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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 induces proliferation and decreases apoptotic cell death in CPCs.Human c-kit+ CPCs were serum starved for 24 hr and labeled with BrdU in the presence or absence of SCF for 24 hr only for the indicated day (A) or continuously for 3 days (B). Cells were stained with anti-BrdU antibody, and the positive cells were expressed as the percentage of total nuclei. C, CPCs were grown in serum free media for 3 days. Activities of caspases 3 and 7 were measured and normalized to the untreated control. Values presented are mean ± SEM. *, p<0.05.
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pone.0140798.g002: c-kit activation induces proliferation and decreases apoptotic cell death in CPCs.Human c-kit+ CPCs were serum starved for 24 hr and labeled with BrdU in the presence or absence of SCF for 24 hr only for the indicated day (A) or continuously for 3 days (B). Cells were stained with anti-BrdU antibody, and the positive cells were expressed as the percentage of total nuclei. C, CPCs were grown in serum free media for 3 days. Activities of caspases 3 and 7 were measured and normalized to the untreated control. Values presented are mean ± SEM. *, p<0.05.

Mentions: In order to test whether the observed increase in the number of surviving cells was due to the mitogenic effect of SCF/c-kit signaling, we performed a BrdU labeling assay [46]. For this assay, we cultured the cells under serum starvation in the presence or absence of SCF and incubated them with BrdU for 24 hr starting at different time points. When compared to the control, SCF increased the BrdU labeling index in CPCs during days 2 and 3 of the treatment, suggesting that SCF treatment promotes proliferation in CPCs (Fig 2A). However, the difference between the two treatment groups did not reach statistical significance. Next, we treated the cells with BrdU continuously for 3 days with or without SCF. With continuous BrdU supply and SCF treatment, the percentage of BrdU-positive CPCs increased to 10.2 ± 3.9% (SCF-treated group) from 3.9 ± 3.4% (untreated control group), significantly raising the rate of proliferation by more than 2-fold (Fig 2B). This demonstrates that SCF acts as a mitogen for CPCs under conditions that restrict their growth.


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 induces proliferation and decreases apoptotic cell death in CPCs.Human c-kit+ CPCs were serum starved for 24 hr and labeled with BrdU in the presence or absence of SCF for 24 hr only for the indicated day (A) or continuously for 3 days (B). Cells were stained with anti-BrdU antibody, and the positive cells were expressed as the percentage of total nuclei. C, CPCs were grown in serum free media for 3 days. Activities of caspases 3 and 7 were measured and normalized to the untreated control. Values presented are mean ± SEM. *, p<0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4608800&req=5

pone.0140798.g002: c-kit activation induces proliferation and decreases apoptotic cell death in CPCs.Human c-kit+ CPCs were serum starved for 24 hr and labeled with BrdU in the presence or absence of SCF for 24 hr only for the indicated day (A) or continuously for 3 days (B). Cells were stained with anti-BrdU antibody, and the positive cells were expressed as the percentage of total nuclei. C, CPCs were grown in serum free media for 3 days. Activities of caspases 3 and 7 were measured and normalized to the untreated control. Values presented are mean ± SEM. *, p<0.05.
Mentions: In order to test whether the observed increase in the number of surviving cells was due to the mitogenic effect of SCF/c-kit signaling, we performed a BrdU labeling assay [46]. For this assay, we cultured the cells under serum starvation in the presence or absence of SCF and incubated them with BrdU for 24 hr starting at different time points. When compared to the control, SCF increased the BrdU labeling index in CPCs during days 2 and 3 of the treatment, suggesting that SCF treatment promotes proliferation in CPCs (Fig 2A). However, the difference between the two treatment groups did not reach statistical significance. Next, we treated the cells with BrdU continuously for 3 days with or without SCF. With continuous BrdU supply and SCF treatment, the percentage of BrdU-positive CPCs increased to 10.2 ± 3.9% (SCF-treated group) from 3.9 ± 3.4% (untreated control group), significantly raising the rate of proliferation by more than 2-fold (Fig 2B). This demonstrates that SCF acts as a mitogen for CPCs under conditions that restrict their growth.

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