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WO6-OR-4 THE RELATIONSHIP BETWEEN ENDOTHELIAL PROGENITOR CELLS, THEIR APOPTOTIC MICROPARTICLES AND THE FRAMINGHAM RISK
Workshops WO6 Atherosclerosis and progenitor cells
6 WO5-OR-6
COMMON VARIANT OF FARNESOID-X-RECEPTOR GENE ENHANCES THE RESPONSE TO LIPID-LOWERING THERAPY
A. Nohara 1 , H. Tada 2 , S. Katsuda 2 , M. Kawashiri 2 , A. Inazu 3 , J. Kobayashi 1 , M. Yamagishi 2 , M. Mabuchi 1 . 1 Department of Lipidology,Kanazawa University Graduate School of Medical Science, Kanazawa,Ishikawa, Japan; 2 Division of Cardiovascular Medicine, Kanazawa University Graduate School of Medical Science, Kanazawa,Ishikawa, Japan; 3 School of Health Science, Kanazawa University, Kanazawa,Ishikawa, Japan Bile acid receptor FXR has crucial roles in cholesterol conversion into bile acids and in its recycling through many target genes that may affect cholesterol levels. We have identified common polymorphism -1g->t in FXR gene in Japanese population, and hypothesized that this polymorphism could affect lipid-lowering therapy response. Methods and Results: Total 276 patients (M/F=147/129) suspected CAD were enrolled. FXR -1g->t genotypes were determined by PCRRFLP, and -1t allele frequency was 0.32. Lipid-lowering drugs (statins 92%) were prescribed in 113 patients (M/F=46/67), and baseline Tcho level was not different with FXR genotype [gg(n=65) vs. gt+tt(n=48) = 253 ± 42 vs. 251 ± 41(mg/dl; mean ± SD)]. With lipid-lowering therapy, -1t carriers showed stronger reduction in Tcho [gg vs. gt+tt = 211 ± 33(-16%) vs. 182 ± 31(-27%) mg/dl, p<0.0001], and in LDL-C [125 ± 34(-22%) vs. 100 ± 25(-39%) mg/dl, p<0.0001]. There was no difference in triglycerides reduction or HDL-C increase. Conclusion: These results demonstrated that common genetic variant of FXR gene showed considerable impact on lipid-lowering therapy, probably through modulation of genes involved in cholesterol metabolism such as CYP7A1. Whether this variant could affect long-term clinical course should further be sought.
WO6 ATHEROSCLEROSIS AND PROGENITOR CELLS WO6-OR-3
ENDOTHELIAL PROGENITOR CELLS AND ENDOTHELIAL-DERIVED MICROPARTICLES IN HYPERTRIGLYCERIDEMIA: THE EFFECT OF THE METABOLIC SYNDROME
M.R. Mannarino, M. Pirro, F. Bagaglia, C. Menecali, R. Paltriccia, G. Schillaci, E. Mannarino. Internal Medicine, Angiology and Arteriosclerosis Diseases, Dept. of Clinical and Experimental Medicine, Perugia, Italy Background and aims: Endothelial microparticles (EMPs) represent a reliable marker of endothelial injury, whereas endothelial progenitor cells (EPCs) play a role in the reparation of the injured endothelium. Impaired renovation of the injured endothelium by EPCs contributes to atherosclerosis development. Hypertriglyceridemia is commonly found in association with other disturbances of the metabolic syndrome, and it is a significant pro-atherogenic condition. We investigated whether hypertriglyceridemia has an influence on the number of circulating EMPs and EPCs. Moreover, we studied whether the metabolic syndrome has a synergistic influence with hypertriglyceridemia on endothelial damage and reparation by EPCs. Methods: The number of circulating EPCs (% of lymphocytes) and EMPs was quantified by FACS analysis in 50 patients with hypertriglyceridemia and in 20 normolipidemic controls. Hypertriglyceridemic patients were divided in 2 groups according to the presence or the absence of the metabolic syndrome. Results: Levels of circulating EPCs (0.006±0.004 vs 0.012±0.007, p=0.01) were halved whereas those of EMPs (830±360 n/microL vs 417±208 n/microL, p=0.002) were doubled in patients with hypertriglyceridemia compared with levels measured in normolipidemic subjects. The presence of the metabolyc syndrome in hypertriglyceridemic patients increased the number of EMPs (1049±590 n/microL) but failed to further reduce the number of EPCs. Conclusion: Hypertriglyceridemia is a significant contributor to endothelial injury and impaired endothelial reparation both in the presence and in the absence of the metabolic syndrome. The latter has a synergistic pro-atherogenic effect with hypertriglyceridemia by increasing the fragmentation into circulation of mature endothelial cells.
WO6-OR-4
THE RELATIONSHIP BETWEEN ENDOTHELIAL PROGENITOR CELLS, THEIR APOPTOTIC MICROPARTICLES AND THE FRAMINGHAM RISK
M. Pirro, F. Bagaglia, C. Menecali, M.R. Mannarino, G. Vaudo, G. Schillaci, E. Mannarino. Internal Medicine, Angiology and Arteriosclerosis Diseases, Dept. of Clinical and Experimental Medicine, Perugia, Italy Background and aims: Exposure to cardiovascular risk factors causes the release of pro-atherogenic microparticles (MPs) from vascular cells and reduces the number of the atheroprotective endothelial progenitor cells (EPCs). We investigated whether MPs shedding from EPCs are detectable in cultures of EPCs and in the circulation of subjects with various degrees of cardiovascular risk. We also investigated the relationship of EPCs-derived MPs to cardiovascular risk factors and aortic stiffness, a marker of cardiovascular risk and impaired vascular repair by EPCs. Methods: We estimated the 10-year Framingham risk score in 105 individuals with various degrees of cardiovascular risk and measured aortic stiffness, as well as the number of circulating EPCs and EPCs-derived MPs (CD34+/KDR+) by FACS analysis. Release of apoptotic CD34+/KDR+ MPs was tested in cultures of EPCs exposed to incremental concentrations of the pro-apoptotic hydrogen-peroxide. Results: The number of annexin-V positive EPCs-derived MPs increased from 1473/mL after vehicle exposure to 5719/mL after 1.5 mM hydrogen-peroxide exposure. The Framingham risk was associated with EPCs (r=-0.47, p<0.001) and CD34+/KDR+ MPs (r=0.56, p<0.001). Framingham risk score (beta=0.40; p<0.001) and EPCs number (beta=0.32; p=0.001) were independently associated with EPCs-derived MPs. Low EPCs (r=-0.59, p<0.001) and high CD34+/KDR+ microparticle levels (r=0.57, p<0.001) were also predictors of aortic stiffness, independent of the Framingham risk. Conclusions: EPCs undergo fragmentation into microparticles when exposed to a pro-apoptotic milieu. Increased microparticle shedding from EPCs may reduce circulating EPCs levels and may thus contribute to increase aortic stiffness beside traditional risk factors. WO6-OR-5
PROANGIOGENIC ACTIVITY OF PROGENITOR CELLS ISOLATED FROM HUMAN ADIPOSE TISSUE
A. Dembinska-Kiec 1 , A. Balwierz 1 , A. Polus 1 , L. Kaczmarek 2 , J. Pryjma 3 . 1 Dept. of Clinical Biochemistry, Jagiellonian University Medical College, Krakow, Poland; 2 Dept. of Molecular and Cellular Neurobiology, Laboratory of Molecular Neurobiology, Nencki Institute, Warsaw, Poland; 3 Dept. of Immunology, Faculty of Biotechnology, Jagiellonian University, Krakow, Poland Introduction: Adipose tissue contains a population stromal vascular fraction (SVF), cells, which differentiate to several lineages. Development of a microvascular network precedes the adipogenesis. Aim of the study was to confirm the proangiogenic properties of the human subcutaneous adipose tissue SVF cells. Methods: SVF cells isolated according to Hauner‘s were cultured in proangiogenic (EBM with 2% FCS), or proadipogenic (DMEM plus hydrocortisone, insulin, transferrin, triiodothyronine after stimulation with dexamethasone, IBMX, insulin) medium. Phenotypes were characterized by flow cytometry or expression of genes characteristic for endothelial progenitors (CD34, CD31, vWF, eNOS, CXCR4, CXCR12, Jagged, Notch4) as well as for preadipocytes (PPARgamma1, PPARgamma2, CEBPalpha, CD36, LPL, ap2, adiponectin, leptin) (real-time PCR). Assay of proliferation and migration was performed. Tubulogenesis of co-cultured the GFP transfected SVF with HUVEC, was measured in 3D matrigel model. Differentiation to adipocyte was confirmed by the red-Oil staining. Results: Growing of SVF cells in serum-free hormone-supplemented medium or incubation of cells in serum-containing medium changes the cell from adipogenic to proangiogenic phenotype. Such SVF cells proliferate, migrate and form the tube-like structure in in vitro 3D model of angiogenesis. Conclusion: The differentiation of SVF cells towards endothelium is stimulated by the presence of human serum and argues for the deep influence of the environment on the differentiation of progenitors towards adipose versus proangiogenic cells. Sponsored by MNiI grant PBZ-MIN-005/P04/2002/5.
76th Congress of the European Atherosclerosis Society, June 10–13, 2007, Helsinki, Finland
6 WO5-OR-6
COMMON VARIANT OF FARNESOID-X-RECEPTOR GENE ENHANCES THE RESPONSE TO LIPID-LOWERING THERAPY
A. Nohara 1 , H. Tada 2 , S. Katsuda 2 , M. Kawashiri 2 , A. Inazu 3 , J. Kobayashi 1 , M. Yamagishi 2 , M. Mabuchi 1 . 1 Department of Lipidology,Kanazawa University Graduate School of Medical Science, Kanazawa,Ishikawa, Japan; 2 Division of Cardiovascular Medicine, Kanazawa University Graduate School of Medical Science, Kanazawa,Ishikawa, Japan; 3 School of Health Science, Kanazawa University, Kanazawa,Ishikawa, Japan Bile acid receptor FXR has crucial roles in cholesterol conversion into bile acids and in its recycling through many target genes that may affect cholesterol levels. We have identified common polymorphism -1g->t in FXR gene in Japanese population, and hypothesized that this polymorphism could affect lipid-lowering therapy response. Methods and Results: Total 276 patients (M/F=147/129) suspected CAD were enrolled. FXR -1g->t genotypes were determined by PCRRFLP, and -1t allele frequency was 0.32. Lipid-lowering drugs (statins 92%) were prescribed in 113 patients (M/F=46/67), and baseline Tcho level was not different with FXR genotype [gg(n=65) vs. gt+tt(n=48) = 253 ± 42 vs. 251 ± 41(mg/dl; mean ± SD)]. With lipid-lowering therapy, -1t carriers showed stronger reduction in Tcho [gg vs. gt+tt = 211 ± 33(-16%) vs. 182 ± 31(-27%) mg/dl, p<0.0001], and in LDL-C [125 ± 34(-22%) vs. 100 ± 25(-39%) mg/dl, p<0.0001]. There was no difference in triglycerides reduction or HDL-C increase. Conclusion: These results demonstrated that common genetic variant of FXR gene showed considerable impact on lipid-lowering therapy, probably through modulation of genes involved in cholesterol metabolism such as CYP7A1. Whether this variant could affect long-term clinical course should further be sought.
WO6 ATHEROSCLEROSIS AND PROGENITOR CELLS WO6-OR-3
ENDOTHELIAL PROGENITOR CELLS AND ENDOTHELIAL-DERIVED MICROPARTICLES IN HYPERTRIGLYCERIDEMIA: THE EFFECT OF THE METABOLIC SYNDROME
M.R. Mannarino, M. Pirro, F. Bagaglia, C. Menecali, R. Paltriccia, G. Schillaci, E. Mannarino. Internal Medicine, Angiology and Arteriosclerosis Diseases, Dept. of Clinical and Experimental Medicine, Perugia, Italy Background and aims: Endothelial microparticles (EMPs) represent a reliable marker of endothelial injury, whereas endothelial progenitor cells (EPCs) play a role in the reparation of the injured endothelium. Impaired renovation of the injured endothelium by EPCs contributes to atherosclerosis development. Hypertriglyceridemia is commonly found in association with other disturbances of the metabolic syndrome, and it is a significant pro-atherogenic condition. We investigated whether hypertriglyceridemia has an influence on the number of circulating EMPs and EPCs. Moreover, we studied whether the metabolic syndrome has a synergistic influence with hypertriglyceridemia on endothelial damage and reparation by EPCs. Methods: The number of circulating EPCs (% of lymphocytes) and EMPs was quantified by FACS analysis in 50 patients with hypertriglyceridemia and in 20 normolipidemic controls. Hypertriglyceridemic patients were divided in 2 groups according to the presence or the absence of the metabolic syndrome. Results: Levels of circulating EPCs (0.006±0.004 vs 0.012±0.007, p=0.01) were halved whereas those of EMPs (830±360 n/microL vs 417±208 n/microL, p=0.002) were doubled in patients with hypertriglyceridemia compared with levels measured in normolipidemic subjects. The presence of the metabolyc syndrome in hypertriglyceridemic patients increased the number of EMPs (1049±590 n/microL) but failed to further reduce the number of EPCs. Conclusion: Hypertriglyceridemia is a significant contributor to endothelial injury and impaired endothelial reparation both in the presence and in the absence of the metabolic syndrome. The latter has a synergistic pro-atherogenic effect with hypertriglyceridemia by increasing the fragmentation into circulation of mature endothelial cells.
WO6-OR-4
THE RELATIONSHIP BETWEEN ENDOTHELIAL PROGENITOR CELLS, THEIR APOPTOTIC MICROPARTICLES AND THE FRAMINGHAM RISK
M. Pirro, F. Bagaglia, C. Menecali, M.R. Mannarino, G. Vaudo, G. Schillaci, E. Mannarino. Internal Medicine, Angiology and Arteriosclerosis Diseases, Dept. of Clinical and Experimental Medicine, Perugia, Italy Background and aims: Exposure to cardiovascular risk factors causes the release of pro-atherogenic microparticles (MPs) from vascular cells and reduces the number of the atheroprotective endothelial progenitor cells (EPCs). We investigated whether MPs shedding from EPCs are detectable in cultures of EPCs and in the circulation of subjects with various degrees of cardiovascular risk. We also investigated the relationship of EPCs-derived MPs to cardiovascular risk factors and aortic stiffness, a marker of cardiovascular risk and impaired vascular repair by EPCs. Methods: We estimated the 10-year Framingham risk score in 105 individuals with various degrees of cardiovascular risk and measured aortic stiffness, as well as the number of circulating EPCs and EPCs-derived MPs (CD34+/KDR+) by FACS analysis. Release of apoptotic CD34+/KDR+ MPs was tested in cultures of EPCs exposed to incremental concentrations of the pro-apoptotic hydrogen-peroxide. Results: The number of annexin-V positive EPCs-derived MPs increased from 1473/mL after vehicle exposure to 5719/mL after 1.5 mM hydrogen-peroxide exposure. The Framingham risk was associated with EPCs (r=-0.47, p<0.001) and CD34+/KDR+ MPs (r=0.56, p<0.001). Framingham risk score (beta=0.40; p<0.001) and EPCs number (beta=0.32; p=0.001) were independently associated with EPCs-derived MPs. Low EPCs (r=-0.59, p<0.001) and high CD34+/KDR+ microparticle levels (r=0.57, p<0.001) were also predictors of aortic stiffness, independent of the Framingham risk. Conclusions: EPCs undergo fragmentation into microparticles when exposed to a pro-apoptotic milieu. Increased microparticle shedding from EPCs may reduce circulating EPCs levels and may thus contribute to increase aortic stiffness beside traditional risk factors. WO6-OR-5
PROANGIOGENIC ACTIVITY OF PROGENITOR CELLS ISOLATED FROM HUMAN ADIPOSE TISSUE
A. Dembinska-Kiec 1 , A. Balwierz 1 , A. Polus 1 , L. Kaczmarek 2 , J. Pryjma 3 . 1 Dept. of Clinical Biochemistry, Jagiellonian University Medical College, Krakow, Poland; 2 Dept. of Molecular and Cellular Neurobiology, Laboratory of Molecular Neurobiology, Nencki Institute, Warsaw, Poland; 3 Dept. of Immunology, Faculty of Biotechnology, Jagiellonian University, Krakow, Poland Introduction: Adipose tissue contains a population stromal vascular fraction (SVF), cells, which differentiate to several lineages. Development of a microvascular network precedes the adipogenesis. Aim of the study was to confirm the proangiogenic properties of the human subcutaneous adipose tissue SVF cells. Methods: SVF cells isolated according to Hauner‘s were cultured in proangiogenic (EBM with 2% FCS), or proadipogenic (DMEM plus hydrocortisone, insulin, transferrin, triiodothyronine after stimulation with dexamethasone, IBMX, insulin) medium. Phenotypes were characterized by flow cytometry or expression of genes characteristic for endothelial progenitors (CD34, CD31, vWF, eNOS, CXCR4, CXCR12, Jagged, Notch4) as well as for preadipocytes (PPARgamma1, PPARgamma2, CEBPalpha, CD36, LPL, ap2, adiponectin, leptin) (real-time PCR). Assay of proliferation and migration was performed. Tubulogenesis of co-cultured the GFP transfected SVF with HUVEC, was measured in 3D matrigel model. Differentiation to adipocyte was confirmed by the red-Oil staining. Results: Growing of SVF cells in serum-free hormone-supplemented medium or incubation of cells in serum-containing medium changes the cell from adipogenic to proangiogenic phenotype. Such SVF cells proliferate, migrate and form the tube-like structure in in vitro 3D model of angiogenesis. Conclusion: The differentiation of SVF cells towards endothelium is stimulated by the presence of human serum and argues for the deep influence of the environment on the differentiation of progenitors towards adipose versus proangiogenic cells. Sponsored by MNiI grant PBZ-MIN-005/P04/2002/5.
76th Congress of the European Atherosclerosis Society, June 10–13, 2007, Helsinki, Finland