J. Trad Wadsworth

Serum Protein Profiles to Identify Head and Neck Cancer

Purpose: New and more consistent biomarkers of head and neck squamous cell carcinoma (HNSCC) are needed to improve early detection of disease and to monitor successful patient management. The purpose of this study was to determine whether a new proteomic technology could correctly identify protein expression profiles for cancer in patient serum samples. Experimental Design: Surface-enhanced laser desorption/ionization-time of flight-mass spectrometry ProteinChip system was used to screen for differentially expressed proteins in serum from 99 patients with HNSCC and 102 normal controls. Protein peak clustering and classification analyses of the surface-enhanced laser desorption/ionization spectral data were performed using the Biomarker Wizard and Biomarker Patterns software (version 3.0), respectively (Ciphergen Biosystems, Fremont, CA). Results: Several proteins, with masses ranging from 2,778 to 20,800 Da, were differentially expressed between HNSCC and the healthy controls. The serum protein expression profiles were used to develop and train a classification and regression tree algorithm, which reliably achieved a sensitivity of 83.3% and a specificity of 100% in discriminating HNSCC from normal controls. Conclusions: We propose that this technique has potential for the development of a screening test for the detection of HNSCC.

Differential Capture of Serum Proteins for Expression Profiling and Biomarker Discovery in Pre‐ and Posttreatment Head and Neck Cancer Samples

Introduction: A long‐term goal of our group is to develop proteomic‐based approaches to the detection and use of protein biomarkers for improvement in diagnosis, prognosis, and tailoring of treatment for head and neck squamous cell cancer (HNSCC). We have previously demonstrated that protein expression profiling of serum can identify multiple protein biomarker events that can serve as molecular fingerprints for the assessment of HNSCC disease state and prognosis.

Reactivation of Suppressed RhoB is a Critical Step for the Inhibition of Anaplastic Thyroid Cancer Growth

Anaplastic thyroid carcinoma (ATC) is a highly aggressive form of the disease for which new therapeutic options are desperately needed. Previously, we showed that the high-affinity peroxisome proliferator-activated receptor gamma (PPARgamma) agonist, RS5444, inhibits cell proliferation of ATC cells via induction of the cyclin-dependent kinase inhibitor p21(WAF1/CIP1) (p21). We show here that up-regulation of RhoB is a critical step in PPARgamma-mediated activation of p21-induced cell stasis. Using multiple independently derived ATC cell lines, we found that treatment with RS5444 leads to the up-regulation of RhoB and subsequent activation of p21, and that silencing of RhoB by RNAi blocks the ability of RS5444 to induce p21 and to inhibit cell proliferation. Our results show that transcriptional regulation of RhoB by the nuclear transcription factor PPARgamma is responsible for the induction of p21 mRNA and protein. We further implicate RhoB as a key signaling effector for the growth inhibition of ATC, as treatment with a histone deacetylase inhibitor shown to increase RhoB expression in lung cancer cells caused the up-regulation of RhoB in ATC cells accompanied by increased expression of p21 and inhibition of cell proliferation; this effect occurred even in ATC cells that were unresponsive to RS5444 due to a lack of expression of PPARgamma. Our results implicate RhoB as a novel intermediate in critical signaling pathways and as an additional target for therapeutic intervention in ATC.

Serum, salivary and tissue proteomics for discovery of biomarkers for head and neck cancers.

Initial clinically oriented applications of emerging proteomic technologies that aim to identify biomarkers for head and neck squamous cell carcinoma diagnostics have yielded promising results. The development of new proteomic diagnostics remains critical for the early detection of head and neck squamous cell carcinoma at more treatable stages. Prognostic markers for disease recurrence and treatment sensitivities are also required. In this overview of current biomarker identification strategies for head and neck squamous cell carcinoma, different combinations of mass spectrometry platforms, laser capture microscopy and 2D gel electrophoresis procedures are summarized as applied to readily available clinical specimens (tissue, blood and saliva). Issues related to assay reproducibility, management of large data sets and future improvements in clinical proteomics are also addressed.

Detailed Molecular Fingerprinting of Four New Anaplastic Thyroid Carcinoma Cell Lines and Their Use for Verification of RhoB as a Molecular Therapeutic Target

CONTEXT Anaplastic thyroid carcinoma (ATC) is a highly aggressive carcinoma in need of therapeutic options. One critical component of drug discovery is the availability of well-characterized cell lines for identification of molecular mechanisms related to tumor biology and drug responsiveness. Up to 42% of human thyroid cancer cell lines are redundant or not of correct tissue origin, and a comprehensive analysis is currently nonexistent. Mechanistically, RhoB has been identified as a novel molecular target for ATC therapy. OBJECTIVE The aim was to develop four ATC cell lines detailing genetic, molecular, and phenotypic characteristics and to test five classes of drugs on the cell lines to determine whether they inhibited cell proliferation in a RhoB-dependent fashion. DESIGN Four cell lines were derived from ATC tumors. Short tandem DNA repeat and mutational status of the originating tumors and cell lines were performed along with molecular and phenotypic characterizations. Compounds were tested for growth inhibition and ability to up-regulate RhoB. RESULTS Cell line authenticity was confirmed by DNA short tandem repeat analysis. Each proved unique regarding expression of thyroid markers, oncogene status, amplified and deleted genes, and proliferative growth rates. FTI-277, GGTI-286, lovastatin, romidepsin, and UCN-01 up-regulated RhoB and inhibited cell proliferation in a dose-responsive fashion with only romidepsin and FTI-277 being RhoB dependent. CONCLUSIONS Molecular descriptions of thyroid lines were matched to the originating tumors, setting a new standard for cell line characterization. Furthermore, suppressed RhoB is implicated as a molecular target for therapy against ATC because five classes of drugs up-regulate RhoB and inhibit growth dose-responsively.

Tube Placement: A Prospective, Randomized Double-Blind Study

Bilateral myringotomy with tympanostomy tube placement is the second most frequently performed pediatric surgical procedure, next to circumcision. Postoperative pain relief for children undergoing this procedure has been an ongoing concern. The authors undertook a prospective, randomized, double‐blind, placebo‐controlled clinical study in 200 consecutive children to investigate the efficacy of oral acetaminophen, acetaminophen with codeine, ibuprofen, and placebo administered preoperatively in relieving postoperative pain in children undergoing this procedure. All children received topical analgesia consisting of antibiotic eardrops mixed with 4% lidocaine intraoperatively. There was no significant difference in postoperative pain score between the four groups ( P > 0.4447). Thus it is likely that the intraoperative administration of antibiotic eardrops mixed with 4% lidocaine is all that is required to alleviate postoperative pain in children undergoing myringotomy with tympanostomy tube placement. Preoperative oral analgesics are apparently of little added benefit.

Effectiveness of surgeon interpretation of technetium tc 99m sestamibi scans in localizing parathyroid adenomas.

OBJECTIVES To evaluate the ability of surgeons to predict the laterality of parathyroid adenomas from technetium Tc 99m sestamibi scans and compare their results with the radiologic interpretations. DESIGN Retrospective medical record review with single- blinded review of sestamibi scans. SETTING Tertiary care academic medical center PATIENTS The study population included 110 consecutive parathyroidectomy cases from January 2001 to June 2004. Inclusion criteria were biochemically proven primary hyperparathyroidism, with documented serum hypercalcemia and elevated parathyroid hormone levels. Only cases due to a single adenoma that were cured with a single surgical procedure were included. INTERVENTION All patients underwent resection of a parathyroid adenoma following a preoperative sestamibi localization study and serum calcium and parathyroid hormone level analysis. MAIN OUTCOME MEASURE Adenoma location was determined from a review of operative and pathological reports. Two head and neck surgeons performed a blinded review of all scans, and their findings were compared with the radiology reports. RESULTS Of 82 adenomas, 51 (62%) were correctly lateralized in the radiology report, while the other 31 were interpreted as normal scans. The sensitivity and specificity of the radiologic interpretations for parathyroid adenomas in all patients with primary hyperparathyroidism were 62% and 83%, respectively. The scan interpretation of the 2 surgeons produced accurate lateralization of 91% and 91% of these single adenomas. Of the 31 single adenoma scans read as normal by the radiologist, the surgeons correctly lateralized 22 of 29 (76%) and 21 of 28 (75%) of the adenomas. CONCLUSION The review of sestamibi scans by surgeons allows accurate localization of parathyroid adenomas that may not be identified by standard radiologic interpretations.

Impact of oncogene panel results on surgical management of cytologically indeterminate thyroid nodules

The impact of oncogene panel results on the surgical management of indeterminate thyroid nodules (ITNs) is currently unknown.

Postoperative Use of Vasopressors in Head and Neck Microvascular Reconstruction

Objectives: Microvascular free tissue reconstruction is overall successful in the treatment of patients with disfiguring wounds resulting from ablative cancer resection, trauma, and osteo(radio)necrosis. Flap failure can be devastating for patients and health care providers. The use of sympathomimetic drugs (vasopressors) in microvascular surgery is controversial; however, current research fails to demonstrate adverse outcomes when intraoperative vasopressors are used. Conversely, intraoperative fluid administration greater than 7 L has been associated with major postoperative complications. The aims of the study were: (1) determine the efficacy of postoperative vasopressor use in free flap patients; (2) compare length of hospital stay in patients who receive vasopressors postoperatively and those who do not. Methods: In this retrospective pilot study, we investigated the use of a postoperative hypotension treatment protocol at our institution, implemented in June 2013. Thirty-nine patients underwent free flap reconstruction of the head and neck secondary to malignancy. Outcome measures included flap viability and length of hospital stay. Results: Analysis of this ongoing study reveals that 23% of patients received peripherally-active vasopressors within the first 3 postoperative days. There were no flap failures in either group. We found no statistically significant difference in the rate of surgical reexploration in patients who received vasopressors postoperatively compared to patients who did not (Fisher’s exact test, P value .12). Conclusions: The length of hospital stay is not statistically significantly different when vasopressors are used postoperatively. The implementation of a formalized hypotension treatment protocol in the postoperative setting does not adversely affect the outcome of free flap survival or length of hospital stay.

Submandibular (Undescended) Ectopic Parathyroid Glands: Elusive to Surgeons and Localization Studies:

Objectives: High cervical parathyroid adenomas, known as undescended adenomas, occur with a frequency of less than 0.1%, but account for 7% of failed parathyroidectomies. Such cases are scarce in the otolaryngology-head and neck surgery literature. Preoperative localization of undescended adenomas using Technetium 99-c sestamibi scintigraphy (sestamibi) is difficult due to uptake and retention of sestamibi by salivary gland tissue. Methods: We present a review of the literature and two surgeons’ experience involving undescended parathyroid glands complicating the treatment of patients with primary (cases 2,3) and secondary (case 1) hyperparathyroidism. In case 1, a 52-year-old dialysis-dependent male experienced recurrent secondary hyperparathyroidism, requiring three operations over two years to excise all parathyroid glands. The right inferior gland was localized to the submandibular region prior to the final operation. In case 2, two sestamibi–single-photon emission computed tomography scans were proven incorrect when a contralateral undescended adenoma was located in a 54-year-old female at surgical exploration. In case 3, a 49-year-old male with a non-localizing scan underwent a failed neck exploration, complicated by acute pancreatitis postoperatively. The surgeon’s suspicion of an undescended adenoma was confirmed by computed tomography; the adenoma was excised. Results: When sestamibi scans are inconclusive, the contour and symmetry of uptake in and around the submandibular glands should be scrutinized. Thallium-pertechnetate subtraction scanning (TPSS) as a second line modality may be useful. Undescended glands must be considered during an unyielding, thorough parathyroid exploration. Conclusions: Surgeons should consider the possibility of undescended adenoma, and use the aforementioned localization techniques when sestamibi scans are inconclusive.