Amelioration of collagen‐induced arthritis in rats by adenovirus‐mediated PTEN gene transfer

Abstract

Objective. The phosphatidylinositol 3-kinase (PI 3-kinase)/Akt pathway is known to be activated in rheumatoid arthritis (RA) synovial tissue, which impacts cell growth, proliferation, survival, and migration. Phosphatase and tensin homolog deleted from chromosome 10 (PTEN) functions as a negative regulator of PI 3-kinase signaling, thus blocking Akt activation. The aim of this study was to examine the effect of PTEN gene transfer in rats with collagen-induced arthritis (CIA).Methods. Adenoviral vectors encoding human PTEN (AdPTEN) or ␤-galactosidase (AdLacZ) were injected intraarticularly into rats with CIA, and their treatment responses were monitored by measures of clinical, radiographic, and histologic changes. The expression of phosphorylated Akt, total Akt, vascular endothelial growth factor (VEGF), proinflammatory cytokines, and chemokines, as well as the extent of microvessel density in the ankle joints were determined.Results. AdPTEN treatment reduced Akt phosphorylation and decreased VEGF production in human RA synovial fibroblasts. Compared with AdLacZ treatment of the rats with CIA, AdPTEN treatment significantly reduced ankle circumference, articular index scores, radiography scores, and histology scores, and also decreased microvessel density and levels of VEGF and interleukin-1␤. Furthermore, PTEN gene transfer led to down-regulation of Akt activation and increased apoptosis in the ankle joints.Conclusion. This study is the first to demonstrate the in vivo effect of intraarticular gene delivery of PTEN on amelioration of arthritis symptoms in rats with CIA, which involved antiangiogenic, antiproliferative, and antiinflammatory effects of PTEN via inhibition of the PI 3-kinase/Akt signaling pathway. Our findings also implicate the PI 3-kinase/Akt pathway as a therapeutic target for the treatment of RA or other inflammatory diseases.

Intra-articular lentivirus-mediated delivery of galectin-3 shRNA and galectin-1 gene ameliorates collagen-induced arthritis

“…Differences in articular index scores between two treatment groups were compared by repeated-measures analysis of variance (ANOVA) using SAS software version 9.1 (GLM program; SAS Institute, Cary, NC, USA). 9 Differences in the remaining data between two groups were compared with Student's t-test. P-values less than 0.05 were considered significant.…”

“…Sections of paraffin-embedded joint specimens were stained with hematoxylin and eosin and immunostained with rabbit anti-von Willebrand factor (Dako, Carpinteria, CA, USA) or anti-CD3 (BD Biosciences, San Diego, CA, USA) antibodies as described previously. [6][7][8][9] Assay of antigen-mediated activation-induced T-cell death in the lymph node cells transduced with lentiviral vectors Rats with CIA were sacrificed on day 18. Cells obtained from the inguinal lymph nodes were infected with Lt.Gal1, Lt.GFP or Lt.shGal3 at an multiplicity of infection of 10 in the presence of polybrene (8 mg ml À1 ) and cultured for 72 h. Cells were then treated with denatured type II collagen (100 mg ml À1 ) for 24 h. They were stained with propidium iodide and analyzed for the proportions of dead cells by flow cytometry using a FACScan (Becton Dickinson, San Jose, CA, USA).…”

“…injection into ankle joints of rats with CIA. [6][7][8][9] Such proof of concept in animal models of RA provides important validation of the pre-clinical miniature, and has led to the elucidation of putative roles for certain molecules in the pathogenesis of inflamed joints.…”

Gene Therapy Outcomes in Experimental Models of Inflammatory Arthritis

“…Further, T-cells recovered from the STAT-4 deficient mice produced less IL-6, TNF-and IL-17. In another interesting study, adenoviral vectors encoding PTEN (AdPTEN) or -galactosidase (AdLacZ) were injected directly into the joints of rats with CIA (Wang et al, 2008). AdPTEN was shown to reduce ankle circumference, articular cartilage degradation, radiographic and histology scores compared to rats with CIA injected with AdLacZ.…”

“…In addition, Ad.ExTek protected mice with CIA against bony erosions, suppressed neoangiogenesis, reduced the level of RANKL, but did not suppress anti-Type II collagen antibody production. However, a search of the PubMed literature since 2007 did not reveal any current studies directed at targeting angiogenesis via either Ang-1 or Tie-2 gene therapy suggesting that a greater focus has been placed on defining a common thread such as PI3K/Akt/PTEN/mTor pathway activation as a way of regulating neoangiogenesis in CIA (Wang et al, 2008). The failure of chondrocytes to respond to insulin-like growth factor-1 (IGF-1) has been proposed as one of the mechanisms that underlie the inefficient repair of articular cartilage www.intechopen.com in arthritis (van de Loo et al, 2008).…”