Analysis of migratory and prosurvival pathways induced by the homeostatic chemokines CCL19 and CCL21 in B-cell chronic lymphocytic leukemia

Cuesta-Mateos, Carlos; López-Giral, Sonia; Alfonso-Pérez, Manuel; de Soria, Valle Gómez García; Loscertales, Javier; Guasch-Vidal, Sara; Beltrán, Amada Elia; Zapata, Juan M.; Muñoz-Calleja, Cecilia

doi:10.1016/j.exphem.2010.05.003

« Previous Next »Experimental Hematology
Volume 38, Issue 9 , Pages 756-764.e4, September 2010

Analysis of migratory and prosurvival pathways induced by the homeostatic chemokines CCL19 and CCL21 in B-cell chronic lymphocytic leukemia

Carlos Cuesta-Mateos
- Department of Immunology, Hospital Universitario de La Princesa, Madrid, Spain
Sonia López-Giral
- Department of Immunology, Hospital Universitario de La Princesa, Madrid, Spain
Manuel Alfonso-Pérez
- Department of Immunology, Hospital Universitario de La Princesa, Madrid, Spain
Valle Gómez García de Soria
- Department of Hematology, Hospital Universitario de La Princesa, Madrid, Spain
Javier Loscertales
- Department of Hematology, Hospital Universitario de La Princesa, Madrid, Spain
Sara Guasch-Vidal
- Department of Immunology, Hospital Universitario de La Princesa, Madrid, Spain
Amada Elia Beltrán
- Department of Immunology, Hospital Universitario de La Princesa, Madrid, Spain
Juan M. Zapata
- Instituto de Investigaciones Biomédicas “Alberto Sols,” CSIC/UAM, Madrid, Spain
Cecilia Muñoz-Calleja
- Department of Immunology, Hospital Universitario de La Princesa, Madrid, Spain
- Offprint requests to: Cecilia Muñoz-Calleja, M.D., Ph.D., Department of Immunology, Hospital Universitario de La Princesa, C/Diego de León 62, Madrid 28006, Spain

Received 11 August 2009; received in revised form 5 May 2010; accepted 6 May 2010. published online 20 May 2010.

Objective

The CCR7 chemokine receptor has been reported to promote homing of B-cell chronic lymphocytic leukemia (CLL) cells into lymph nodes and support their survival, but the mechanisms mediating these effects are largely unknown. We investigated the role of different signaling pathways triggered by CCR7 engagement by its ligands, the chemokines CCL19 and CCL21, in the control of CLL migration and survival.

Materials and Methods

Chemotaxis and apoptosis assays were performed in the presence of pharmacologic inhibitors and genetic mutants of the phosphatidylinositol-3-OH kinase (PI3K), Rho guanosine triphosphatase, and mitogen-activated protein kinase (MAPK) signaling cascades to assess the role of these pathways on primary CLL migration and survival in response to CCR7 activation. Kinase activation was determined by immunoblotting and pull-down experiments.

Results

CLL chemotactic activity induced by CCL19 or CCL21 was markedly reduced by inhibitors of PI3K and the Rho effector molecule Rho-associated coiled-coil forming protein kinases (ROCK), and also by the expression of dominant negative forms of PI3K and RhoA, whereas constitutively activated PI3K and RhoA mutants strongly promoted CLL migration. In contrast, MAPKs were not significantly involved in CLL migration to CCL19/CCL21. Conversely, extracellular signal-regulated kinase and c-Jun-N-terminal kinase, along with PI3K, had a role in CCR7-mediated CLL cell survival. Biochemical experiments confirmed that CCL19/21 induced PI3K-dependent phosphorylation of Akt/protein kinase B, activation of the Rho/Rho-associated coiled-coil forming protein kinases/myosin light chain pathway and MAPKs phosphorylation.

Conclusions

The role of PI3K, Rho guanosine triphosphatases, and MAPKs in CCR7-mediated CLL cells migration and survival suggests that these signal transduction pathways could represent promising targets for CLL therapy.