Preclinical Efficacy of Nimotuzumab, an Anti-Egfr Monoclonal Antibody as a Single Agent Therapy in Human GBM u87mg Xenografts — Oak Academic Publishing
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Preclinical Efficacy of Nimotuzumab, an Anti-Egfr Monoclonal Antibody as a Single Agent Therapy in Human GBM u87mg Xenografts
Center of Molecular Immunology; Havana, Cuba
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Center of Molecular Immunology; Havana, Cuba
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Max Delbrück Center for Molecular Medicine. Berlin, Germany
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Max Delbrück Center for Molecular Medicine. Berlin, Germany
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Center of Molecular Immunology; Havana, Cuba
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Center of Molecular Immunology; Havana, Cuba
1 Center of Molecular Immunology; Havana, Cuba
2 Center of Molecular Immunology; Havana, Cuba
3 Max Delbrück Center for Molecular Medicine. Berlin, Germany
4 Max Delbrück Center for Molecular Medicine. Berlin, Germany
Background: The poor prognosis of patients with high-grade glioma multiform (GBM) has led investigators to the search of new therapeutic strategies. Current treatment includes surgery when possible, radiotherapy and chemotherapy. Molecular-targeted therapies are in the process of clinical testing, and promising agents include monoclonal antibodies. Our study examined the antitumor activity of three different single therapies in nude mice bearing both subcutaneous and orthotopic brain xenografts of the U87MG human GBM cell line. Methods: Cell culture, Histology, Immunohistochemistry, Animal experiments, Statistical analysis. Results: Different groups of treatment included nimotuzumab, a humanized monoclonal antibody that inhibits the EGFR tyrosine kinase activity, or total body irradiation, or the chemotherapeutic agent temozolomide (TMZ). For the control group animals received saline solution instead of the antibody. For the subcutaneous model, only nimotuzumab or TMZ produced a significant delay in tumor growth. In the intracranial model, unlike TMZ, the systemic administration of the antibody did not reduce the tumor growth, despite both therapies inhibited the formation of microsatellites in the brain of mice. The antitumor activity of nimotuzumab was accompanied by a decrease in the microvessel density and the proliferative activity of tumor cells. TMZ only inhibited the tumor cell proliferation but not the formation of new tumor-associated microvessels in xenografts. For radiation therapy, neither antiproliferative nor antiangiogenic activity was found, in accordance with the lack of antitumor activity. Only nimotuzumab reduced the frequency of chemo and radioresistant CD133+ population. Conclusion: Our results illustrate the potential efficacy of nimotuzumab as a single agent against an EGFR-amplified human GBM, a tumor resistant to the therapy with all well-known forms of treatment.
R. Stupp, W. P. Mason, M. J. van den Bent, M. Weller, B. Fisher, M. J. Taphoorn, K. Belanger, A. A. Brandes, C. Marosi, U. Bogdahn, J. Curschmann, R. C. Janzer, S. K. Ludwin, T. Gorlia, A. Allgeier, D. Lacombe, J. G. Cairncross, E. Eisenhauer and R. O. Mirimanoff, “Radiotherapy plus Concomitant and Adjuvant Temozolomide for Glioblastoma,” The New England Journal of Medicine, Vol. 352, No. 10, 2005, pp. 987-996. doi:10.1056/NEJMoa043330
E. G. Van Meir, C. G. Hadjipanayis, A. D. Norden, H. K. Shu, P. Y. Wen and J. J. Olson, “Exciting New Advances in Neuro-Oncology: The Avenue to a Cure for Malignant Glioma,” CA: A Cancer Journal for Clinicians, Vol. 60, No. 3, 2010, pp. 166-193. doi:10.3322/caac.20069
L. G. Feun, N. Savaraj and H. J. Landy, “Drug Resistance in Brain Tumors,” Journal of Neuro-Oncology, Vol. 20, No. 2, 1994, pp. 165-176. doi:10.1007/BF01052726
J. N. Rich, “Cancer Stem Cells in Radiation Resistance,” Cancer Research, Vol. 67, No. 19, 2007, pp. 8980-8984. doi:10.1158/0008-5472.CAN-07-0895
M. D. Prados and C. Russo, “Chemotherapy of Brain Tumors,” Seminars in Surgical Oncology, Vol. 14, No. 1, 1998, pp. 88-95. doi:10.1002/(SICI)1098-2388(199801/02)14:1 3.0.CO;2-5
D. Beier, S. Rohrl, D. R. Pillai, S. Schwarz, L. A. Kunz-Schughart, P. Leukel, M. Proescholdt, A. Brawanski, U. Bogdahn, A. Trampe-Kieslich, B. Giebel, J. Wischhusen, G. Reifenberger, P. Hau and C. P. Beier, “Temozolomide Preferentially Depletes Cancer Stem Cells in Glioblastoma,” Cancer Research, Vol. 68, No. 14, 2008, pp. 5706-5015. doi:10.1158/0008-5472.CAN-07-6878
H. Hauch, M. Sajedi and J. E. Wolff, “Treatment Arms Summarizing Analysis of 220 High-Grade Glioma Studies,” Anticancer Research, Vol. 25, No. 5, 2005, pp. 3585-3590.
T. C. Ramos, J. Figueredo, M. Catala, S. Gonzalez, J. C. Selva, T. M. Cruz, C. Toledo, S. Silva, Y. Pestano, M. Ramos, I. Leonard, O. Torres, P. Marinello, R. Perez and A. Lage, “Treatment of High-Grade Glioma Patients with the Humanized Anti-Epidermal Growth Factor Receptor (EGFR) Antibody h-R3: Report from a Phase I/II Trial,” Cancer Biology & Therapy, Vol. 5, No. 4, 2006, pp. 375-379. doi:10.4161/cbt.5.4.2522
P. K. Julka, S. Mallick, T. Puri, S. Goyal, N. Joshi, N. Gupta and G. K. Rath, “Feasibility and Safety of Combining Nimotuzumab with Temozolomide and Radiotherapy in Adult Patients with Glioblastoma: An Indian Clinical Experience,” Journal of Clinical Oncology, Vol. 28, No. 15, 2010.
U. Bode, S. Buchen, M. Warmuth-Metz, T. Pietsch, F. Bach and G. Fleischhack, “Final Report of a Phase II Trial of Nimotuzumab in the Treatment of Refractory and Relapsed High-Grade Gliomas in Children and Adolescents,” Journal of Clinical Oncology, Vol. 25, No. 18, 2007.
A. Diaz Miqueli, J. Rolff, M. Lemm, I. Fichtner, R. Perez and E. Montero, “Radiosensitisation of U87MG Brain Tumours by Anti-Epidermal Growth Factor Receptor Monoclonal Antibodies,” British Journal of Cancer, Vol. 100, No. 6, 2009, pp. 950-958. doi:10.1038/sj.bjc.6604943
C. Mateo, E. Moreno, K. Amour, J. Lombardero, W. Harris and R. Perez, “Humanization of a Mouse Monoclonal Antibody That Blocks the Epidermal Growth Factor Receptor: Recovery of Antagonistic Activity,” Immunotechnology, Vol. 3, No. 1, 1997, pp. 71-81. doi:10.1016/S1380-2933(97)00065-1
“United Kingdom Coordinating Committee on Cancer Research (UKCCCR) Guidelines for the Welfare of Animals in Experimental Neoplasia (Second Edition),” British Journal of Cancer, Vol. 77, No. 1, 1998, pp. 1-10.
A. M. Mansour, J. Drevs, N. Esser, F. M. Hamada, O. A. Badary, C. Unger, I. Fichtner, and F. Kratz, “A New Approach for the Treatment of Malignant Melanoma: Enhanced Antitumor Efficacy of an Albumin-Binding Doxorubicin Prodrug That Is Cleaved by Matrix Metalloproteinase 2,” Cancer Research, Vol. 63, No. 14, 2003, pp. 4062-4066.
S. C. Yu, Y. F. Ping, L. Yi, Z. H. Zhou, J. H. Chen, X. H. Yao, L. Gao, J. M. Wang and X. W. Bian, “Isolation and Characterization of Cancer Stem Cells from a Human Glioblastoma Cell Line U87,” Cancer Letters, Vol. 265, No. 1, 2008, pp. 124-134. doi:10.1016/j.canlet.2008.02.010
B. Kaur, F. W. Khwaja, E. A. Severson, S. L. Matheny, D. J. Brat and E. G. Van Meir, “Hypoxia and the Hypoxia-Inducible-Factor Pathway in Glioma Growth and Angiogenesis,” Journal of Neuro-Oncology, Vol. 7, No. 2, 2005, pp. 134-153. doi:10.1215/S1152851704001115
T. Crombet-Ramos, J. Rak, R. Perez and A. Viloria-Petit, “Antiproliferative, Antiangiogenic and Proapoptotic Activity of h-R3: A Humanized Anti-EGFR Antibody,” International Journal of Cancer, Vol. 101, No. 6, 2002, pp. 567-575. doi:10.1002/ijc.10647
J. Folkman, “Angiogenesis: An Organizing Principle for Drug Discovery,” Nature Reviews Drug Discovery, Vol. 6, No. 4, 2007, pp. 273-286. doi:10.1038/nrd2115
S. K. Singh, C. Hawkins, I. D. Clarke, J. A. Squire, J. Bayani, T. Hide, R. M. Henkelman, M. D. Cusimano and P. B. Dirks, “Identification of Human Brain Tumour Initiating Cells,” Nature, Vol. 432, No. 7015, 2004, pp. 396-401. doi:10.1038/nature03128
S. Bao, Q. Wu, R. E. McLendon, Y. Hao, Q. Shi, A. B. Hjelmeland, M. W. Dewhirst, D. D. Bigner and J. N. Rich, “Glioma Stem Cells Promote Radioresistance by Preferential Activation of the DNA Damage Response,” Nature, Vol. 444, No. 7120, 2006, pp. 756-760. doi:10.1038/nature05236
S. Bao, Q. Wu, S. Sathornsumetee, Y. Hao, Z. Li, A. B. Hjelmeland, Q. Shi, R. E. McLendon, D. D. Bigner and J. N. Rich, “Stem Cell-Like Glioma Cells Promote Tumor Angiogenesis through Vascular Endothelial Growth Factor,” Cancer Research, Vol. 66, No. 16, 2006, pp. 7843-7848. doi:10.1158/0008-5472.CAN-06-1010
A. Eramo, L. Ricci-Vitiani, A. Zeuner, R. Pallini, F. Lotti, G. Sette, E. Pilozzi, L. M. Larocca, C. Peschle and R. De Maria, “Chemotherapy Resistance of Glioblastoma Stem Cells,” Cell Death & Differentiation, Vol. 13, No. 7, 2006, pp. 1238-1241. doi:10.1038/sj.cdd.4401872
G. Liu, X. Yuan, Z. Zeng, P. Tunici, H. Ng, I. R. Abdulkadir, L. Lu, D. Irvin, K. L. Black and J. S. Yu, “Analysis of Gene Expression and Chemoresistance of CD133+ Cancer Stem Cells in Glioblastoma,” Molecular Cancer, Vol. 5, 2006, p. 67. doi:10.1186/1476-4598-5-67
N. Shinojima, K. Tada, S. Shiraishi, T. Kamiryo, M. Kochi, H. Nakamura, K. Makino, H. Saya, H. Hirano, J. Kuratsu, K. Oka, Y. Ishimaru and Y. Ushio, “Prognostic Value of Epidermal Growth Factor Receptor in Patients with Glioblastoma Multiforme,” Cancer Research, Vol. 63, No. 20, 2003, pp. 6962-6970.
J. J. Vredenburgh, A. Desjardins, J. E. Herndon II, J. M. Dowell, D. A. Reardon, J. A. Quinn, J. N. Rich, S. Sathornsumetee, S. Gururangan, M. Wagner, D. D. Bigner, A. H. Friedman and H. S. Friedman, “Phase II Trial of Bevacizumab and Irinotecan in Recurrent Malignant Glioma,” Clinical Cancer Research, Vol. 13, No. 4, 2007, pp. 1253-1259. doi:10.1158/1078-0432.CCR-06-2309
E. S. Kim, A. Serur, J. Huang, C. A. Manley, K. W. McCrudden, J. S. Frischer, S. Z. Soffer, L. Ring, T. New, S. Zabski, J. S. Rudge, J. Holash, G. D. Yancopoulos, J. J. Kandel and D. J. Yamashiro, “Potent VEGF Blockade Causes Regression of Coopted Vessels in a Model of Neuroblastoma,” Proceedings of the National Academy of Sciences of the United States of America, Vol. 99, No. 17, 2002, pp. 11399-11404. doi:10.1073/pnas.172398399
P. S. Steeg, “Angiogenesis Inhibitors: Motivators of Metastasis?” Nature Medicine, Vol. 9, No. 7, 2003, pp. 822-823. doi:10.1038/nm0703-822
G. P. Pidgeon, M. P. Barr, J. H. Harmey, D. A. Foley and D. J. Bouchier-Hayes, “Vascular Endothelial Growth Factor (VEGF) Upregulates BCL-2 and Inhibits Apoptosis in Human and Murine Mammary Adenocarcinoma Cells,” British Journal of Cancer, Vol. 85, No. 2, 2001, pp. 273-278. doi:10.1054/bjoc.2001.1876
L. M. Ellis, “Epidermal Growth Factor Receptor in Tumor Angiogenesis,” Hematology/Oncology Clinics of North America, Vol. 18, No. 5, 2004, pp. 1007-1021. doi:10.1016/j.hoc.2004.06.002
P. L. Penar, S. Khoshyomn, A. Bhushan and T. R. Tritton, “Inhibition of Epidermal Growth Factor Receptor-Associated Tyrosine Kinase Blocks Glioblastoma Invasion of the Brain,” Neurosurgery, Vol. 40, No. 1, 1997, pp. 141-151.
K. Lamszus, M. A. Brockmann, C. Eckerich, P. Bohlen, C. May, U. Mangold, R. Fillbrandt and M. Westphal, “Inhibition of Glioblastoma Angiogenesis and Invasion by Combined Treatments Directed against Vascular Endothelial Growth Factor Receptor-2, Epidermal Growth Factor Receptor, and Vascular Endothelial-Cadherin,” Clinical Cancer Research, Vol. 11, No. 13, 2005, pp. 4934-4940. doi:10.1158/1078-0432.CCR-04-2270
H. D. Hemmati, I. Nakano, J. A. Lazareff, M. Masterman-Smith, D. H. Geschwind, M. Bronner-Fraser and H. I. Kornblum, “Cancerous Stem Cells Can Arise from Pediatric Brain Tumors,” Proceedings of the National Academy of Sciences of the United States of America, Vol. 100, No. 25, 2003, pp. 15178-15183. doi:10.1073/pnas.2036535100
S. K. Singh, I. D. Clarke, M. Terasaki, V. E. Bonn, C. Hawkins, J. Squire and P. B. Dirks, “Identification of a Cancer Stem Cell in Human Brain Tumors,” Cancer Research, Vol. 63, No. 18, 2003, pp. 5821-5828.
C. Calabrese, H. Poppleton, M. Kocak, T. L. Hogg, C. Fuller, B. Hamner, E. Y. Oh, M. W. Gaber, D. Finklestein, M. Allen, A. Frank, I. T. Bayazitov, S. S. Zakharenko, A. Gajjar, A. Davidoff and R. J. Gilbertson, “A Perivascular Niche for Brain Tumor Stem Cells,” Cancer Cell, Vol. 11, No. 1, 2007, pp. 69-82. doi:10.1016/j.ccr.2006.11.020
A. Diaz Miqueli, R. Blanco, B. Garcia, T. Badia, A. E. Batista, R. Alonso and E. Montero, “Biological Activity in Vitro of Anti-Epidermal Growth Factor Receptor Monoclonal Antibodies with Different Affinities,” Hybridoma (Larchmt), Vol. 26, No. 6, 2007, pp. 423-431. doi:10.1089/hyb.2007.0516
Y. Akashi, I. Okamoto, T. Iwasa, T. Yoshida, M. Suzuki, E. Hatashita, Y. Yamada, T. Satoh, M. Fukuoka, K. Ono and K. Nakagawa, “Enhancement of the Antitumor Activity of Ionising Radiation by Nimotuzumab, a Humanised Monoclonal Antibody to the Epidermal Growth Factor Receptor, in Non-Small Cell Lung Cancer Cell Lines of Differing Epidermal Growth Factor Receptor Status,” British Journal of Cancer, Vol. 98, No. 4, 2008, pp. 749-755. doi:10.1038/sj.bjc.6604222
M. O. Rodriguez, T. C. Rivero, R. C. Bahi, C. R. Muchuli, M. A. Bilbao, E. N. Vinageras, J. Alert, J. J. Galainena, E. Rodriguez, E. Gracias, B. Mulen, B. Wilkinson, E. L. de Armas, K. Perez, I. Pineda, M. Frometa, I. Leonard, V. Mullens, C. Viada, P. Luaces, O. Torres, N. Iznaga and T. Crombet, “Nimotuzumab plus Radiotherapy for Unresectable Squamous-Cell Carcinoma of the Head and Neck,” Cancer Biology & Therapy, Vol. 9, No. 5, 2010, pp. 343-349. doi:10.4161/cbt.9.5.10981
B. J. Denny, R. T. Wheelhouse, M. F. Stevens, L. L. Tsang and J. A. Slack, “NMR and Molecular Modeling Investigation of the Mechanism of Activation of the Antitumor Drug Temozolomide and Its Interaction with DNA,” Biochemistry, Vol. 33, No. 31, 1994, pp. 9045-9051. doi:10.1021/bi00197a003
S. D’Atri, L. Tentori, P. M. Lacal, G. Graziani, E. Pagani, E. Benincasa, G. Zambruno, E. Bonmassar and J. Jiricny, “Involvement of the Mismatch Repair System in temozolomide-Induced Apoptosis,” Molecular Pharmacology, Vol. 54, No. 2, 1998, pp. 334-341.
M. E. Hegi, A. C. Diserens, T. Gorlia, M. F. Hamou, N. de Tribolet, M. Weller, J. M. Kros, J. A. Hainfellner, W. Mason, L. Mariani, J. E. Bromberg, P. Hau, R. O. Mirimanoff, J. G. Cairncross, R. C. Janzer and R. Stupp, “MG- MT Gene Silencing and Benefit from Temozolomide in Glioblastoma,” The New England Journal of Medicine, Vol. 352, No. 10, 2005, pp. 997-1003. doi:10.1056/NEJMoa043331
M. Ramos-Suzarte, N. Rodriguez, J. P. Oliva, N. Iznaga-Escobar, A. Perera, A. Morales, N. Gonzalez, M. Cordero, L. Torres, G. Pimentel, M. Borron, J. Gonzalez, O. Torres, T. Rodriguez and R. Perez, “99mTc-Labeled Antihuman Epidermal Growth Factor Receptor Antibody in Patients with Tumors of Epithelial Origin: Part III. Clinical Trials Safety and Diagnostic Efficacy,” Journal of Nuclear Medicine, Vol. 40, No. 5, 1999, pp. 768-775.
W. K. Boland and G. Bebb, “Nimotuzumab: A Novel Anti-EGFR Monoclonal Antibody That Retains Anti-EGFR Activity While Minimizing Skin Toxicity,” Expert Opinion on Biological Therapy, Vol. 9, No. 9, 2009, pp. 1199-1206. doi:10.1517/14712590903110709
G. Bebb, W. Boland and B. Melosky, “Don’t Jump to Rash Conclusions,” Cancer Biology & Therapy, Vol. 11, No. 7, 2011, pp. 639-641. doi:10.4161/cbt.11.7.14920