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

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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.