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C-Kit Promotes Growth and Migration of Human Cardiac Progenitor Cells via the PI3K-AKT and MEK-ERK Pathways.

Vajravelu BN, Hong KU, Al-Maqtari T, Cao P, Keith MC, Wysoczynski M, Zhao J, Moore JB, Bolli R - PLoS ONE (2015)

Bottom Line: SCF treatment led to a significant increase in cell survival and a reduction in cell death under serum depletion conditions.In addition, SCF significantly promoted CPC migration in vitro.These results imply that the efficiency of CPC homing to the injury site as well as their survival after transplantation may be improved by modulating the activity of c-kit.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Cardiology, Department of Medicine, University of Louisville, Louisville, KY 40202, United States of America.

ABSTRACT
A recent phase I clinical trial (SCIPIO) has shown that autologous c-kit+ cardiac progenitor cells (CPCs) improve cardiac function and quality of life when transplanted into patients with ischemic heart disease. Although c-kit is widely used as a marker of resident CPCs, its role in the regulation of the cellular characteristics of CPCs remains unknown. We hypothesized that c-kit plays a role in the survival, growth, and migration of CPCs. To test this hypothesis, human CPCs were grown under stress conditions in the presence or absence of SCF, and the effects of SCF-mediated activation of c-kit on CPC survival/growth and migration were measured. SCF treatment led to a significant increase in cell survival and a reduction in cell death under serum depletion conditions. In addition, SCF significantly promoted CPC migration in vitro. Furthermore, the pro-survival and pro-migratory effects of SCF were augmented by c-kit overexpression and abrogated by c-kit inhibition with imatinib. Mechanistically, c-kit activation in CPCs led to activation of the PI3K and the MAPK pathways. With the use of specific inhibitors, we confirmed that the SCF/c-kit-dependent survival and chemotaxis of CPCs are dependent on both pathways. Taken together, our findings suggest that c-kit promotes the survival/growth and migration of human CPCs cultured ex vivo via the activation of PI3K and MAPK pathways. These results imply that the efficiency of CPC homing to the injury site as well as their survival after transplantation may be improved by modulating the activity of c-kit.

No MeSH data available.


Related in: MedlinePlus

c-kit activation does not rescue CPCs from oxidative stress.A, Human c-kit+ CPCs were pre-treated with SCF for 24 hr in serum free media and subjected to 8 μM DMNQ treatment. B, Cells pre-treated with or without SCF received 1 hr treatment in 0.5 mM H2O2, followed by media change with or without SCF. Insulin was used as a positive control. Cell viability was assessed after 3 days using PrestoBlueTM. *, p<0.05 compared to the untreated control, #, p<0.05 compared to the H2O2 only control. NS, no statistical significance. Values presented are mean ± SEM. *, p<0.05.
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pone.0140798.g003: c-kit activation does not rescue CPCs from oxidative stress.A, Human c-kit+ CPCs were pre-treated with SCF for 24 hr in serum free media and subjected to 8 μM DMNQ treatment. B, Cells pre-treated with or without SCF received 1 hr treatment in 0.5 mM H2O2, followed by media change with or without SCF. Insulin was used as a positive control. Cell viability was assessed after 3 days using PrestoBlueTM. *, p<0.05 compared to the untreated control, #, p<0.05 compared to the H2O2 only control. NS, no statistical significance. Values presented are mean ± SEM. *, p<0.05.

Mentions: Next, we tested if SCF can prevent cell death induced by other stress conditions, such as oxidative stress, in CPCs. For this, CPCs were pre-treated with SCF for 24 hr followed by treatment with an oxidative stress inducer, dimethoxy-naphthoquinone (DMNQ) or H2O2. Of note, DMNQ is a quinone that induces superoxide anion formation by redox cycling [47], whereas H2O2 undergoes the Fenton reaction in the presence of iron and produces reactive oxygen species, causing cell injury and death [48]. As shown in Fig 3A, DMNQ treatment led to a significant decrease in the number of cells. However, pre-treating the cells with SCF failed to prevent the cell death induced by DMNQ treatment (Fig 3A). Similarly, SCF treatment of CPCs was not able to prevent or attenuate the cell death following H2O2-induced oxidative stress (Fig 3B). In contrast, insulin pre-treatment (a positive control) was able to rescue CPCs from H2O2-induced oxidative stress (Fig 3C). These results indicate that while SCF-induced activation of c-kit promotes survival of CPCs grown under serum depletion, it does not recue CPCs subjected to oxidative stress.


C-Kit Promotes Growth and Migration of Human Cardiac Progenitor Cells via the PI3K-AKT and MEK-ERK Pathways.

Vajravelu BN, Hong KU, Al-Maqtari T, Cao P, Keith MC, Wysoczynski M, Zhao J, Moore JB, Bolli R - PLoS ONE (2015)

c-kit activation does not rescue CPCs from oxidative stress.A, Human c-kit+ CPCs were pre-treated with SCF for 24 hr in serum free media and subjected to 8 μM DMNQ treatment. B, Cells pre-treated with or without SCF received 1 hr treatment in 0.5 mM H2O2, followed by media change with or without SCF. Insulin was used as a positive control. Cell viability was assessed after 3 days using PrestoBlueTM. *, p<0.05 compared to the untreated control, #, p<0.05 compared to the H2O2 only control. NS, no statistical significance. Values presented are mean ± SEM. *, p<0.05.
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Related In: Results  -  Collection

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pone.0140798.g003: c-kit activation does not rescue CPCs from oxidative stress.A, Human c-kit+ CPCs were pre-treated with SCF for 24 hr in serum free media and subjected to 8 μM DMNQ treatment. B, Cells pre-treated with or without SCF received 1 hr treatment in 0.5 mM H2O2, followed by media change with or without SCF. Insulin was used as a positive control. Cell viability was assessed after 3 days using PrestoBlueTM. *, p<0.05 compared to the untreated control, #, p<0.05 compared to the H2O2 only control. NS, no statistical significance. Values presented are mean ± SEM. *, p<0.05.
Mentions: Next, we tested if SCF can prevent cell death induced by other stress conditions, such as oxidative stress, in CPCs. For this, CPCs were pre-treated with SCF for 24 hr followed by treatment with an oxidative stress inducer, dimethoxy-naphthoquinone (DMNQ) or H2O2. Of note, DMNQ is a quinone that induces superoxide anion formation by redox cycling [47], whereas H2O2 undergoes the Fenton reaction in the presence of iron and produces reactive oxygen species, causing cell injury and death [48]. As shown in Fig 3A, DMNQ treatment led to a significant decrease in the number of cells. However, pre-treating the cells with SCF failed to prevent the cell death induced by DMNQ treatment (Fig 3A). Similarly, SCF treatment of CPCs was not able to prevent or attenuate the cell death following H2O2-induced oxidative stress (Fig 3B). In contrast, insulin pre-treatment (a positive control) was able to rescue CPCs from H2O2-induced oxidative stress (Fig 3C). These results indicate that while SCF-induced activation of c-kit promotes survival of CPCs grown under serum depletion, it does not recue CPCs subjected to oxidative stress.

Bottom Line: SCF treatment led to a significant increase in cell survival and a reduction in cell death under serum depletion conditions.In addition, SCF significantly promoted CPC migration in vitro.These results imply that the efficiency of CPC homing to the injury site as well as their survival after transplantation may be improved by modulating the activity of c-kit.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Cardiology, Department of Medicine, University of Louisville, Louisville, KY 40202, United States of America.

ABSTRACT
A recent phase I clinical trial (SCIPIO) has shown that autologous c-kit+ cardiac progenitor cells (CPCs) improve cardiac function and quality of life when transplanted into patients with ischemic heart disease. Although c-kit is widely used as a marker of resident CPCs, its role in the regulation of the cellular characteristics of CPCs remains unknown. We hypothesized that c-kit plays a role in the survival, growth, and migration of CPCs. To test this hypothesis, human CPCs were grown under stress conditions in the presence or absence of SCF, and the effects of SCF-mediated activation of c-kit on CPC survival/growth and migration were measured. SCF treatment led to a significant increase in cell survival and a reduction in cell death under serum depletion conditions. In addition, SCF significantly promoted CPC migration in vitro. Furthermore, the pro-survival and pro-migratory effects of SCF were augmented by c-kit overexpression and abrogated by c-kit inhibition with imatinib. Mechanistically, c-kit activation in CPCs led to activation of the PI3K and the MAPK pathways. With the use of specific inhibitors, we confirmed that the SCF/c-kit-dependent survival and chemotaxis of CPCs are dependent on both pathways. Taken together, our findings suggest that c-kit promotes the survival/growth and migration of human CPCs cultured ex vivo via the activation of PI3K and MAPK pathways. These results imply that the efficiency of CPC homing to the injury site as well as their survival after transplantation may be improved by modulating the activity of c-kit.

No MeSH data available.


Related in: MedlinePlus