Computational Protein Design to Re-Engineer Stromal Cell-Derived Factor-1α (SDF) Generates an Effective and Translatable Angiogenic Polypeptide Analog

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Departmental Papers (Chemistry)
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Angiogenic Proteins
Animals
Cardiac Output
Cell Movement
Cells, Cultured
Chemokine CXCL12
Computational Biology
Coronary Vessels
Endothelial Cells
Male
Mice
Mice, Inbred Strains
Models, Animal
Myocardial Infarction
Neovascularization, Physiologic
Protein Engineering
Rats
Rats, Wistar
Stem Cells
Stroke Volume
Angiogenic Proteins
Animals
Cardiac Output
Cell Movement
Cells
Cultured
Chemokine CXCL12
Computational Biology
Coronary Vessels
Endothelial Cells
Male
Mice
Mice
Inbred Strains
Models
Animal
Myocardial Infarction
Neovascularization
Physiologic
Protein Engineering
Rats
Rats
Wistar
Stem Cells
Stroke Volume
Biochemistry
Organic Chemistry
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Hiesinger, William
Atluri, Pavan
Marotta, Nicole A
Frederick, John R
Fitzpatrick, J Raymond
McCormick, Ryan C
Muenzer, Jeffrey R
Yang, Elaine C
Levit, Rebecca D
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Abstract

BACKGROUND: Experimentally, exogenous administration of recombinant stromal cell-derived factor-1α (SDF) enhances neovasculogenesis and cardiac function after myocardial infarction. Smaller analogs of SDF may provide translational advantages including enhanced stability and function, ease of synthesis, lower cost, and potential modulated delivery via engineered biomaterials. In this study, computational protein design was used to create a more efficient evolution of the native SDF protein. METHODS AND RESULTS: Protein structure modeling was used to engineer an SDF polypeptide analog (engineered SDF analog [ESA]) that splices the N-terminus (activation and binding) and C-terminus (extracellular stabilization) with a diproline segment designed to limit the conformational flexibility of the peptide backbone and retain the relative orientation of these segments observed in the native structure of SDF. Endothelial progenitor cells (EPCs) in ESA gradient, assayed by Boyden chamber, showed significantly increased migration compared with both SDF and control gradients. EPC receptor activation was evaluated by quantification of phosphorylated AKT, and cells treated with ESA yielded significantly greater phosphorylated AKT levels than SDF and control cells. Angiogenic growth factor assays revealed a distinct increase in angiopoietin-1 expression in the ESA- and SDF-treated hearts. In addition, CD-1 mice (n=30) underwent ligation of the left anterior descending coronary artery and peri-infarct intramyocardial injection of ESA, SDF-1α, or saline. At 2 weeks, echocardiography demonstrated a significant gain in ejection fraction, cardiac output, stroke volume, and fractional area change in mice treated with ESA compared with controls. CONCLUSIONS: Compared with native SDF, a novel engineered SDF polypeptide analog (ESA) more efficiently induces EPC migration and improves post-myocardial infarction cardiac function and thus offers a more clinically translatable neovasculogenic therapy.

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2011-09-13
Journal title
Circulation
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