Preclinical Toxicology and Biodistribution Studies of Recombinant Adeno-Associated Virus 1 Human Acid α-Glucosidase

作者：Thomas J. Conlon, Kirsten E. Erger, Stacy Porvasnik, Travis L. Cossette, Cheryl Roberts, Lynn A. Combee, Saleem Islam, Jeffry S. Kelley, Denise Cloutier, Nathalie Clément, Corinne R. Abernathy, Barry J. Byrne · 发表于：Human Gene Therapy Clinical Development · 年份：2013 · DOI：10.1089/humc.2013.147 · 被引用次数：21 · 研究领域：Virus-based gene therapy research、Cytomegalovirus and herpesvirus research、Herpesvirus Infections and Treatments

A biodistribution and toxicology study was performed to test the acute toxicities of intradiaphragmatic injection of a recombinant adeno-associated virus (rAAV) 2/1-human acid alpha-Glucosidase (hGAA) driven by a cytomegalovirus (CMV) promoter (rAAV1-CMV-hGAA) in New Zealand white rabbits and in the rodent Pompe disease model by injecting at the right quadriceps. Studies performed using fluoroscopy and AAV2-GFP demonstrated spread upon intradiaphragmatic injection, and the ability of AAV to infect and express acid α-glucosidase (GAA) throughout the diaphragm. For the preclinical study, 10 rabbits (5 male, 5 female) were divided into two groups, vehicle control (Lactated Ringer's) and test article (1.5×1012 vector genomes [vg] rAAV1-CMV-hGAA), and euthanized on day 21. After direct visualization, the left hemidiaphragm was injected at three locations. There was up to a 2,500-fold increase in circulating anti-AAV1 antibodies directed to the vector capsids. In addition, up to an 18-fold increase in antibodies against the GAA protein was generated. Injection sites maintained up to 1.0×105 vg/μg genomic DNA (gDNA), while uninjected sites had up to 1.0×104 vg/μg gDNA. Vector DNA was present in blood at 24 hr postinjection at up to 1.0×106 vg/μg gDNA, followed by a decrease to 1.0×103 vg/μg gDNA at euthanization on day 21. Nominal amounts of vector DNA were present in peripheral organs, including the brain, spinal cord, gonads, and skeletal muscle. Upon histopathological examination...