Overexpression of protein kinase C ɛ improves retention and survival of transplanted mesenchymal stem cells in rat acute myocardial infarction.

H He,X-H Liu,Y-J Zeng,Z-H Zhao,R Wang,F-S Han

doi:10.1038/cddis.2015.417

H He, X-H Liu + Show 4 more

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https://doi.org/10.1038/cddis.2015.417

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Abstract

We assessed the effects of protein kinase C ɛ (PKCɛ) for improving stem cell therapy for acute myocardial infarction (AMI). Primary mesenchymal stem cells (MSCs) were harvested from rat bone marrow. PKCɛ-overexpressed MSCs and control MSCs were transplanted into infarct border zones in a rat AMI model. MSCs and PKCɛ distribution and expression of principal proteins involved in PKCɛ signaling through the stromal cell-derived factor 1 (SDF-1)/CXC chemokine receptor type 4 (CXCR4) axis and the phosphatidylinositol 3 kinase (PI3K)/protein kinase B (AKT) pathway were analyzed by immunofluorescence and western blot 1 day after transplantation. Echocardiographic measurements and histologic studies were performed at 4 weeks after transplantation, and MSC survival, expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), transforming growth factor β (TGFβ), cardiac troponin I (cTnI), von Willebrand factor (vWF), smooth muscle actin (SMA) and factor VIII and apoptosis in infarct border zones were assessed. Rat heart muscles retained more MSCs and SDF-1, CXCR4, PI3K and phosphorylated AKT increased with PKCɛ overexpression 1 day after transplantation. MSC survival and VEGF, bFGF, TGFβ, cTnI, vWF, SMA and factor VIII expression increased in animals with PKCɛ-overexpressed MSCs at 4 weeks after transplantation and cardiac dysfunction and remodeling improved. Infarct size and apoptosis decreased as well. Inhibitory actions of CXCR4 or PI3K partly attenuated the effects of PKCɛ. Activation of PKCɛ may improve retention, survival and differentiation of transplanted MSCs in myocardia. Augmentation of PKCɛ expression may enhance the therapeutic effects of stem cell therapy for AMI.

Highlights

1 (SDF-1)/CXC chemokine receptor type 4 (CXCR4) axis has an important role during migration, proliferation and survival of stem cells, but using this knowledge to improve homing and survival of therapeutic stem cells has not been successful
Analysis by real-time polymerase chain reaction (PCR), western blot and immunocytochemistry indicated that expression of protein kinase C ε (PKCε) in mesenchymal stem cells (MSCs)-PKCε-green fluorescent protein (GFP) increased significantly compared with MSCs-GFP (Figure 1), verifying that PKCε-overexpressed MSCs were successfully constructed
Rats died in all treatment groups, giving numbers as follows: one for sham; four for acute myocardial infarction (AMI); two for MSCs; two for MSCs-GFP, one for MSCs-PKCε, two for AMI +MSCs-PKCε-GFP+AMD3100 group (AMD3100) and three for the LY294002 group

Summary

Introduction

1 (SDF-1)/CXC chemokine receptor type 4 (CXCR4) axis has an important role during migration, proliferation and survival of stem cells, but using this knowledge to improve homing and survival of therapeutic stem cells has not been successful. Previous studies[9,10,11] suggest that protein kinase C ε (PKCε) is essential for signal transduction for ischemic cardioprotection, but whether it has an effect on stem cell retention and survival and what mechanism underlies this effect is uncertain. We know that SDF-1 increased significantly in mesenchymal stem cells (MSCs) after treatment with PKC activator and decreased after treatment with a PKCε inhibitor in preliminary experiments, and our latest work indicates that activating PKCε improves migration and paracrine function of MSCs in vitro.[12] we suggest that PKCε overexpression in transplanted bone marrow MSCs (BMMSCs) would improve retention and survival of MSC’s and improve cardiac function and remodeling in a rat AMI model

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