TY - JOUR
T1 - Neovasculogenic Therapy to Augment Perfusion and Preserve Viability in Ischemic Cardiomyopathy
AU - Atluri, Pavan
AU - Liao, George P.
AU - Panlilio, Corinna M.
AU - Hsu, Vivian M.
AU - Leskowitz, Matthew J.
AU - Morine, Kevin J.
AU - Cohen, Jeffrey E.
AU - Berry, Mark F.
AU - Suarez, Erik E.
AU - Murphy, Danielle A.
AU - Lee, William M.F.
AU - Gardner, Timothy J.
AU - Sweeney, H. Lee
AU - Woo, Y. Joseph
N1 - Funding Information:
This work was supported in part by grants from the NIH National Heart Lung and Blood Institute/Thoracic Surgery Foundation for Research and Education HL072812 (Y. Joseph Woo, MD), American Heart Association 0465519U (Y. Joseph Woo, MD), NIH-Training Grant in Cardiovascular Biology and Medicine T32 HL07843 (Mark F. Berry, MD, and Pavan Atluri, MD), and National Institutes of Health, National Heart Lung Blood Institute – Ruth L. Kirschstein National Research Service Award, Individual Fellowship 1 F32 HL79769-01 (Pavan Atluri, MD).
PY - 2006/5
Y1 - 2006/5
N2 - Background: Ischemic cardiomyopathy is a global health concern with limited therapy. We recently described endogenous revascularization utilizing granulocyte-macrophage colony stimulating factor (GMCSF) to induce endothelial progenitor cell (EPC) production and intramyocardial stromal cell-derived factor-1α (SDF) as a specific EPC chemokine. The EPC-mediated neovascularization and enhancement of myocardial function was observed. In this study we examined the regional biologic mechanisms underlying this therapy. Methods: Lewis rats underwent left anterior descending coronary artery (LAD) ligation and developed ischemic cardiomyopathy over 6 weeks. Three weeks after ligation, the animals received either subcutaneous GMCSF and intramyocardial SDF injections or saline injections as control. Six weeks after LAD ligation circulating EPC density was studied by flow cytometry. Quadruple immunofluorescent vessel staining for mature, proliferating vasculature was performed. Confocal angiography was utilized to identify fluorescein lectin-lined vessels to assess perfusion. Ischemia reversal was studied by measuring myocardial adenosine triphosphate (ATP) levels. Myocardial viability was assayed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling detection of apoptosis and quantitation of myofilament density. Results: The GMCSF/SDF therapy enhanced circulating leukocyte (13.1 ± 4.5 × 106 vs 3.1 ± 0.5 × 106/cc, p = 0.001, n = 6) and EPC (14.2 ± 6.6 vs 2.2 ± 2.1/cc, p = 0.001, n = 6) concentrations. Tetraimmunofluorescent labeling demonstrated enhanced stable vasculature with this therapy (39.2 ± 8.1 vs 25.4 ± 5.1%, p = 0.006, n = 7). Enhanced perfusion was shown by confocal microangiography of borderzone lectin-labeled vessels (28.2 ± 5.4 vs 11.5 ± 3.0 vessels/high power field [hpf], p = 0.00001, n = 10). Ischemia reversal was demonstrated by enhanced cellular ATP levels in the GMCSF/SDF borderzone myocardium (102.5 ± 31.0 vs 26.9 ± 4.1 nmol/g, p = 0.008, n = 5). Borderzone cardiomyocyte viability was noted by decreased apoptosis (3.2 ± 1.4% vs 5.4 ± 1.0%,p = 0.004, n = 10) and enhanced cardiomyocyte density (40.0 ± 5.6 vs 27.0 ± 6 myofilaments/hpf, p = 0.01, n=10). Conclusions: Endogenous revascularization for ischemic cardiomyopathy utilizing GMCSF EPC upregulation and SDF EPC chemokinesis upregulates circulating EPCs, enhances vascular stability, and augments myocardial function by enhancing perfusion, reversing cellular ischemia, and increasing cardiomyocyte viability.
AB - Background: Ischemic cardiomyopathy is a global health concern with limited therapy. We recently described endogenous revascularization utilizing granulocyte-macrophage colony stimulating factor (GMCSF) to induce endothelial progenitor cell (EPC) production and intramyocardial stromal cell-derived factor-1α (SDF) as a specific EPC chemokine. The EPC-mediated neovascularization and enhancement of myocardial function was observed. In this study we examined the regional biologic mechanisms underlying this therapy. Methods: Lewis rats underwent left anterior descending coronary artery (LAD) ligation and developed ischemic cardiomyopathy over 6 weeks. Three weeks after ligation, the animals received either subcutaneous GMCSF and intramyocardial SDF injections or saline injections as control. Six weeks after LAD ligation circulating EPC density was studied by flow cytometry. Quadruple immunofluorescent vessel staining for mature, proliferating vasculature was performed. Confocal angiography was utilized to identify fluorescein lectin-lined vessels to assess perfusion. Ischemia reversal was studied by measuring myocardial adenosine triphosphate (ATP) levels. Myocardial viability was assayed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling detection of apoptosis and quantitation of myofilament density. Results: The GMCSF/SDF therapy enhanced circulating leukocyte (13.1 ± 4.5 × 106 vs 3.1 ± 0.5 × 106/cc, p = 0.001, n = 6) and EPC (14.2 ± 6.6 vs 2.2 ± 2.1/cc, p = 0.001, n = 6) concentrations. Tetraimmunofluorescent labeling demonstrated enhanced stable vasculature with this therapy (39.2 ± 8.1 vs 25.4 ± 5.1%, p = 0.006, n = 7). Enhanced perfusion was shown by confocal microangiography of borderzone lectin-labeled vessels (28.2 ± 5.4 vs 11.5 ± 3.0 vessels/high power field [hpf], p = 0.00001, n = 10). Ischemia reversal was demonstrated by enhanced cellular ATP levels in the GMCSF/SDF borderzone myocardium (102.5 ± 31.0 vs 26.9 ± 4.1 nmol/g, p = 0.008, n = 5). Borderzone cardiomyocyte viability was noted by decreased apoptosis (3.2 ± 1.4% vs 5.4 ± 1.0%,p = 0.004, n = 10) and enhanced cardiomyocyte density (40.0 ± 5.6 vs 27.0 ± 6 myofilaments/hpf, p = 0.01, n=10). Conclusions: Endogenous revascularization for ischemic cardiomyopathy utilizing GMCSF EPC upregulation and SDF EPC chemokinesis upregulates circulating EPCs, enhances vascular stability, and augments myocardial function by enhancing perfusion, reversing cellular ischemia, and increasing cardiomyocyte viability.
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U2 - 10.1016/j.athoracsur.2005.12.015
DO - 10.1016/j.athoracsur.2005.12.015
M3 - Article
C2 - 16631663
AN - SCOPUS:33646835383
SN - 0003-4975
VL - 81
SP - 1728
EP - 1736
JO - Annals of Thoracic Surgery
JF - Annals of Thoracic Surgery
IS - 5
ER -