Pnpla3 silencing with antisense oligonucleotides ameliorates nonalcoholic steatohepatitis and fibrosis in Pnpla3 I148M knock-in mice

Daniel Lindén, Andrea Ahnmark, Piero Pingitore, Ester Ciociola, Ingela Ahlstedt, Anne-Christine Andréasson, Kavitha Sasidharan, Katja Madeyski-Bengtson, Magdalena Zurek, Rosellina M. Mancina, Anna Lindblom, Mikael Bjursell, Gerhard Böttcher, Marcus Ståhlman, Mohammad Bohlooly-Y, William G. Haynes, Björn Carlsson, Mark Graham, Richard Lee, Sue Murray, Luca Valenti, Sanjay Bhanot, Peter Åkerblad, Stefano Romeo

Nonalcoholic fatty liver disease (NAFLD) is becoming the leading cause of chronic liver disease worldwide. The progression of NAFLD has a strong genetic component. A polymorphism in the patatin-like phospholipase domain-containing 3 (PNPLA3) gene resultig in a change from isoleucine (I) to methionine (M) at position 148 of the protein exerts the largest effect on NAFLD progression identified to date. Lindén and colleagues show that Pnpla3 silencing exerts a beneficial effect on liver fat accumulation, inflammation, and fibrosis that is more pronounced in the presence of the Pnpla3 I148M mutation.

Objective: Nonalcoholic fatty liver disease (NAFLD) is becoming a leading cause of advanced chronic liver disease. The progression of NAFLD, including nonalcoholic steatohepatitis (NASH), has a strong genetic component, and the most robust contributor is the patatin-like phospholipase domain-containing 3 (PNPLA3) rs738409 encoding the 148M protein sequence variant. We hypothesized that suppressing the expression of the PNPLA3 148M mutant protein would exert a beneficial effect on the entire spectrum of NAFLD.

Methods: We examined the effects of liver-targeted GalNAc3-conjugated antisense oligonucleotide (ASO)-mediated silencing of Pnpla3 in a knock-in mouse model in which we introduced the human PNPLA3 I148M mutation.

Results: ASO-mediated silencing of Pnpla3 reduced liver steatosis (p = 0.038) in homozygous Pnpla3 148M/M knock-in mutant mice but not in wild-type littermates fed a steatogenic high-sucrose diet. In mice fed a NASH-inducing diet, ASO-mediated silencing of Pnpla3 reduced liver steatosis score and NAFLD activity score independent of the Pnpla3 genotype, while reductions in liver inflammation score (p = 0.018) and fibrosis stage (p = 0.031) were observed only in the Pnpla3 knock-in 148M/M mutant mice. These responses were accompanied by reduced liver levels of Mcp1 (p = 0.026) and Timp2 (p = 0.007) specifically in the mutant knock-in mice. This may reduce levels of chemokine attracting inflammatory cells and increase the collagenolytic activity during tissue regeneration.

Conclusion: This study provides the first evidence that a Pnpla3 ASO therapy can improve all features of NAFLD, including liver fibrosis, and suppress the expression of a strong innate genetic risk factor, Pnpla3 148M, which may open up a precision medicine approach in NASH.