Professor University of North Carolina, Chapel Hill Chapel Hill, North Carolina, United States
Abstract: The transition at birth, marked by increased circulatory demands and rapid growth, necessitates extensive remodeling of the heart’s structure, function, and metabolism. This transformation requires precise spatial and temporal coordination among diverse cardiac cell types, yet the intrinsic and extrinsic regulatory mechanisms driving these changes remain incompletely understood. Here, we generated a single-cell-resolution temporal and spatial atlas of postnatal hearts by coupling time-course single-nucleus RNA sequencing with in situ single-cell spatial imaging analysis. Our integrative approach not only provided a spatial map of the postnatal heart’s transcriptome at the single-cell level but also unveiled the dynamic of localized intrinsic and extrinsic regulatory mechanisms driving postnatal heart development. To further interrogate intrinsic and extrinsic regulators’ function within the native cardiac environment in a high-throughput manner, we developed an in vivo Probe-based Indel-detectable Perturb-seq (PIP-seq) platform. Applying PIP-seq to postnatal cardiomyocyte maturation uncovered key regulatory nodal points underlying postnatal cardiomyocyte maturation. Through these efforts, we have not only created a single-cell-resolution temporal and spatial atlas of postnatal hearts but also demonstrated the functional importance of cell-cell communication during postnatal heart development. Importantly, our PIP-seq opens up new possibilities for research, no longer limiting to a single gene but allowing for exploring a network of genes’ function without sacrificing in vivo physiological context.
