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Research on human nail tissue has been limited by the restricted access to fresh specimen. Here, we studied transcriptome profiles of human nail units using polydactyly specimens. Single-cell RNAseq with 11,541 cells from 4 extra digits revealed nail-specific mesenchymal and epithelial cell populations, characterized by  RSPO4  (major gene in congenital anonychia) and  SPINK6 , respectively. In situ RNA hybridization demonstrated the localization of  RSPO4, MSX1 and WIF1  in onychofibroblasts suggesting the activation of WNT signaling.  BMP-5  was also expressed in onychofibroblasts implicating the contribution of BMP signaling.  SPINK6  expression distinguished the nail-specific keratinocytes from epidermal keratinocytes.  RSPO4   +   onychofibroblasts were distributed at close proximity with  LGR6  +  nail matrix, leading to WNT/β-catenin activation. In addition, we demonstrated  RSPO4  was overexpressed in the fibroblasts of onychomatricoma and  LGR6  was highly expressed at the basal layer of the overlying epithelial component, suggesting that onychofibroblasts may play an important role in the pathogenesis of onychomatricoma.

Single-cell RNA sequencing of human nail unit defines RSPO4 onychofibroblasts and SPINK6 nail epithelium