ZFN-mediated in vivo gene editing in hepatocytes leads to supraphysiologic α-Gal A activity and effective substrate reduction in Fabry mice.

Autor:	Pagant S; Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA., Huston MW; Sangamo Therapeutics, Inc., Brisbane, CA 94005, USA., Moreira L; Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA., Gan L; Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA., St Martin S; Sangamo Therapeutics, Inc., Brisbane, CA 94005, USA., Sproul S; Sangamo Therapeutics, Inc., Brisbane, CA 94005, USA., Holmes MC; Sangamo Therapeutics, Inc., Brisbane, CA 94005, USA., Meyer K; Sangamo Therapeutics, Inc., Brisbane, CA 94005, USA., Wechsler T; Sangamo Therapeutics, Inc., Brisbane, CA 94005, USA., Desnick RJ; Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA., Yasuda M; Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA. Electronic address: makiko.yasuda@mssm.edu.
Jazyk:	angličtina
Zdroj:	Molecular therapy : the journal of the American Society of Gene Therapy [Mol Ther] 2021 Nov 03; Vol. 29 (11), pp. 3230-3242. Date of Electronic Publication: 2021 Mar 26.
DOI:	10.1016/j.ymthe.2021.03.018
Abstrakt:	Fabry disease, a lysosomal storage disorder resulting from the deficient activity of α-galactosidase A (α-Gal A), is characterized by cardiac, renal, and/or cerebrovascular disease due to progressive accumulation of the enzyme's substrates, globotriaosylceramide (Gb3) and globotriaosylsphingosine (Lyso-Gb3). We report here the preclinical evaluation of liver-targeted in vivo genome editing using zinc-finger nuclease (ZFN) technology to insert the human α-galactosidase A (hGLA) cDNA into the albumin "safe harbor" locus of Fabry mice, thereby generating an albumin-α-Gal A fusion protein. The mature α-Gal A protein is secreted into the circulation for subsequent mannose-6-phosphate receptor-mediated tissue uptake. Donor vector optimization studies showed that replacing the hGLA cDNA signal peptide sequence with that of human iduronate 2-sulfatase (IDS) achieved higher transgene expression. Intravenous adeno-associated virus (AAV) 2/8-mediated co-delivery of the IDS-hGLA donor and ZFNs targeting the albumin locus resulted in continuous, supraphysiological plasma and tissue α-Gal A activities, which essentially normalized Gb3 and Lyso-Gb3 levels in key tissues of pathology. Notably, this was achieved with <10% of hepatocytes being edited to express hGLA, occurring mostly via non-homologous end joining (NHEJ) rather than homology-directed repair (HDR). These studies indicate that ZFN-mediated in vivo genome editing has the potential to be an effective one-time therapy for Fabry disease. Competing Interests: Declaration of interests R.J.D. is a consultant to Genzyme-Sanofi and Sangamo Therapeutics, Inc. He owns founder stock in Amicus Therapeutics and options for Sangamo Therapeutics, Inc. and receives royalties from Genzyme-Sanofi. R.J.D. and M.Y. received a research grant from Sangamo Therapeutics, Inc. to perform these studies. M.W.H., S.S.M., S.S., K.M., M.C.H., and T.W. are full-time employees and/or shareholders of Sangamo Therapeutics, Inc. (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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