A small-molecule antagonist of CXCR4 inhibits intracranial growth of primary brain tumors
Abstract
The vast majority of brain tumors in adults exhibit glial characteristics. Brain tumors in children are diverse: Many have neuronal characteristics, whereas others have glial features. Here we show that activation of the Gi protein-coupled receptor CXCR4 is critical for the growth of both malignant neuronal and glial tumors. Systemic administration of CXCR4 antagonist AMD 3100 inhibits growth of intracranial glioblastoma and medulloblastoma xenografts by increasing apoptosis and decreasing the proliferation of tumor cells. This reflects the ability of AMD 3100 to reduce the activation of extracellular signal-regulated kinases 1 and 2 and Akt, all of which are pathways downstream of CXCR4 that promote survival, proliferation, and migration. These studies (i) demonstrate that CXCR4 is critical to the progression of diverse brain malignances and (ii) provide a scientific rationale for clinical evaluation of AMD 3100 in treating both adults and children with malignant brain tumors.
Malignant brain tumors are a major cause of cancer-related morbidity and mortality in both adults and children. Each year ≈15,000 patients die despite the best available multimodal therapy (1). The genetics of common adult and pediatric CNS malignancies, such as glioblastoma multiforme (GBM) and medulloblastoma, suggest that enhanced proliferation, increased resistance to apoptosis, and increased cell migration are all involved in the progression and maintenance of a cancerous state (2, 3). Thus, specific biological agents that simultaneously target all these processes may provide therapeutic alternatives to conventional cytotoxic therapies.
Chemokines, secreted factors initially described as regulators of leukocyte trafficking (4), are now known to have far-reaching influence in the development and functioning of many tissues. A critical role in the developing brain for the chemokine stromal cell-derived factor 1α (CXCL12) and its receptor, CXCR4, became evident when targeted gene deletion of either resulted in significant abnormalities in cerebellar development (5, 6). Cardinal features of the mutant animals were the mislocalization and failed proliferation of cerebellar granule precursor cells (GPCs). These phenotypic changes in GPCs reflect two effects of CXCL12, a chemotactic effect (7-9) and the ability of CXCL12 to synergize with Sonic hedgehog (Shh) in the promotion of GPC proliferation (7). The capacity of CXCL12 and CXCR4 to regulate proliferation and migration of neural precursor cells raises the possibility that these molecules might be therapeutic targets in malignancies arising from CNS progenitors.
CXCR4 expression in the brain is not limited to GPCs. Additional neuronal populations (10), astrocytes (11-13), and adult GBM all express CXCR4 (14-16). Here we report that CXCR4 mRNA and protein are expressed at high levels in brain tumors of both neuronal and astrocytic lineage. The ligand CXCL12 is expressed in tumor-associated blood vessels and/or tumor cells, suggesting a paracrine relationship for CXCR4 activation in vivo. In vitro, CXCL12 exerts proliferative, antiapoptotic, and chemotactic effects on both GBM and medulloblastoma cell lines. In vivo, systemic administration of AMD 3100, a small-molecule inhibitor of CXCR4 (17), decreased growth of GBM and medulloblastoma xenografts. Direct antitumor effects of AMD 3100 were evident in reduced activation of extracellular signal-regulated kinases 1 and 2 (Erk 1/2) and Akt and increased rates of apoptosis in both tumor types. Together these studies identify CXCR4 signaling as a critical component of brain tumor biology and demonstrate that small-molecule inhibition of CXCR4 has significant antineoplastic activity. AMD 3100 is well tolerated in human studies (18); thus, these findings could rapidly lead to clinical trials for malignant brain tumors.
Click here to view.Acknowledgments
We thank N. Warrington, E. Tanner, R. Wright, M. Pazyra, E. Berry, T. Zolotarev, and E. Lin for technical assistance; Dr. P. Febbo for assistance with genomic analysis; Dr. A. Annuar for the gift of AMD 3100; Dr. O. Gilchrist for the gift of Shh protein; and Drs. G. Bridger, G. Calandra, J. Kim, C. Stiles, B. Rollins, and S. Pomeroy for helpful discussions. This work was supported by grants from the National Institutes of Health (to J.B.R., A.L.K., R.S.K., A.D.L., and R.A.S.), the Goldhirsh Foundation (to J.B.R. and A.L.K.), the Claudia Adams Barr Program (to A.L.K. and R.A.S.), the Whitaker Foundation and the Brigham Radiology Research and Education Fund (to Y.S. and K.S.), and the Stop and Shop Family Pediatric Brain Tumor Program (to M.W.K.).
Notes
Abbreviations: GBM, glioblastoma multiforme; GPC, granule precursor cell; Shh, Sonic hedgehog; Erk 1/2, extracellular signal-regulated kinases 1 and 2; TUNEL, terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling; PECAM, platelet endothelial cell adhesion molecules; SFM, serum-free media.
References
- 1. Ries, L. A. G., Eisner, M. P., Kosary, C. L., Hankey, B. F., Miller, B. A., Clegg, L. & Edwards, B. K., eds. (2001) SEER Cancer Statistics Review (Natl. Cancer Inst., Bethesda).
- 2. Kleihues, P., Burger, P. C., Collins, V. P., Newcomb, E. W., Ohgaki, H. & Cavenee, W. K. (2000) in World Health Organization Classification of Tumours: Pathology and Genetics of Tumours of the Central Nervous System, eds. Kleihues, P. C. & Cavenee, W. K. (Int. Agency for Res. on Cancer, Lyon, France), pp. 29-39.
- 3. Giangaspero, F., Bigner, S. H., Kleihues, P., Pietsch, T. & Trojanowski, J. Q. (2000) in World Health Organization Classification of Tumours: Pathology and Genetics of Tumours of the Central Nervous System, eds. Kleihues, P. C. & Cavenee, W. K. (Int. Agency for Res. on Cancer, Lyon, France), pp. 129-137.
- 4. Luster, A. D. (1998) N. Engl. J. Med.338, 436-445. [[PubMed]
- 5. Ma, Q., Jones, D., Borghesani, P. R., Segal, R. A., Nagasawa, T., Kishimoto, T., Bronson, R. T. & Springer, T. A. (1998) Proc. Natl. Acad. Sci. USA95, 9448-9453.
- 6. Zou, Y. R., Kottmann, A. H., Kuroda, M., Taniuchi, I. & Littman, D. R. (1998) Nature393, 595-599. [[PubMed]
- 7. Klein, R. S., Rubin, J. B., Gibson, H. D., DeHaan, E. N., Alvarez-Hernandez, X., Segal, R. A. & Luster, A. D. (2001) Development (Cambridge, U.K.)128, 1971-1981. [PubMed]
- 8. Zhu, Y., Yu, T., Zhang, X. C., Nagasawa, T., Wu, J. Y. & Rao, Y. (2002) Nat. Neurosci.5, 719-720.
- 9. Lu, Q., Sun, E. E., Klein, R. S. & Flanagan, J. G. (2001) Cell105, 69-79. [[PubMed]
- 10. Stumm, R. K., Rummel, J., Junker, V., Culmsee, C., Pfeiffer, M., Krieglstein, J., Hollt, V. & Schulz, S. (2002) J. Neurosci.22, 5865-5878.
- 11. Han, Y., Wang, J., He, T. & Ransohoff, R. M. (2001) Brain Res.888, 1-10. [[PubMed]
- 12. Dorf, M. E., Berman, M. A., Tanabe, S., Heesen, M. & Luo, Y. (2000) J. Neuroimmunol.111, 109-121. [[PubMed]
- 13. Bajetto, A., Bonavia, R., Barbero, S., Piccioli, P., Costa, A., Florio, T. & Schettini, G. (1999) J. Neurochem.73, 2348-2357. [[PubMed]
- 14. Rempel, S. A., Dudas, S., Ge, S. & Gutierrez, J. A. (2000) Clin. Cancer Res.6, 102-111. [[PubMed]
- 15. Sehgal, A., Keener, C., Boynton, A. L., Warrick, J. & Murphy, G. P. (1998) J. Surg. Oncol.69, 99-104. [[PubMed]
- 16. Zhou, Y., Larsen, P. H., Hao, C. & Yong, V. W. (2002) J. Biol. Chem.277, 49481-49487. [[PubMed]
- 17. Gerlach, L. O., Skerlj, R. T., Bridger, G. J. & Schwartz, T. W. (2001) J. Biol. Chem.276, 14153-14160. [[PubMed]
- 18. Hendrix, C. W., Flexner, C., MacFarland, R. T., Giandomenico, C., Fuchs, E. J., Redpath, E., Bridger, G. & Henson, G. W. (2000) Antimicrob. Agents Che-mother.44, 1667-1673.
- 19. Sun, Y., Zhou, J., Stayner, C., Munasinghe, J., Shen, X., Beier, D. R. & Albert, M. S. (2002) Comp. Med.52, 433-438. [[PubMed]
- 20. Pomeroy, S. L., Tamayo, P., Gaasenbeek, M., Sturla, L. M., Angelo, M., McLaughlin, M. E., Kim, J. Y., Goumnerova, L. C., Black, P. M., Lau, C., et al. (2002) Nature415, 436-442. [[PubMed]
- 21. Klein, R. S., Williams, K. C., Alvarez-Hernandez, X., Westmoreland, S., Force, T., Lackner, A. A. & Luster, A. D. (1999) J. Immunol.163, 1636-1646. [[PubMed]
- 22. Rezaie, P., Trillo-Pazos, G., Everall, I. P. & Male, D. K. (2002) Glia37, 64-75. [[PubMed]
- 23. Goodrich, L. V., Milenkovic, L., Higgins, K. M. & Scott, M. P. (1997) Science277, 1109-1113. [[PubMed]
- 24. Evans, D. G., Farndon, P. A., Burnell, L. D., Gattamaneni, H. R. & Birch, J. M. (1991) Br. J. Cancer64, 959-961.
- 25. Zhao, Q., Kho, A., Kenney, A. M., Yuk Di, D. I., Kohane, I. & Rowitch, D. H. (2002) Proc. Natl. Acad. Sci. USA99, 5704-5709.
- 26. Hatse, S., Princen, K., Bridger, G., De Clercq, E. & Schols, D. (2002) FEBS Lett.527, 255-262. [[PubMed]
- 27. Bajetto, A., Barbero, S., Bonavia, R., Piccioli, P., Pirani, P., Florio, T. & Schettini, G. (2001) J. Neurochem.77, 1226-1236. [[PubMed]
- 28. Suzuki, Y., Rahman, M. & Mitsuya, H. (2001) J. Immunol.167, 3064-3073. [[PubMed]
- 29. Rehemtulla, A., Stegman, L. D., Cardozo, S. J., Gupta, S., Hall, D. E., Contag, C. H. & Ross, B. D. (2000) Neoplasia2, 491-495.
- 30. Vooijs, M., Jonkers, J., Lyons, S. & Berns, A. (2002) Cancer Res.62, 1862-1867. [[PubMed]
- 31. Vorechovsky, I., Tingby, O., Hartman, M., Stromberg, B., Nister, M., Collins, V. P. & Toftgard, R. (1997) Oncogene15, 361-366. [[PubMed]
- 32. Lee, T. H., Avraham, H., Lee, S. H. & Avraham, S. (2002) J. Biol. Chem.277, 10445-10451. [[PubMed]
- 33. Helbig, G., Christopherson, K. W., II, Bhat-Nakshatri, P., Kumar, S., Kishimoto, H., Miller, K. D., Broxmeyer, H. E. & Nakshatri, H. (2003) J. Biol. Chem.278, 21631-21638. [[PubMed]
- 34. Barbero, S., Bonavia, R., Bajetto, A., Porcile, C., Pirani, P., Ravetti, J. L., Zona, G. L., Spaziante, R., Florio, T. & Schettini, G. (2003) Cancer Res.63, 1969-1974. [[PubMed]
- 35. Libura, J., Drukala, J., Majka, M., Tomescu, O., Navenot, J. M., Kucia, M., Marquez, L., Peiper, S. C., Barr, F. G., Janowska-Wieczorek, A. & Ratajczak, M. Z. (2002) Blood100, 2597-2606. [[PubMed]
- 36. Geminder, H., Sagi-Assif, O., Goldberg, L., Meshel, T., Rechavi, G., Witz, I. P. & Ben-Baruch, A. (2001) J. Immunol.167, 4747-4757. [[PubMed]
- 37. Muller, A., Homey, B., Soto, H., Ge, N., Catron, D., Buchanan, M. E., McClanahan, T., Murphy, E., Yuan, W., Wagner, S. N., et al. (2001) Nature410, 50-56. [[PubMed]
- 38. Schrader, A. J., Lechner, O., Templin, M., Dittmar, K. E., Machtens, S., Mengel, M., Probst-Kepper, M., Franzke, A., Wollensak, T., Gatzlaff, P., et al. (2002) Br. J. Cancer86, 1250-1256.
- 39. Koshiba, T., Hosotani, R., Miyamoto, Y., Ida, J., Tsuji, S., Nakajima, S., Kawaguchi, M., Kobayashi, H., Doi, R., Hori, T., et al. (2000) Clin. Cancer Res.6, 3530-3535. [[PubMed]
- 40. Kijima, T., Maulik, G., Ma, P. C., Tibaldi, E. V., Turner, R. E., Rollins, B., Sattler, M., Johnson, B. E. & Salgia, R. (2002) Cancer Res.62, 6304-6311. [[PubMed]
- 41. Payne, A. S. & Cornelius, L. A. (2002) J. Invest. Dermatol.118, 915-922. [[PubMed]
- 42. Durig, J., Schmucker, U. & Duhrsen, U. (2001) Leukemia15, 752-756. [[PubMed]
- 43. Scherer, H. J. (1938) Am. J. Cancer34, 333-351. [PubMed]
- 44. Davies, D. C. (2002) J. Anat.200, 639-646.
- 45. Seitz, R. J. & Wechsler, W. (1987) Acta Neuropathol.73, 145-152. [[PubMed]