Hayashita-Kinoh Hiromi, Guillermo Posadas-Herrera, Nitahara-Kasahara Yuko, Kuraoka Mutsuki, Okada Hironori, Chiyo Tomoko, Takeda Shin'ichi, Okada Takashi
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.
Department of Molecular Therapy, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Kodaira, Tokyo 187-8502, Japan.
Mol Ther Methods Clin Dev. 2020 Nov 17;20:133-141. doi: 10.1016/j.omtm.2020.11.003. eCollection 2021 Mar 12.
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.
杜氏肌营养不良症（DMD）是一种与肌营养不良蛋白基因突变相关的严重先天性疾病。使用重组腺相关病毒（rAAV）补充肌营养不良蛋白有望成为治疗DMD的方法，尽管高剂量rAAV全身给药可能会引发与载体相关的一般毒性，如肝损伤、神经毒性和生殖系传播。在此，我们用rAAV9转导联合多能间充质基质细胞（MSC）注射治疗正常或患肌营养不良症的犬，以研究在免疫调节条件下表达微肌营养不良蛋白（μDys）的rAAV的治疗效果。将骨髓来源的MSC、rAAV-CMV-μDys和rAAV-CAG-荧光素酶（Luc）注入一只年轻的患肌营养不良症犬的颈静脉，以诱导μDys的全身表达。首次注射一周后，该犬接受第二次静脉注射MSC，次日，将rAAV静脉注射到同一只犬体内。用MSC预处理后全身注射rAAV9可改善正常和患肌营养不良症犬的基因转移。在注射rAAV-μDys的患肌营养不良症犬中，营养不良表型显著改善，这表明包括免疫调节在内的改良rAAV-μDys治疗可诱导成功的长期转基因表达，从而改善营养不良表型。
