CRISPR/Cas9-based double-strand oligonucleotide insertion strategy corrects metabolic abnormalities in murine glycogen storage disease type-Ia
- PMID: 37467014
- PMCID: PMC10796839
- DOI: 10.1002/jimd.12660
CRISPR/Cas9-based double-strand oligonucleotide insertion strategy corrects metabolic abnormalities in murine glycogen storage disease type-Ia
Abstract
Glycogen storage disease type-Ia (GSD-Ia), characterized by impaired blood glucose homeostasis, is caused by a deficiency in glucose-6-phosphatase-α (G6Pase-α or G6PC). Using the G6pc-R83C mouse model of GSD-Ia, we explored a CRISPR/Cas9-based double-strand DNA oligonucleotide (dsODN) insertional strategy that uses the nonhomologous end-joining repair mechanism to correct the pathogenic p.R83C variant in G6pc exon-2. The strategy is based on the insertion of a short dsODN into G6pc exon-2 to disrupt the native exon and to introduce an additional splice acceptor site and the correcting sequence. When transcribed and spliced, the edited gene would generate a wild-type mRNA encoding the native G6Pase-α protein. The editing reagents formulated in lipid nanoparticles (LNPs) were delivered to the liver. Mice were treated either with one dose of LNP-dsODN at age 4 weeks or with two doses of LNP-dsODN at age 2 and 4 weeks. The G6pc-R83C mice receiving successful editing expressed ~4% of normal hepatic G6Pase-α activity, maintained glucose homeostasis, lacked hypoglycemic seizures, and displayed normalized blood metabolite profile. The outcomes are consistent with preclinical studies supporting previous gene augmentation therapy which is currently in clinical trials. This editing strategy may offer the basis for a therapeutic approach with an earlier clinical intervention than gene augmentation, with the additional benefit of a potentially permanent correction of the GSD-Ia phenotype.
Keywords: genome editing; glucose-6-phosphatase-α; glycogen storage disease type-Ia mouse model; lipid nanoparticles; nonhomologous end-joining.
© 2023 SSIEM. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.
Conflict of interest statement
CONFLICT OF INTEREST
Ananya Samanta, Nelson George, Irina Arnaoutova, Hung-Dar Chen, Brian C. Mansfield, and Janice Y. Chou declare that they have no conflicts of interest.
Christopher Hart and Troy Carlo are former employees of CRISPR Therapeutics, Inc., and own shares of CRISPR Therapeutics stock.
Similar articles
-
Correction of metabolic abnormalities in a mouse model of glycogen storage disease type Ia by CRISPR/Cas9-based gene editing.Mol Ther. 2021 Apr 7;29(4):1602-1610. doi: 10.1016/j.ymthe.2020.12.027. Epub 2020 Dec 23. Mol Ther. 2021. PMID: 33359667 Free PMC article.
-
Prevention of hepatocellular adenoma and correction of metabolic abnormalities in murine glycogen storage disease type Ia by gene therapy.Hepatology. 2012 Nov;56(5):1719-29. doi: 10.1002/hep.25717. Epub 2012 Aug 27. Hepatology. 2012. PMID: 22422504 Free PMC article.
-
Glycogen storage disease type Ia mice with less than 2% of normal hepatic glucose-6-phosphatase-α activity restored are at risk of developing hepatic tumors.Mol Genet Metab. 2017 Mar;120(3):229-234. doi: 10.1016/j.ymgme.2017.01.003. Epub 2017 Jan 10. Mol Genet Metab. 2017. PMID: 28096054 Free PMC article.
-
Type I glycogen storage diseases: disorders of the glucose-6-phosphatase/glucose-6-phosphate transporter complexes.J Inherit Metab Dis. 2015 May;38(3):511-9. doi: 10.1007/s10545-014-9772-x. Epub 2014 Oct 7. J Inherit Metab Dis. 2015. PMID: 25288127 Review.
-
Emerging roles of autophagy in hepatic tumorigenesis and therapeutic strategies in glycogen storage disease type Ia: A review.J Inherit Metab Dis. 2021 Jan;44(1):118-128. doi: 10.1002/jimd.12267. Epub 2020 Jul 2. J Inherit Metab Dis. 2021. PMID: 32474930 Review.
Cited by
-
Targeted Gene Insertion: The Cutting Edge of CRISPR Drug Development with Hemophilia as a Highlight.BioDrugs. 2024 May;38(3):369-385. doi: 10.1007/s40259-024-00654-5. Epub 2024 Mar 15. BioDrugs. 2024. PMID: 38489061 Free PMC article. Review.
References
-
- Chou JY, Matern D, Mansfield BC, Chen YT. Type I glycogen storage diseases: disorders of the glucose-6-phosphatase complex. Curr Mol Med. 2002;2(2):121–143. - PubMed
-
- Chou JY, Jun HS, Mansfield BC. Type I glycogen storage diseases: Disorders of the glucose-6-phosphatase/glucose-6-phosphate transporter complexes. J Inherit Metab Dis. 2015;38(3):511–519. - PubMed
-
- Greene HL, Slonim AE, O’Neill JA Jr, Burr IM. Continuous nocturnal intragastric feeding for management of type 1 glycogen-storage disease. N Engl J Med. 1976;294(8):423–425. - PubMed
-
- Chen YT, Cornblath M, Sidbury JB. Cornstarch therapy in type I glycogen storage disease. N Engl J Med. 1984;310(3):171–175. - PubMed
MeSH terms
Substances
Supplementary concepts
Grants and funding
LinkOut - more resources
Full Text Sources