Thrombospondin 2 is a key determinant of fibrogenesis in non‐alcoholic fatty liver disease

Abstract Objective Hepatic overexpression of the thrombospondin 2 gene (THBS2) and elevated levels of circulating thrombospondin 2 (TSP2) have been observed in patients with chronic liver disease. This study aimed to identify the specific cells expressing THBS2/TSP2 in non‐alcoholic fatty liver disease (NAFLD) and investigate the underlying mechanism behind THBS2/TSP2 upregulation. Design Comprehensive NAFLD liver gene datasets, including single‐cell RNA sequencing (scRNA‐seq), in‐house NAFLD liver tissue, and LX‐2 cells derived from human hepatic stellate cells (HSCs), were analysed using a combination of computational biology, genetic, immunological, and pharmacological approaches. Results Analysis of the genetic dataset revealed the presence of 1433 variable genes in patients with advanced fibrosis NAFLD, with THBS2 ranked among the top 2 genes. Quantitative polymerase chain reaction (qPCR) examination of NAFLD livers showed a significant correlation between THBS2 expression and fibrosis stage ( r = .349, p < .001). In support of this, scRNA‐seq data and in situ hybridization demonstrated that the THBS2 gene was highly expressed in HSCs of NAFLD patients with advanced fibrosis. Pathway analysis of the gene dataset revealed THBS2 expression to be associated with the transforming growth factor beta (TGFβ) pathway and collagen gene activation. Moreover, the activation of LX‐2 cells with TGFβ increased THBS2/TSP2 and collagen expression independently of the TGFβ‐SMAD2/3 pathway. THBS2 gene knockdown significantly decreased collagen expression in LX‐2 cells. Conclusions THBS2/TSP2 is highly expressed in HSCs and plays a role in regulating fibrogenesis in NAFLD patients. THBS2/TSP2 may therefore represent a potential target for anti‐fibrotic therapy in NAFLD.