Carol G. Shores

Molecular classification of head and neck squamous cell carcinomas using patterns of gene expression.

The prognostication of head and neck squamous cell carcinoma (HNSCC) is largely based upon the tumor size and location and the presence of lymph node metastases. Here we show that gene expression patterns from 60 HNSCC samples assayed on cDNA microarrays allowed categorization of these tumors into four distinct subtypes. These subtypes showed statistically significant differences in recurrence-free survival and included a subtype with a possible EGFR-pathway signature, a mesenchymal-enriched subtype, a normal epithelium-like subtype, and a subtype with high levels of antioxidant enzymes. Supervised analyses to predict lymph node metastasis status were approximately 80% accurate when tumor subsite and pathological node status were considered simultaneously. This work represents an important step toward the identification of clinically significant biomarkers for HNSCC.

Molecular subtypes in head and neck cancer exhibit distinct patterns of chromosomal gain and loss of canonical cancer genes.

Head and neck squamous cell carcinoma (HNSCC) is a frequently fatal heterogeneous disease. Beyond the role of human papilloma virus (HPV), no validated molecular characterization of the disease has been established. Using an integrated genomic analysis and validation methodology we confirm four molecular classes of HNSCC (basal, mesenchymal, atypical, and classical) consistent with signatures established for squamous carcinoma of the lung, including deregulation of the KEAP1/NFE2L2 oxidative stress pathway, differential utilization of the lineage markers SOX2 and TP63, and preference for the oncogenes PIK3CA and EGFR. For potential clinical use the signatures are complimentary to classification by HPV infection status as well as the putative high risk marker CCND1 copy number gain. A molecular etiology for the subtypes is suggested by statistically significant chromosomal gains and losses and differential cell of origin expression patterns. Model systems representative of each of the four subtypes are also presented.

CANCER CACHEXIA SYNDROME IN HEAD AND NECK CANCER PATIENTS: PART I. DIAGNOSIS, IMPACT ON QUALITY OF LIFE AND SURVIVAL, AND TREATMENT

Cancer cachexia is a debilitating, wasting condition that affects many cancer patients, including those with head and neck cancer. The overall incidence of cancer cachexia is quite high for some types of cancer, and cachexia will be the main cause of death for more than 20% of all cancer patients. This syndrome uniquely challenges patients with head and neck cancer. This article outlines the diagnosis of cancer cachexia, reviews its impact on patient quality of life (QOL) and survival, and updates the reader on potential therapies that may suppress it.

ras Mutations and expression in head and neck squamous cell carcinomas

Mutational activation and overexpression of the family of ras proto‐oncogenes have been associated with many human tumors. The role of mutations of H‐ras, K‐ras, and N‐ras, as well as expression of the respective protein products (p21s) in normal mucosa, dysplastic mucosa, and squamous cell carcinomas (SCCs) of the head and neck has not been fully described. In our study, 51 tumors (40 paraffin embedded and 11 fresh frozen) were examined to determine if mutational activation of ras is an important molecular event in head and neck SCC. Analyses of codons 12, 13, and 61 of H‐ras, K‐ras, and N‐ras revealed no mutations, suggesting that mutational activation of ras is not important in the majority of head and neck SCCs.

Use, accuracy, and implications for patient management of [18F]-2-fluorodeoxyglucose-positron emission/computerized tomography for head and neck tumors.

Objectives/Hypothesis: Positron emission tomography (PET) has shown promise for early detection and accurate staging of cancer patients. A limited number of studies suggest PET/computed tomography (CT) may improve these variables; however, no published study has specifically evaluated clinical outcomes with PET/CT for head and neck (HN) tumors. The current study evaluates the use, accuracy, and implications for patient management of PET/CT scans in patients with HN tumors.

Results of a pilot study of the effects of celecoxib on cancer cachexia in patients with cancer of the head, neck, and gastrointestinal tract

Animal models suggest that cyclooxygenase‐2 (COX‐2) inhibitors may be beneficial in suppressing cancer cachexia. We investigated the effect of short‐course celecoxib on body composition, inflammation, and quality of life (QOL) in patients with cancer cachexia in a phase II clinical pilot trial.

BIIB021, a novel Hsp90 inhibitor, sensitizes head and neck squamous cell carcinoma to radiotherapy.

Heat shock protein 90 (Hsp90) is a molecular chaperone that promotes the conformational maturation of numerous client proteins, many of which play critical roles in tumor cell growth and survival. The ansamycin‐based Hsp90 inhibitor 17‐allylamino‐17‐demethoxygeldanamycin (17‐AAG) is currently in Phase III clinical testing. However, 17‐AAG is difficult to formulate and associated with dose‐limited toxicity issues. A fully synthetic and bioavailable Hsp90 inhibitor, BIIB021, was evaluated for antitumor activity in a variety of head and neck squamous cell carcinoma (HNSCC) cell lines and HNSCC xenograft models, either as a single agent or in combination with fractionated radiation and the results were compared with that of 17‐AAG. BIIB021 showed strong antitumor activity, comparable with, and in certain instances, superior to 17‐AAG. BIIB021 enhanced the in vitro radiosensitivity of HNSCCA cell lines with a corresponding reduction in the expression of key radioresponsive proteins, increased apoptotic cells and enhance G2 arrest. In xenograft studies, BIIB021 exhibited a strong antitumor effect outperforming 17‐AAG, either as a single agent and or in combination with radiation, thereby improved the efficacy of radiation. These results suggest that this synthetic and bioavailable Hsp90 inhibitor affects multiple pathways involved in tumor development and progression in the HNSCC setting and may represent a better strategy for the treatment of HNSCC patients, either as a monotherapy or a radiosensitizer. Furthermore, it also demonstrates the benefits of using preclinical models of chemosensitization to radiotherapy to explore clinically relevant radiation dosing schemes.

The effectiveness of salvage surgery after the failure of primary concomitant chemoradiation in head and neck cancer

OBJECTIVE: To determine survival outcomes and locoregional control rates in patients with locoregional head and neck squamous cell cancer (HNSCC) who failed primary concomitant chemoradiation (CRT) intended for cure and underwent attempted surgical salvage. STUDY DESIGN AND SETTING: Design was a nonrandomized retrospective cohort study. Of 204 patients with HNSCC who received primary concomitant chemoradiation intended for cure between 1995 and 2004, 38 recurred and underwent attempted salvage surgery at a tertiary care academic center. RESULTS: Among the 38 patients undergoing surgical salvage, 12- and 24-month overall survival rates were 60 percent and 27 percent. Locoregional control at 24 months was 42 percent. Lower survival was seen with initial N3 disease (P = 0.0115). Overall surgical morbidity was 24 percent. CONCLUSION/SIGNIFICANCE: The results of salvage surgery after failed chemoradiation for HNSCC are poor. Those with N3 disease fare least well. Patients should be well informed about the realistic chances of cure and potential morbidity of surgery.

Defining Cancer Cachexia in Head and Neck Squamous Cell Carcinoma

Purpose: Cancer cachexia is a devastating and understudied illness in patients with head and neck squamous cell carcinoma (HNSCC). The primary objective was to identify clinical characteristics and serum levels of cytokines and cachexia-related factors in patients with HNSCC. The secondary objective was to detect the occurrence of cytokine and cachexia-related factor gene expression in HNSCC tumors. Experimental Design: For the primary objective, cross-sectional data were obtained from prospectively recruited patients identified as cachexia cases and matching cachexia-free controls. For the secondary objective, a retrospective cohort design with matched controls was used. Results: Clinical characteristics associated with cancer cachexia in HNSCC were T4 status (P = 0.01), increased C-reactive protein (P = 0.01), and decreased hemoglobin (P < 0.01). Exploratory multiplex analysis of serum cytokine levels found increased interleukin (IL)-6 (P = 0.04). A highly sensitive ELISA confirmed the multiplex result for increased IL-6 in cachectic patients (P = 0.02). Quality of life was substantially reduced in patients with cachexia compared with noncachectic patients (P < 0.01). All tumors of HNSCC patients both with and without cachexia expressed RNA for each cytokine tested and the cachexia factor lipid-mobilizing factor. There were no statistically significant differences between the cytokine and cachexia factor RNA expression of cachectic and noncachectic patients (each P > 0.05). No tumors expressed the cachexia factor proteolysis-inducing factor. Conclusion: We have identified clinical characteristics and pathophysiologic mechanisms associated with cancer cachexia in a carefully defined population of patients with HNSCC. The data suggest that the acute-phase response and elevated IL-6 are associated with this complex disease state. We therefore hypothesize that IL-6 may represent an important therapeutic target for HNSCC patients with cancer cachexia.

Immunohistochemical Distinction of Follicular Thyroid Adenomas and Follicular Carcinomas

OBJECTIVES To use immunohistochemical (IHC) evaluation of proteins encoded by genes that were differentially expressed in follicular thyroid adenomas (FAs) vs follicular thyroid carcinomas (FTCs) to distinguish benign vs malignant follicular thyroid lesions. Multiple gene microarray studies suggest that benign and malignant follicular thyroid neoplasms have different gene expression profiles. DESIGN Immunohistochemical analysis of thyroid neoplasms, including FA (n = 62), FTC (n = 62), and follicular variant of papillary thyroid carcinoma (n = 58), using tissue microarrays. We evaluated antibodies galectin-3, autotaxin, intestinal trefoil factor 3 (TFF3), extracellular matrix metalloproteinase inducer (EMMPRIN), and growth arrest and DNA damage-inducible protein 153 (GADD153). We analyzed data for quantitative differences in IHC intensity and the percentage of positive cells between FAs and combined follicular carcinomas. Sensitivity and specificity analysis are reported, along with a dual-protein clinical algorithm. SETTING Academic tertiary care center. PATIENTS Adults with known follicular and papillary thyroid lesions that were surgically resected during the past 15 years. MAIN OUTCOME MEASURES Sensitivity and specificity of individual and combined antibodies for detecting benign from malignant lesions. RESULTS Quantitative analysis showed IHC validation of the gene expression differences noted in previously published microarray reports. A significantly higher percentage of FTC cells stained with galectin-3, EMMPRIN, and GADD153. Galectin-3 and EMMPRIN also showed a significantly higher intensity of staining in FTC cells. Compared with malignant lesions, TFF3 stained a greater cell percentage in FAs. Galectin-3 (sensitivity, 0.72; specificity, 0.62) and EMMPRIN (sensitivity, 0.63; specificity, 0.49) had the most promising diagnostic potential with a dual-protein sensitivity of 0.80 and specificity of 0.70. Autotaxin and GADD153 had overall higher sensitivities (0.88 and 0.82, respectively) but very poor specificities (0.02 and 0.21, respectively). CONCLUSIONS Protein expression data validate the pooled gene expression results that differentiate FTC from FA. Our results show promise for multiple-protein IHC analysis algorithms and their diagnostic ability. Future studies should focus on clinical translation of these molecular differences for the diagnosis of follicular thyroid neoplasms.