PNPLA3 downregulation exacerbates the fibrotic response in human hepatic stellate cells.
Brian RadyTakahiro NishioDebanjan DharXiao LiuMark ErionTatiana KisselevaDavid A BrennerAlessandro PocaiPublished in: PloS one (2021)
Non-alcoholic steatohepatitis (NASH) results, in part, from the interaction of metabolic derangements with predisposing genetic variants, leading to liver-related complications and mortality. The strongest genetic determinant is a highly prevalent missense variant in patatin-like phospholipase domain-containing protein 3 (PNPLA3 p.I148M). In human liver hepatocytes PNPLA3 localizes to the surface of lipid droplets where the mutant form is believed to enhance lipid accumulation and release of pro-inflammatory cytokines. Less is known about the role of PNPLA3 in hepatic stellate cells (HSCs). Here we characterized HSC obtained from patients carrying the wild type (n = 8 C/C) and the heterozygous (n = 6, C/G) or homozygous (n = 6, G/G) PNPLA3 I148M and investigated the effect of genotype and PNPLA3 downregulation on baseline and TGF-β-stimulated fibrotic gene expression. HSCs from all genotypes showed comparable baseline levels of PNPLA3 and expression of the fibrotic genes α-SMA, COL1A1, TIMP1 and SMAD7. Treatment with TGF-β increased PNPLA3 expression in all 3 genotypes (~2-fold) and resulted in similar stimulation of the expression of several fibrogenic genes. In primary human HSCs carrying wild-type (WT) PNPLA3, siRNA treatment reduced PNPLA3 mRNA by 79% resulting in increased expression of α-SMA, Col1a1, TIMP1, and SMAD7 in cells stimulated with TGF-β. Similarly, knock-down of PNPLA3 in HSCs carrying either C/G or G/G genotypes resulted in potentiation of TGF-β induced expression of fibrotic genes. Knockdown of PNPLA3 did not impact fibrotic gene expression in the absence of TGF-β treatment. Together, these data indicate that the presence of the I148M PNPLA3 mutation in HSC has no effect on baseline activation and that downregulation of PNPLA3 exacerbates the fibrotic response irrespective of the genotype.
Keyphrases
- poor prognosis
- transforming growth factor
- gene expression
- wild type
- induced apoptosis
- systemic sclerosis
- idiopathic pulmonary fibrosis
- binding protein
- genome wide
- epithelial mesenchymal transition
- cell proliferation
- dna methylation
- cell cycle arrest
- endothelial cells
- signaling pathway
- oxidative stress
- liver injury
- type diabetes
- drug induced
- long non coding rna
- endoplasmic reticulum stress
- copy number
- big data
- deep learning
- diabetic rats
- anti inflammatory
- pluripotent stem cells