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NFIB is a governor of epithelial–melanocyte stem cell behaviour in a shared niche

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Abstract

Adult stem cells reside in specialized niches where they receive environmental cues to maintain tissue homeostasis. In mammals, the stem cell niche within hair follicles is home to epithelial hair follicle stem cells and melanocyte stem cells, which sustain cyclical bouts of hair regeneration and pigmentation1,2,3,4. To generate pigmented hairs, synchrony is achieved such that upon initiation of a new hair cycle, stem cells of each type activate lineage commitment2,5. Dissecting the inter-stem-cell crosstalk governing this intricate coordination has been difficult, because mutations affecting one lineage often affect the other. Here we identify transcription factor NFIB as an unanticipated coordinator of stem cell behaviour. Hair follicle stem-cell-specific conditional targeting of Nfib in mice uncouples stem cell synchrony. Remarkably, this happens not by perturbing hair cycle and follicle architecture, but rather by promoting melanocyte stem cell proliferation and differentiation. The early production of melanin is restricted to melanocyte stem cells at the niche base. Melanocyte stem cells more distant from the dermal papilla are unscathed, thereby preventing hair greying typical of melanocyte stem cell differentiation mutants. Furthermore, we pinpoint KIT-ligand as a dermal papilla signal promoting melanocyte stem cell differentiation. Additionally, through chromatin-immunoprecipitation with high-throughput-sequencing and transcriptional profiling, we identify endothelin 2 (Edn2) as an NFIB target aberrantly activated in NFIB-deficient hair follicle stem cells. Ectopically induced Edn2 recapitulates NFIB-deficient phenotypes in wild-type mice. Conversely, endothelin receptor antagonists and/or KIT blocking antibodies prevent precocious melanocyte stem cell differentiation in the NFIB-deficient niche. Our findings reveal how melanocyte and hair follicle stem cell behaviours maintain reliance upon cooperative factors within the niche, and how this can be uncoupled in injury, stress and disease states.

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Figure 1: Conditional Nfib targeting in hair follicle stem cells does not perturb hair cycle or follicle architecture.
Figure 2: NFIB loss enhances melanocyte stem-cell self-renewal and perturbs melanocyte stem-cells activity in the hair follicle stem cell niche.
Figure 3: Premature transfer of pigment promotes apoptotic cell death in hair follicle stem cells in the NFIB-deficient niche.
Figure 4: RNA-seq and ChIP-seq analyses identify Edn2 as a direct NFIB-regulated gene mediating inter-stem cell crosstalk.

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Gene Expression Omnibus

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ChIP-seq data have been deposited in the Gene Expression Omnibus database under accession number GSE42900.

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Acknowledgements

We thank Y.-C. Hsu, T. Chen, B. Keyes, S. E. Williams, A. Rodriguez-Folgueras and other Fuchs laboratory members for intellectual input and suggestions; L. Polak and N. Stokes for breeding of mouse lines and conducting in utero lentiviral injections; V. J. Hearing for providing anti-DCT, TYRP1 and TYR antibodies. We also thank Rockefeller facilities: Comparative Bioscience Center (AAALAC accredited) for care and husbandry care of mice in accordance with National Institutes of Health (NIH) guidelines; Bioimaging Center for advice on image acquisition; Flow Cytometry facility for FACS sorting. E.F. is an investigator of the Howard Hughes Medical Institute. This work was supported by grants from the NIH to E.F. (R01-AR050452 and R01-AR31737) and R.M.G. (R01-HL080624), and a CAREER grant to O.E. from the National Science Foundation (DB1054964).

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C.-Y.C. performed all experiments; H.A.P. performed the ultrastructural analyses; E.G.G. and O.E. performed the bioinformatic analyses; G.G. carried out the initial characterization of NFIB expression during mouse development; R.M.G. provided the conditional Nfibfl/fl mice; E.F. supervised the project; E.F. and C.-Y.C. wrote the manuscript.

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Correspondence to Elaine Fuchs.

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Chang, CY., Pasolli, H., Giannopoulou, E. et al. NFIB is a governor of epithelial–melanocyte stem cell behaviour in a shared niche. Nature 495, 98–102 (2013). https://doi.org/10.1038/nature11847

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