Persistent Expression of FLAG-tagged Micro-dystrophin in Nonhuman Primates Following Intramuscular and Vascular Delivery

被引：76

作者：

Rodino-Klapac, Louise R. ^{[1
,2
]}

Montgomery, Chrystal L. ^{[1
,2
]}

Bremer, William G. ^{[2
,3
]}

Shontz, Kimberly M. ^{[1
,2
]}

Malik, Vinod ^{[1
,2
]}

Davis, Nancy ^{[1
,2
]}

Sprinkle, Spencer ^{[1
,2
]}

Campbell, Katherine J. ^{[2
,3
]}

Sahenk, Zarife ^{[1
,2
,4
]}

Clark, K. Reed ^{[1
,2
]}

Walker, Christopher M. ^{[2
,3
]}

Mendell, Jerry R. ^{[1
,2
,4
]}

Chicoine, Louis G. ^{[1
,2
]}

机构：

[1] Nationwide Childrens Hosp, Res Inst, Ctr Gene Therapy, Columbus, OH 43205 USA

[2] Ohio State Univ, Dept Pediat, Nationwide Childrens Hosp, Columbus, OH 43210 USA

[3] Nationwide Childrens Hosp, Res Inst, Ctr Vaccines & Immun, Columbus, OH 43205 USA

[4] Ohio State Univ, Dept Neurol, Nationwide Childrens Hosp, Columbus, OH 43210 USA

来源：

MOLECULAR THERAPY | 2010年 / 18卷 / 01期

基金：

美国国家卫生研究院;

关键词：

DUCHENNE MUSCULAR-DYSTROPHY; RECOMBINANT ADENOASSOCIATED VIRUS; GENE-THERAPY; MDX MICE; T-CELLS; MUSCLE; VECTORS; TRANSDUCTION; DIAPHRAGM; PROTEIN;

D O I：

10.1038/mt.2009.254

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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.

引用

页码：109 / 117

页数：9

共 27 条

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