Session Information
Date: Monday, November 9, 2015
Title: Rheumatoid Arthritis - Small Molecules, Biologics and Gene Therapy Poster II
Session Type: ACR Poster Session B
Session Time: 9:00AM-11:00AM
Background/Purpose: The current treatment for patients with rheumatoid arthritis (RA) consists of biological drugs, often in combination with methotrexate. However, some patients fail to respond to biological therapy in one or multiple joints. We are therefore aiming to develop a gene therapy for local treatment of affected joints. By using disease-inducible promoters to drive expression, the joints will only produce biological drugs during a disease flare.
Methods: Microarrays on joint tissue of 20 RA patients and 7 patients without joint disease were analyzed to find genes that are upregulated in the joint during active disease.
The promoter of the CXCL10 gene was isolated from human cDNA and cloned into a lentiviral vector containing the firefly luciferase reporter gene. THP-1 cells were transduced with the lentiviral constructs and stimulated for 6 hours with pro-inflammatory stimuli, 64 RA patient sera and 63 control sera to determine the inducibility of the promoters. The luciferase reporter gene was replaced by IL-10 and the expression of recombinant IL-10 was measured under control of the CXCL10promoter. The production of TNFα, IL-1β, IL-8 and MCP-1 by lipopolysaccharide (LPS) stimulated synovial cells were measured by multiplex ELISA assay.
Results: The microarrays showed a 10-fold upregulation of CXCL10, an interferon inducible gene known to be expressed by many cell types. Primary cells obtained from patient synovium were transduced with the CXCL10-luciferase reporter vector and stimulated with pro-inflammatory lipopolysaccharide (LPS) or with TNFα. This resulted in a 3.3- and 2.3-fold upregulation of the luciferase signal respectively. The CXCL10 promoter was also activated by serum from RA patients and could significantly distinguish between RA serum and healthy donor serum (P=0.017). The CXCL10 promoter in THP-1 monocytes showed a 33.7-fold upregulation after 8h stimulation with TNFα. The promoter activity declined after 96h, but was re-inducible by a second TNFα-stimulation. This shows the temporal responsiveness of the CXCL10promoter to inflammatory signals and the vigilant state under basal conditions. For therapeutic testing, the luciferase reporter gene was replaced by the anti-inflammatory Interleukin-10 (IL-10) gene. RA synovial cells transduced with the CXCL10-IL10 lentivirus produced significantly less pro-inflammatory cytokines (TNFα, IL-1β, IL-8 and MCP-1) after LPS stimulation compared to control virus. This was observed in multiple patients. These results show the functionality of the CXCL10-IL10 lentivirus in the desired synovial target cells.
Conclusion: Our CXCL10 regulated IL-10 gene construct shows good responsiveness to inflammatory stimuli and inhibits inflammatory cytokine production by RA synovial cells. CXCL10-regulated IL10 overexpression can thus provide inflammation-inducible local gene therapy suitable for joints with persistent RA that are refractory to treatment.
To cite this abstract in AMA style:
Broeren MGA, Bennink MB, Arntz OJ, van den Berg W, van de Loo FAJ. Disease-Regulated Expression of Anti-Inflammatory Interleukin-10 for the Treatment of Rheumatoid Arthritis [abstract]. Arthritis Rheumatol. 2015; 67 (suppl 10). https://acrabstracts.org/abstract/disease-regulated-expression-of-anti-inflammatory-interleukin-10-for-the-treatment-of-rheumatoid-arthritis/. Accessed .« Back to 2015 ACR/ARHP Annual Meeting
ACR Meeting Abstracts - https://acrabstracts.org/abstract/disease-regulated-expression-of-anti-inflammatory-interleukin-10-for-the-treatment-of-rheumatoid-arthritis/