Cells and gene expression programs in the adult human heart

Monika Litvinukova; Carlos Talavera-Lopez; Henrike Maatz; Daniel Reichart; Catherine L. Worth; Eric L. Lindberg; Masatoshi Kanda; Krzysztof Polanski; Eirini S. Fasouli; Sara Samari; Kenny Roberts; Elizabeth Tuck; Matthias Heinig; Daniel DeLaughter; Barbara McDonough; Hiroko Wakimoto; Joshua M. Gorham; Emily Nadelmann; Krishnaa T. Mahbubani; Kourosh Saeb-Parsy; Giannino Patone; Joseph J Boyle; Hongbo Zhang; Hao Zhang; Anissa Viveiros; Gavin Oudit; Omer Bayraktar; J. G. Seidman; Christine Seidman; Michela Noseda; Norbert Hubner; Sarah A. Teichmann

3 . CC-BY-NC 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.03.024075 doi: bioRxiv preprint While previous studies using a combination of conventional bulk genomics and microscopy have hinted at the cellular complexity of the myocardium, limitations of these techniques have allowed definition of very few distinct cell populations 4 . Bulk RNA-seq analysis is unable to assign gene expression to defined cell subpopulations, light microscopy fails to define features beyond morphology of cell subpopulations and immunostainings are limited to the analysis of few markers at once. Moreover, the large size of cardiomyocytes (length/width:~100/25µM) limits the unbiased capture of single cells requiring analyses of single nuclei transcriptomics to ensure a comprehensive approach. Here, we present a broad transcriptomic census of multiple regions of the adult human heart. We profiled RNA expression of both single cells and nuclei, capturing them from six distinct cardiac anatomical regions. We also analysed the spatial distribution of selected cell populations using multiplex smFISH imaging with RNAscope probes. Our anatomically defined resolved adult human heart cell census provides a reference framework for studies directed towards understanding the cellular and molecular drivers that enable functional plasticity in response to varying physiological conditions in the normal heart, and will inform the heart's responses to disease.