Intrinsic Apoptosis Phenotype and Susceptibility to Squamous Cell Carcinoma of the Oral Cavity
Co-Principal Investigator(s): Qingyi Wei, M.D., Ph.D.; Erich M. Sturgis, M.D.; Randal S. Weber, M.D.
Only a fraction of smokers and drinkers develop squamous cell carcinoma of the oral cavity (SCCOC), suggesting that genetic susceptibility plays a role in the etiology of SCCOC. Our previous studies have established that host-cell DNA repair capacity and related genetic variations, as measured by polymorphisms, are susceptibility factors for SCCOC. To date, no published studies have investigated apoptotic capacity as a susceptibility factor for SCCOC. There are at least two known apoptotic pathways, intrinsic and extrinsic, that lead to cell death. While the extrinsic pathway of apoptosis is primarily important in the immunological mechanisms of antigen-induced cell death, the intrinsic or mitochondrial pathway of apoptosis is activated in response to unrepaired DNA damage. Therefore, we hypothesize that genetically determined capacity of the intrinsic apoptotic pathway is associated with risk of SCCOC.
In this new proposal, we have the following specific aims:
- To establish a comprehensive database for 600 prospectively accrued (including 300 recruited in the previous SPORE project), newly diagnosed SCCOC cases and 600 hospital-based and genetically unrelated controls (including 300 recruited in an R01-supported project), frequency-matched by age, sex, ethnicity/race and county of residence.
- To determine differences in phenotypes of the established intrinsic apoptosis capacity between 300 cases and 300 controls. We will test the hypothesis that CPT-induced apoptotic capacity is lower in SCCOC cases than in controls and is modified by known risk factors such as smoking and alcohol use.
- To determine the associations between variant apoptotic genotypes and risk of SCCOC in the 600 cases and 600 controls that will have genotyping data for the case-control analysis, which will generate new hypotheses for further validation by future larger studies and the INHANCE group.
- To study the correlations between apoptotic phenotypes and functional polymorphisms (genetic variants) in 300 cases and 300 controls. We will test the hypothesis that intrinsic apoptotic capacity is correlated with (predicted by) common functional genetic variants of selected apoptotic genes in 300 cases and 300 controls that will have apoptotic phenotype data.
Our long-term goal is to identify effective biomarkers for risk assessment and at-risk individuals who can be targeted for primary prevention and early detection of SCCOC.
Cancer Risk Assessment in Patients with Oral Premalignant Lesions: An Integrative Approach
Co-Principal Investigator(s): Xifeng Wu, M.D., Ph.D.; Scott M. Lippman, M.D.
This proposal is built upon the epidemiologic and molecular genetic findings from our 20-year experience in translational research in oral premalignant lesions (OPLs), a hallmark of oral cancer development. Although the etiology of OPLs is not fully understood, these lesions are often associated with tobacco and alcohol exposures; however, it is unclear why only a fraction of exposed individuals with OPL subsequently develop oral cancer. We have previously shown that OPL patients with certain genetic backgrounds are more susceptible to developing oral cancer, and that certain molecular abnormalities that present in OPL target tissues predict a higher rate of malignant transformation of these lesions, independent of clinicopathologic parameters. These predictive markers include OPL histology and the combined biomarker scores of chromosomal polysomy, p53 protein expression and loss of heterozygosity (LOH) at 3p and 9p in OPL tissues. We have demonstrated a successful strategy for comprehensive cancer risk assessment in OPL patients by combining conventional medical/ demographic variables and a panel of biomarkers; however, all of these studies have utilized small sample sizes. Therefore, we propose to extend our studies by utilizing the largest available resource of OPL specimens that allows us the unprecedented opportunity to study 350 patients with OPLs, prospectively followed, to parallel our investigation of genetic markers in surrogate and target tissues, critical molecular candidates and global genetic aberrations and expression profiles to identify predictors of oral cancer risk in these patients. Using this unique resource, we will integrate clinical, epidemiologic and genetic data to build a quantitative risk assessment model for oral cancer development.
Our Specific Aims include:
- To assess susceptibility markers in surrogate tissue (lymphocytes).
- To assess global genomic and transcriptomic abnormalities as well as critical candidate molecules in OPL target tissues.
- To complete comprehensive analyses of data from clinical, epidemiologic, surrogate-target tissue, genotype-phenotype, global-candidate biomarker panels with patients’ clinical outcomes to build a risk assessment model for oral cancer development.
The identification of subgroups of OPL patients at higher risk for oral cancer development will allow the development of individually tailored, mechanism-based chemopreventive interventions and strategies for more effective screening and treatment.
Predictors of Resistance to Dual VEGFR / EGFR Targeted Therapy of Head and Neck Cancer
Co-Principal Investigator(s): Gordon Mills, M.D., Ph.D.; Jeffrey N. Myers, M.D., Ph.D.; Vassiliki A. Papadimitrakopoulou, M.D.; John V. Heymach, M.D., Ph.D.
Epidermal growth factor receptor (EGFR)-targeted therapy in combination with radiation has been demonstrated to improve locoregional control and survival of patients with head and neck squamous cell carcinoma (HNSCC) in a randomized phase III clinical trial. Based on the encouraging pre-clinical studies of dual EGFR/vascular endothelial growth factor receptor (VEGFR) inhibition explored during the first five years of this SPORE, we are now investigating EGFR-VEGFR co-targeting using ZD6474 and radiation in the same clinical setting. Although this approach carries an enormous therapeutic potential, it is very likely that resistance to inhibition of these pathways will emerge as a potential obstacle to be overcome in clinical practice. The main goal of this Project 3 is to identify the potential pathways of resistance to ZD6474 employing high throughput methodology, consisting of tissue proteomics (reverse phase protein arrays [RPPA], and phosphorylated receptor tyrosine kinase arrays [p-RTK]) and cytokine and angiogenic factors (CAF) profiling.
We will apply these three methods in both the pre-clinical (in vitro and in vivo models) and clinical settings (archival and prospectively collected samples from an ongoing trial of ZD6474 plus chemoradiation in patients with locoregionally advanced HNSCC) through the following specific aims:
- To determine the sensitivity of HNSCC cell lines to ZD6474 and identify signatures of in vitro resistance to the drug.
- To determine in vivo resistance signatures to the drug using orthotopic xenograft models subjected to treatment with ZD6474 +/- radiation.
- To identify signatures predictive of poor outcome in archival samples obtained from three cohorts of patients and develop a predictive model combining molecular signature information from pre-clinical aims and archival specimens. This overall predictive model will be prospectively evaluated in the context of the ongoing study of ZD6474 plus chemoradiation.
This strategy will allow us to:
- Identify potential pathways of resistance to be targeted in future (pre)clinical studies.
- Validate pre-clinical models for identifying resistance to targeted agents.
- Develop high-throughput technology-based predictive models that will reflect both prognosis and resistance to dual EGFR and VEGFR inhibition and can inform the design of future therapeutic strategies.
Targeting EGFR and the IGF Axis for Therapy of Head and Neck Squamous Cell Carcinoma
Co-Principal Investigator(s): Ho-Young Lee, Ph.D.; Bonnie S. Glisson, M.D.
Signaling through the insulin-like growth factor 1 receptor (IGF-1R) is increasingly recognized as a precursor to and a potentiator of the malignant phenotype in preclinical models. The axis of proteins involved is complex, including two ligands, six binding proteins, IGF-1R and IGF-2R, a non-signaling receptor, and two adaptor proteins. Circulating levels or tumor expression of IGF axis proteins has been correlated with risk of malignancy or poor prognosis of established cancer. IGF-1R interacts with other cellular receptors including epidermal growth factor receptor (EGFR), a validated drug target in HNSCC. We have observed that activation of IGF-1R signaling in cell lines promotes proliferation, survival, angiogenesis and invasion. Further, our data indicate that activation of IGF-1R can mediate resistance to EGF-R-targeted therapy and vice versa, suggesting an advantage to co-targeting EGFR and IGF-1R. Thus, we hypothesize that co-targeting IGF-1R and EGFR will be more successful than inhibition of either pathway alone in therapy of HNSCC. In this project, we will assess IGF axis protein expression and downstream proteins in tumor microarrays from patients resected for SCC of the oral tongue (Aim 1). Correlation of biomarkers with disease-specific survival will be explored, especially as regards the negative impact of nodal spread and extracapsular invasion, an invasive phenotype. We will study cooperation of IGF-1R and EGFR in mediating proliferative, angiogenic and invasive activities of HNSCC lines in vitro (Aim 2). Antitumor effects of the anti-IGF-1R humanized monoclonal antibody (IMC-A12) will be studied alone or with anti-EGFR antibody cetuximab, using multiple complementary models, including a highly metastatic cell line, in orthotopic tongue xenograft models (Aim 3). The activity of IMC-A12 as monotherapy or combined with cetuximab will be defined in a randomized trial for patients with recurrent HNSCC (Aim 4), targeting a 50% improvement in progression-free survival compared to cetuximab alone. Tissue and blood-based biomarkers will be assessed and correlations with clinical outcomes explored. This proposal builds on our results implicating IGF axis signaling in potentiating the malignant phenotype of HNSCC and promising data co-targeting EGFR and IGF-1R in therapy of HNSCC. Insights to be gained from this work, both preclinical and clinical aims, have the potential to facilitate targeting therapy to those patients most likely to benefit.
Co-Leader(s): Scott M. Lippman, M.D.; Jeffrey N. Myers, M.D., Ph.D.; Xifeng Wu, M.D., Ph.D.
Core A provides oversight and support of all SPORE projects and activities; ensures compliance with all general, governmental and National Cancer Institute (NCI) / National Institutes of Health (NIH) regulations and requirements; coordinates communications and consultations with the NCI program official and other NCI staff in preparing all required reports and publications; coordinates all SPORE-related meetings and communications of the Executive Committee, Internal and External Advisory Boards, the Biostatistics and Data Management Core (Core B) and the Tissue Procurement, Pathology and Biomarker Core (Core C); and other key functions. Core A oversees the annual budget/rebudget development, distribution and oversight of grant funds; supports the Executive Committee and advisory boards in scheduling and coordinating regularly scheduled meetings and ad hoc consultations; coordinates the annual solicitation, review and award process for the Career Development and the Developmental Research Programs; serves as the primary liaison in developing potential InterSPORE collaborations and with the NCI SPORE program representatives; and interacts on a regular basis with Core directors and project leaders to review progress, identify problems and resolve issues hampering successful achievement of their aims.
Through the Administrative Core, the SPORE leadership oversees the activation and accrual of our SPORE clinical trials, including compliance with requirements, quarterly accrual report submissions to the Executive Committee (and more frequently should concerns arise), quarterly presentations by the PIs of the SPORE trials to the Executive Committee on accrual progress, annual review of all trials by the Internal and External Advisory Boards and a requirement to address accrual deficiencies within six months or risk study closure or replacement.
Biostatistics and Data Management Core
Leader: J. Jack Lee, Ph.D.
Based on a strong track record of providing biostatistical support and data management infrastructure for translational research, the Biostatistics and Data Management Core (BDMC) is a comprehensive, multi-lateral resource for acquiring, managing and integrating data, designing clinical and basic science experiments, performing statistical analysis, building comprehensive risk assessment models, developing innovative statistical methodology and publishing the research results generated from this SPORE. BDMC has significantly enhanced the database infrastructure for the MD Anderson Head / Neck Tissue Bank and the Oracle Clinical Database for two clinical trials. We have developed and posted on-line the first common data element (CDE) for the head and neck tissue bank. Based on the CDEs, we built a comprehensive, Web-based data management system for the tissue bank that now stores over 338,000 tissue samples (fresh, paraffin, slide or fluid samples) from more than 29,000 patients. The Tissue Bank Data Management System also provides extensive query and reporting capabilities. In addition, in collaboration with staff at NCI, we have developed two clinical trial data management systems, based on Oracle Clinical, which allows for remote data capture and is ideal for expanding for inter-SPORE collaborations. We will continue to ensure the incorporation of sound experimental design principles within each project that will increase the clarity and enhance interpretability of study results. Each project will be provided with tailored analyses, accompanied by novel statistical development as necessary, to reveal apparent and hidden relationships among data. The BDMC will provide expertise in the design of an integrated data management system to facilitate communication among all projects and cores. This process includes prospective data collection, data quality control, data security, data management, data integration and assurance of patient confidentiality. The BDMC will continue to provide statistical support for the entire Head and Neck SPORE, including design of in vitro and in vivo experiments, clinical trials, sample size/power calculations, statistical data analysis, preparation of statistical reports for clinical trials and collaborating in the development of related publications.
Leader: Adel K. El-Naggar, M.D., Ph.D.
The Tissue Procurement, Pathology and Biomarkers Core (Core C) provides investigators at MD Anderson Cancer Center and other collaborating institutions with high-quality tissue samples from patients with head and neck cancer and oral premalignancies. The Head and Neck Tissue Bank, directed by Core director, Dr. Adel El-Naggar, now contains more than 338,000 specimens from more than 29,000 patients. Standardized and centralized procedures have been established for tissue procurement, quality control, processing and storage, as well as distribution of samples to individual investigators. In addition, Core C provides centralized, specialized services such as tissue microarray (TMA) construction, tissue microdissection, qualitative and quantitative biomarker analyses both in tissue [e.g., immunohistochemistry (IHC), Western blotting] and non-tissue (e.g., blood-based cytokine profiles) specimens, nucleic acid extractions, development and maintenance of cell lines. These services and expertise provided by Core C were a critical component of the past SPORE research activities, leading to over 158 publications during the past five years, with 78 of these publications co-authored by Dr. El-Naggar. This Core has been at the forefront of multi-institutional efforts to standardize pathologic terminology and criteria for reporting and evaluating pathologic specimens and analyses, and developing and integrating common data elements of our Tissue Bank database with the NCI’s CaBIG™ Initiative.
Specific aims of the Core include:
- To obtain, process and maintain a repository of premalignant, tumorous and matching non-tumorous specimens and related specimens, and to distribute and track utilization of specimens.
- To maintain and expand innovative and unique tissue and blood-based resources and histopathologic services.
- To perform and interpret various tissue- and blood-based methodologies with rigorous quality control, maximize tissue utilization and ensure the successful completion of each project’s aims.
- To foster and support inter-SPORE and other NCI/NIH interactions and collaborations.
The Core will continue to ensure the highest quality tissue samples and related analyses will be provided to all HN SPORE investigators, as well as to collaborating researchers participating in other NIH/NCI-sponsored programs.