Unbiased mapping of non-myocyte cell (NMC) populations in the human heart – Cellular composition & single cell transcriptome analysis considering age, anatomical location and cardiac pathologies

心脏非肌细胞（NMC）群体对心脏发育、稳态和各种心脏疾病有重要作用。因此，NMCs是创新再生策略的有趣目标。然而，在数量和体积方面，心脏中NMCs的比例及其各自的基因表达谱仍然存在争议，因为它们的分析受到细胞鉴定和分离困难的阻碍。在应用项目中，它的目的是，第一次，分析NMC组成和NMC转录组的出生后的“人”的心脏，在一个公正的方式，由国家的最先进的单细胞技术，允许规避上述问题。我们试图分析来自具有不同心脏诊断的新生儿、婴儿和成人患者的不同解剖位置的心脏活检的单细胞悬液。我们的总体目标是（I）鉴定人心脏内NMC的细胞组成，和（II）创建hNMC群体的分子基因表达图谱。这些目标将在四个工作包（WP）中解决。(WP1)使用新鲜收获的人心脏组织建立单细胞RNA测序（sc-RNAseq）技术。(WP2)Sc-RNAseq的30人心脏组织样品的定义的定位，年龄组和心脏疾病。(WP3)根据（a）心脏内的定位，（B）出生后发育年龄和（c）基础诊断，鉴定hNMC组成和基因表达谱的差异。(WP4)hNMCs中心脏成纤维细胞亚群的定位、年龄和疾病的定义我们假设，通过使用最先进的sc-RNAseq，首次可以定义人类心脏中不同NMC群体的可重复和明确区分的分子表型。因此，该项目将导致受上述变量影响的hNMC群体的详细绘图，这是开发新疗法和更好地理解某些心脏疾病的病理生理学的不可或缺的先决条件。

Cardiac non-myocyte cell (NMC) populations significantly contribute to cardiac development, homeostasis and various cardiac diseases. Thus, NMCs are interesting targets for innovative regenerative strategies. The proportion of NMCs in the heart, however, with respect to number and volume, and their respective gene expression profiles still remain controversial since their analysis has been hindered by difficulties in cell identification and isolation. In the applied project it is intended, for the first time, to analyse NMC composition and NMC transcriptomes of the postnatal “human” heart in an unbiased fashion by state-of-the-art single cell technology which allows circumventing the above mentioned problems. We seek to analyse single cell suspensions from cardiac biopsies of different anatomical locations of neonatal, infant and adult patients with different cardiac diagnosis. Our overall aim is (I) to identify the cellular composition of NMCs within the human heart and, (II) to create a molecular gene expression map for hNMC populations. These objectives will be addressed in four work packages (WP). (WP1) Establishment of the single cell RNA sequencing (sc-RNAseq) technology using freshly harvested human cardiac tissue. (WP2) Sc-RNAseq of 30 human cardiac tissue samples of defined localizations, age classes and cardiac diseases. (WP3) Identification of differences in the hNMC composition and gene expression profiles depending on (a) localization within the heart, (b) postnatal developmental age and (c) underlying diagnosis. (WP4) Definition of cardiac fibroblast subpopulations within the hNMCs with respect to localization, age and disease.We hypothesize that by using state-of-the-art sc-RNAseq, for the first time, a reproducible and clearly distinguished molecular phenotype of different NMC populations within the human heart could be defined. This project will therefore result in a detailed mapping of hNMC populations influenced by the above mentioned variables which is an indispensable prerequisite for the development of new therapies and a better pathophysiologic understanding of certain cardiac diseases.