In vivo prime editing rescues alternating hemiplegia of childhood in mice

Alexander A Sousa¹, Markus Terrey², Holt A Sakai¹, Christine Q Simmons³, Elena Arystarkhova⁴, Natalia S Morsci⁵, Laura C Anderson², Jun Xie⁶, Fabian Suri-Payer⁷, Linda C Laux⁸, Emmanuel Roze⁹, Sylvie Forlani¹⁰, Guangping Gao⁶, Simon Frost⁵, Nina Frost⁵, Kathleen J Sweadner⁴, Alfred L George Jr³, Cathleen M Lutz¹¹, David R Liu¹²

Affiliations

¹ Merkin Institute of Transformative Technologies in Healthcare, The Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA 02142, USA; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA; Howard Hughes Medical Institute, Harvard University, Cambridge, MA 02142, USA.
² Rare Disease Translational Center, The Jackson Laboratory, Bar Harbor, ME 04609, USA.
³ Department of Pharmacology, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.
⁴ Department of Neurosurgery, Massachusetts General Hospital, Boston, MA 02129, USA; Harvard Medical School, Boston, MA 02115, USA.
⁵ RARE Hope, Washington, DC, USA.
⁶ Department of Genetics & Cellular Medicine, Horae Gene Therapy Center, Li Weibo Institute for Rare Diseases Research, UMass Chan Medical School, University of Massachusetts, Worcester, MA 01605, USA; Department of Microbiology, UMass Chan Medical School, University of Massachusetts, Worcester, MA 01605, USA.
⁷ Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA; Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA 02142, USA.
⁸ Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA; Ann & Robert H. Lurie Children's Hospital of Chicago, Chicago, IL 60611, USA.
⁹ Sorbonne University, Inserm, CNRS, Paris Brain Institute, APHP, Institute of Neurology, 75005 Paris, France.
¹⁰ Sorbonne University, Inserm, CNRS, Paris Brain Institute, APHP, Hôpital de la Pitié Salpêtrière, 75013 Paris, France.
¹¹ Rare Disease Translational Center, The Jackson Laboratory, Bar Harbor, ME 04609, USA; JAX Center for Precision Genetics, The Jackson Laboratory, Bar Harbor, ME 04609, USA. Electronic address: cat.lutz@jax.org.
¹² Merkin Institute of Transformative Technologies in Healthcare, The Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA 02142, USA; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA; Howard Hughes Medical Institute, Harvard University, Cambridge, MA 02142, USA. Electronic address: drliu@fas.harvard.edu.

PMID: 40695277
DOI: 10.1016/j.cell.2025.06.038

Free article

In vivo prime editing rescues alternating hemiplegia of childhood in mice

Alexander A Sousa et al. Cell. 2025.

Free article

. 2025 Aug 7;188(16):4275-4294.e23.

doi: 10.1016/j.cell.2025.06.038. Epub 2025 Jul 21.

Authors

Affiliations

¹ Merkin Institute of Transformative Technologies in Healthcare, The Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA 02142, USA; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA; Howard Hughes Medical Institute, Harvard University, Cambridge, MA 02142, USA.
² Rare Disease Translational Center, The Jackson Laboratory, Bar Harbor, ME 04609, USA.
³ Department of Pharmacology, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.
⁴ Department of Neurosurgery, Massachusetts General Hospital, Boston, MA 02129, USA; Harvard Medical School, Boston, MA 02115, USA.
⁵ RARE Hope, Washington, DC, USA.
⁶ Department of Genetics & Cellular Medicine, Horae Gene Therapy Center, Li Weibo Institute for Rare Diseases Research, UMass Chan Medical School, University of Massachusetts, Worcester, MA 01605, USA; Department of Microbiology, UMass Chan Medical School, University of Massachusetts, Worcester, MA 01605, USA.
⁷ Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA; Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA 02142, USA.
⁸ Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA; Ann & Robert H. Lurie Children's Hospital of Chicago, Chicago, IL 60611, USA.
⁹ Sorbonne University, Inserm, CNRS, Paris Brain Institute, APHP, Institute of Neurology, 75005 Paris, France.
¹⁰ Sorbonne University, Inserm, CNRS, Paris Brain Institute, APHP, Hôpital de la Pitié Salpêtrière, 75013 Paris, France.
¹¹ Rare Disease Translational Center, The Jackson Laboratory, Bar Harbor, ME 04609, USA; JAX Center for Precision Genetics, The Jackson Laboratory, Bar Harbor, ME 04609, USA. Electronic address: cat.lutz@jax.org.
¹² Merkin Institute of Transformative Technologies in Healthcare, The Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA 02142, USA; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA; Howard Hughes Medical Institute, Harvard University, Cambridge, MA 02142, USA. Electronic address: drliu@fas.harvard.edu.

PMID: 40695277
DOI: 10.1016/j.cell.2025.06.038

Abstract

Alternating hemiplegia of childhood (AHC) is a neurodevelopmental disorder with no disease-modifying treatment. Mutations in ATP1A3, encoding an Na⁺/K⁺ ATPase subunit, cause 70% of AHC cases. Here, we present prime editing (PE) and base editing (BE) strategies to correct ATP1A3 and Atp1a3 mutations in human cells and in two AHC mouse models. We used PE and BE to correct five prevalent ATP1A3 mutations with 43%-90% efficiency. AAV9-mediated in vivo PE corrects Atp1a3 D801N and E815K in the CNS of two AHC mouse models, yielding up to 48% DNA correction and 73% mRNA correction in bulk brain cortex. In vivo PE rescued clinically relevant phenotypes, including restoration of ATPase activity; amelioration of paroxysmal spells, motor defects, and cognition deficits; and dramatic extension of animal lifespan. This work suggests a potential one-time PE treatment for AHC and establishes the ability of PE to rescue a neurological disease in animals.

Keywords: CRISPR-Cas9; alternating hemiplegia of childhood; genetic therapy; genome editing; neurological disorder; prime editing.

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Conflict of interest statement

Declaration of interests A.A.S., H.A.S., and D.R.L. have filed patent applications on prime editing and base editing approaches to treat alternating hemiplegia of childhood. D.R.L. is a co-founder and equity holder of Beam Therapeutics, Prime Medicine, Pairwise Plants, and nChromaBio, companies that use or deliver gene editing or epigenome modulating agents.

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In vivo prime editing rescues alternating hemiplegia of childhood in mice

Affiliations

In vivo prime editing rescues alternating hemiplegia of childhood in mice

Authors

Affiliations

Abstract

Conflict of interest statement

MeSH terms

Substances

Supplementary concepts

Grants and funding

LinkOut - more resources

Full Text Sources

Molecular Biology Databases