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Adeno-associated virus 8-mediated gene therapy for choroideremia: preclinical studies in in vitro and in vivo models.

The journal of gene medicine (2014-06-26)
Aaron Black, Vidyullatha Vasireddy, Daniel C Chung, Albert M Maguire, Rajashekhar Gaddameedi, Tania Tolmachova, Miguel Seabra, Jean Bennett
ABSTRACT

Choroideremia (CHM) is a slowly progressive X-linked retinal degeneration that results in a loss of photoreceptors, retinal pigment epithelium and choroid. CHM, the gene implicated in choroideremia, encodes Rab escort protein-1 (REP-1), which is involved in the post-translational activation via prenylation of Rab proteins. We evaluated AAV8.CBA.hCHM, a recombinant adeno-associated virus serotype 8 (rAAV8) vector, which targets retinal cells efficiently, for both therapeutic effect and safety in vitro and in vivo in a murine model. In vitro studies included western blot analyses and prenylation assays. In vivo studies included ophthalmoscopy, pupillometry, histology and immunofluorescence analysis. Infection with AAV8.CBA.hCHM induced the expression of REP-1 protein in a dose-responsive fashion. Transduction with AAV8.CBA.hCHM reverses the biochemical and pathogenetic defects in CHM both in vitro and in vivo and showed no safety concerns in the in vivo investigations performed in the present study. AAV8 is a promising vector for human clinical gene therapy trials for choroideremia.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-(−)-Glucose, ≥99%
Sigma-Aldrich
Anti-MRP1 antibody ,Mouse monoclonal, clone QCRL-4, purified from hybridoma cell culture
Sigma-Aldrich
Anti-CHM antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution
Sigma-Aldrich
Anti-MSH3 (center) antibody produced in rabbit, saturated ammonium sulfate (SAS) precipitated, buffered aqueous solution
Sigma-Aldrich
Anti-MRP1 antibody,Mouse monoclonal, clone QCRL-1, purified from hybridoma cell culture