Gene expression profiling in circulating endothelial cells from systemic sclerosis patients shows an altered control of apoptosis and angiogenesis that is modified by iloprost infusion.
BACKGROUND
Circulating endothelial cells are increased in patients affected by systemic sclerosis (SSc) and their number strongly correlates with vascular damage. The effects of iloprost in systemic sclerosis are only partially known. We aimed at studying the gene expression profile of circulating endothelial cells and the effects of iloprost infusion and gene expression in patients with systemic sclerosis.


METHODS
We enrolled 50 patients affected by systemic sclerosis, 37 patients without and 13 patients with digital ulcers. Blood samples were collected from all patients before and 72 hours after either a single day or five days eight hours iloprost infusion. Blood samples were also collected from 50 sex- and age-matched healthy controls. Circulating endothelial cells and endothelial progenitors cells were detected in the peripheral blood of patients with systemic sclerosis by flow cytometry with a four-colour panel of antibodies. Statistical analysis was performed with the SPSS 16 statistical package.Circulating endothelial cells were then isolated from peripheral blood by immunomagnetic CD45 negative selection for the gene array study.


RESULTS
The number of both circulating endothelial cells and progenitors was significantly higher in patients affected by systemic sclerosis than in controls and among patients in those with digital ulcers than in patients without them. Circulating endothelial cells and progenitors number increased after iloprost infusion. Gene array analysis of endothelial cells showed a different transcriptional profile in patients compared to controls. Indeed, patients displayed an altered expression of genes involved in the control of apoptosis and angiogenesis. Iloprost infusion had a profound impact on endothelial cells gene expression since the treatment was able to modulate a very high number of transcripts.


CONCLUSIONS
We report here that circulating endothelial cells in patients with systemic sclerosis show an altered expression of genes involved in the control of apoptosis and angiogenesis. Moreover we describe that iloprost infusion has a strong effect on endothelial cells and progenitors since it is able to modulate both their number and their gene expression profile.
Tinazzi
Elisa
E
Department of Medicine, University of Verona, le LA Scuro, Verona, Italy. elisatinazzi@virgilio.it
Dolcino
Marzia
M
Puccetti
Antonio
A
Rigo
Antonella
A
Beri
Ruggero
R
Valenti
Maria Teresa
MT
Corrocher
Roberto
R
Lunardi
Claudio
C
D017209:Apoptosis / Q000187:drug effects / Q000235:genetics / Q000276:immunology; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D005260:Female; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D006801:Humans; D016285:Iloprost / Q000008:administration & dosage*; D008297:Male; D009389:Neovascularization, Pathologic / Q000188:drug therapy / Q000235:genetics / Q000276:immunology; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards; D012595:Scleroderma, Systemic / Q000188:drug therapy / Q000235:genetics / Q000276:immunology; D012883:Skin Ulcer / Q000188:drug therapy / Q000235:genetics / Q000276:immunology; D014665:Vasodilator Agents / Q000008:administration & dosage
D014665:Vasodilator Agents; D016285:Iloprost