ILK induces cardiomyogenesis in the human heart

Alexandra Traister, Shabana Aafaqi, Stephane Masse, Xiaojing Dai, Mark Li, Aleksander Hinek, Kumaraswamy Nanthakumar, Gregory Hannigan, John G Coles

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Abstract

Background: Integrin-linked kinase (ILK) is a widely conserved serine/threonine kinase that regulates diverse signal transduction pathways implicated in cardiac hypertrophy and contractility. In this study we explored whether experimental overexpression of ILK would up-regulate morphogenesis in the human fetal heart. Methodology/Principal Findings: Primary cultures of human fetal myocardial cells (19a??22 weeks gestation) yielded scattered aggregates of cardioblasts positive for the early cardiac lineage marker nk62.5 and containing nascent sarcomeres. Cardiac cells in colonies uniformly expressed the gap junction protein connexin 43 (C643) and displayed a spectrum of differentiation with only a subset of cells exhibiting the late cardiomyogenic marker troponin T (cTnT) and evidence of electrical excitability. Adenovirus-mediated overexpression of ILK potently increased the number of new aggregates of primitive cardioblasts (p,0.001). The number of cardioblast colonies was significantly decreased (p,0.05) when ILK expression was knocked down with ILK targeted siRNA. Interestingly, overexpression of the activation resistant ILK mutant (ILKR211A) resulted in much greater increase in the number of new cell aggregates as compared to overexpression of wild-type ILK (ILKWT). The cardiomyogenic effects of ILKR211A and ILKWT were accompanied by concurrent activation of bcatenin (p,0.001) and increase expression of progenitor cell marker islet-1, which was also observed in lysates of transgenic mice with cardiac-specific over-expression of ILKR211A and ILKWT. Finally, endogenous ILK expression was shown to increase in concert with those of cardiomyogenic markers during directed cardiomyogenic differentiation in human embryonic stem cells (hESCs). Conclusions/Significance: In the human fetal heart ILK activation is instructive to the specification of mesodermal precursor cells towards a cardiomyogenic lineage. Induction of cardiomyogenesis by ILK overexpression bypasses the requirement of proximal PI3K activation for transduction of growth factor- and b1-integrin-mediated differentiation signals. Altogether, our data indicate that ILK represents a novel regulatory checkpoint during human cardiomyogenesis.
Original languageEnglish
Article numbere37802
Number of pages14
JournalPLoS ONE
Volume7
Issue number5
DOIs
Publication statusPublished - 2012

Cite this

Traister, A., Aafaqi, S., Masse, S., Dai, X., Li, M., Hinek, A., Nanthakumar, K., Hannigan, G., & Coles, J. G. (2012). ILK induces cardiomyogenesis in the human heart. PLoS ONE, 7(5), [e37802]. https://doi.org/10.1371/journal.pone.0037802