title: Adeno-associated virus type 5-mediated intraarticular administration of tumor necrosis factor small interfering RNA improves collagen-induced arthritis.
abstract: OBJECTIVE
  RNA interference (RNAi) is a powerful tool for sequence-specific gene silencing, and interest in its application in human diseases is growing. Given the success of recent strategies for administering gene therapy in rheumatoid arthritis using recombinant vectors such as adeno-associated virus type 5 (rAAV5) for optimized intraarticular gene transfer, we undertook the present study to determine the feasibility of using rAAV5-mediated RNAi-based therapy in arthritis.
  
  
  METHODS
  We developed rAAV5 vectors expressing short hairpin small interfering RNA (shRNA) against tumor necrosis factor alpha (TNFalpha) under H1 promoter, and carrying the enhanced green fluorescent protein (eGFP) reporter gene under cytomegalovirus promoter (rAAV5-shTNF). TNFalpha gene silencing was validated in vitro with mouse macrophages. Mice with collagen-induced arthritis were injected in the ankle and knee joints, at disease onset, with either rAAV5-shTNF or control rAAV5-eGFP vectors (5 x 10(9) particles). Arthritis severity was assessed clinically and histologically, and immunologic response was examined. Local and systemic transgene expression was monitored using quantitative reverse transcriptase-polymerase chain reaction, immunohistochemical analysis, and enzyme-linked immunosorbent assay.
  
  
  RESULTS
  After a single injection of rAAV5-shTNF into inflamed joints, local TNFalpha gene silencing provided rapid and long-term suppression of arthritis progression and reduced joint damage compared with that observed in control groups. Treatment with rAAV5-shTNF was associated with decreased proliferation and interferon-gamma production by antigen-stimulated T cells from draining lymph nodes, and the potency of this treatment was similar to that observed with other treatment strategies targeting TNFalpha at the protein level, either locally or systemically.
  
  
  CONCLUSIONS
  Our data present the first proof-of-concept for the application of rAAV5-mediated RNAi-based gene therapy for local blockade of inflammation in experimental arthritis.
authors.affiliation: INSERM, U844, Hôpital Saint Eloi, and Université Montpellier I, Montpellier, France.
authors.forename: Maroun
authors.initials: M
authors.lastname: Khoury
authors.forename: Gabriel
authors.initials: G
authors.lastname: Courties
authors.forename: Sylvie
authors.initials: S
authors.lastname: Fabre
authors.forename: Carine
authors.initials: C
authors.lastname: Bouffi
authors.forename: Christian A
authors.initials: CA
authors.lastname: Seemayer
authors.forename: Margriet J
authors.initials: MJ
authors.lastname: Vervoordeldonk
authors.forename: Paul P
authors.initials: PP
authors.lastname: Tak
authors.forename: Christian
authors.initials: C
authors.lastname: Jorgensen
authors.forename: Florence
authors.initials: F
authors.lastname: Apparailly
mesh_terms: D000818:Animals; D001169:Arthritis, Experimental / Q000628:therapy*; D000229:Dependovirus; D015536:Down-Regulation; D004797:Enzyme-Linked Immunosorbent Assay; D020868:Gene Silencing / Q000502:physiology; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D007150:Immunohistochemistry; D007270:Injections, Intra-Articular; D051379:Mice; D034622:RNA Interference / Q000502:physiology*; D034741:RNA, Small Interfering / Q000008:administration & dosage*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics* / Q000378:metabolism
chemical_list: D034741:RNA, Small Interfering; D014409:Tumor Necrosis Factor-alpha; D049452:Green Fluorescent Proteins