Primary Alcohol‐Activated Human and Mouse Hepatic Stellate Cells Share Similarities in Gene‐Expression Profiles

Abstract

Alcoholic liver disease (ALD) is a leading cause of cirrhosis in the United States, which is characterized by extensive deposition of extracellular matrix proteins and formation of a fibrous scar. Hepatic stellate cells (HSCs) are the major source of collagen type 1 producing myofibroblasts in ALD fibrosis. However, the mechanism of alcohol‐induced activation of human and mouse HSCs is not fully understood. We compared the gene‐expression profiles of primary cultured human HSCs (hHSCs) isolated from patients with ALD (n = 3) or without underlying liver disease (n = 4) using RNA‐sequencing analysis. Furthermore, the gene‐expression profile of ALD hHSCs was compared with that of alcohol‐activated mHSCs (isolated from intragastric alcohol‐fed mice) or CCl4‐activated mouse HSCs (mHSCs). Comparative transcriptome analysis revealed that ALD hHSCs, in addition to alcohol‐activated and CCl4‐activated mHSCs, share the expression of common HSC activation (Col1a1 [collagen type I alpha 1 chain], Acta1 [actin alpha 1, skeletal muscle], PAI1 [plasminogen activator inhibitor‐1], TIMP1 [tissue inhibitor of metalloproteinase 1], and LOXL2 [lysyl oxidase homolog 2]), indicating that a common mechanism underlies the activation of human and mouse HSCs. Furthermore, alcohol‐activated mHSCs most closely recapitulate the gene‐expression profile of ALD hHSCs. We identified the genes that are similarly and uniquely up‐regulated in primary cultured alcohol‐activated hHSCs and freshly isolated mHSCs, which include CSF1R (macrophage colony‐stimulating factor 1 receptor), PLEK (pleckstrin), LAPTM5 (lysosmal‐associated transmembrane protein 5), CD74 (class I transactivator, the invariant chain), CD53, MMP9 (matrix metallopeptidase 9), CD14, CTSS (cathepsin S), TYROBP (TYRO protein tyrosine kinase‐binding protein), and ITGB2 (integrin beta‐2), and other genes (compared with CCl4‐activated mHSCs). Conclusion: We identified genes in alcohol‐activated mHSCs from intragastric alcohol‐fed mice that are largely consistent with the gene‐expression profile of primary cultured hHSCs from patients with ALD. These genes are unique to alcohol‐induced HSC activation in two species, and therefore may become targets or readout for antifibrotic therapy in experimental models of ALD.

Nonalcoholic Steatohepatitis and HCC in a Hyperphagic Mouse Accelerated by Western Diet

“…RNA sequencing was performed at La Jolla Institute for Immunology and described previously. 40 Sequenced reads were aligned to the mouse genome (National Center for Biotechnology Information, MGSCv38, mm10) using STAR (Spliced Transcripts Alignment to a Reference). 41 Read quantification was performed with RSEM3 v1.3.0 (University of Wisconsin, Madison, WI) and Ensembl annotation (Gencode v19, Wellcome Genome Campus, Hinxton, Cambridgeshire, UK).…”

“…RNA sequencing was performed at La Jolla Institute for Immunology and described previously. 40 Sequenced reads were aligned to the mouse genome (National Center for Biotechnology Information, MGSCv38, mm10) using STAR Q41…”

Proceeding of the Ronald G. Thurman Memorial Symposium 2020

“…However, the mechanism of alcohol-induced activation of human and mouse HSCs is not fully understood 6) . To validate mouse model of toxic fibrosis, primary alcohol-activated human HSCs (hHSCs, isolated from patients with ALD or without underlying liver disease) and mouse HSCs (mHSCs, from mice subjected to carbon tetrachloride (CCl4), isolated from intragastric alcohol-fed or pair-fed mice), and their gene expression profiles were compared using RNA-Seq 7,8) . Comparative transcriptome analysis revealed that both, alcohol-activated hHSCs and mHSCs, and CCl4-injured mHSCs share expression of Col1a1, Acta1, PAI-1, TIMP1 and LOXL2 9) .…”

Genomics of Human Fibrotic Diseases: Disordered Wound Healing Response

“…While abstinence from alcohol has proven beneficial at reversing fibrosis [ 106 ], the underlying genomic changes in alcoholic cirrhosis are poorly understood. In one study, mice and human cultured HSCs were compared using single-cell RNA-seq; CSF1R, PLEK, LAPTM5, CD74, CD53, MMP9, CD14, CTSS, TYROBP, and ITGB2 were upregulated in both alcohol-induced mouse HSCs and alcohol liver disease HSCs [ 107 ]. GWAS studies of a Mestizo Mexican and European Caucasian cohorts found the SNP rs738409 in PNPLA3 to be linked with alcoholic liver disease, similar to NAFLD [ 108 , 109 ].…”