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 MurrayLuca Valenti, Sanjay Bhanot, Peter Åkerblad, Stefano Romeo

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

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 GalNAc 3 -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.

Original languageEnglish
Pages (from-to)49-61
Number of pages13
JournalMolecular Metabolism
Volume22
DOIs
Publication statusPublished - Apr 1 2019

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Antisense Oligonucleotides
Fibrosis
Phospholipases
Fatty Liver
Liver
Diet
Precision Medicine
Non-alcoholic Fatty Liver Disease
Mutant Proteins
Chemokines
Liver Cirrhosis
Sucrose
Liver Diseases
Regeneration
Chronic Disease
Genotype
Inflammation
Mutation

Keywords

  • Fibrosis
  • Liver
  • NAFLD
  • NASH
  • PNPLA3

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Pnpla3 silencing with antisense oligonucleotides ameliorates nonalcoholic steatohepatitis and fibrosis in Pnpla3 I148M knock-in mice. / Lindén, Daniel; Ahnmark, Andrea; Pingitore, Piero; Ciociola, Ester; Ahlstedt, Ingela; Andréasson, Anne Christine; Sasidharan, Kavitha; Madeyski-Bengtson, Katja; Zurek, Magdalena; Mancina, Rosellina M.; Lindblom, Anna; Bjursell, Mikael; Böttcher, Gerhard; Ståhlman, Marcus; Bohlooly-Y, Mohammad; Haynes, William G.; Carlsson, Björn; Graham, Mark; Lee, Richard; Murray, Sue; Valenti, Luca; Bhanot, Sanjay; Åkerblad, Peter; Romeo, Stefano.

In: Molecular Metabolism, Vol. 22, 01.04.2019, p. 49-61.

Research output: Contribution to journalArticle

Lindén, D, Ahnmark, A, Pingitore, P, Ciociola, E, Ahlstedt, I, Andréasson, AC, Sasidharan, K, Madeyski-Bengtson, K, Zurek, M, Mancina, RM, Lindblom, A, Bjursell, M, Böttcher, G, Ståhlman, M, Bohlooly-Y, M, Haynes, WG, Carlsson, B, Graham, M, Lee, R, Murray, S, Valenti, L, Bhanot, S, Åkerblad, P & Romeo, S 2019, 'Pnpla3 silencing with antisense oligonucleotides ameliorates nonalcoholic steatohepatitis and fibrosis in Pnpla3 I148M knock-in mice', Molecular Metabolism, vol. 22, pp. 49-61. https://doi.org/10.1016/j.molmet.2019.01.013
Lindén, Daniel ; Ahnmark, Andrea ; Pingitore, Piero ; Ciociola, Ester ; Ahlstedt, Ingela ; Andréasson, Anne Christine ; Sasidharan, Kavitha ; Madeyski-Bengtson, Katja ; Zurek, Magdalena ; Mancina, Rosellina M. ; Lindblom, Anna ; Bjursell, Mikael ; Böttcher, Gerhard ; Ståhlman, Marcus ; Bohlooly-Y, Mohammad ; Haynes, William G. ; Carlsson, Björn ; Graham, Mark ; Lee, Richard ; Murray, Sue ; Valenti, Luca ; Bhanot, Sanjay ; Åkerblad, Peter ; Romeo, Stefano. / Pnpla3 silencing with antisense oligonucleotides ameliorates nonalcoholic steatohepatitis and fibrosis in Pnpla3 I148M knock-in mice. In: Molecular Metabolism. 2019 ; Vol. 22. pp. 49-61.
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abstract = "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 GalNAc 3 -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.",
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T1 - Pnpla3 silencing with antisense oligonucleotides ameliorates nonalcoholic steatohepatitis and fibrosis in Pnpla3 I148M knock-in mice

AU - Lindén, Daniel

AU - Ahnmark, Andrea

AU - Pingitore, Piero

AU - Ciociola, Ester

AU - Ahlstedt, Ingela

AU - Andréasson, Anne Christine

AU - Sasidharan, Kavitha

AU - Madeyski-Bengtson, Katja

AU - Zurek, Magdalena

AU - Mancina, Rosellina M.

AU - Lindblom, Anna

AU - Bjursell, Mikael

AU - Böttcher, Gerhard

AU - Ståhlman, Marcus

AU - Bohlooly-Y, Mohammad

AU - Haynes, William G.

AU - Carlsson, Björn

AU - Graham, Mark

AU - Lee, Richard

AU - Murray, Sue

AU - Valenti, Luca

AU - Bhanot, Sanjay

AU - Åkerblad, Peter

AU - Romeo, Stefano

PY - 2019/4/1

Y1 - 2019/4/1

N2 - 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 GalNAc 3 -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.

AB - 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 GalNAc 3 -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.

KW - Fibrosis

KW - Liver

KW - NAFLD

KW - NASH

KW - PNPLA3

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