Correction of a urea cycle defect after ex vivo gene editing of human hepatocytes.

Autor:	Zabulica M; Department of Laboratory Medicine, Karolinska Institutet, 141 52 Huddinge, Sweden., Srinivasan RC; Department of Laboratory Medicine, Karolinska Institutet, 141 52 Huddinge, Sweden., Akcakaya P; Discovery Sciences, BioPharmaceuticals R&D Unit, AstraZeneca, Gothenburg, Sweden., Allegri G; Division of Metabolism and Children's Research Center, University Children's Hospital, Zürich, Switzerland., Bestas B; Discovery Sciences, BioPharmaceuticals R&D Unit, AstraZeneca, Gothenburg, Sweden., Firth M; Discovery Sciences, BioPharmaceuticals R&D Unit, AstraZeneca, Cambridge, UK., Hammarstedt C; Department of Laboratory Medicine, Karolinska Institutet, 141 52 Huddinge, Sweden., Jakobsson T; Department of Laboratory Medicine, Karolinska Institutet, 141 52 Huddinge, Sweden., Jakobsson T; Department of Clinical Sciences Intervention and Technology, Karolinska Institutet, Stockholm, Sweden., Ellis E; Department of Clinical Sciences Intervention and Technology, Karolinska Institutet, Stockholm, Sweden., Jorns C; Department of Clinical Sciences Intervention and Technology, Karolinska Institutet, Stockholm, Sweden., Makris G; Division of Metabolism and Children's Research Center, University Children's Hospital, Zürich, Switzerland., Scherer T; Division of Metabolism and Children's Research Center, University Children's Hospital, Zürich, Switzerland., Rimann N; Division of Metabolism and Children's Research Center, University Children's Hospital, Zürich, Switzerland., van Zuydam NR; Department of Quantitative Biology, Discovery Sciences, R&D BioPharmaceuticals, AstraZeneca, Gothenburg, Sweden., Gramignoli R; Department of Laboratory Medicine, Karolinska Institutet, 141 52 Huddinge, Sweden., Forslöw A; Discovery Sciences, BioPharmaceuticals R&D Unit, AstraZeneca, Gothenburg, Sweden., Engberg S; Discovery Sciences, BioPharmaceuticals R&D Unit, AstraZeneca, Gothenburg, Sweden., Maresca M; Discovery Sciences, BioPharmaceuticals R&D Unit, AstraZeneca, Gothenburg, Sweden., Rooyackers O; Department of Clinical Sciences Intervention and Technology, Karolinska Institutet, Stockholm, Sweden., Thöny B; Division of Metabolism and Children's Research Center, University Children's Hospital, Zürich, Switzerland., Häberle J; Division of Metabolism and Children's Research Center, University Children's Hospital, Zürich, Switzerland., Rosen B; Discovery Sciences, BioPharmaceuticals R&D Unit, AstraZeneca, Cambridge, UK., Strom SC; Department of Laboratory Medicine, Karolinska Institutet, 141 52 Huddinge, Sweden. Electronic address: stephen.strom7.4@gmail.com.
Jazyk:	angličtina
Zdroj:	Molecular therapy : the journal of the American Society of Gene Therapy [Mol Ther] 2021 May 05; Vol. 29 (5), pp. 1903-1917. Date of Electronic Publication: 2021 Jan 21.
DOI:	10.1016/j.ymthe.2021.01.024
Abstrakt:	Ornithine transcarbamylase deficiency (OTCD) is a monogenic disease of ammonia metabolism in hepatocytes. Severe disease is frequently treated by orthotopic liver transplantation. An attractive approach is the correction of a patient's own cells to regenerate the liver with gene-repaired hepatocytes. This study investigates the efficacy and safety of ex vivo correction of primary human hepatocytes. Hepatocytes isolated from an OTCD patient were genetically corrected ex vivo, through the deletion of a mutant intronic splicing site achieving editing efficiencies >60% and the restoration of the urea cycle in vitro. The corrected hepatocytes were transplanted into the liver of FRGN mice and repopulated to high levels (>80%). Animals transplanted and liver repopulated with genetically edited patient hepatocytes displayed normal ammonia, enhanced clearance of an ammonia challenge and OTC enzyme activity, as well as lower urinary orotic acid when compared to mice repopulated with unedited patient hepatocytes. Gene expression was shown to be similar between mice transplanted with unedited or edited patient hepatocytes. Finally, a genome-wide screening by performing CIRCLE-seq and deep sequencing of >70 potential off-targets revealed no unspecific editing. Overall analysis of disease phenotype, gene expression, and possible off-target editing indicated that the gene editing of a severe genetic liver disease was safe and effective. Competing Interests: Declaration of interests S.C.S. has stock in Yecuris (no Yecuris animals were used in these studies). (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu