Spatial transcriptomics identifies a translayer architecture of pyroptosis-related transcription in systemic sclerosis skin

Oryoji, D.; Doi, G.; Fujimoto, S.; Nishimura, N.; Otsuka, K.; Kuwahara, A.; Ayano, M.; Kimoto, Y.; Akashi, K.; Niiro, H.; Mitoma, H.

doi:10.64898/2026.05.03.722547

bioRxiv · 10.64898/2026.05.03.722547

Spatial transcriptomics identifies a translayer architecture of pyroptosis-related transcription in systemic sclerosis skin

Oryoji, D.·Doi, G.·Fujimoto, S.·Nishimura, N.·Otsuka, K.·Kuwahara, A.·Ayano, M.·Kimoto, Y.·Akashi, K.·Niiro, H.·Mitoma, H.

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Abstract

Anatomical-layer enrichment does not establish cellular attribution, so correspondence with major-cell-class expression must be tested gene by gene. Systemic sclerosis (SSc) skin is informative because inflammasome- and pyroptosis-related molecules have been reported across epidermal, fibroblast-associated and vascular contexts. Spatial transcriptomics of SSc and control skin, with same-donor single-nucleus profiles from 10 SSc donors, revealed gene-specific correspondence. Across 13 discovery sections, NLRP1, PYCARD and CASP4 were epidermally biased, whereas GSDMD was dermally biased. An independent 10-section SSc cohort supported epidermal NLRP1 (q = 0.0078) and PYCARD (q = 0.043), with directional, non-significant support for CASP4 and GSDMD. Despite epidermal NLRP1 bias, endothelial class-pseudobulk exceeded keratinocyte class-pseudobulk in all 10 donors. Restricted three-class reconstruction reproduced the NLRP1 direction in 1 and 0 of 10 donors under the two fixed dermal weightings and in 0-8 of 10 across a post hoc sweep of the dermal endothelial share; PYCARD and GSDMD were concordant in most donors at every weighting, and CASP4 in no more than 4 of 10. Thus, tissue layer and major cell class are gene-specifically non-equivalent axes of spatial interpretation.

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Oryoji, D., Doi, G., Fujimoto, S., Nishimura, N., Otsuka, K., Kuwahara, A., Ayano, M., Kimoto, Y., Akashi, K., Niiro, H., Mitoma, H.. 2026-05-06. Spatial transcriptomics identifies a translayer architecture of pyroptosis-related transcription in systemic sclerosis skin. https://doi.org/10.64898/2026.05.03.722547

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