Postinfarct active cardiac-targeted delivery of erythropoietin by liposomes with sialyl Lewis X repairs infarcted myocardium in rabbits.

We investigated the effect of cardiac-targeting erythropoietin (EPO)-encapsulated liposomes with sialyl Lewis(X) (SLX) on myocardial infarct (MI) size, left ventricular (LV) remodeling and function, and its molecular mechanism for repairing infarcted myocardium. In rabbits, MI was induced by 30 min of coronary occlusion followed by reperfusion. EPO-encapsulated liposomes with SLX (L-EPO group), EPO-encapsulated liposomes without SLX (L-EPO without SLX group), liposomes with SLX without EPO (L group), or saline (saline group) were intravenously administered immediately after MI. MI sizes and numbers of microvessels were assessed 14 days after MI. Prosurvival proteins and signals were assessed by Western blot analysis 2 and 14 days after MI. Confocal microscopy and electron microscopy showed the specific accumulation of liposomes with SLX in the infarcted myocardium. MI and cardiac fibrosis areas were significantly smaller in the L-EPO group than in the other groups. LV function and remodeling were improved in the L-EPO group. The number of CD31-positive microvessels was significantly greater in the L-EPO group than in the other groups. Higher expressions of EPO receptors, phosphorylated (p)Akt, pERK, pStat3, VEGF, Bcl-2, and promatrix metalloproteinase-1 were observed in the infarct area in the L-EPO group than in the other groups. EPO-encapsulated liposomes with SLX selectively accumulated in the infarct area, reduced MI size, and improved LV remodeling and function through activation of prosurvival signals and by exerting antifibrotic and angiogenic effects. EPO-encapsulated liposomes with SLX may be a promising strategy for active targeting treatment of acute MI.

[1]  C. Heeschen,et al.  Erythropoietin is a potent physiologic stimulus for endothelial progenitor cell mobilization. , 2003, Blood.

[2]  B. Seed,et al.  Endothelial leukocyte adhesion molecule 1: an inducible receptor for neutrophils related to complement regulatory proteins and lectins. , 1989, Science.

[3]  K. Maruyama,et al.  Enhanced tumor targeting of doxorubicin by ganglioside GM1-bearing long-circulating liposomes. , 1993, Journal of drug targeting.

[4]  R. Visse,et al.  Matrix Metalloproteinases Regulation and Dysregulation in the Failing Heart , 2002 .

[5]  Avid,et al.  THE EFFECTS OF NORMAL AS COMPARED WITH LOW HEMATOCRIT VALUES IN PATIENTS WITH CARDIAC DISEASE WHO ARE RECEIVING HEMODIALYSIS AND EPOETIN , 2000 .

[6]  Y. Taniyama,et al.  Potential contribution of a novel antifibrotic factor, hepatocyte growth factor, to prevention of myocardial fibrosis by angiotensin II blockade in cardiomyopathic hamsters. , 2000, Circulation.

[7]  G. Wright,et al.  Erythropoietin receptor expression in adult rat cardiomyocytes is associated with an acute cardioprotective effect for recombinant erythropoietin during ischemia‐reperfusion injury , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[8]  Richard B. Thompson,et al.  Cardioprotective Effects of Erythropoietin in the Reperfused Ischemic Heart , 2004, Journal of Biological Chemistry.

[9]  E. Creemers,et al.  Matrix Metalloproteinase Inhibition After Myocardial Infarction: A New Approach to Prevent Heart Failure? , 2001, Circulation research.

[10]  G. Takemura,et al.  Preventive Effect of Erythropoietin on Cardiac Dysfunction in Doxorubicin-Induced Cardiomyopathy , 2006, Circulation.

[11]  R. Burrow,et al.  Heart , 2013, Feline Soft Tissue and General Surgery.

[12]  N. Yamazaki.  Analysis of the carbohydrate-binding specificity of lectin-conjugated lipid vesicles which interact with polysaccharide fragments☆ , 1989 .

[13]  J. Paulson,et al.  Reduction of rat myocardial ischemia and reperfusion injury by sialyl Lewis x oligosaccharide and anti-rat P-selectin antibodies. , 1996, Glycobiology.

[14]  G. Takemura,et al.  Reduction of inflammatory cytokine expression and oxidative damage by erythropoietin in chronic heart failure. , 2006, Cardiovascular research.

[15]  R. Bolli,et al.  An essential role of the JAK-STAT pathway in ischemic preconditioning , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[16]  N. Maulik,et al.  An essential role of the antioxidant gene Bcl-2 in myocardial adaptation to ischemia: an insight with antisense Bcl-2 therapy. , 2001, Antioxidants & redox signaling.

[17]  H. Gabius,et al.  Neoglycoprotein-liposome and lectin-liposome conjugates as tools for carbohydrate recognition research. , 1994, Methods in enzymology.

[18]  R. Kloner,et al.  Administration of Erythropoietin Fails to Improve Long-Term Healing or Cardiac Function After Myocardial Infarction in the Rat , 2005, Journal of cardiovascular pharmacology.

[19]  Hiroshi Yokoyama,et al.  Biomedical nanotechnology for active drug delivery systems by applying sugar-chain molecular functions , 2005 .

[20]  D. Vestweber,et al.  Mechanisms that regulate the function of the selectins and their ligands. , 1999, Physiological reviews.

[21]  A. Ohtsuka,et al.  Development of an active targeting liposome encapsulated with high-density colloidal gold for transmission electron microscopy. , 2011, Journal of electron microscopy.

[22]  K. Maruyama,et al.  Prolonged circulation time in vivo of large unilamellar liposomes composed of distearoyl phosphatidylcholine and cholesterol containing amphipathic poly(ethylene glycol). , 1992, Biochimica et biophysica acta.

[23]  Yechezkel Barenholz,et al.  Pharmacokinetics of Pegylated Liposomal Doxorubicin , 2003, Clinical pharmacokinetics.

[24]  M. Hirai,et al.  Accumulation of liposome with Sialyl Lewis X to inflammation and tumor region: application to in vivo bio-imaging. , 2007, Biochemical and biophysical research communications.

[25]  A. Khorana,et al.  Risk factors for chemotherapy‐associated venous thromboembolism in a prospective observational study , 2005, Cancer.

[26]  G. Yannarelli,et al.  High-dose Erythropoietin Has No Long-term Protective Effects in Sheep with Reperfused Myocardial Infarction , 2006, Journal of cardiovascular pharmacology.

[27]  Richard B. Thompson,et al.  A novel protective effect of erythropoietin in the infarcted heart. , 2003, The Journal of clinical investigation.

[28]  MasazumiArai,et al.  Acceleration of the Healing Process and Myocardial Regeneration May Be Important as a Mechanism of Improvement of Cardiac Function and Remodeling by Postinfarction Granulocyte Colony–Stimulating Factor Treatment , 2004 .

[29]  J. Isner,et al.  VEGF contributes to postnatal neovascularization by mobilizing bone marrow‐derived endothelial progenitor cells , 1999, The EMBO journal.

[30]  K. Sugamura,et al.  Important Role of Erythropoietin Receptor to Promote VEGF Expression and Angiogenesis in Peripheral Ischemia in Mice , 2007, Circulation research.

[31]  A. Gabizon,et al.  Sterically stabilized liposomes: improvements in pharmacokinetics and antitumor therapeutic efficacy. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[32]  S. Nielsen,et al.  Lack of acute cardioprotective effect from preischaemic erythropoietin administration in a porcine coronary occlusion model , 2005, Clinical physiology and functional imaging.

[33]  Y. Tabata,et al.  Post-infarct treatment with an erythropoietin-gelatin hydrogel drug delivery system for cardiac repair. , 2008, Cardiovascular research.

[34]  A. Buda,et al.  A sialyl Lewis(x)-containing carbohydrate reduces infarct size: role of selectins in myocardial reperfusion injury. , 1996, The American journal of physiology.

[35]  R. Guyton,et al.  Inhibition of myocardial injury by ischemic postconditioning during reperfusion: comparison with ischemic preconditioning. , 2003, American journal of physiology. Heart and circulatory physiology.

[36]  Santhosh Palani,et al.  Positive receptor feedback during lineage commitment can generate ultrasensitivity to ligand and confer robustness to a bistable switch. , 2008, Biophysical journal.

[37]  M. Mocanu,et al.  Postconditioning: A Form of “Modified Reperfusion” Protects the Myocardium by Activating the Phosphatidylinositol 3-Kinase–Akt Pathway , 2004, Circulation research.

[38]  A. Gabizon,et al.  Liposome formulations with prolonged circulation time in blood and enhanced uptake by tumors. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[39]  Pietro Ghezzi,et al.  Recombinant human erythropoietin protects the myocardium from ischemia-reperfusion injury and promotes beneficial remodeling , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[40]  M. Mocanu,et al.  Ischemic preconditioning protects by activating prosurvival kinases at reperfusion. , 2005, American journal of physiology. Heart and circulatory physiology.

[41]  S. Vyas,et al.  Ligand-receptor-mediated drug delivery: an emerging paradigm in cellular drug targeting. , 2001, Critical reviews in therapeutic drug carrier systems.

[42]  E. Lakatta,et al.  Cardioprotection by Recombinant Human Erythropoietin Following Acute Experimental Myocardial Infarction: Dose Response and Therapeutic Window , 2005, Cardiovascular Drugs and Therapy.

[43]  A R Nissenson,et al.  The effects of normal as compared with low hematocrit values in patients with cardiac disease who are receiving hemodialysis and epoetin. , 1998, The New England journal of medicine.

[44]  S. S. Zhang,et al.  Cardiomyocyte-restricted knockout of STAT3 results in higher sensitivity to inflammation, cardiac fibrosis, and heart failure with advanced age , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[45]  E. Brogi,et al.  Therapeutic angiogenesis. A single intraarterial bolus of vascular endothelial growth factor augments revascularization in a rabbit ischemic hind limb model. , 1994, The Journal of clinical investigation.