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Persistent expression of FLAG-tagged micro dystrophin in nonhuman primates following intramuscular and vascular delivery.


ABSTRACT: Animal models for Duchenne muscular dystrophy (DMD) have species limitations related to assessing function, immune response, and distribution of micro- or mini-dystrophins. Nonhuman primates (NHPs) provide the ideal model to optimize vector delivery across a vascular barrier and provide accurate dose estimates for widespread transduction. To address vascular delivery and dosing in rhesus macaques, we have generated a fusion construct that encodes an eight amino-acid FLAG epitope at the C-terminus of micro-dystrophin to facilitate translational studies targeting DMD. Intramuscular (IM) injection of AAV8.MCK.micro-dys.FLAG in the tibialis anterior (TA) of macaques demonstrated robust gene expression, with muscle transduction (50-79%) persisting for up to 5 months. Success by IM injection was followed by targeted vascular delivery studies using a fluoroscopy-guided catheter threaded through the femoral artery. Three months after gene transfer, >80% of muscle fibers showed gene expression in the targeted muscle. No cellular immune response to AAV8 capsid, micro-dystrophin, or the FLAG tag was detected by interferon-gamma (IFN-gamma) enzyme-linked immunosorbent spot (ELISpot) at any time point with either route. In summary, an epitope-tagged micro-dystrophin cassette enhances the ability to evaluate site-specific localization and distribution of gene expression in the NHP in preparation for vascular delivery clinical trials.

SUBMITTER: Rodino-Klapac LR 

PROVIDER: S-EPMC2839222 | biostudies-literature | 2010 Jan

REPOSITORIES: biostudies-literature

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Persistent expression of FLAG-tagged micro dystrophin in nonhuman primates following intramuscular and vascular delivery.

Rodino-Klapac Louise R LR   Montgomery Chrystal L CL   Bremer William G WG   Shontz Kimberly M KM   Malik Vinod V   Davis Nancy N   Sprinkle Spencer S   Campbell Katherine J KJ   Sahenk Zarife Z   Clark K Reed KR   Walker Christopher M CM   Mendell Jerry R JR   Chicoine Louis G LG  

Molecular therapy : the journal of the American Society of Gene Therapy 20091110 1


Animal models for Duchenne muscular dystrophy (DMD) have species limitations related to assessing function, immune response, and distribution of micro- or mini-dystrophins. Nonhuman primates (NHPs) provide the ideal model to optimize vector delivery across a vascular barrier and provide accurate dose estimates for widespread transduction. To address vascular delivery and dosing in rhesus macaques, we have generated a fusion construct that encodes an eight amino-acid FLAG epitope at the C-terminu  ...[more]

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