Spinella Amelia, Lo Tartaro Domenico, Gibellini Lara, de Pinto Marco, Pinto Valentina, Bonetti Elisa, Lolli Francesca, Lattanzi Melba, Lumetti Federica, Amati Gabriele, De Santis Giorgio, Cossarizza Andrea, Salvarani Carlo, Giuggioli Dilia
Scleroderma Unit and Rheumatology Unit, Medical School, University of Modena and Reggio Emilia, University Hospital of Modena Policlinico of Modena, Modena, Italy.
Department of Medical and Surgical Sciences of Children and Adults, University of Modena and Reggio Emilia, Modena, Italy.
J Scleroderma Relat Disord. 2023 Jun;8(2):151-166. doi: 10.1177/23971983221130145. Epub 2022 Oct 17.
Systemic sclerosis is characterized by endothelial dysfunction, autoimmunity abnormalities, and fibrosis of the skin and internal organs. The pathogenetic mechanisms underlying systemic sclerosis vasculopathy are still not clarified. A complex cellular and extracellular network of interactions has been studied, but it is currently unclear what drives the activation of fibroblasts/myofibroblasts and the extracellular matrix deposition.
Using RNA sequencing, the aim of the work was to identify potential functional pathways implied in systemic sclerosis pathogenesis and markers of endothelial dysfunction and fibrosis in systemic sclerosis patients. RNA-sequencing analysis was performed on RNA obtained from biopsies from three systemic sclerosis patients and three healthy controls enrolled in our University Hospital. RNA was used to generate sequencing libraries that were sequenced according to proper transcriptomic analyses. Subsequently, we performed gene set enrichment analysis of differentially expressed genes on the entire list of genes that compose the RNA-sequencing expression matrix.
Gene set enrichment analysis revealed that healthy controls were characterized by gene signatures related to stromal stem cells proliferation, cytokine-cytokine receptor interaction, macrophage-enriched metabolic network, whereas systemic sclerosis tissues were enriched in signatures associated with keratinization, cornification, retinoblastoma 1 and tumor suppressor 53 signaling.
According to our data, RNA-sequencing and pathway analysis revealed that systemic sclerosis subjects display a discrete pattern of gene expression associated with keratinization, extracellular matrix generation, and negative regulation of angiogenesis and stromal stem cells proliferation. Further analysis on larger numbers of patients is needed; however, our findings provide an interesting framework for the development of biomarkers useful to explore potential future therapeutic approaches.
系统性硬化症的特征是内皮功能障碍、自身免疫异常以及皮肤和内脏器官纤维化。系统性硬化症血管病变的发病机制仍未阐明。虽然已经研究了一个复杂的细胞和细胞外相互作用网络，但目前尚不清楚是什么驱动成纤维细胞/肌成纤维细胞的激活和细胞外基质沉积。
本研究旨在通过RNA测序确定系统性硬化症发病机制中潜在的功能途径以及系统性硬化症患者内皮功能障碍和纤维化的标志物。对取自我校医院的三名系统性硬化症患者和三名健康对照的活检组织所获得的RNA进行RNA测序分析。RNA用于生成测序文库，并根据适当的转录组分析进行测序。随后，我们对构成RNA测序表达矩阵的整个基因列表中的差异表达基因进行基因集富集分析。
基因集富集分析显示，健康对照的特征是与基质干细胞增殖、细胞因子-细胞因子受体相互作用、富含巨噬细胞的代谢网络相关的基因特征，而系统性硬化症组织则富含与角质化、角化、视网膜母细胞瘤1和肿瘤抑制因子53信号相关的特征。
根据我们的数据，RNA测序和通路分析显示，系统性硬化症患者表现出与角质化、细胞外基质生成以及血管生成和基质干细胞增殖的负调控相关的离散基因表达模式。需要对更多患者进行进一步分析；然而，我们的研究结果为开发有助于探索未来潜在治疗方法的生物标志物提供了一个有趣的框架。
