TY - JOUR
T1 - Synthetic mRNA rescues very long-chain acyl-CoA dehydrogenase deficiency in patient fibroblasts and a murine model
AU - Zhao, Xue Jun
AU - Mohsen, AI Walid
AU - Mihalik, Stephanie
AU - Solo, Keaton
AU - Aliu, Ermal
AU - Shi, Huifang
AU - Basu, Shakuntala
AU - Kochersperger, Catherine
AU - Van't Land, Clinton
AU - Karunanidhi, Anuradha
AU - Coughlan, Kimberly A.
AU - Siddiqui, Summar
AU - Rice, Lisa M.
AU - Hillier, Shawn
AU - Guadagnin, Eleonora
AU - Giangrande, Paloma H.
AU - Martini, Paolo G.V.
AU - Vockley, Jerry
N1 - Funding Information:
We thank the staff from the Rangos Flow Cytometry Core Facility, University of Pittsburgh, for help with FACS. We thank Lorna Cropcho, Biochemical Genetics, UPMC Children's Hospital of Pittsburgh, for both acylcarnitine standards solutions and technical advice and methods transfer to the API4000 mass spectrometer. Seahorse Analyzer and LC/MS/MS services were performed in collaboration with the Rangos Metabolic Core Facility, University of Pittsburgh. This work was supported by Moderna Therapeutics, Inc. and JV and A-WM were supported in part by NIH grant R01 DK78755.
Funding Information:
We thank the staff from the Rangos Flow Cytometry Core Facility, University of Pittsburgh, for help with FACS. We thank Lorna Cropcho, Biochemical Genetics, UPMC Children's Hospital of Pittsburgh, for both acylcarnitine standards solutions and technical advice and methods transfer to the API4000 mass spectrometer. Seahorse Analyzer and LC/MS/MS services were performed in collaboration with the Rangos Metabolic Core Facility, University of Pittsburgh. This work was supported by Moderna Therapeutics, Inc., and JV and A-WM were supported in part by NIH grant R01 DK78755.
Publisher Copyright:
© 2022
PY - 2023/1
Y1 - 2023/1
N2 - 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.
AB - 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.
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U2 - 10.1016/j.ymgme.2022.106982
DO - 10.1016/j.ymgme.2022.106982
M3 - Article
C2 - 36580829
AN - SCOPUS:85144934716
SN - 1096-7192
VL - 138
JO - Molecular Genetics and Metabolism
JF - Molecular Genetics and Metabolism
IS - 1
M1 - 106982
ER -
