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

基本信息

批准号：
10242231
负责人：
Edward Lau
金额：
$ 24.45万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
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/10242231
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

项目摘要

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.

人心积极释放蛋白质使者进入细胞外空间。最近的工作暗示这些分泌的蛋白质（称为心脏代动力学）可以介导至重要角色的细胞串扰在心肌病和心力衰竭的发展中。虽然几种古典心脏因子，例如亚钠肽已经广泛表征分类人类心脏细胞分泌的蛋白质。使用蛋白质组学和人类诱导的多能干细胞（HIPSC）模型，我们最近通过从hipsc衍生的心肌细胞，心脏成纤维细胞中识别和对比分泌分子和内皮细胞。结果表明，释放了大量候选心脏。来自每种细胞类型，包括许多分泌的蛋白质，在心血管中具有未表征的功能系统。此外，我们发现携带扩张的心肌细胞的分泌模式发生了广泛的变化正常细胞上的心肌病因果变异。在这些发现的基础上，我们在R00阶段的目标现在要（i）：确定在成纤维细胞串扰中起作用的心肌细胞分泌的蛋白质并预测他们调节的生物过程；（ii）优先考虑与疾病相关的心脏动物并验证其作用在受体成纤维细胞基因表达上。为了实现这些目标，我们将采用作为我们的HIPSC模型建立的计算，单细胞测序和蛋白质组学策略作为我们的基础。如果成功，拟议的研究有可能发现许多新颖的分泌蛋白质及其在人类心脏细胞中的功能，并阐明了细胞通信在心肌病的发展和进展。