Arthritis & Rheumatism, Volume 62,
November 2010 Abstract Supplement
Abstracts of the American College of
Rheumatology/Association of Rheumatology Health Professionals
Annual Scientific Meeting
Atlanta, Georgia November 6-11, 2010.
A Novel Role for Inducible Fucosyltransferase2 as a Regulator of Angiogenesis.
Amin1, Mohammad A., Ruth2, Jeffrey H., Tsou2, Pei-suen, Campbell2, Phillip L., Marotte2, Hubert, Lee2, SolHee, Nallasivam2, Sivakumar
Fucosyltransferases (Futs) are involved in the synthesis of fucosylated glycans and blood group antigens. A number of studies have shown the role of some Futs in angiogenesis and inflammation. The importance of angiogenesis is well established in diseases such as rheumatoid arthritis (RA). This is a novel study which demonstrates that Fut2, heretofore not known to regulate angiogenesis, is cytokine inducible in human dermal microvascular endothelial cells (HMVECs) and plays an essential role in inflammatory angiogenesis.
We performed reverse transcriptase polymerase chain reaction (RT-PCR) and quantitative PCR to determine Fut2 mRNA expression in HMVECs after stimulation with interleukin-1b (IL-1b). To determine the role of signaling molecules in Fut2 mRNA expression, we stimulated HMVECs with IL-1b in the presence or absence of chemical signaling inhibitors and collected RNA to examine the role of signaling molecules in Fut2 expression. We harvested endothelial cells (ECs) from Fut2 null and wild type (wt) mice and performed Matrigel tube formation assays in vitro. We also used Matrigel plug and the inflammatory sponge granuloma angiogenesis assays in vivo to elucidate the significance of Fut2 in angiogenesis by employing Fut2 null and wt mice. To evaluate the mechanism by which Fut2 contributes to angiogenesis, we stimulated Fut2 null and wt mouse ECs with IL-1b, and performed enzyme linked immunosorbent (ELISA) assays to determine if Fut2 deficient ECs produced lower amounts of angiogenic factors than their wt counterparts.
IL-1b enhanced Fut2 mRNA expression in HMVECs compared to nonstimulated cells with a maximum increase at 60 minutes. Phosphatidylinositol 3 kinase and p38 were critical in HMVEC IL-1b stimulated Fut2 mRNA expression. Fut2 null ECs exhibited a significant 2 fold decrease in tube formation compared to wt ECs, indicating that Fut2 may be an important factor in angiogenesis in vitro. To determine whether Fut2 contributed to angiogenesis in vivo, we performed mouse Matrigel plug and inflammatory sponge granuloma angiogenesis assays using Fut2 null and wt mice. Fut2 null mice were deficient in angiogenesis in both the Matrigel plug and the sponge granuloma compared to wt mice (p<0.05). To determine the mechanism of impaired angiogenesis in the absence of Fut2, ECs harvested from Fut2 null and wt mice were stimulated with IL-1b. Vascular endothelial growth factor (VEGF) was less in Fut2 null ECs at both the mRNA and protein levels, suggesting a novel role for Fut2 as regulator of angiogenesis.
These data suggest an intriguing role for Fut2 as regulator of angiogenesis. Fut2 may be a potential therapeutic target controlling production of angiogenic factors such as VEGF in angiogenesis-dependent diseases such as RA.
To cite this abstract, please use the following information:
Amin, Mohammad A., Ruth, Jeffrey H., Tsou, Pei-suen, Campbell, Phillip L., Marotte, Hubert, Lee, SolHee, et al; A Novel Role for Inducible Fucosyltransferase2 as a Regulator of Angiogenesis. [abstract]. Arthritis Rheum 2010;62 Suppl 10 :1504