Multi-Omics Approach to Identify Cardiokines in Human iPSC Models

识别人类 iPSC 模型中心肌因子的多组学方法

• 批准号: 10204224
• 负责人: Edward Lau
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-20 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10204224
• 关键词: Address; Adopted; Biological Assay; Biological Process; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cardiovascular system; Catalogs; Cell model; Cells; Communication; Computational Technique; Contracts; Data; Development; Dilated Cardiomyopathy; Disease; Disease Pathway; Endothelial Cells; Extracellular Space; Fibroblasts; Foundations; Gene Expression; Genes; Goals; Health; Heart; Heart Diseases; Heart failure; Human; In Vitro; Ligands; Light; Maps; Mass Spectrum Analysis; Mediating; Modeling; Multiomic Data; Muscle Cells; Natriuretic Peptides; Normal Cell; Nucleic Acids; Pattern; Phase; Physiological; Play; Process; Proteins; Proteomics; Research; Role; Signal Transduction; Work; causal variant; cell type; experimental study; extracellular; induced pluripotent stem cell; interest; multiple omics; novel; protein function; receptor; single cell sequencing; single-cell RNA sequencing; stem cell model

The human heart actively releases protein messengers into the extracellular space. Recent work suggests that these secreted proteins, termed cardiokines, can mediate cellular crosstalk that plays important roles in the development of cardiomyopathies and heart failure. Although several classical cardiokines such as the natriuretic peptides have been extensively characterized, there have been few efforts to systematically catalog the proteins secreted by human cardiac cells. Using proteomics and human induced pluripotent stem cell (hiPSC) models, we recently created a draft map of the human cardiac secretome through identifying and contrasting secreted molecules from hiPSC-derived cardiomyocytes, cardiac fibroblasts, and endothelial cells. The results revealed a surprisingly large number of candidate cardiokines released from each cell type including many secreted proteins with uncharacterized function in the cardiovascular system. Moreover, we found broad changes in the secretome patterns of cardiomyocytes carrying dilated cardiomyopathy causal variants over normal cells. Building on these findings, our aims in the R00 phase are now to (i): identify cardiomyocyte secreted proteins that function in fibroblast crosstalk and predict the biological processes they regulate; and (ii) prioritize disease-relevant cardiokines and validate their effects on recipient fibroblast gene expression. To achieve these aims, we will employ a combination of computational, single-cell sequencing, and proteomics strategies building on hiPSC models as our foundation. If successful, the proposed research has the potential to uncover a number of novel secreted proteins and their function in human cardiac cells, and shed light on the role of cellular communications in the development and progression of cardiomyopathies.

人类心脏主动释放蛋白质信使到细胞外空间。最近的研究表明 这些分泌的蛋白质，称为心脏因子，可以介导细胞串扰，发挥重要作用， 心肌病和心力衰竭的发展。虽然一些经典的心脏因子， 利钠肽已被广泛表征，但很少有人系统地 对人类心脏细胞分泌的蛋白质进行分类。利用蛋白质组学和人类诱导多能 在干细胞（hiPSC）模型中，我们最近通过以下方法创建了人类心脏分泌组的草图： 鉴定和对比来自hiPSC衍生的心肌细胞，心脏成纤维细胞， 和内皮细胞。结果显示，释放的候选心血管因子数量惊人 包括许多在心血管疾病中具有未知功能的分泌蛋白质 系统此外，我们发现携带扩张型心肌细胞的分泌组模式发生了广泛的变化， 心肌病致病变异体超过正常细胞。基于这些发现，我们在R 00阶段的目标 我们现在要做的是（i）：鉴定在成纤维细胞串扰中起作用的心肌细胞分泌蛋白，并预测心肌细胞的凋亡。 它们调节的生物过程;以及（ii）优先考虑疾病相关的心脏因子并验证其效果 对受体成纤维细胞基因表达的影响。为达致这些目标，我们会采取以下措施： 计算，单细胞测序和蛋白质组学策略建立在hiPSC模型上， 基金会如果成功的话，这项研究有可能揭示一些新的分泌性的 蛋白质及其在人类心脏细胞中的功能，并阐明了细胞通讯在心脏疾病中的作用。 心肌病的发生和进展。