Dynamic regulation of chromatin accessibility during melanocyte stem cell activation

Abstract Melanocyte stem cells (McSCs) of the hair follicle are necessary for hair pigmentation and can serve as melanoma cells of origin when harboring cancer‐driving mutations. McSCs can be released from quiescence, activated, and undergo differentiation into pigment‐producing melanocytes during the hair cycle or due to environmental stimuli, such as ultraviolet‐B (UVB) exposure. However, our current understanding of the mechanisms regulating McSC stemness, activation, and differentiation remains limited. Here, to capture the differing possible states in which murine McSCs can exist, we sorted melanocyte nuclei from quiescent (telogen) skin, skin actively producing hair shafts (anagen), and skin exposed to UVB. With these sorted nuclei, we then utilized single‐nucleus assay for transposase‐accessible chromatin with high‐throughput sequencing (snATAC‐seq) and characterized three melanocyte lineages: quiescent McSCs (qMcSCs), activated McSCs (aMcSCs), and differentiated melanocytes (dMCs) that co‐exist in all three skin conditions. Furthermore, we successfully identified differentially accessible genes and enriched transcription factor binding motifs for each melanocyte lineage. Our findings reveal potential gene regulators that determine these melanocyte cell states and provide new insights into how aMcSC chromatin states are regulated differently under divergent intrinsic and extrinsic cues. We also provide a publicly available online tool with a user‐friendly interface to explore this comprehensive dataset, which will provide a resource for further studies on McSC regulation upon natural or UVB‐mediated stem cell activation.