Arthritis & Rheumatism, Volume 63,
November 2011 Abstract Supplement
Abstracts of the American College of
Rheumatology/Association of Rheumatology Health Professionals
Annual Scientific Meeting
Chicago, Illinois November 4-9, 2011.
Analysis of Endothelium from Systemic Lupus Erythematosus Patients Demonstrates a Marked Interferon Inducible Signature and an Associated Decrease in Transforming Growth Factor Beta Signaling Genes.
Goldenberg1, Diana, Olferiev1, Mikhail, Onat2, Duygu, Harxhi2, Ante, Andrade1, Danieli, Crow1, Mary K., Colombo2, Paolo
Endothelial dysfunction may promote premature atherosclerosis in patients with systemic lupus erythematosus (SLE). Using a novel approach of endothelial sampling coupled with microarray analysis of amplified endothelial RNA, we have previously shown upregulation of interferon inducible genes in SLE patients compared with healthy controls. Herein, we used quantitative real time polymerase chain reaction (RT-PCR) and quantitative immunofluorescence microscopy i) to further validate our initial interferon findings and ii) to investigate the interaction between the interferon and transforming growth factor beta (TGFb) signaling pathways.
Fourteen patients with SLE (age 30.9 ± 9.4 yrs, SLEDAI score 6.2; range 020) and 12 age-matched healthy subjects were studied. Endothelial cells were collected from arm veins using endovascular wires then separated using magnetic beads coated with endothelial specific antibodies. Analysis of the microarray profiles was performed on amplified RNA using GeneSpring GX 11 software. Student T test analysis was performed and differentially expressed genes with >1.5 fold expression and p<0.05 were selected. Microarray findings were further validated using RT-PCR and quantitative immunofluorescence microscopy (Scion Image Software 4.0).
Microarray analysis showed that several interferon-inducible genes (i.e. IFIT3, IFI44L, IFI6, MX2, IFITM1, OAS1 and OAS2) were upregulated and that several genes in the TGFb signaling pathway (i.e. TGFBR2, TGFBR3, SMAD2 and SMAD3) were downregulated in SLE patients compared with controls. RT-PCR confirmed upregulation of IFIT3 and downregulation of TGFBR2, TGFBR3 and SMAD2 in the SLE patients. Immunofluorescence analysis demonstrated increased protein levels of IFIT3 and ISG15 in SLE patients compared with controls.
We used a novel in-vivo approach of venous endothelial sampling coupled with gene expression and protein analysis to study the molecular events that promote endothelial dysfunction in SLE. Our results indicate an opposite expression pattern of interferon and TGFb signaling pathways in the venous endothelium of SLE patients, the former being upregulated and the latter downregulated. These findings further characterize processes that appear relevant to the pathobiology of endothelial dysfunction in SLE.
To cite this abstract, please use the following information:
Goldenberg, Diana, Olferiev, Mikhail, Onat, Duygu, Harxhi, Ante, Andrade, Danieli, Crow, Mary K., et al; Analysis of Endothelium from Systemic Lupus Erythematosus Patients Demonstrates a Marked Interferon Inducible Signature and an Associated Decrease in Transforming Growth Factor Beta Signaling Genes. [abstract]. Arthritis Rheum 2011;63 Suppl 10 :1438