ENDOTHELIAL PROGENITOR CELLS AS A NOVEL BIOMARKER OF VASCULAR HEALTH
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1 ENDOTHELIAL PROGENITOR CELLS AS A NOVEL BIOMARKER OF VASCULAR HEALTH I Jialal, MD, PhD. FRCPath.DABCC Robert E. Stowell Chair in Experimental Pathology Professor of Pathology and Medicine Director, Laboratory for Atherosclerosis and Metabolic Research UC Davis Medical Center Sacramento, CA
2 Steps in the Progression of the Atherosclerotic Plaque Libby Nature 2002
3 enos-no
4 Endothelial Dysfunction Diabetes Hypertension Smoking Dyslipidemia enos Prostacyclin Endothelin-1 CAMS PAI-1
5 ENDOTHELIAL CELL NO Reduced enos activity, stability, espression and phosphorylation, enos uncoupling ENDOTHELIAL CELL NO ROS Reduced enos activity, stability, espression and phosphorylation, enos uncoupling ROS No Bioavailability Apoptosis and senescene Vascular homeostasis NORMAL ENDOTHELIAL FUNCTION Aging and other cardiovascular risk factors Disrupted Vascular Homeostasis IMPAIRED ENDOTHELIAL FUNCTION Normal numbers and functional activity Circulating Endothelial Progenitor Cell Reduced mobilisation, survival and homing Reduced numbers and impaired functional activity Circulating Endothelial Progenitor Cell Craenenbroeck and Conraads. Microvasc Res. 2010
6 CARDIOVASCULAR RISK FACTORS Advanced age, hypertension, obesity diabetes, smoking, hyperlipidemia, sedentarity ENDOTHELIAL DYSFUNTION ENDOTHEILIAL PROGENATOR CELLS Reduced circulating numbers Impaired anglogenic capacity Physical Activity Healthy Diet Smoking cessasion Weight Reduction Stress Reduction Craenenbroeck and Conraads. Microvasc Res. 2010
7 Schematic drawing of conditions influencing the liberation of EPCs from the bone marrow, and its therapeutical application in various cardiovascular diseases Mobius-Winkler et al. Cytometry, 2009
8 Endothelial Progenitor Cells Werner and Nickenig. JCEM, 2006
9 EPC Measurement Flow Cytometry Cell Culture: Colony forming units (CFUs) Migration (VEGF, SDF) Adhesion (Fibronectin) Tubule Formation (DiI-EPC and HAEC) In-vivo (Hind limb ischemia)
10 Schematic drawing of the isolation and characterization of endothelial progenitor cells starting with circulating blood Mobius-Winkler et al. Cytometry, 2009
11 The two main protocols for culture of putative EPCs. On the left, the 5-day CFU-EC assay, which is based on pre-plating total mononuclear cells (MNC) and re-plating nonattaching cells. On the right, the prolonged culture protocol needed to isolate late ( true ) EPCs (Endothelial Colony Forming Cells, ECFC). Fadini et al. Atherosclerosis, 2008
12 Mean cell count of diverse progenitor cell phenotypes obtained in a total of 439 subjects using FITC-conjugated anti-cd34, PEconjugated anti-kdr and APC-conjugated anti-cd133 Fadini et al. Atherosclerosis, 2008
13 Technical Hints for Analyses of EPCs Fadini et al. Atherosclerosis, 2008
14 Relation between the Number of Endothelial Progenitor Cells and Endothelial Function Hill J et al. N Engl J Med 2003
15 Association between Cardiovascular Risk Factors and Endothelial- Progenitor-Cell Colony Counts Hill J et al. N Engl J Med 2003
16 Effect of risk factors on CD34/KDR positive cells Vasa, M. et al. Circ Res 2001
17 Effect of risk factors on migration of EPCs Vasa, M. et al. Circ Res 2001
18 Cumulative Event-free Survival in an Analysis of Death from Cardiovascular Causes at 12 Months, According to Levels of Circulating CD34+KDR+ Endothelial Progenitor Cells at the Time of Enrollment Werner N et al. N Engl J Med 2005
19 Number of Colony Forming Units Endothelial Cells and Outcome P = Werner and Nickenig. JCEM, 2006
20 Median EPC counts by severity of CAD CFU Us P < /1 Vessel Multivessel Median Kunz et al. Am Heart J 2006
21 Multivariable predictors of CAD severity Kunz et al. Am Heart J 2006
22 Endothelial progenitor cell (EPC) colony-forming units (A) and migration (B) in healthy, sedentary young, middle-aged, and older men Hoetzer, G. L. et al. J Appl Physiol, 2007
23 Event-free survival according to levels of circulating CD34+KDR+-EPCs defined by ROC curve analysis Schmidt-Locke, C. et al. Circulation 2005
24 EPC and MACE Crude, Disease Activity-Adjusted, and Risk Factor-Adjusted Relative Risks of a First Major Cardiovascular Event in Patients With Low EPC Count. Variable HR for MACE (95% Cl) Crude relative risk 6.3 ( ) Disease activity-adjusted relative 4.2 ( ) risk Risk factor-and disease activityadjusted 3.9 ( ) MACE indicates major adverse cardiovascular event. P Schmidt-Lucke, C. et al. Circulation 2005
25 The Northern Manhattan Study (NOMAS) Flow Media ated Dilation(%) ± ± 3.70 P < No MetS (n = 442) MetS (n = 377) Suzuki et al. Am Heart J 2008
26 Control (n=30) MetS (n=45) P value Age (yrs) 49 ± ± Females/Males 25/6 35/ Waist circumference (cm) 94 ± ± Weight (kg) 85±22 102± BMI (kg/m2) 30±6 36± Systolic BP(mmHg) 121 ± ± Diastolic BP (mmhg) 74 ± 8 82 ± Fasting Glucose (mg/dl) 90 ± 7 101± Total Cholesterol (mg/dl) 185 ± ± HDL-Cholesterol (mg/dl) 65 ± ± LDL-Cholesterol (mg/dl) 114 ± ± Triglycerides (mg/dl) 75 (62,95) 134 (106,172) Fasting insulin (uiu/ml) 8.6 ± ± HOMA-IR 1.7 (0.9, 2.8) 3.3 (2.3, 5.8) hscrp (mg/l) 1.3 (0.5,3) 3.5(1.6,5.7) Data are summarized as mean ± SD except for hscrp and triglycerides which are presented as median (25% percentile, 75% percentile) Jialal et al. Atherosclerosis, 2010
27 Assays of EPC Functionality
28 Enumeration of EPCs by FACS Surface Expression (MFI) CD34+ PC *p<0.001 CD34+/KDR+ EPC *p< Control (n=30) MetS (n=46) Control (n=30) MetS (n=46) Jialal et al. Atherosclerosis, 2010
29 CFU in Control and Metabolic Syndrome Subjects Number of CFUs/5 hpf Control MetS *p<0.001 (n=25) (n=43) Jialal et al. Atherosclerosis, 2010
30 Vasculogenic capacity of EPCs in Control and MetS Tubules (/5 hpf) *p< Control (n=15) MetS (n=15) Jialal et al. Atherosclerosis, 2010
31 Migration assay in Control and Metabolic Syndrome Subjects 1000 Migratio on (%) Control (n=16) MetS (n=36) Jialal et al. Atherosclerosis, 2010
32 EPC Mobilizing Factors **P<0.01 pg/m ml ***P<0.001 Control (n=38) MetS (n=36) 0 G-CSF SCF Jialal et al. (Submitted), 2010
33 EPC Mobilizing Factors pg/m ml VEGF* *P<0.05 Control (n=38) MetS (n=36) Jialal et al. 2011
34 EPC Mobilizing Factors EPC Mobilizing Factors **P< pg/ml 300 Control (n=38) MetS (n=36) **P<0.01 *P<0.05 ***P< SCF-sR VEGF G-CSF SCF Jialal et al. A.J.Cardiol. 2010
35 EPC Mobilizing Factors **P<0.01 ng/m ml Control (n=38) MetS (n=36) MMP-9
36 Drugs that modulate Endothelial Progenitor cells Statins Insulin Oestrogens ACE-inhibitors PPAR-Gamma-agonists
37 Acknowledgements Grants: ADA, NIH S. Devaraj C. Duncan-Staley M. Kaur
38 Cumulative Event-free Survival in Analysis of a First Major Cardiovascular Event (Myocardial Infarction, Hospitalization, Revascularization, or Cardiovascular Death) at 12 Months, According to Levels of Circulating CD34+KDR+ Endothelial Progenitor Cells at the Time of Enrollment Werner N et al. N Engl J Med 2005
39 Effect of risk factors on number of EPCs Vasa, M. et al. Circ Res 2001
40 CD133+ Endothelial Progenitor Cells and Outcome Werner and Nickenig. JCEM, 2006
41 Craenenbroeck and Conraads. Microvasc Res. 2010
42 Fadini and Avogaro. Diabetes Obes Metab. 2010
43 Everaert B. et al. Int J Cardiol. 2010
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