stains; Stem Cell Development; Structure; Supplementation; Translations; WNT Signaling Pathway; cardiogenesis; cell determination; cell injury; cell type; congenital heart disorder; differential expression; differentiation protocol; heart cell; high risk; human model; human stem cells; human tissue; induced pluripotent stem cell; insight; interest; maternal diabetes; maternal hyperglycemia; mitochondrial metabolism; mortality; mouse model; offspring; oxidative damage; response; single-cell RNA sequencing; stem cell differentiation; stem cells; therapeutic evaluation; transcriptome sequencing; transcriptomics; two-dimensional

Congenital heart defects (CHD) are the most common type of birth defect, with cardiac malformation resulting from abnormal heat development contributed by genetic and/or environmental risk factors. The association of maternal diabetes with increased offspring CHD incidence is proposed to be attributed to factors including glucose imbalance and metabolic dysregulation. Murine studies on hyperglycemia have been beneficial in uncovering the phenotypic effects of maternal hyperglycemia on embryonic development, with reports that high glucose conditions are associated with general suppression of WNT signaling, apoptosis, and excessive reactive oxygen species (ROS) production. However, little is known about the effects of hyperglycemia on human stem cells in regard to cardiomyocyte determination, metabolism, and functionality. Additionally, there is little reports of these alterations using induced pluripotent stemcells (iPSCs), which provide an opportunityto observe cardiac progenitor cell (CPC) heterogeneity, multicellular crosstalk, and differentiated cell functionality at key development timepoints. We hypothesize that high glucose environments interfere with normal cardiac differentiation to cause altered cell lineage determination in iPSCs and iPSC-derived cardiomyocytes (iPSC- CMs). We propose use of immunofluorescence assays and staining to quantify mitochondrial metabolism, mitochondrial ROS production, and apoptosis marker amounts. Our goal is to investigate the impact of high glucose on human cardiomyocyte differentiation and crosstalk signaling abnormalities during 3D stem cell differentiation that could lead to impaired cardiac development. We will investigate differentially expressed genes at the single-cell resolutionin early and late differentiation in responseto high glucose doses in iPSC populations. Should our transcriptional profiling comparisons yield iPSC and iPSC-CM expression differences, the resulting phenotypes would provide insight into cardiac cell determination during hyperglycemic conditions. Understanding the hyperglycemia-response mechanisms is important in the onset of maternal hyperglycemia- associated CHD. These mechanisms would be further investigated through looking at cell lineage population composition in organoids and analyze response per lineage to narrow down what structures of the mature heart are affected.

先天性心脏缺陷（CHD）是最常见的先天缺陷类型，导致心脏畸形 由遗传和/或环境风险因素造成的异常热量发展。协会 提议随着后代冠心病发病率增加的母亲糖尿病归因于包括 葡萄糖失衡和代谢失调。关于高血糖的鼠研究对 发现母体高血糖对胚胎发育的表型作用，有报道称 葡萄糖条件与一般抑制Wnt信号传导，凋亡和反应性过多有关 氧（ROS）产生。但是，对于高血糖对人茎的影响知之甚少 关于心肌细胞确定，代谢和功能的细胞。此外，几乎没有报告 使用诱导的多能干细胞（IPSC）进行的这些变化，这提供了观察心脏的机会 祖细胞（CPC）异质性，多细胞串扰和钥匙处分化的细胞功能 开发时间点。我们假设高葡萄糖环境会干扰正常的心脏 在IPSC和IPSC衍生的心肌细胞中导致细胞谱系测定改变的分化（IPSC- CMS）。我们建议使用免疫荧光测定和染色来量化线粒体代谢， 线粒体ROS产生和凋亡标记量。我们的目标是调查高 3D干细胞期间人类心肌细胞分化和串扰信号异常的葡萄糖 差异化可能导致心脏发展受损。我们将研究差异表达的基因 在IPSC种群中，在Responseto高葡萄糖剂量的早期和晚分化的单细胞分辨率下。 我们的转录分析比较是否会产生IPSC和IPSC-CM表达差异，结果 表型将在高血糖条件下对心脏细胞测定的见解。 了解高血糖 - 响应机制在母体高血糖的发作中很重要 相关的冠心。这些机制将通过查看细胞谱系种群进一步研究 器官中的组成并分析每个谱系的反应，以缩小成熟心脏的结构 受到影响。