Article Text

PDF
AB0084 Non-canonical nf-kappab signaling induces a pro-angiogenic response in rheumatoid arthritis synovial fibroblasts and endothelial cells
  1. C. X. Maracle1,2,
  2. A. R. Noort1,2,
  3. K. P. van Zoest1,2,
  4. S. W. Tas1,2
  1. 1Divison of Clinical Immunology and Rheumatology
  2. 2Department of Experimental Immunology, Academic Medical Center/University of Amsterdam, Amsterdam, Netherlands

Abstract

Background In rheumatoid arthritis (RA) leukocytes migrate to the joints and gain entry into the synovial tissue through increased permeability of local endothelial cells[1]. These immune cells activate synovial fibroblasts (SF) that in turn promote inflammation and cartilage destruction[2]. RASF also contribute to angiogenesis, which allows for further leukocyte entry into the inflamed synovium, essentially generating a positive feedback loop, and is regarded as a switch from acute to chronic inflammation[3]. Based on previous work done by our group, the non-canonical nuclear factor kappaB (NF-kB) pathway, with its main regulator NF-kB inducing kinase (NIK) may play a central role in this process[4].

Objectives To determine the effect of non-canonical NF-kB signaling on pro-angiogenic gene expression in RASF and on the angiogenic potential of human umbilical vein endothelial cells (HUVEC).

Methods RASF were stimulated with lymphotoxin α1β2 (LT) or LIGHT to activate non-canonical NF-kB signaling, and/or TNF to selectively activate the canonical NF-kB pathway. To effectively block the non-canonical pathway, a dominant negative NIK expressing adenovirus (Ad.NIKdn) was used in comparison with Ad.GFP as a control vector. Changes in pro-angiogenic gene expression were measured by RT-PCR. Furthermore, to determine the effect on EC proliferation, RASF and HUVEC were co-cultured in the presence or absence of LT, LIGHT, TNF or VEGF. EC proliferation was visualized by immunohistochemical staining of CD31 and scored semi-quantitatively.

Results Gene expression analysis of RASF revealed an increase in mRNA levels of VCAM-1 and IL-6 after stimulation with LT, LIGHT and TNF. Increased expression of IL-8 and MMP-3 was also observed in cells treated with both TNF and LT or LIGHT. These levels were attenuated in cells transduced with Ad.NIKdn prior to stimulation, indicating that the increased expression levels were at least in part non-canonical NF-kB dependent. The pro-angiogenic factors CCL2 and bFGF were expressed continuously by RASF regardless of stimulation. In the co-culture, proliferation levels of EC increased under all stimulation conditions, with LIGHT inducing almost a 2-fold increase (p<0.05), which was comparable to VEGF (p<0.05).

Conclusions RASF contribute to synovial angiogenesis through the expression of adhesion molecules, cytokines, chemokines, matrix remodeling enzymes and growth factors. We demonstrate that the non-canonical NF-kB pathway plays an important role in this process by regulating pro-angiogenic genes and promoting EC proliferation. Further investigation of this pathway could lead to novel non-canonical NF-kB blocking therapeutics that inhibit angiogenesis in RA and halt disease progression.

  1. McInnes IB, Schett G. The pathogenesis of rheumatoid arthritis. N Engl J Med, 2011 Dec 8; 365(23): 2205-19.

  2. Bottini N, Firestein GS. Duality of fibroblast-like synoviocytes in RA: passive responders and imprinted aggressors. Nat. Rev. Rheumatol, 2012 Nov; 9, 24–33.

  3. Szekanec Z, Besenyei T, Paragh G, Koch AE. Angiogenesis in rheumatoid arthritis. Autoimmunity, 2009 Nov; 42(7): 563-73.

  4. Noort AR, van Zoest KPM, Koolwijk P, Tak PP, Tas SW. NF-kappaB Inducing Kinase (NIK) is a key regulator of inflammation-induced angiogenesis. Arthritis Rheum, 2012 Oct; 64(10): S683.

Disclosure of Interest None Declared

Statistics from Altmetric.com

Request permissions

If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.