Expansion of normal and leukemic human hematopoietic stem/progenitor cells requires Rac-mediated interaction with stromal cells

Objective. To determine the involvement of Rac signaling in self-renewal and expansion on bone marrow stroma of normal CD34(+) cells vs leukemic CD34(+) cells from acute myeloid leukemia (AML) patients.

Materials and Methods. Rac signaling was modulated by retroviral introduction of Rac1-N17, Rac1-V12, or by using the Rac inhibitor NSC23766. In long-term MS5 cocultures (leukemic) expansion, migration, adhesion, and presence of stem/progenitor cells were monitored in both normal as well as leukemic CD34(+) cells.

Results. Inhibition of Rac signaling impaired migration and adhesion of cord blood (CB) CD34(+) cells on MS5 stroma. Long-term inhibition of Rac during a 5-week coculture period on stroma prevented association of hematopoietic progenitors with the bone marrow stromal cells and resulted in a dramatic decrease in the primitive stem cell frequency (long-term culture-initiating cell) in a dose-dependent manner. Many of these phenotypes were reversed in the presence of activated Rac1-V12, including improved migration toward, and association with, MS5 cells. CD34(+) AML cells were characterized by elevated levels of Rac activity (five of seven patients) and enhanced migration and adhesion to MS5 bone marrow stroma as compared to CB CD34(+) cells. A dramatic decrease was observed in the formation of leukemic cobblestone area-forming cells as well as strongly diminished clonal expansion in the presence of the Rac inhibitor NSC23766.

Conclusion. Our data indicate that Rac signal transduction is required for the maintenance and expansion of both normal as well as leukemic stem/progenitor cells by mediating their interaction with stromal cells. (c) 2007 International Society for Experimental Hematology. Published by Elsevier Inc.