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

先天性心脏病是最常见的出生缺陷类型， 由遗传和/或环境风险因素引起的异常发热。联盟 母亲糖尿病与后代CHD发病率增加被认为归因于以下因素， 葡萄糖失衡和代谢失调。对高血糖症的小鼠研究有益于 揭示了母体高血糖对胚胎发育的表型影响，有报道称， 葡萄糖条件与WNT信号传导、细胞凋亡和过度反应性的一般抑制有关。 氧物种（ROS）的产生。然而，关于高血糖对人类干细胞的影响知之甚少 细胞在心肌细胞决定、代谢和功能方面的作用。此外，很少有报道称 使用诱导多能干细胞（iPSC）进行这些改变，这为观察心脏 祖细胞（CPC）异质性，多细胞串扰和分化的细胞功能是关键 发展时间点。我们假设高糖环境干扰了正常的心脏功能， 诱导分化以引起iPSC和iPSC衍生的心肌细胞（iPSC-1）中改变的细胞谱系决定。 CM）。我们建议使用免疫荧光分析和染色来量化线粒体代谢， 线粒体ROS产生和凋亡标志物量。我们的目标是调查高血压的影响 葡萄糖对人心肌细胞分化及3D干细胞过程中串扰信号异常的影响 分化，可能导致心脏发育受损。我们将研究差异表达的基因 在iPSC群体中，在早期和晚期分化中对高葡萄糖剂量的反应中的单细胞分辨率。 如果我们的转录谱比较产生iPSC和iPSC-CM表达差异，那么结果将是： 表型将提供对高血糖状况期间心脏细胞决定的深入了解。 了解高血糖反应机制对母体高血糖的发生非常重要- 相关CHD这些机制将通过观察细胞谱系群体来进一步研究 类器官中的组成，并分析每个谱系的反应，以缩小成熟心脏的结构 受到影响。

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.