Adeno-associated virus production of soluble tumor necrosis factor receptor neutralizes tumor necrosis factor alpha and reduces arthritis

• Published in: Human gene therapy Vol. 11; no. 17; p. 2431
• Main Authors: Zhang, H G, Xie, J, Yang, P, Wang, Y, Xu, L, Liu, D, Hsu, H C, Zhou, T, Edwards, 3rd, C K, Mountz, J D
• Format: Journal Article
• Language: English
• Published: United States 20.11.2000
• Subjects: Animals; Antigens, CD - genetics; Antigens, CD - metabolism; Arthritis - pathology; Arthritis - therapy; Arthritis, Experimental - therapy; Arthritis, Rheumatoid - pathology; Cells, Cultured; Collagenases - drug effects; Collagenases - metabolism; Dependovirus - genetics; Female; Fibroblasts - virology; Genetic Therapy - methods; Humans; L Cells (Cell Line); Mice; Mice, Transgenic; Muscles - virology; Receptors, Tumor Necrosis Factor - genetics; Receptors, Tumor Necrosis Factor - metabolism; Receptors, Tumor Necrosis Factor, Type I; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Synovitis - pathology; Synovitis - therapy; Toxicity Tests; Tumor Necrosis Factor-alpha - metabolism
• ISSN: 1043-0342
• DOI: 10.1089/104303400750038525

The major limitation of adenovirus is its association with induction of an inflammatory response and relatively short-term production of the gene therapy transgene product. Adeno-associated virus (AAV) is a 4.68-kb single-strand DNA virus that contains ITRs for viral replication and a packaging signal, and also has been engineered to contain therapeutic genes up to 5 kb in length. Transduction of recombinant AAV (rAAV) results in low inflammatory response and long-term expression. We have cloned a low-immunogenic form of human sTNFRI (sTNFRI2.6D) into AAV (rAAVsTNFRI). This vector was analyzed for its ability to transfect and neutralize the effect of TNF-alpha on primary rheumatoid arthritis synovial fibroblast (RASFs). The rAAVsTNFRI was transduced into the cells at 1.8 x 10(1), 1.8 x 10(2), and 1.8 x 10(3) viral particles per cell. There was greater than 90% neutralization of TNF-alpha at 1.8 x 10(3) viral particles/cell. There was a significant decrease in the synovial cell hyperplasia and cartilage and bone destruction in human TNF-alpha transgenic mice treated intraarticularly with rAAVsTNFRI. These results indicate that the low-immunogenic and long-term expressing vector, rAAVsTNFRI, can be used to deliver the soluble TNF-alpha in vitro and in vivo and effectively reduce the severity of arthritis.