The interplay among different cells in a tissue is essential for maintaining homeostasis. Although disease states have been traditionally attributed to individual cell types, increasing evidence and new therapeutic options have demonstrated the primary role of multicellular functions to understand health and disease, opening new avenues to understand pathogenesis and develop new treatment strategies. We recently described the cellular composition and dynamics of the human oral mucosa; however, the spatial arrangement of cells is needed to better understand a morphologically complex tissue. Here, we link single-cell RNA sequencing, spatial transcriptomics, and high-resolution multiplex fluorescence in situ hybridisation to characterise human oral mucosa in health and oral chronic inflammatory disease. We deconvolved expression for resolution enhancement of spatial transcriptomic data and defined highly specialised epithelial and stromal compartments describing location-specific immune programs. Furthermore, we spatially mapped a rare pathogenic fibroblast population localised in a highly immunogenic region, responsible for lymphocyte recruitment through CXCL8 and CXCL10 and with a possible role in pathological angiogenesis through ALOX5AP. Collectively, our study provides a comprehensive reference for the study of oral chronic disease pathogenesis.

Centre for Craniofacial and Regenerative Biology Faculty of Dentistry Oral and Craniofacial Sciences King's College London London United Kingdom

Department of Endodontics Faculty of Dentistry Oral and Craniofacial Sciences King's College London London United Kingdom

Department of Periodontology Faculty of Dentistry Oral and Craniofacial Sciences King's College London London United Kingdom

Laboratory of Odontogenesis and Osteogenesis Institute of Animal Physiology and Genetics Brno Czech Republic

NIHR BRC Genomics Research Platform Guy's and St Thomas' NHS Foundation Trust King's College London School of Medicine Guy's Hospital London United Kingdom

Unilever R and D Colworth Science Park Sharnbrook United Kingdom

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Mapping Small Extracellular Vesicle Secretion Potential in Healthy Human Gingiva Using Spatial Transcriptomics

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