Tree shrews as a new animal model for systemic sclerosis research

Leting Zheng¹ Shu Yuan Chen¹ Qiulin Wu² Xi Li³ Wen Zeng¹ Fei Dong⁴ Weiwei An⁵ Fang Qin¹ Ling Lei¹ Cheng Zhao^1*

• ¹Department of Rheumatology and Immunology, First Affiliated Hospital, Guangxi Medical University, China
• ²Department of General Surgery, The Second Affiliated Hospital of Guangxi Medical University, China
• ³Department of Clinical Laboratory, First Affiliated Hospital of Guangxi Medical University, China
• ⁴Department of Rheumatology and Immunology, The First Affiliated Hospital of China Medical University, China
• ⁵Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Guangxi Medical University, China

Objective: Systemic sclerosis (SSc) is a chronic systemic disease characterized by immune dysregulation and fibrosis for which there is no effective treatment. Animal models is crucial for advancing SSc research. Tree shrews are genetically, anatomically, and immunologically closer to humans than rodents. Thus, the tree shrew model provides a unique opportunity for translational research in SSc.In this study, a SSc tree shrew model was constructed by subcutaneous injection of different doses of bleomycin (BLM) for 21 days. We assessed the degree of inflammation and fibrosis in the skin and internal organs, and antibodies in serum. Furthermore, RNA sequencing and a series of bioinformatics analyses were performed to analyze the transcriptome changes, hub genes and immune infiltration in the skin tissues of BLM induced SSc tree shrew models. Multiple sequence alignment was utilized to analyze the conservation of selected target genes across multiple species. Results: Subcutaneous injection of BLM successfully induced a SSc model in tree shrew. This model exhibited inflammation and fibrosis in skin and lung, and some Justified Formatted[WPS_1605205607]: Justified Formatted[WPS_1605205607]: Justified Formatted[WPS_1605205607]:developed esophageal fibrosis and secrum autoantibodies including antinuclear antibodies and anti-scleroderma-70 antibody. Using RNA sequencing, we compiled skin transcriptome profiles in SSc tree shrew models. 90 differentially expressed genes (DEGs) were identified, which were mainly enriched in the PPAR signaling pathway, tyrosine metabolic pathway, p53 signaling pathway, ECM receptor interaction and glutathione metabolism, all of which are closely associated with SSc.Immune infiltration analysis identified 20 different types of immune cells infiltrating the skin of the BLM-induced SSc tree shrew models and correlations between those immune cells. By constructing a protein-protein interaction (PPI) network, we identified 10 hub genes that were significantly highly expressed in the skin of the SSc models compared to controls. Furthermore, these genes were confirmed to be highly conserved in tree shrews, humans and mice.This study for the first time comfirmed that tree shrew model of SSc can be used as a novel and promising experimental animal model to study the pathogenesis and translational research in SSc.