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A novel gene editing system to treat both Tay–Sachs and Sandhoff diseases

Abstract

The GM2-gangliosidoses are neurological diseases causing premature death, thus developing effective treatment protocols is urgent. GM2-gangliosidoses result from deficiency of a lysosomal enzyme β-hexosaminidase (Hex) and subsequent accumulation of GM2 gangliosides. Genetic changes in HEXA, encoding the Hex α subunit, or HEXB, encoding the Hex β subunit, causes Tay–Sachs disease and Sandhoff disease, respectively. Previous studies have showed that a modified human Hex µ subunit (HEXM) can treat both Tay–Sachs and Sandhoff diseases by forming a homodimer to degrade GM2 gangliosides. To this end, we applied this HEXM subunit in our PS813 gene editing system to treat neonatal Sandhoff mice. Through AAV delivery of the CRISPR system, a promoterless HEXM cDNA will be integrated into the albumin safe harbor locus, and lysosomal enzyme will be expressed and secreted from edited hepatocytes. 4 months after the i.v. of AAV vectors, plasma MUGS and MUG activities reached up to 144- and 17-fold of wild-type levels (n = 10, p < 0.0001), respectively. More importantly, MUGS and MUG activities in the brain also increased significantly compared with untreated Sandhoff mice (p < 0.001). Further, HPLC-MS/MS analysis showed that GM2 gangliosides in multiple tissues, except the brain, of treated mice were reduced to normal levels. Rotarod analysis showed that coordination and motor memory of treated mice were improved (p < 0.05). Histological analysis of H&E stained tissues showed reduced cellular vacuolation in the brain and liver of treated Sandhoff mice. These results demonstrate the potential of developing a treatment of in vivo genome editing for Tay–Sachs and Sandhoff patients.

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Fig. 1: Construct design and gRNA validation by Surveyor assay.
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Fig. 5: Histological analysis showed that cellular vacuolation was reduced in the brain and liver of treated Sandhoff mice.
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Acknowledgements

The authors would like to thank Dr Michael Benneyworth, Mouse Behavior Core, University of Minnesota for technical assistance in behavior tests.

Funding

LO is a fellow of the Lysosomal Disease Network (U54NS065768). The Lysosomal Disease Network is a part of the Rare Diseases Clinical Research Network (RDCRN), an initiative of the Office of Rare Diseases Research (ORDR), and NCATS. This consortium is funded through a collaboration between NCATS, the National Institute of Neurological Disorders and Stroke (NINDS), and the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK).

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Correspondence to Li Ou.

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LO and CBW are inventors of a pending patent (PCT/US2018/065747) based on the PS gene editing system. All other authors declare no conflict of interest.

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Ou, L., Przybilla, M.J., Tăbăran, AF. et al. A novel gene editing system to treat both Tay–Sachs and Sandhoff diseases. Gene Ther 27, 226–236 (2020). https://doi.org/10.1038/s41434-019-0120-5

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