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Thymosin β4 activates integrin-linked kinase and promotes cardiac cell migration, survival and cardiac repair

Abstract

Heart disease is a leading cause of death in newborn children and in adults. Efforts to promote cardiac repair through the use of stem cells hold promise but typically involve isolation and introduction of progenitor cells. Here, we show that the G-actin sequestering peptide thymosin β4 promotes myocardial and endothelial cell migration in the embryonic heart and retains this property in postnatal cardiomyocytes. Survival of embryonic and postnatal cardiomyocytes in culture was also enhanced by thymosin β4. We found that thymosin β4 formed a functional complex with PINCH and integrin-linked kinase (ILK), resulting in activation of the survival kinase Akt (also known as protein kinase B). After coronary artery ligation in mice, thymosin β4 treatment resulted in upregulation of ILK and Akt activity in the heart, enhanced early myocyte survival and improved cardiac function. These findings suggest that thymosin β4 promotes cardiomyocyte migration, survival and repair and the pathway it regulates may be a new therapeutic target in the setting of acute myocardial damage.

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Figure 1: Thymosin β4 is expressed in specific cardiac cell types during development.
Figure 2: Thymosin β4 is secreted and promotes cardiac cell migration and survival.
Figure 3: Thymosin β4 forms a functional complex with PINCH and ILK resulting in phosphorylation of Akt.
Figure 4: Thymosin β4 treatment after coronary ligation improves myocardial function in vivo.
Figure 5: Thymosin β4 promotes survival and alters scar formation after coronary artery ligation in mice.

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Acknowledgements

The authors wish to thank A. L. Goldstein for his advice and RegeneRx Biopharmaceuticals Inc. for providing the synthetic thymosin β4 protein; J. Richardson and the histopathology core for histological support; G. A. Adams for technical asssistance; E. N. Olson and members of the Srivastava laboratory for discussions and critical review; and S. Johnson and J. E. Marquette for graphical help and suggestions. D.S. was supported by grants from the NHLBI/NIH, March of Dimes Birth Defects Foundation, American Heart Association and Donald W. Reynolds Clinical Cardiovascular Research Center.

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Correspondence to Deepak Srivastava.

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Supplementary information

Supplementary Movie 1

Rare beating foci in cells migrating from cardiac explants. (MP4 744 kb)

Supplementary Movie 2

Frequent beating foci in cells migrating from cardiac explants after thymosin β4 treatment. (MP4 429 kb)

Supplementary Methods

Additional methods used in the experiments, including: embryonic T7 phage display cDNA library; T7 phage biopanning; and ELISA confirmation assay. (DOC 21 kb)

Supplementary Movie Legends

Legends to accompany the Supplementary Movies (DOC 20 kb)

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Bock-Marquette, I., Saxena, A., White, M. et al. Thymosin β4 activates integrin-linked kinase and promotes cardiac cell migration, survival and cardiac repair. Nature 432, 466–472 (2004). https://doi.org/10.1038/nature03000

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