Enhancement of Adenovirus-Mediated Gene Delivery to Rheumatoid Arthritis Synoviocytes and Synovium by Fiber Modifications:Role of Arginine-Glycine-Aspartic Acid (RGD)-and Non-RGD-Binding Integrins

摘要

Rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS) do not express the coxsackie-adenovirus (Ad) receptor and are poorly permissive to Ad serotype 5 (Ad5).Genetically modified,coxsackie-Ad receptor-independent Ad5 vectors were studied for gene delivery in human RA FLS and synovium explants and murine collagen-induced arthritis.Short-fiber Ad5 vectors with seven fiber shaft repeats Ad5GFP-R7-knob,Ad5GFP-R7-arginine-glycine-aspartic acid (RGD) (RGD-liganded),and Ad5GFP DELTAknob (knob-deleted) were compared with Ad5GFP-FiWT,a conventional wild-type (WT) Ad5 vector.Gene transfer by Ad5GFP-R7-knob and Ad5GFP-R7-RGD was 40- to 50-fold and 25-fold higher,respectively,than Ad5GFP-FiWT in FLS.Ad5GFP DELTAknob was more efficacious than its knob-bearing version Ad5GFP-R7-knob in FLS transduction.Virus attachment and entry required RGD-and LDV-binding integrins including alpha_v,alpha_v beta_3,alpha_v beta_5,and beta_1.Ad5GFP-R7-knob infection of FLS was partially neutralized by synovial fluid (SF),but remained 30- to 40-fold higher than Ad5GFP-FiWT in the presence of SF.Ad5GFP DELTAknob was partially neutralized by SF at low virus input,but escaped viral neutralization by SF at higher virus input.Gene transfer to human synovium ex vivo explants and murine collagen-induced arthritis in vivo was also more efficient with short fiber-modified vectors (with and without the knob domain) than Ad5GFPFiWT.Gene transfer by short fiber-modified vectors was enhanced by inflammatory cytokines in vitro and in the presence of inflammation in murine synovium in vivo.Our data indicated that the highly efficient gene delivery RA was mediated by RGD- and non-RGD-binding integrins and enhanced by inflammation.Short fiber modifications with knob ablation may be a strategy to enhance gene delivery,reducing vector dose and vector-induced inflammation and toxicity.