J. Silvio Gutkind, Ph.D., Chief
Cancer of the head and neck area is the sixth most common neoplastic disease in the developed world, representing perhaps one the most serious health problem of this part of the body based on annual morbidity and mortality rates. This form of cancer share many common traits with those from other tissues. However, a number of issues pertaining to diagnosis, etiology, tumor promotion, genetic and environmental risk factors are unique and distinct, thus meriting a high-impact research effort in this area. The molecular and etiological factors involved in the development of head and neck tumors are largely unknown.
The Branch conducts basic, translational, clinical and epidemiological studies into the etiology, treatment and prevention of oral cancers and premalignant lesions. The Branch conducts basic studies in those traits common to all cancers as well as those unique to cancers of the head and neck. This information can be utilized to develop successful strategies to diagnose, treat and prevent cancers in the oral cavity and the head and neck area.
General Program Statement
OPCB scientists work on several complementary aspects of cancer cell biology in an effort to understand the molecular basis for malignant transformation, as well as to use this knowledge to develop molecular markers of disease progression and novel therapeutic approaches for oral malignancies. OPCB has strategically initiated programs and recruited staff in areas which are relevant to oral cancer biology, including receptor-mediated mitogenic signaling, cell cycle regulation and tumor suppressor gene biology, human papillomavirus research, animal and cellular models for squamous cell carcinogenesis, mechanisms controlling programmed cell death, molecular diagnostics and evaluation of molecular biomarkers for squamous cell carcinogenesis. These approaches will afford a better understanding of normal epithelial cell biology, and will likely broaden the horizon in developing oral tumor markers and treatments. In addition, we have established a drug evaluation program, and plan to initiate clinical studies aimed at identifying novel strategies for treatment of head and neck cancers, partnering with other intra- and extramural research teams and private organizations. A number of research efforts at the OPCB are described below.
Examples of Branch Research Projects
Molecular mechanisms controlling normal and aberrant cell growth
There are many complex signaling steps between the initial triggering of cell surface receptors for mitogens and the final biological response. As subtle alterations in molecules involved in proliferative pathways can lead to unregulated cell growth, including cancer, it is believed that the complete elucidation of the mechanisms controlling cell proliferation and DNA-synthesis is central to understanding, and thus controlling, malignancy. Members of OPCB have focused on the basic molecular mechanisms underpinning communication between the membrane and the nucleus. This effort, together with those of other laboratories, has led to the identification of multiple kinase cascades connecting different types of cell surface receptors to key molecules, including nuclear transcription factors. Ongoing and future studies are expected to help elucidate how signals beginning at the level of the cell surface (growth factor receptors, antiproliferative receptors, cell-cell and cell-matrix contacts, environmental signals, etc.) are integrated in space and time to control the orderly progression through the cell cycle. This knowledge will ultimately be useful for understanding normal and aberrant cell growth, and for identifying novel targets for therapeutic intervention in neoplastic disease.
Molecular mechanisms involved in squamous cell carcinogenesis
Carcinogenesis is a multistep process involving the activation of cellular protooncogenes and inactivation of tumor suppressor genes. In spite of the high frequency of oral cancers, little is known about the molecular events which contribute to disease initiation and progression. Recently, it has been found that 40-50% of squamous cell carcinomas harbor inactivating mutations in the p53 tumor suppressor gene. However, there is limited information on tumor-causing genes expressed in oral cancers. Current efforts to identify oncogenic sequences expressed in squamous carcinoma cells will contribute to the understanding of the basic mechanisms of neoplastic conversion in oral cancer, and will provide useful new biomarkers of disease progression.
Development of animal models for squamous carcinogenesis
One of the major limitations in the area of oral cancer research is the lack of suitable animal models to test the validity of current genetic models of tumorigenesis, and to explore the effectiveness of treatment modalities or chemopreventive approaches. The final goal of this project is the development of oral models (for example, rat oral squamous cell carcinoma models, transgenic mice targeting oncogenes to oral mucosa, HPV transgenics treated with oral carcinogens, animal and human oral cell lines) to generate tumor tissues at various stages of neoplastic progression for evaluating marker expression, biochemical changes, and treatment strategies. This may allow us to recapitulate in animal models all the molecular events believed to lead to squamous cell carcinomas in humans, and will facilitate the search for alternative oral cancer treatments.
Oral cancers result from progressive genetic changes leading to malignancy in a multistep process. If molecular markers representing early and late events can be defined, it would be possible to identify persons at high risk of oral cancer, namely, those with lesions progressing through the premalignant state. Such biochemical markers heralding malignancy would facilitate the monitoring of recurrence and expedite the evaluation of chemopreventing agents, a number of which are ready for testing in clinical trials. Current efforts to identify molecular and cellular markers of oral tumorigenesis focus on the analyzing markers of proliferation, squamous differentiation, and genetic changes in genes believed to play a role in the carcinogenic process. Additional approaches involve the study of molecules whose levels of expression change during tumor progression, using cellular RNAs derived from normal, dysplastic, and malignant keratinocytes. The identity of the differentially expressed transcripts will then be established using the nucleotide information generated by the Human Genome Project and by private efforts. This work is expected to help identify gene products that determine the transformed and/or the metastatic phenotype, as well as additional molecules that, without playing an obvious role in the neoplastic process, can nevertheless be used as clinically useful markers of squamous carcinogenesis.
Development of new therapeutic approaches
The ultimate goal of this project is to develop novel therapies aimed at improving the quality of life and life expectancy of oral cancer patients. The recent use of high throughput screening procedures followed by computer assisted molecular modeling and chemical synthesis of lead compounds has tremendously accelerated the discovery of small molecule inhibitors of signaling and cell cycle controlling proteins. Several such drug candidates are now becoming available. These drugs will be evaluated for activity, efficacy and toxicity in biologically relevant in vitro and in vivo models of squamous cell carcinogenesis. Cell systems and animal models described above are expected to be of significant importance for this drug evaluation, aimed at the launching of a clinical oral cancer program. The use of gene transfer approaches to prevent neoplastic conversion of premalignant lesions and to treat oral cancer will also be explored, with emphasis on the evaluation of new genetically engineered viruses and viral vectors for their ability to express genes in oral tissues and for their therapeutic potential in squamous cell carcinoma.
Gutkind J.S., Novotny E., Brann M.R., and Robbins K.C. Muscarinic acetylcholine receptor subtypes as agonist dependent oncogenes. Proc. Natl. Acad. Sci. USA, 88:4703-4707, 1991.
Crespo P., Xu N., Simonds W.F., and Gutkind J.S. Ras-dependent activation of MAP kinase pathway mediated by G-protein bg subunits. Nature, 369:418-420, 1994.
Coso O., Chiariello M., Yu J.-C., Crespo P., Teramoto, H., Xu N., Miki T., and Gutkind J.S. The small GTP-binding proteins Rac1 and Cdc42 regulate the activity of the JNK (SAPK) signaling pathway. Cell, 81:1137-1148, 1995.
Lopez-Ilasaca M., Crespo P., Pellici P.G., Gutkind J.S.*, and Wetzker R. Linkage of G protein-coupled receptors to the MAPK signaling pathway through PI 3-kinase g. Science, 275:394-397, 1997.
Park MH, Lee YB and Joe YA (1997) Hypusine is essential for eukaryotic cell proliferation. Biol. Signals 6: 115-123
Joe YA, Wolff EC, Lee YB and Park MH (1997) Enzyme-substrate intermediate at a specific lysine residue is required for deoxyhypusine synthesis: the role of Lys 329 in human deoxyhypusine synthase J. Biol. Chem. 272: 32679-326785
Crespo P., Schuebel K.E., Ostrom A.A., Gutkind J.S., and Bustelo X.R. Phosphotyrosine-dependent activation of Rac-1 GDP/GTP exchange by the vav proto-oncogene product. Nature, 385:169-172, 1997.
Fromm C., Coso O.A., Montaner S., Xu N., and Gutkind J.S. The small GTP-binding protein Rho links G protein-coupled receptors and Ga12 to the serum response element and to cellular transformation. Proc. Natl. Acad. Sci. U.S.A., 94:10098-10103, 1997.
Park MH, Joe YA and Kang KR (1998) Deoxyhypusine synthase activity is essential for cell viability in the yeast Saccharomyces cerevisiae J. Biol. Chem. 273: 1677-1683
Patel V., Senderowicz A.M., Pinto D., Ensley J., Igishi T., Sausville E.A., and Gutkind J.S. Flavopiridol, a novel CDK inhibitor, suppresses the growth of squamous cell carcinomas by inducing apoptosis. J. Clin. Invest., 102:1674-1681, 1998.
Leethanakul C., Patel V., Gillespie J., Shillitoe E., Kellman R.M., Ensley J.E., Limwongse V., Emmert-Buck M.R., Krizman D.V., and Gutkind J.S. Gene expression profiles in squamous cell Carcinomas of the oral cavity: Use of Laser Capture Microdissection for the construction and analysis of stage-specific cDNA libraries. Oral Oncology, 36:474-483, 2000.
Leethanakul C., Patel V., Gillespie J., Pallante M., Ensley J.F., Liotta L.A., Emmert-Buck M., and Gutkind J.S. Distinct pattern of expression of differentiation and growth-related genes in squamous cell carcinomas of the head and neck revealed by the use of laser capture microdissection and cDNA arrays. Oncogene, 19:3220-3224, 2000.
Wolff EC, Wolff J and Park MH (2000): Deoxyhypusine synthase generates and uses bound NADH in a transient hydride transfer mechanism. J. Biol. Chem. 275, 9170-9177
Chiariello M., Marinissen M.J., and Gutkind J.S. Regulation of c-myc expression by PDGF through Rho GTPases. Nature Cell Biol., 3:580-586, 2001.
Clement PMJ, Hanauske-Abel HM, Wolff EC, Kleinman HK and Park MH (2002) The antifungal drug ciclopirox inhibits deoxyhypusine and proline hydroxylation, endothelial cell growth and angiogenesis in vitro. Int. J. Cancer 100, 491-498
Patel V., Lahusen T., Leethanakul C., Igishi T., Kremer M., Quintanilla-Martinez L., Ensley J.F., Sausville E.A., Gutkind J.S., and Senderowicz A.M. Antitumor activity of UCN-01 in carcinomas of the head and neck is associated with altered expression of cyclin D3 and p27KIP1. Clin. Cancer Res. 8:3549-3560, 2002.
Sriuranpong V., Park J.I., Amornphimoltham P., Patel P., Nelkin B.D., and Gutkind J.S. EGFR-independent constitutive activation of STAT3 in head and neck squamous cell carcinoma is mediated by the autocrine/paracrine stimulation of the IL6/gp130 cytokine system. Cancer Research, 63:2948-2956, 2003.
Engelholm, L. H., List, K., Netzel-Arnett, S., Cukierman, E., Mitola, D. J., Aaronson, H., Kjøller, L., Larsen, J. K., Yamada, K. M., Strickland, D. S., Holmback, K., Danø, K., Birkedal-Hansen, H., Behrendt, N., and Bugge T. H. uPARAP/Endo180 is essential for cellular uptake of collagen and promotes fibroblast collagen adhesion. J. Cell Biol., 160: 1009-1015, 2003.
Montaner S., Sodhi A., Molinolo A., Bugge T.H., Sawai E.T., He Y., Li Y., Ray P.E., and Gutkind J.S. Endothelial infection with KSHV genes in vivo reveals that vGPCR initiates Kaposi's sarcomagenesis and can promote the tumorigenic potential of viral latent genes. Cancer Cell, 3:23-36, 2003.
List, K., Szabo, R., Wertz, P.W., Segre, J., Haudenschild, C. C., Kim, S.-Y., and Bugge, T. H. Loss of proteolytically processed filaggrin caused by epidermal deletion of Matriptase/MT-SP1. J. Cell Biol., 163; 901-910, 2003.
Liu, S., Aaronson, H., Mitola, D., Leppla, S. H., and Bugge, T. H. Potent anti-tumor activity of a urokinase-activated engineered anthrax toxin. Proc. Natl. Acad. Sci. (USA), 100: 657-662, 2003.
Park J-H, Wolff EC, Folk JE and Park MH (2003) Reversal of the deoxyhypusine synthesis reaction: Generation of spermidine or homospermidine from deoxyhypusine by deoxyhypusine synthase. J. Biol. Chem. 278, 32683-32691
Jeon GA, Lee J-S, Patel V, Gutkind JS, Thorgeirsson S, Kim EC, Chu I-S and Park MH (2004) Global gene expression profile of human head and neck squamous carcinoma cell lines. Int. J. Cancer 112, 249-258
Umland TC, Wolff EC, Park MH and Davies DR (2004) A new crystal structure of deoxyhypusine synthase reveals the configuration of the active site enzyme and of an enzyme:NAD:inhibitor ternary complex. J. Biol. Chem. 279, 28697-28705
Hobson, J. P., Netzel-Arnett, S., Szabo, R., Rehault, S. M., Church F. C., Strickland, D. K., Lawrence, D. A., Antalis, T. M. and Bugge, T. H. Mouse DESC1 is located within a cluster of seven DESC1-like genes and encodes a type II transmembrane serine protease that forms serpin inhibitory complexes. J. Biol. Chem., 279: 46981-46994, 2004.
Amornphimoltham P., Sriuranpong V., Patel V., Benavides F., Conti C.J., Sauk J., Sausville E.A., Molinolo A.A., and Gutkind J.S., Persistent activation of the Akt pathway in head and neck squamous cell carcinoma: a potential target for UCN-01, Clin. Cancer Research, 10:4029-37, 2004.
Marinissen M.J., Chiariello M., Tanos T., Bernard O., Narumiya S., and Gutkind J.S. The small GTP-binding protein RhoA regulates c-Jun by a ROCK-JNK signaling axis, Mol. Cell, 14:29-41, 2004.
Sriuranpong V., Mutirangura A., Gillespie J.W., Patel V., Amornphimoltham P., Molinolo A.A., Kerekhanjanarong V., Supanakorn S., Supiyaphun P., Rangdaeng S., Voravud N., and Gutkind J.S. Global gene expression profile of nasopharyngeal carcinoma by laser capture microdissection and cDNA microarrays. Clin. Cancer Research, 10:4944-58, 2004.
Vitale-Cross L., Amornphimoltham P., Fisher G., Molinolo A.A., and Gutkind J.S. Conditional expression of K-ras in an epithelial compartment that includes the stem-cells is sufficient to promote squamous cell carcinogenesis. Cancer Res., 64:8804-8807, 2004.
Castellone M.D., Teramoto H., Williams B.O., Druey K.M., and Gutkind J.S. Prostaglandin E2 promotes colon cancer cell growth through a novel Gs-axin-b-catenin signaling axis. Science, 310:1504-1510, 2005.
Szabo, R., Netzel-Arnett, S., Hobson, J. P., Antalis, T. M., and Bugge, T. H. Matriptase-3 is a novel phylogenetically preserved membrane-anchored serine protease with broad serpin reactivity. Biochemistry: 390, 231-242.
Baker H., Patel V., Molinolo A.A., Myers J.N., El-Naggar A.K., Gutkind J.S., and Hancock W.S. Proteome-wide analysis of head and neck squamous cell carcinomas using laser-capture microdissection and tandem mass-spectrometry, Oral Oncol., 41:183-199, 2005.
Amornphimoltham P., Patel V., Sodhi A., Nikitakis N.G., Sauk J.J., Sausville E.A., Molinolo A.A., and Gutkind J.S. mTOR, a molecular target in squamous cell carcinomas of the head and neck. Cancer Research, 65:9953-9961, 2005.
List, K., Szabo, R., Molinolo, A., Sriuranpong, V., Redeye, V., Burke B., Murdock, T., Nielsen, B.S., Gutkind, J.S., and Bugge, T. H. Deregulated matriptase causes ras-independent multistage carcinogenesis and promotes ras-mediated malignant transformation. Genes & Dev., 19: 1934-1950, 2005.
Curino, A. C., Nielsen, B. S., Engelholm, L. H., Molinolo, A. A., Behrendt, N., and Bugge, T. H. Intracellular collagen degradation mediated by uPARAP/Endo180 is a major pathway of extracellular matrix turnover during malignancy. J. Cell Biol. 169:977-986, 2005.
Liu, S., Redeye, V., Kuremsky, J. G., Bugge, T. H., and Leppla, S. H. Tumor targeting by anthrax toxin proteins that use intermolecular complementation to require simultaneous activation by two tumor-enriched cell-surface proteases. Nature Biotechnology, 23: 725-730, 2005.
Park, J-H, Aravind, L, Wolff, EC, Kaevel, J, Kim, YS and Park MH (2006) Molecular cloning, expression and structural prediction of deoxyhypusine hydroxylase: a novel HEAT-repeat-containing metalloenzyme, Proc. Natl. Acad. Sci. USA, 103, 51-56.