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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 promotes migration of CPCs.Boyden chamber assay. Human c-kit+ CPCs were plated in the upper chamber, and the indicated growth factors were used as a chemoattractant in the lower chamber. After 24 hr, the migrated cells were fixed, stained with propidium iodide, and counted. A, Comparison of the pro-migratory effect of SCF and VEGF. B, Representative images showing the cells that migrated towards the indicated growth factors. C, CPCs transduced with lentivirus expressing mCherry (control) or c-kit were compared for their chemotaxis towards SCF in the presence (+) or absence (-) of 0.5 mM imatinib. Values presented are mean ± SEM. *, p<0.05.
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pone.0140798.g004: c-kit activation promotes migration of CPCs.Boyden chamber assay. Human c-kit+ CPCs were plated in the upper chamber, and the indicated growth factors were used as a chemoattractant in the lower chamber. After 24 hr, the migrated cells were fixed, stained with propidium iodide, and counted. A, Comparison of the pro-migratory effect of SCF and VEGF. B, Representative images showing the cells that migrated towards the indicated growth factors. C, CPCs transduced with lentivirus expressing mCherry (control) or c-kit were compared for their chemotaxis towards SCF in the presence (+) or absence (-) of 0.5 mM imatinib. Values presented are mean ± SEM. *, p<0.05.

Mentions: For successful engraftment of the donor cells following transplantation, homing of the CPCs to the recipient myocardium and the injured area is required. SCF/c-kit signaling has been implicated in stimulating migration of various c-kit-expressing cell types [40, 49, 50]. Therefore, we sought to investigate whether SCF/c-kit signaling contributes to migration of CPCs. To test this, we implemented the Boyden chamber assay and used SCF as a chemo attractant. CPCs migrated towards the SCF gradient, and such chemotactic effect of SCF was comparable to that of VEGF (Fig 4A and 4B), which has previously been shown to promote migration of CPCs [51]. To further confirm this finding, we overexpressed c-kit (or mCherry as a control) in CPCs using lentivirus and performed the same migration assay. As expected, upon c-kit overexpression and SCF treatment, the number of migrated cells was further increased compared to the mCherry-expressing control CPCs (Fig 4C). Moreover, inhibition of c-kit with imatinib abolished the pro-migratory effect of SCF/c-kit signaling (Fig 4C), suggesting that the chemotactic effect of SCF on CPCs are indeed mediated through activation of the c-kit receptor.


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 promotes migration of CPCs.Boyden chamber assay. Human c-kit+ CPCs were plated in the upper chamber, and the indicated growth factors were used as a chemoattractant in the lower chamber. After 24 hr, the migrated cells were fixed, stained with propidium iodide, and counted. A, Comparison of the pro-migratory effect of SCF and VEGF. B, Representative images showing the cells that migrated towards the indicated growth factors. C, CPCs transduced with lentivirus expressing mCherry (control) or c-kit were compared for their chemotaxis towards SCF in the presence (+) or absence (-) of 0.5 mM imatinib. 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.g004: c-kit activation promotes migration of CPCs.Boyden chamber assay. Human c-kit+ CPCs were plated in the upper chamber, and the indicated growth factors were used as a chemoattractant in the lower chamber. After 24 hr, the migrated cells were fixed, stained with propidium iodide, and counted. A, Comparison of the pro-migratory effect of SCF and VEGF. B, Representative images showing the cells that migrated towards the indicated growth factors. C, CPCs transduced with lentivirus expressing mCherry (control) or c-kit were compared for their chemotaxis towards SCF in the presence (+) or absence (-) of 0.5 mM imatinib. Values presented are mean ± SEM. *, p<0.05.
Mentions: For successful engraftment of the donor cells following transplantation, homing of the CPCs to the recipient myocardium and the injured area is required. SCF/c-kit signaling has been implicated in stimulating migration of various c-kit-expressing cell types [40, 49, 50]. Therefore, we sought to investigate whether SCF/c-kit signaling contributes to migration of CPCs. To test this, we implemented the Boyden chamber assay and used SCF as a chemo attractant. CPCs migrated towards the SCF gradient, and such chemotactic effect of SCF was comparable to that of VEGF (Fig 4A and 4B), which has previously been shown to promote migration of CPCs [51]. To further confirm this finding, we overexpressed c-kit (or mCherry as a control) in CPCs using lentivirus and performed the same migration assay. As expected, upon c-kit overexpression and SCF treatment, the number of migrated cells was further increased compared to the mCherry-expressing control CPCs (Fig 4C). Moreover, inhibition of c-kit with imatinib abolished the pro-migratory effect of SCF/c-kit signaling (Fig 4C), suggesting that the chemotactic effect of SCF on CPCs are indeed mediated through activation of the c-kit receptor.

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