In vivo base editing rescues Hutchinson-Gilford progeria syndrome in mice.

Autor: Koblan LW; Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of Harvard and MIT, Cambridge, MA, USA.; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.; Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA., Erdos MR; National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA., Wilson C; Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of Harvard and MIT, Cambridge, MA, USA.; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.; Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA., Cabral WA; National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA., Levy JM; Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of Harvard and MIT, Cambridge, MA, USA.; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.; Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA., Xiong ZM; National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA., Tavarez UL; National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA., Davison LM; Division of Cardiovascular Medicine, Vanderbilt University Medical Center, Nashville, TN, USA., Gete YG; Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD, USA., Mao X; Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD, USA., Newby GA; Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of Harvard and MIT, Cambridge, MA, USA.; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.; Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA., Doherty SP; Division of Cardiovascular Medicine, Vanderbilt University Medical Center, Nashville, TN, USA., Narisu N; National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA., Sheng Q; Department of Biostatistics, Vanderbilt University Medical Center, Nashville, TN, USA., Krilow C; National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA., Lin CY; Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.; Therapeutic Innovation Center, Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, USA.; Kronos, Bio Inc., Cambridge, MA, USA., Gordon LB; Hasbro Children's Hospital, Alpert Medical School of Brown University, Providence, RI, USA.; Boston Children's Hospital, Harvard Medical School, Boston, MA, USA., Cao K; Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD, USA., Collins FS; National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA. francis.collins3@nih.gov., Brown JD; Division of Cardiovascular Medicine, Vanderbilt University Medical Center, Nashville, TN, USA. jonathan.d.brown@vumc.org., Liu DR; Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of Harvard and MIT, Cambridge, MA, USA. drliu@fas.harvard.edu.; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA. drliu@fas.harvard.edu.; Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA. drliu@fas.harvard.edu.
Jazyk: angličtina
Zdroj: Nature [Nature] 2021 Jan; Vol. 589 (7843), pp. 608-614. Date of Electronic Publication: 2021 Jan 06.
DOI: 10.1038/s41586-020-03086-7
Abstrakt: Hutchinson-Gilford progeria syndrome (HGPS or progeria) is typically caused by a dominant-negative C•G-to-T•A mutation (c.1824 C>T; p.G608G) in LMNA, the gene that encodes nuclear lamin A. This mutation causes RNA mis-splicing that produces progerin, a toxic protein that induces rapid ageing and shortens the lifespan of children with progeria to approximately 14 years 1-4 . Adenine base editors (ABEs) convert targeted A•T base pairs to G•C base pairs with minimal by-products and without requiring double-strand DNA breaks or donor DNA templates 5,6 . Here we describe the use of an ABE to directly correct the pathogenic HGPS mutation in cultured fibroblasts derived from children with progeria and in a mouse model of HGPS. Lentiviral delivery of the ABE to fibroblasts from children with HGPS resulted in 87-91% correction of the pathogenic allele, mitigation of RNA mis-splicing, reduced levels of progerin and correction of nuclear abnormalities. Unbiased off-target DNA and RNA editing analysis did not detect off-target editing in treated patient-derived fibroblasts. In transgenic mice that are homozygous for the human LMNA c.1824 C>T allele, a single retro-orbital injection of adeno-associated virus 9 (AAV9) encoding the ABE resulted in substantial, durable correction of the pathogenic mutation (around 20-60% across various organs six months after injection), restoration of normal RNA splicing and reduction of progerin protein levels. In vivo base editing rescued the vascular pathology of the mice, preserving vascular smooth muscle cell counts and preventing adventitial fibrosis. A single injection of ABE-expressing AAV9 at postnatal day 14 improved vitality and greatly extended the median lifespan of the mice from 215 to 510 days. These findings demonstrate the potential of in vivo base editing as a possible treatment for HGPS and other genetic diseases by directly correcting their root cause.
Databáze: MEDLINE
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