Improved transduction of canine X-linked muscular dystrophy with rAAV9-microdystrophin via multipotent MSC pretreatment

Hiromi Hayashita-Kinoh; Posadas-Herrera Guillermo; Yuko Nitahara-Kasahara; Mutsuki Kuraoka; Hironori Okada; Tomoko Chiyo; Shin'ichi Takeda; Takashi Okada

doi:10.1016/j.omtm.2020.11.003

Improved transduction of canine X-linked muscular dystrophy with rAAV9-microdystrophin via multipotent MSC pretreatment

Mol Ther Methods Clin Dev. 2020 Nov 17:20:133-141. doi: 10.1016/j.omtm.2020.11.003. eCollection 2021 Mar 12.

Authors

Hiromi Hayashita-Kinoh^{1

2}, Posadas-Herrera Guillermo^{1

2}, Yuko Nitahara-Kasahara^{3

2}, Mutsuki Kuraoka², Hironori Okada², Tomoko Chiyo², Shin'ichi Takeda², Takashi Okada^{1

2}

Affiliations

¹ Division of Molecular and Medical Genetics, Center for Gene and Cell Therapy, The Institute of Medical Science, The University of Tokyo, Tokyo 108-8631, Japan.
² Department of Molecular Therapy, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Kodaira, Tokyo 187-8502, Japan.
³ Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo 113-8632, Japan.

Abstract

Duchenne muscular dystrophy (DMD) is a severe congenital disease associated with mutation of the dystrophin gene. Supplementation of dystrophin using recombinant adeno-associated virus (rAAV) has promise as a treatment for DMD, although vector-related general toxicities, such as liver injury, neurotoxicity, and germline transmission, have been suggested in association with the systemic delivery of high doses of rAAV. Here, we treated normal or dystrophic dogs with rAAV9 transduction in conjunction with multipotent mesenchymal stromal cell (MSC) injection to investigate the therapeutic effects of an rAAV expressing microdystrophin (μDys) under conditions of immune modulation. Bone-marrow-derived MSCs, rAAV-CMV-μDys, and a rAAV-CAG-luciferase (Luc) were injected into the jugular vein of a young dystrophic dog to induce systemic expression of μDys. One week after the first injection, the dog received a second intravenous injection of MSCs, and on the following day, rAAV was intravenously injected into the same dog. Systemic injection of rAAV9 with MSCs pretreatment improves gene transfer into normal and dystrophic dogs. Dystrophic phenotypes significantly improved in the rAAV-μDys-injected dystrophic dog, suggesting that an improved rAAV-μDys treatment including immune modulation induces successful long-term transgene expression to improve dystrophic phenotypes.

Keywords: AAV; DMD; DMD dog model; MSC; immune modulation; microdystrophin.