Gα₁₂ overexpression induced by miR-16 dysregulation contributes to liver fibrosis by promoting autophagy in hepatic stellate cells

Background & aims: Hepatic stellate cells (HSCs) have a role in liver fibrosis. Guanine nucleotide-binding α-subunit 12 (Gα₁₂) converges signals from G-protein-coupled receptors whose ligand levels are elevated in the environment during liver fibrosis; however, information is lacking on the effect of Gα₁₂ on HSC trans-differentiation. This study investigated the expression of Gα₁₂ in HSCs and the molecular basis of the effects of its expression on liver fibrosis.

Methods: Gα₁₂ expression was assessed by immunostaining, and immunoblot analyses of mouse fibrotic liver tissues and primary HSCs. The role of Gα₁₂ in liver fibrosis was estimated using a toxicant injury mouse model with Gα₁₂ gene knockout and/or HSC-specific Gα₁₂ delivery using lentiviral vectors, in addition to primary HSCs and LX-2 cells using microRNA (miR) inhibitors, overexpression vectors, or adenoviruses. miR-16, Gα₁₂, and LC3 were also examined in samples from patients with fibrosis.

Results: Gα₁₂ was overexpressed in activated HSCs and fibrotic liver, and was colocalised with desmin. In a carbon tetrachloride-induced fibrosis mouse model, Gα₁₂ ablation prevented increases in fibrosis and liver injury. This effect was attenuated by HSC-specific lentiviral delivery of Gα₁₂. Moreover, Gα₁₂ activation promoted autophagy accompanying c-Jun N-terminal kinase-dependent ATG12-5 conjugation. In addition, miR-16 was found to be a direct inhibitor of the de novo synthesis of Gα₁₂. Modulations of miR-16 altered autophagy in HSCs. In a fibrosis animal model or patients with severe fibrosis, miR-16 levels were lower than in their corresponding controls. Consistently, cirrhotic patient liver tissues showed Gα₁₂ and LC3 upregulation in desmin-positive areas.

Conclusions: miR-16 dysregulation in HSCs results in Gα₁₂ overexpression, which activates HSCs by facilitating autophagy through ATG12-5 formation. This suggests that Gα₁₂ and its regulatory molecules could serve as targets for the amelioration of liver fibrosis.

Lay summary: Guanine nucleotide-binding α-subunit 12 (Gα₁₂) is upregulated in activated hepatic stellate cells (HSCs) as a consequence of the dysregulation of a specific microRNA that is abundant in HSCs, facilitating the progression of liver fibrosis. This event is mediated by c-Jun N-terminal kinase-dependent ATG12-5 formation and the promotion of autophagy. We suggest that Gα₁₂ and its associated regulators could serve as new targets in HSCs for the treatment of liver fibrosis.