title: Cyclosporin A differentially inhibits multiple steps in VEGF induced angiogenesis in human microvascular endothelial cells through altered intracellular signaling.
abstract: The immunosuppressive agent cyclosporin A (CsA), a calcineurin inhibitor which blocks T cell activation has provided the pharmacologic foundation for organ transplantation. CsA exerts additional effects on non-immune cell populations and may adversely effect microvascular endothelial cells, contributing to chronic rejection, a long-term clinical complication and significant cause of mortality in solid-organ transplants, including patients with small bowel allografts. Growth of new blood vessels, or angiogenesis, is a critical homeostatic mechanism in organs and tissues, and regulates vascular populations in response to physiologic requirements. We hypothesized that CsA would inhibit the angiogenic capacity of human gut microvessels. Primary cultures of human intestinal microvascular endothelial cells (HIMEC) were used to evaluate CsA's effect on four in vitro measures of angiogenesis, including endothelial stress fiber assembly, migration, proliferation and tube formation, in response to the endothelial growth factor VEGF. We characterized the effect of CsA on intracellular signaling mechanisms following VEGF stimulation. CsA affected all VEGF induced angiogenic events assessed in HIMEC. CsA differentially inhibited signaling pathways which mediated distinct steps of the angiogenic process. CsA blocked VEGF induced nuclear translocation of the transcription factor NFAT, activation of p44/42 MAPK, and partially inhibited JNK and p38 MAPK. CsA differentially affected signaling cascades in a dose dependent fashion and completely blocked expression of COX-2, which was integrally linked to HIMEC angiogenesis. These data suggest that CsA inhibits the ability of microvascular endothelial cells to undergo angiogenesis, impairing vascular homeostatic mechanisms and contributing to the vasculopathy associated with chronic rejection.
authors.affiliation: Department of Surgery, Medical College of Wisconsin, Milwaukee, WI, 53226, USA. prafiee@mcw.edu
authors.forename: Parvaneh
authors.initials: P
authors.lastname: Rafiee
authors.forename: Jan
authors.initials: J
authors.lastname: Heidemann
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Ogawa
authors.forename: Nathan A
authors.initials: NA
authors.lastname: Johnson
authors.forename: Pamela J
authors.initials: PJ
authors.lastname: Fisher
authors.forename: Mona S
authors.initials: MS
authors.lastname: Li
authors.forename: Mary F
authors.initials: MF
authors.lastname: Otterson
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Johnson
authors.forename: David G
authors.initials: DG
authors.lastname: Binion