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and Stem Cell Factor/kit Ligand Share Signaling Pathways in Hemopoietic Progenitors: A Potential Mechanism for Cooperative Induction of Chemotaxis1
Divisions of Experimental Medicine and Hematology/Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215
Stromal cell-derived factor (SDF-1
), the ligand for CXCR4, is a
chemokine that acts as a potent chemoattractant for hemopoietic
progenitor cells. Stem cell factor/kit ligand (SCF/KL),
an early acting cytokine, has recently been reported to enhance the
chemotaxis induced by SDF-1. However, very little is known about
downstream signaling events following these receptor-ligand
interactions. To investigate these events, we utilized a model
progenitor cell line, CTS, which expresses both the CXCR4 and
c-kit receptors. We observed strong Ca2+
mobilization and enhancement of chemotaxis following treatment with
SDF-1 or SCF/KL. A combination of these factors enhanced this
chemotaxis in CTS cells as well as in CD34+ bone marrow
cells. Prior treatment of CTS cells with pertussis toxin inhibited the
SDF-1-induced chemotaxis, suggesting that SDF-1 signaling
involves a pertussis-sensitive Gi-coupled protein. SDF-1
treatment resulted in a rapid phosphorylation of the focal adhesion
molecules RAFTK (related adhesion focal tyrosine kinase), paxillin, and
p130cas, which then declined within
minutes. SCF/KL alone or in combination with SDF-1 induced a rapid
and sustained effect on phosphorylation of these substrates. SDF-1
treatment resulted in a rapid and robust activation of p44/42
mitogen-activated protein kinase compared with the relatively weak and
delayed effect of SCF/KL treatment. Interestingly, a delayed but
sustained activation of mitogen-activated protein kinase activation was
observed when the factors were used in combination. Such cooperativity
in downstream signaling pathways may explain the enhanced chemotaxis of
progenitors observed with SDF-1 in combination with
SCF/KL.
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