title: Gene therapy with an improved doxycycline-regulated plasmid encoding a tumour necrosis factor-alpha inhibitor in experimental arthritis.
abstract: Inhibition of tumour necrosis factor (TNF)-alpha with biological molecules has proven an effective treatment for rheumatoid arthritis, achieving a 20% improvement in American College of Rheumatology score in up to 65% of patients. The main drawback to these and many other biological treatments has been their expense, which has precluded their widespread application. Biological molecules could alternatively be delivered by gene therapy as the encoding DNA. We have developed novel plasmid vectors termed pGTLMIK and pGTTMIK, from which luciferase and a dimeric TNF receptor II (dTNFR) are respectively expressed in a doxycycline (Dox)-regulated manner. Regulated expression of luciferase from the self-contained plasmid pGTLMIK was examined in vitro in a variety of cell lines and in vivo following intramuscular delivery with electroporation in DBA/1 mice. Dox-regulated expression of luciferase from pGTLMIK of approximately 1,000-fold was demonstrated in vitro, and efficient regulation was observed in vivo. The vector pGTTMIK encoding dTNFR was delivered by the same route with and without administration of Dox to mice with collagen-induced arthritis. When pGTTMIK was delivered after the onset of arthritis, progression of the disease in terms of both paw thickness and clinical score was inhibited when Dox was also administered. Vectors with similar regulation characteristics may be suitable for clinical application.
authors.affiliation: Bone and Joint Research Unit, Barts and The London, Queen Mary's School of Medicine and Dentistry, Charterhouse Square, University of London, London, UK. d.j.gould@qmul.ac.uk
authors.forename: David
authors.initials: D
authors.lastname: Gould
authors.forename: Nasim
authors.initials: N
authors.lastname: Yousaf
authors.forename: Rewas
authors.initials: R
authors.lastname: Fatah
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Subang
authors.forename: Yuti
authors.initials: Y
authors.lastname: Chernajovsky
mesh_terms: D000818:Animals; D001169:Arthritis, Experimental / Q000188:drug therapy* / Q000235:genetics; D019556:COS Cells; D002522:Chlorocebus aethiops; D004318:Doxycycline / Q000008:administration & dosage*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D006801:Humans; D051379:Mice; D008811:Mice, Inbred DBA; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000235:genetics
chemical_list: D014409:Tumor Necrosis Factor-alpha; D004318:Doxycycline