Inhibition of cartilage destruction by double gene transfer of IL-1Ra and IL-10 involves the activin pathway

Neumann, Elena; Judex, Martin; Kullmann, Frank; Grifka, Joachim; Robbins, Paul D.; Pap, Thomas; Evans, Christopher H.; Schölmerich, Jürgen; Müller‐Ladner, Ulf

doi:10.1038/sj.gt.3301811

Cited by 62 publications

(58 citation statements)

References 77 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Underlining this concept, double gene transfer of interleukin-1 receptor antagonist (IL-1Ra) and IL-10 has been demonstrated to inhibit both cartilage invasion and the perichondrocytic cartilage degradation in the SCID mouse model of RA. 31 In line with our results targeting CL, in malignant cell lines, it could be shown that cDNA fragments inserted in antisense direction into an expression vector reduced both the mRNA levels and the enzymatic activity of CL in vitro. Of interest, the transduced cells showed also a decreased potential for tumor growth in Balb/c mice in vivo.…”

Section: Ribozymes Decrease Cartilage Destruction In Ra J Schedel supporting

confidence: 75%

Targeting cathepsin L (CL) by specific ribozymes decreases CL protein synthesis and cartilage destruction in rheumatoid arthritis

et al. 2004

Self Cite

View full text Add to dashboard Cite

The present study was undertaken to examine whether ribozymes cleaving specifically cathepsin L (CL) mRNA are able to decrease the synthesis of CL protease in rheumatoid arthritis synovial fibroblasts (RA-SF) and thereby reduce the invasiveness into cartilage both in vitro and in the SCID mouse coimplantation model of RA. Two different ribozymes that cleave CL mRNA specifically at positions 533 (RzCL533) and 790 (RzCL790) were generated. Using retroviral gene transfer, RA-SF were transduced with the ribozyme constructs or the empty vector. To examine the effect of the ribozymes on the mRNA level, quantitative analysis for CL mRNA was performed using real-time PCR. For evaluation on the protein level, ELISA using specific anti-CL antibodies was performed. In addition, transduced RA-SF were examined in vitro in a three-dimensional destruction assay evaluating their ability to degrade extracellular matrix produced by human chondrocytes. Matrix destruction was monitored by the release of soluble glycosaminoglycans (sGAG). Using the in vivo SCID mouse coimplantation model of RA, RzCL533-transduced RA-SF and control cells were coimplanted with human cartilage for 60 days. After being killed, invasion of RA-SF into the cartilage was evaluated by using a semiquantitative score. Transduction of RA-SF with RzCL533 and RzCL790 ribozymes decreased significantly the expression of CL mRNA to 44% (range 25-62%) and 20% (range 1-43%), respectively, when compared to mocktransduced cells. The protein concentration of CL in the cell culture supernatants of transduced RA-SF was decreased from 16.0 ng/ml in the mock constructs to 4.1 and 8.2 ng/ml (mean), respectively. Using the in vitro cartilage destruction assay, the release of sGAG decreased to 46 and 60%, respectively, after 14 days when compared to mocktransduced cells. In the SCID mouse coimplantation model of RA, RzCL533-transduced RA-SF revealed a significant lower cartilage invasion when compared to mock and untransduced cells. Using retroviral gene transfer, ribozymes cleaving CL mRNA inhibit specifically the synthesis of this matrix-degrading enzyme and reduce cartilage destruction in in vitro and in vivo models. Our study therefore suggests that ribozymes targeting CL could be a novel and efficient tool to inhibit joint destruction in RA.

show abstract

Section: Ribozymes Decrease Cartilage Destruction In Ra J Schedel supporting

confidence: 75%

Targeting cathepsin L (CL) by specific ribozymes decreases CL protein synthesis and cartilage destruction in rheumatoid arthritis

et al. 2004

Self Cite

View full text Add to dashboard Cite

show abstract

“…All patients gave informed consent and fulfilled the criteria of the American College of Rheumatology (38,39). Following enzymatic digestion, primary synovial fibroblasts were isolated and cultured in supplemented DMEM as described previously (40). Human mononuclear cells were isolated from peripheral blood using Vacutainer Cell Preparation Tubes (BD Biosciences), washed, and resuspended in DMEM ϩ 5% fetal calf serum.…”

Section: Methodsmentioning

confidence: 99%

Interleukin-17 (IL-17) and IL-1 Activate Translation of Overlapping Sets of mRNAs, Including That of the Negative Regulator of Inflammation, MCPIP1

Dhamija

Winzen

Doerrie

et al. 2013

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background:The role of translational control mechanisms in gene expression during inflammation is incompletely understood. Results:The proinflammatory cytokines IL-1 and IL-17 activate translation of certain mRNAs, including that of MCPIP1, a negative regulator of inflammation. Conclusion: Translational activation of MCPIP1 contributes to changes in gene expression induced by IL-1 and IL-17. Significance: Translational control may determine physiological and pathological consequences of inflammation.

show abstract

“…Over the last decade the anti-infl ammatory potential of IL-10 in arthritis has been proposed through in vivo animal studies indicating that this immunoregulatory cytokine (IL-10) might be a promising candidate for gene therapy in the infl amed joint (Lechman et al 1999(Lechman et al , 2003Neumann et al 2002;Van de Loo and van den Berg, 2002;Gelse et al 2003;Zhang et al 2004;Kuroda et al 2006). Only differentiated chondrocytes can be used for chondrocyte implantation to produce a stable cartilage-specifi c matrix and it remains unclear whether high concentrations of IL-10 might affect the differentiated phenotype of chondrocytes.…”

Section: Discussionmentioning

confidence: 99%

“…Several experimental in vivo studies revealed that the application of recombinant IL-10 or IL-10 overexpression vectors in the joint reduced infl ammation (Joosten et al 1997;Lechman et al 1999;Neumann et al 2002;Van de Loo and van den Berg, 2002;Zhang et al 2004;Kuroda et al 2006). However, the direct effects of IL-10 on articular chondrocyte homeostasis remain still unclear.…”

Section: Introductionmentioning

confidence: 99%

Cartilage-Specific Matrix and Integrin Expression in Three-Dimensional Articular Chondrocyte Cultures Overexpressing Human Interleukin-10

Müller

John

Kohl

et al. 2008

Clinical medicine. Arthritis and musculoskeletal disorders

View full text Add to dashboard Cite

Interleukin (IL)-10 overexpression inhibits joint infl ammation, however the effect of high local concentrations of IL-10 on chondrocyte homeostasis remains unclear. The aim of this study was to determine the effects of IL-10 overexpression on cartilage matrix production in three-dimensional (3D) chondrocyte cultures. Human articular chondrocytes were transduced with adenoviral vectors alone (adv/empty) or by vectors either overexpressing enhanced green fl uorescence protein (adv/EGFP) or human IL-10 (adv/hIL-10) before their transfer to a 3D culture system. Non-transduced chondrocytes were used as controls. The expression of IL-10 or EGFP was confi rmed using ELISA or fl ow cytometry. Chondrocytes synthesis of collagen types II and I, aggrecan, fi bronectin and β1-integrin was determined over a period of 14 days post transduction using fl ow cytometry or immunohistochemistry. adv/EGFP or adv/IL-10 transduced chondrocytes expressed EGFP or secreted IL-10 detectable over the 2 weeks culture period. No suppression of collagen type II, aggrecan or β1-integrin synthesis by IL-10 overexpression was found and the deposition of collagen type I and fi bronectin remained unaffected compared to the controls. IL-10 overexpression does not impair key features of chondrocytes differentiated phenotype (e.g. collagen type II and aggrecan expression) suggesting the potential use of IL-10 for gene therapeutic approaches in the joint.

show abstract

Inhibition of cartilage destruction by double gene transfer of IL-1Ra and IL-10 involves the activin pathway

Cited by 62 publications

References 77 publications

Targeting cathepsin L (CL) by specific ribozymes decreases CL protein synthesis and cartilage destruction in rheumatoid arthritis

Targeting cathepsin L (CL) by specific ribozymes decreases CL protein synthesis and cartilage destruction in rheumatoid arthritis

Interleukin-17 (IL-17) and IL-1 Activate Translation of Overlapping Sets of mRNAs, Including That of the Negative Regulator of Inflammation, MCPIP1

Cartilage-Specific Matrix and Integrin Expression in Three-Dimensional Articular Chondrocyte Cultures Overexpressing Human Interleukin-10

Contact Info

Product

Resources

About