权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Improving cardiovascular disease modeling using human pluripotent stem cell-derived cardiac fibroblasts

使用人类多能干细胞来源的心脏成纤维细胞改善心血管疾病模型

基本信息

批准号：
10472759
负责人：
Charles Matthew Kerr
金额：
$ 3.46万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
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：
2020-09-01 至 2023-05-20
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10472759
关键词：
3-Dimensional Address Adipose tissue Adrenergic Agents Adult Animal Model Biological Models Cardiac Cardiac Myocytes Cardiovascular Diseases Cause of Death Cells Chronic Data Deposition Development Diagnosis Diffusion Disease model Effectiveness Endocardium Endothelial Cells Epicardium Extracellular Matrix Fibroblasts Genes Genetic Genetic Transcription Goals Heart Heart failure Heterogeneity Human In Vitro Infarction Inflammatory Injury Lead Metabolic Metabolism Modeling Morphology Mus Myocardial Infarction Myocardium Organoids Oxygen Patients Population Protein Secretion Protocols documentation RNA analysis Role Source Stromal Cells System Therapeutic Time Umbilical vein United States Universities Ventricular Wisconsin coronary fibrosis cytokine drug testing effectiveness evaluation fetal high risk human RNA sequencing human model human pluripotent stem cell improved in vivo innovation mortality risk mouse model progenitor regenerative therapy response self assembly single-cell RNA sequencing stem cell differentiation stem cells transcriptome sequencing transcriptomics

项目摘要

Project Summary: Annually, there are ~790,000 cases of myocardial infarction (MI) in the United States. Typically, MI progresses into heart failure where patients have a high risk of mortality within 5 years after diagnosis. While animal models provide a valuable model system of MI, interspecies differences lead to inaccurate recapitulation of human myocardium. To address this, our lab originally developed 3D human cardiac organoids through self-assembly of hPSC-CMs, human primary adult cardiac fibroblasts (adult-cFbs), endothelial cells, and stromal cells. Further, we leveraged the oxygen diffusion limitation in 3D human cardiac organoids along with chronic adrenergic stimulation to generate an organotypic model of post-MI hearts. The human cardiac infarct organoids recapitulated transcriptional, structural and functional hallmarks of post-MI myocardium. However, the use of primary, non-myocyte cell populations in our current organoids limit their potential to mimic patient-specific myocardium. To develop human isogenic cardiac organoids, we are collaborating with Dr. Sean Palecek at the University of Wisconsin-Madison to derive cardiac fibroblasts from human pluripotent stem cells (hPSC) to replace adult-cFbs in our cardiac organoid model. Dr. Palecek’s lab has developed expertise to direct hPSC differentiation into cardiac fibroblasts (hPSC-cFbs) in 2 different lineages: epicardial-derived fibroblasts (hPSC-cFb(EpiC)s) and second heart field progenitor-derived fibroblasts (hPSC- cFb(SHFP)s). While both lineages contribute to cardiac fibrosis and are functionally similar, in murine hearts, the epicardium is the predominate source of ventricular cardiac fibroblasts while a small population arise from the endocardium. In addition, the enhanced maturation may be needed for the hPSC-cFb(EpiC)s to replace human adult-cFbs, as our preliminary data that showed that prolonged culture improved cell organization of hPSC-cFb(SHFP)s in cardiac organoids when compared to that of adult-cFb organoids. The central hypothesize of this proposal is that high passage hPSC-cFb(EpiC)s will best replicate adult-cFb transcriptomics and functionality. The proposal is innovative in that, for the first time, we will identify a suitable hPSC-cFb population to replace adult-cFbs to develop an isogenic 3D organotypic model of human myocardium. Our long-term goal is to develop patient-specific cardiac organoids for in vitro disease modeling and drug testing. Accordingly, we will pursue the following two Aims: 1) Determine the effectiveness of high passage hPSC- cFb(EpiC)s to replicate the transcriptomics and functionality of adult cFbs, and 2) Determine the effectiveness of human cardiac organoids composed of high passage hPSC-cFb(EpiC)s in modeling post-MI human myocardium and responsiveness to anti-MI therapeutics. We also will perform single cell RNA-seq to examine the heterogeneity of hPSC-cFb(EpiC)s in response to our infarction protocol. Completion of this study would provide the first step towards an isogenic human myocardium model. The single cell RNA-seq studies will reveal the various roles/subpopulations of cardiac fibroblasts in post-MI human myocardium.

项目总结：美国每年约有79万例心肌梗死（MI）病例。