The online version of this article (https://doi.org/10.1186/s12916-018-1136-1) contains supplementary material, which is available to authorized users.
Non-alcoholic steatohepatitis (NASH) is among the leading causes of liver disease worldwide. It is increasingly recognized that the phenotype of NASH may involve a number of different pathways, of which each could become important therapeutic targets. The aim of this study is to use high resolution mass spectrometry (MS) and phosphoproteomics techniques to assess the serum proteome and hepatic phosphoproteome in subjects with NASH-related fibrosis.
Sixty-seven biopsy-proven NAFLD subjects with frozen sera and liver tissue were included. Reverse phase protein microarray was used to quantify the phosphorylation of key signaling proteins in liver and nano-liquid chromatography (LC)-MS was used to sequence target biomarkers in the serum. An image analysis algorithm was used to quantify the percentage of collagen (% collagen) using computer-assisted morphometry. Using multiple regression models, serum proteomes and phosphorylated hepatic proteins that were independently (p ≤ 0.05) associated with advanced fibrosis (stage ≥ 2) and higher % collagen were assessed.
Phosphorylated signaling pathways in the liver revealed that apoptosis signal-regulating kinase 1, mitogen-activated protein kinase (ASK1-MAPK pathway involving ASK1 S38 (p < 0.02) and p38 MAPK (p = 0.0002)) activated by the inflammatory cytokine interleukin (IL-10) (p < 0.001), were independently associated with higher % collagen. LC-MS data revealed that serum alpha-2 macroglobulin (α2M) (p = 0.0004) and coagulation factor V (p = 0.0127) were independently associated with higher % hepatic collagen.
Simultaneous profiling of serum proteome and hepatic phosphoproteome reveals that the activation of ASK1 S38, p38 MAPK in the liver, and serum α2M and coagulation factor V are independently associated with hepatic collagen deposition in patients with NASH. These data suggest the role of these pathways in the pathogenesis of NASH-related fibrosis as a potential therapeutic target.
Additional file 1: Figure S1. Representative image of collagen quantification after staining with Sirius red. Table S1. Associations or correlations of phosphorylated hepatic proteins with fibrosis stage and higher % collagen deposition in livers of NAFLD patients. Figure S2. Images showing (a) advanced liver fibrosis signaling protein-protein network and (b) higher hepatic percentage collagen protein-protein network. Figure S3. Phosphoproteins and proteomes involved in biological processes and KEGG pathways. Table S2. Pathways associated with advanced fibrosis stage ≥ 2. Table S3. Pathways associated with higher % collagen deposition in the liver. Figure S4. Scatter plots of A. alpha-2 macroglobulin precursor vs. IL-10 (ρ = 0.28, p = 0.02) and ASK1 S83 (ρ = 0.25, p = 0.04). (DOCX 2995 kb)12916_2018_1136_MOESM1_ESM.docx
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- An exploratory study examining how nano-liquid chromatography–mass spectrometry and phosphoproteomics can differentiate patients with advanced fibrosis and higher percentage collagen in non-alcoholic fatty liver disease
Zobair M. Younossi
Zachary D. Goodman
Emanuel F. Petricoin
- BioMed Central
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