Synthetic mRNA rescues very long-chain acyl-CoA dehydrogenase deficiency in patient fibroblasts and a murine model

Xue-Jun Zhao; Ai-Walid Mohsen; Stephanie Mihalik; Keaton Solo; Ermal Aliu; Huifang Shi; Shakuntala Basu; Catherine Kochersperger; Clinton Van't Land; Anuradha Karunanidhi; Kimberly A Coughlan; Summar Siddiqui; Lisa M Rice; Shawn Hillier; Eleonora Guadagnin; Paloma H Giangrande; Paolo G V Martini; Jerry Vockley

doi:10.1016/j.ymgme.2022.106982

Synthetic mRNA rescues very long-chain acyl-CoA dehydrogenase deficiency in patient fibroblasts and a murine model

Mol Genet Metab. 2023 Jan;138(1):106982. doi: 10.1016/j.ymgme.2022.106982. Epub 2022 Dec 23.

Authors

Xue-Jun Zhao¹, Ai-Walid Mohsen², Stephanie Mihalik¹, Keaton Solo¹, Ermal Aliu¹, Huifang Shi¹, Shakuntala Basu¹, Catherine Kochersperger¹, Clinton Van't Land¹, Anuradha Karunanidhi¹, Kimberly A Coughlan³, Summar Siddiqui³, Lisa M Rice³, Shawn Hillier³, Eleonora Guadagnin³, Paloma H Giangrande³, Paolo G V Martini³, Jerry Vockley⁴

Affiliations

¹ Division of Genetic and Genomic Medicine, Department of Pediatrics, University of Pittsburgh, Pittsburgh, PA, USA.
² Division of Genetic and Genomic Medicine, Department of Pediatrics, University of Pittsburgh, Pittsburgh, PA, USA; Department of Human Genetics, University of Pittsburgh, Pittsburgh, PA, USA.
³ Moderna Therapeutics, Inc., Rare Diseases, 200 Technology Square, Cambridge, MA, USA.
⁴ Division of Genetic and Genomic Medicine, Department of Pediatrics, University of Pittsburgh, Pittsburgh, PA, USA; Department of Human Genetics, University of Pittsburgh, Pittsburgh, PA, USA. Electronic address: vockleyg@upmc.edu.

Abstract

Very long-chain acyl-CoA dehydrogenase (VLCAD) deficiency is an inborn error of long chain fatty acid β-oxidation (FAO) with limited treatment options. Patients present with heterogeneous clinical phenotypes affecting predominantly heart, liver, and skeletal muscle. While VLCAD deficiency is a systemic disease, restoration of liver FAO has the potential to improve symptoms more broadly due to increased total body ATP production and reduced accumulation of potentially toxic metabolites. We explored the use of synthetic human VLCAD (hVLCAD) mRNA and lipid nanoparticle encapsulated hVLCAD mRNA (LNP-VLCAD) to generate functional VLCAD enzyme in patient fibroblasts derived from VLCAD deficient patients, mouse embryonic fibroblasts, hepatocytes isolated from VLCAD knockout (Acadvl^-/-) mice, and Acadvl^-/- mice to reverse the metabolic effects of the deficiency. Transfection of all cell types with hVLCAD mRNA resulted in high level expression of protein that localized to mitochondria with increased enzyme activity. Intravenous administration of LNP-VLCAD to Acadvl^-/- mice produced a significant amount of VLCAD protein in liver, which declined over a week. Treated Acadvl^-/- mice showed reduced hepatic steatosis, were more resistant to cold stress, and accumulated less toxic metabolites in blood than untreated animals. Results from this study support the potential for hVLCAD mRNA for treatment of VLCAD deficiency.

Keywords: Acadvl(-/-) mouse; Fatty acid β-oxidation; Fibroblasts; Synthetic mRNA; Very long-chain acyl-CoA dehydrogenase deficiency.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Acyl-CoA Dehydrogenase, Long-Chain*
Animals
Disease Models, Animal
Fibroblasts / metabolism
Humans
Lipid Metabolism, Inborn Errors* / genetics
Lipid Metabolism, Inborn Errors* / therapy
Mice
RNA, Messenger / genetics
RNA, Messenger / metabolism

Substances

Acyl-CoA Dehydrogenase, Long-Chain
RNA, Messenger

Supplementary concepts

VLCAD deficiency

Grants and funding

R01 DK078775/DK/NIDDK NIH HHS/United States