Characterization of Endothelial Cell Subclusters in Localized Scleroderma Skin with Single-Cell RNA Sequencing Identifies NOTCH Signaling Pathway

• Published in: International journal of molecular sciences 2024-10, Vol.25 (19), p.10473
• Main Authors: Hutchins, Theresa, Sanyal, Anwesha, Esencan, Deren, Lafyatis, Robert, Jacobe, Heidi, Torok, Kathryn S
• Format: Article
• Language: English
• Subjects: Adolescent; Adult; Adults; Angiogenesis; Cells; Cellular signal transduction; Child; Connective tissue; Development and progression; endothelial cells; Endothelial Cells - metabolism; Endothelial Cells - pathology; Endothelium; Female; Fibroblasts; Genes; Health aspects; Humans; JAG-NOTCH signaling; Jagged-1 Protein - genetics; Jagged-1 Protein - metabolism; localized scleroderma; Male; morphea; Pediatrics; Physiological aspects; Receptors, Notch - genetics; Receptors, Notch - metabolism; Scleroderma; Scleroderma (Disease); Scleroderma, Localized - genetics; Scleroderma, Localized - metabolism; Scleroderma, Localized - pathology; Sequence Analysis, RNA; Signal Transduction; Single-Cell Analysis; single-cell RNA sequencing; skin; Skin - metabolism; Skin - pathology; Smooth muscle; Systemic scleroderma
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms251910473

Localized scleroderma (LS) is an autoimmune disease characterized by inflammation and fibrosis, leading to severe cutaneous manifestations such as skin hardening, tightness, discoloration, and other textural changes that may result in disability. While LS shares similar histopathologic features and immune-fibroblast interactions with systemic sclerosis (SSc), its molecular mechanisms remain understudied. Endothelial cells (EC) are known to play a crucial role in SSc but have not been investigated in LS. Single-cell RNA sequencing (scRNA-seq) now allows for detailed examination of this cell type in the primary organ of interest for scleroderma, the skin. In this study, we analyzed skin-isolated cells from 27 LS patients (pediatric and adult) and 17 healthy controls using scRNA-seq. Given the known role of EC damage as an initial event in SSc and the histologic and clinical skin similarities to LS, we focused primarily on endothelial cells. Our analysis identified eight endothelial subclusters within the dataset, encompassing both disease and healthy samples. Interaction analysis revealed that signaling from diseased endothelial cells was predicted to promote fibrosis through interaction with and other target genes. We also observed high levels of in arterial endothelial cells and in capillary endothelial cells, indicating the activation of a signaling pathway potentially responsible for epidermal abnormalities and contributing to LS pathogenesis. In summary, our scRNA-seq analysis identified potential disease-propagating endothelial cell clusters with upregulated pathways in LS skin, highlighting their importance in disease progression.