Generation of Quiescent Cardiac Fibroblasts From Human Induced Pluripotent Stem Cells for In Vitro Modeling of Cardiac Fibrosis
Generation of Quiescent Cardiac Fibroblasts From Human Induced Pluripotent Stem Cells for In Vitro Modeling of Cardiac Fibrosis
复制标题
人诱导多能干细胞诱导静止期心肌成纤维细胞的建立及心肌纤维化模型的建立
DOI：
10.1161/circresaha.119.315491
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发表时间：
2019-08-16
影响因子：
20.1
通讯作者：
Wu, Joseph C.
中科院分区：
文献类型：
--
作者：
Zhang, Had;Tian, Lei;Wu, Joseph C.
基本原理：活化的成纤维细胞是响应于损伤而分泌过量的细胞外基质的主要细胞类型，导致病理性纤维化并导致器官衰竭。然而，由于组织特异性成纤维细胞的特征定义不明确和不可用，因此无法获得有效的抗纤维化治疗方案。单细胞RNA测序的最新进展填补了此类知识空白，能够描绘发育轨迹并识别不同器官中组织特异性成纤维细胞的调节途径。目的：本研究旨在使用最近报道的小鼠单细胞RNA测序图谱来定义组织特异性成纤维细胞的转录组谱，并开发一种稳健的化学定义的方案，以从人诱导多能干细胞中获得心脏成纤维细胞（CF），用于心脏纤维化的体外建模和药物筛选。方法和结果：通过分析来自10个选定的小鼠组织的成纤维细胞的单细胞转录组谱，我们确定了不同的组织特异性标记基因，包括定义心脏、肺、气管和膀胱中成纤维细胞身份的转录因子。我们还确定，CF在大的心外膜血统。因此，我们开发了一个强大的化学定义的协议，从人类诱导多能干细胞产生CF。功能研究证实，iPSC衍生的CF保留了静止表型，并且在转录、细胞和功能水平上与初级CF高度相似。我们证明了这种基于细胞的平台对促纤维化和抗纤维化药物都敏感。最后，我们发现，通过心房/脑利钠肽-利钠肽受体-1途径，人诱导多能干细胞衍生的心肌细胞和CF之间的串扰与抑制纤维化有关。结论：这项研究揭示了独特的基因签名，定义成纤维细胞的组织特异性身份。来源于人诱导多能干细胞的真正静止的CFs可以作为一个忠实的体外平台，以更好地了解心脏纤维化的潜在机制和筛选抗纤维化药物。
Rationale: Activated fibroblasts are the major cell type that secretes excessive extracellular matrix in response to injury, contributing to pathological fibrosis and leading to organ failure. Effective anti-fibrotic therapeutic solutions, however, are not available due to the poorly defined characteristics and unavailability of tissue-specific fibroblasts. Recent advances in single-cell RNA-sequencing fill such gaps of knowledge by enabling delineation of the developmental trajectories and identification of regulatory pathways of tissue-specific fibroblasts among different organs. Objective: This study aims to define the transcriptome profiles of tissue-specific fibroblasts using recently reported mouse single-cell RNA-sequencing atlas and to develop a robust chemically defined protocol to derive cardiac fibroblasts (CFs) from human induced pluripotent stem cells for in vitro modeling of cardiac fibrosis and drug screening. Methods and Results: By analyzing the single-cell transcriptome profiles of fibroblasts from 10 selected mouse tissues, we identified distinct tissue-specific signature genes, including transcription factors that define the identities of fibroblasts in the heart, lungs, trachea, and bladder. We also determined that CFs in large are of the epicardial lineage. We thus developed a robust chemically defined protocol that generates CFs from human induced pluripotent stem cells. Functional studies confirmed that iPSC-derived CFs preserved a quiescent phenotype and highly resembled primary CFs at the transcriptional, cellular, and functional levels. We demonstrated that this cell-based platform is sensitive to both pro- and anti-fibrosis drugs. Finally, we showed that crosstalk between human induced pluripotent stem cell-derived cardiomyocytes and CFs via the atrial/brain natriuretic peptide-natriuretic peptide receptor-1 pathway is implicated in suppressing fibrogenesis. Conclusions: This study uncovers unique gene signatures that define tissue-specific identities of fibroblasts. The bona fide quiescent CFs derived from human induced pluripotent stem cells can serve as a faithful in vitro platform to better understand the underlying mechanisms of cardiac fibrosis and to screen anti-fibrotic drugs.