Transcriptomics Identify Thrombospondin‐2 as a Biomarker for NASH and Advanced Liver Fibrosis

Abstract

Background and Aims: Nonalcoholic fatty liver disease (NAFLD) is the most common liver disease worldwide. Nonalcoholic steatohepatitis (NASH), the progressive form of NAFLD, and advanced fibrosis are associated with poor outcomes. We searched for their noninvasive biomarkers.Approach and Results: Global RNA sequencing of liver tissue from 98 patients with biopsy-proven NAFLD was performed. Unsupervised hierarchical clustering well distinguished NASH from nonalcoholic fatty liver (NAFL), and NASH patients exhibited molecular abnormalities reflecting their pathological features. Transcriptomic analysis identified proteins upregulated in NASH and/or advanced fibrosis (stage F3-4), including matricellular glycoprotein thrombospondin-2 (TSP-2), encoded by the thrombospondin 2 (THBS2) gene. The intrahepatic THBS2 expression level showed the highest areas under the receiver operating characteristic curves (AUROCs) of 0.915 and 0.957 for diagnosing NASH and advanced fibrosis, respectively. THBS2 positively correlated with inflammation and ballooning according to NAFLD activity score, serum aspartate aminotransferase and hyaluronic acid (HA) levels, and NAFLD Fibrosis Score (NFS). THBS2 was associated with extracellular matrix and collagen biosynthesis, platelet activation, caspase-mediated cleavage of cytoskeletal proteins, and immune cell infiltration. Serum TSP-2 expression was measured in 213 patients with biopsy-proven NAFLD, was significantly higher in NASH than in NAFL, and increased parallel to fibrosis stage. The AUROCs for predicting NASH and advanced fibrosis were 0.776 and 0.856, respectively, which were comparable to Fibrosis-4 index, serum HA level, and NFS in advanced fibrosis diagnosis. Serum TSP-2 level and platelet count were independent predictors of NASH and advanced Accepted ArticleThis article is protected by copyright. All rights reserved fibrosis. Serum TSP-2 levels could stratify NAFLD patients according to the risk of hepatic complications, including liver cancer and decompensated cirrhotic events.Conclusions: TSP-2 may be a useful biomarker for NASH and advanced fibrosis diagnosis in NAFLD patients.

Diagnostic Modalities of Non-Alcoholic Fatty Liver Disease: From Biochemical Biomarkers to Multi-Omics Non-Invasive Approaches

“…Enriched pathways were found to be related to metabolic processes, including glycolysis/gluconeogenesis and metabolism of glycine, serine and threonine [ 257 ]. THBS2 has recently been proposed by Kozumi et al as a potential biomarker for advanced fibrosis and NASH, as it has been found to be significantly overexpressed in patients with fibrosis, and its circulating protein product TSP-2 (thrombospondin 2) was shown to be an independent predictor of NASH [ 258 ]. THBS2 was found to demonstrate the highest diagnostic accuracy for NASH with an AUROC of 0.96 in comparison with other genes and clinical scores, such as NFS (AUROC 0.88) and FIB-4 (AUROC 0.84) [ 258 ].…”

“…THBS2 has recently been proposed by Kozumi et al as a potential biomarker for advanced fibrosis and NASH, as it has been found to be significantly overexpressed in patients with fibrosis, and its circulating protein product TSP-2 (thrombospondin 2) was shown to be an independent predictor of NASH [ 258 ]. THBS2 was found to demonstrate the highest diagnostic accuracy for NASH with an AUROC of 0.96 in comparison with other genes and clinical scores, such as NFS (AUROC 0.88) and FIB-4 (AUROC 0.84) [ 258 ]. Intrahepatic THBS2 and serum TSP-2 levels were found to be significantly associated with inflammation and hepatocellular ballooning [ 258 ].…”

“…THBS2 was found to demonstrate the highest diagnostic accuracy for NASH with an AUROC of 0.96 in comparison with other genes and clinical scores, such as NFS (AUROC 0.88) and FIB-4 (AUROC 0.84) [ 258 ]. Intrahepatic THBS2 and serum TSP-2 levels were found to be significantly associated with inflammation and hepatocellular ballooning [ 258 ]. Hepatic and circulating mRNA IL32 levels have also been found to be robustly upregulated in NAFLD and to be associated with the severity of the disease, independently from the presence of the PNPLA3 variant [ 259 ].…”

Thrombospondin 2 Promotes IL-6 Production in Osteoarthritis Synovial Fibroblasts via the PI3K/AKT/NF-κB Pathway

“…TSP2 was proposed as prognosis marker in liver fibrosis, 47,48 non-alcoholic fatty liver disease, 49 and different cancers. [50][51][52][53] This study demonstrates that TSP2 is an important player in OA pathogenesis, with a positive correlation observed between TSP2 expression and synovial tissue inflammation, suggesting TSP2 could been developed as novel osteoarthritis marker.…”

Current Options and Future Directions for NAFLD and NASH Treatment

“…However, it is invasive and expensive and may cause improper diagnosis due to sampling bias [ 1 ]. Multi-omics have been used to investigate new non-invasive markers for the diagnosis of NAFLD and advanced liver disease [ 67 , 68 ]. A significant reduction of hepatic fat with advanced liver fibrosis in patients with NASH, even to the point of complete fat loss, burnt-out NASH [ 69 ].…”