Phase I dose-escalating trial of Escherichia coli purine nucleoside phosphorylase and fludarabine gene therapy for advanced solid tumors.

BACKGROUND: The use of Escherichia coli purine nucleoside phosphorylase (PNP) to activate fludarabine has demonstrated safety and antitumor activity during preclinical analysis and has been approved for clinical investigation. PATIENTS AND METHODS: A first-in-human phase I clinical trial (NCT 01310179; IND 14271) was initiated to evaluate safety and efficacy of an intratumoral injection of adenoviral vector expressing E. coli PNP in combination with intravenous fludarabine for the treatment of solid tumors. The study was designed with escalating doses of fludarabine in the first three cohorts (15, 45, and 75 mg/m(2)) and escalating virus in the fourth (10(11)-10(12) viral particles, VP). RESULTS: All 12 study subjects completed therapy without dose-limiting toxicity. Tumor size change from baseline to final measurement demonstrated a dose-dependent response, with 5 of 6 patients in cohorts 3 and 4 achieving significant tumor regression compared with 0 responsive subjects in cohorts 1 and 2. The overall adverse event rate was not dose-dependent. Most common adverse events included pain at the viral injection site (92%), drainage/itching/burning (50%), fatigue (50%), and fever/chills/influenza-like symptoms (42%). Analysis of serum confirmed the lack of systemic exposure to fluoroadenine. Antibody response to adenovirus was detected in two patients, suggesting that neutralizing immune response is not a barrier to efficacy. CONCLUSIONS: This first-in-human clinical trial found that localized generation of fluoroadenine within tumor tissues using E. coli PNP and fludarabine is safe and effective. The pronounced effect on tumor volume after a single treatment cycle suggests that phase II studies are warranted. CLINICALTRIALSGOV IDENTIFIER: NCT01310179.

Fluorescence Imaging for Cancer Screening and Surveillance.

The advent of fluorescence imaging (FI) for cancer cell detection in the field of oncology is promising for both cancer screening and surgical resection. Particularly, FI in cancer screening and surveillance is actively being evaluated in many new clinical trials with over 30 listed on Clinical Trials.gov . While surgical resection forms the foundation of many oncologic treatments, early detection is the cornerstone for improving outcomes and reducing cancer-related morbidity and mortality. The applications of FI are twofold as it can be applied to high-risk patients in addition to those undergoing active surveillance. This technology has the promise of highlighting lesions not readily detected by conventional imaging or physical examination, allowing disease detection at an earlier stage of development. Additionally, there is a persistent need for innovative, cost-effective imaging modalities to ameliorate healthcare disparities and the global burden of cancer worldwide. In this review, we outline the current utility of FI for screening and detection in a range of cancer types.

Pathology-based staging for HPV-positive squamous carcinoma of the oropharynx.

OBJECTIVE: The rapid worldwide rise in incidence of human papillomavirus (HPV)-positive oropharyngeal squamous cell carcinoma (OPSCC) has generated studies confirming this disease as an entity distinct from traditional OPSCC. Based on pathology, surgical studies have revealed prognosticators specific to HPV-positive OPSCC. The current AJCC/UICC staging and pathologic nodal (pN)-classification do not differentiate for survival, demonstrating the need for new, HPV-specific OPSCC staging. The objective of this study was to define a pathologic staging system specific to HPV-positive OPSCC. METHODS: Data were assembled from a surgically-managed, p16-positive OPSCC cohort (any T, any N, M0) of 704 patients from five cancer centers. Analysis was performed for (a) the AJCC/UICC pathologic staging, (b) newly published clinical staging for non-surgically managed HPV-positive OPSCC, and (c) a novel, pathology-based, "HPVpath" staging system that combines features of the primary tumor and nodal metastases. RESULTS: A combination of AJCC/UICC pT-classification and pathology-confirmed metastatic node count (⩽4 versus ⩾5) yielded three groups: stages I (pT1-T2, ⩽4 nodes), II (pT1-T2, ⩾5 nodes; pT3-T4, ⩽4 nodes), and III (pT3-T4, ⩾5 nodes), with incrementally worse prognosis (Kaplan-Meier overall survival of 90%, 84% and 48% respectively). Existing AJCC/UICC pathologic staging lacked prognostic definition. Newly published HPV-specific clinical stagings from non-surgically managed patients, although prognostic, showed lower precision for this surgically managed cohort. CONCLUSIONS: Three loco-regional "HPVpath" stages are identifiable for HPV-positive OPSCC, based on a combination of AJCC/UICC primary tumor pT-classification and metastatic node count. A workable, pathologic staging system is feasible to establish prognosis and guide adjuvant therapy decisions in surgically-managed HPV-positive OPSCC.

Chemotherapy followed by accelerated fractionated radiation for larynx preservation in patients with advanced laryngeal cancer.

PURPOSE: Larynx preservation in advanced, resectable laryngeal cancer may be achieved using induction chemotherapy (CT) followed in responding patients by definitive radiation (RT). To address potential accelerated repopulation of clonogenic tumor cells during the prolonged total treatment time, we studied the feasibility of accelerated fractionated RT after CT. METHODS: Patients with advanced laryngeal cancer received two cycles of cisplatin 100 mg/m2 and fluorouracil (5-Fu) 1,000 mg/m2/d for 5 days. Responding patients received a third cycle after which those who had complete response or tumor down-staging to T1 proceeded with accelerated RT: 70.4 Gy delivered over 5.5 weeks. Patients who achieved a lesser response to CT underwent total laryngectomy and postoperative RT. RESULTS: Thirty-three patients were accrued. Three died during the course of CT and two declined definitive treatment after CT. Twenty-one patients had a major response to CT, 20 of whom received accelerated RT. Median weight loss during RT was 11%. Late severe morbidity was observed in five patients (25%). All four patients who underwent salvage laryngectomy after accelerated RT experienced major postoperative complications. The locoregional failure rate was 25%. The larynx was preserved in 48% of the total study population and in 80% of the patients irradiated according to the study protocol. CONCLUSION: Accelerated RT after CT as delivered in this study may increase both acute and long-term morbidity rates compared with studies using standard RT after CT. It did not seem to improve local/regional tumor control or survival despite stringent patient selection criteria.

Enhanced apoptosis with combination C225/radiation treatment serves as the impetus for clinical investigation in head and neck cancers.

PURPOSE: Epidermal growth factor receptor (EGFr) is overexpressed in a majority of head and neck squamous cell carcinomas, and this overexpression is associated with a poor prognosis. Therefore, EGFr has become the target of investigations aimed at disabling the receptor to determine whether this process leads to improved tumor kill with conventional treatment. MATERIALS AND METHODS: C225 is an anti-EGFr monoclonal antibody that inhibits receptor activity by blocking the ligand binding site. A panel of human head and neck squamous cell carcinoma cell lines was used to study the combination of C225 and radiation. RESULTS: It was determined that the combination of C225 (5 microgram/mL) delivered simultaneously with radiation (3 Gy) resulted in a greater decrement in cellular proliferation than either treatment alone. This reduction in proliferation correlated with reduced EGFr tyrosine phosphorylation and a reduction in phosphorylated signal transducer and activator of transcription-3 (STAT-3) protein (known to protect cells from apoptosis). Also, the decrement in proliferation correlated with increased apoptotic events, thereby indirectly linking C225/radiation-induced regulation of STAT-3 protein to apoptosis. CONCLUSION: This preclinical work serves as important support for the ongoing clinical investigation of C225 and radiotherapy for patients with head and neck carcinomas. The initial results of these clinical studies have been promising.

Selective gene delivery to head and neck cancer cells via an integrin targeted adenoviral vector.

In vivo cancer gene therapy approaches for squamous cell carcinoma of the head and neck (SCCHN) based on adenoviral vector-mediated gene delivery have been limited by the suboptimal efficacy of gene transfer to tumor cells. We hypothesized that this issue was due to deficiency of the primary adenoviral receptor, the coxsackie-adenovirus receptor (CAR), on the tumor targets. Studies of CAR levels on SCCHN cell lines confirmed that their relative refractoriness to the adenoviral vector was based on this deficiency. To circumvent this deficiency, we applied an adenoviral vector targeted to a tumor cell marker characteristic of SCCHN. In this regard, integrins of the alpha2beta1 and alpha3beta1 class are frequently overexpressed in SCCHN. Furthermore, these integrins recognize the RGD peptide motif. On this basis, we applied an adenoviral vector genetically modified to contain such a peptide within the HI loop of the fiber protein as a means to alter viral tropism. Studies confirmed that the CAR-independent gene delivery achieved via this strategy allowed enhanced gene transfer efficiencies to SCCHN tumor cells. Importantly, this strategy could achieve preferential augmentation of gene transfer in tumor cells compared with normal cells. The ability to achieve enhanced and specific gene transfer to tumor cells via adenoviral vectors has important implications for gene therapy strategies for SCCHN and for other neoplasms in general.

Ionized serum calcium levels following combined treatment for cancer of the head and neck.

Thyroid function may be reduced after treatment of cancer of the head and neck, and hypothyroidism is much more common after combination therapy. Whether hypoparathyroidism and subsequent hypocalcemia also occur after such treatment is unknown. Few related studies have been published in which changes in total serum calcium have been studied after cancer treatment with radioactive iodine or external radiation. Twenty-two disease-free head and neck cancer patients were studied, 1 to 3 years after multimodal treatment, to determine if changes in serum ionized calcium levels or thyroid function were present. Our results suggest that parathyroid function, as represented by ionized calcium levels remains normal after multimodality (surgery, radiation and/or chemotherapy) combined treatment.

Molecular differences in mucoepidermoid carcinoma and adenoid cystic carcinoma of the major salivary glands.

OBJECTIVE/HYPOTHESIS: Mucoepidermoid carcinoma (MEC) and adenoid cystic carcinoma (ACC), the most common malignancies of the major salivary glands, are clinically and pathologically different. To determine whether MEC and ACC have different molecular characteristics, we examined the expression of erbB-2, erbB-3, epidermal growth factor receptor (EGFR), and transforming growth factor-alpha (TGF-alpha), important molecular features in other malignancies. STUDY DESIGN/METHODS: Archival tissue sections of 22 MEC and 6 ACC tumors of the major salivary glands were evaluated immunohistochemically for expression of erbB-2, erbB-3, EGRF, and TGF-alpha. A differential immunostaining score, reflecting the difference in immunostaining between carcinoma and uninvolved salivary gland tissue, was calculated for cytoplasmic and membranous staining. RESULTS: Positive immunostaining for all biomarkers was observed in the cytoplasm and membrane of both tumors. However, expression was higher in MEC than in ACC tumors and was statistically significant for cytoplasmic EGFR (P =.009), TGF-alpha (P =.041), and membranous EGFR (P =.004). A significantly higher percentage of MEC cells also demonstrated positive immunostaining for cytoplasmic erbB-3 (P =.022), EGFR (P =.005), membranous erbB-3 (P =.022), and EGFR (P =.013). The differential immunostaining score was significantly higher for MEC compared with uninvolved alveolar tissue and the membranes of uninvolved ductal tissue. There were no statistically positive differential immunostaining scores for ACC. CONCLUSIONS: There is a clear difference in the molecular phenotypes of MEC and ACC. The lack of statistically significant expression in ACC, when compared with similar uninvolved salivary gland tissue, suggests minimal involvement for these molecular structures in the pathogenesis of ACC. Conversely, erbB-2, erbB-3, EGFR, and TGF-alpha may have a role in the development and progression of MEC. These results have therapeutic implications for MEC of the major salivary glands.

Extracapsular spread and the perineural extension of squamous cell cancer in the cervical plexus.

Extracapsular spread of squamous cell carcinoma in cervical lymph nodes is associated with approximately 50% decrease in survival and a twofold increase in regional recurrence. This study examines the hypothesis that increased regional recurrence may be, in part, due to unrecognized microscopic perineural invasion of the nerve rootlets of the cervical plexus. Thirty patients with head and neck squamous cell carcinoma with clinically N+ necks undergoing radical neck dissection were prospectively studied. Neck dissection specimens were evaluated for extracapsular spread, and the cervical plexus rootlets were histologically examined for perineural invasion. The incidence of extracapsular spread was 83% (25 of 30 patients). Only one (4%) of 25 had involvement of the cervical plexus, and this patient had gross as well as microscopic cervical plexus invasion. Microscopic perineural spread of squamous cell carcinoma in the cervical plexus occurs infrequently when extracapsular spread is present. Routine histologic evaluation of cervical rootlets for margins is warranted only when gross tumor is in close proximity to the cervical plexus.

Preoperative vascular imaging for the fibular osteocutaneous flap.

The fibular osteocutaneous free flap has become a well-accepted method of mandibular reconstruction. Aberrations in the blood supply to the foot affect 5% to 7% of the population, and substantial atherosclerotic disease of the lower extremities is often found in elderly individuals, many of whom have been smokers. Therefore, the use of preoperative vascular imaging is justified in all patients scheduled for fibular osteocutaneous free-flap harvest. In a series of 25 consecutive patients clinically judged to be satisfactory candidates for fibular free-flap reconstruction, preoperative arteriograms excluded 4 patients from use of this donor site and determined which leg was used in 2 other patients.

Repair of a complete glottic-subglottic stenosis with a fibular osseocutaneous free flap.

Reconstruction of extensive laryngotracheal stenosis remains a formidable challenge. The ideal reconstructive technique has not been found because of the variability in the complexity and degree of laryngotracheal stenosis and the challenge of wound healing in a contaminated tubular structure. The application of microvascular free-tissue transfer in laryngotracheal reconstruction is limited. We used a fibula osseocutaneous revascularized flap for reconstruction of a complex laryngotracheal stenosis. The clinical course, long-term follow-up, and potential advantages and disadvantages are discussed.

Monitoring of revascularized jejunal autografts.

Free jejunal autografts have been widely used to reconstruct circumferential pharyngeal defects in a single stage. Close postoperative monitoring of the perfusion of the free jejunal autograft is extremely important in achieving a successful outcome. Presented herein is our method of postoperative monitoring of jejunal autografts in which a segment of jejunum pedicled on its connecting mesentery is exteriorized. This method of monitoring was performed in 17 cases, and there were no cases of late graft failure. Of three patients who had undergone this operation before the use of this monitoring technique, one had an unrecognized late graft failure. We present our experience with this simple and reliable method to monitor free jejunal autografts.

Lateral arm free flap in head and neck reconstruction.

The lateral arm fasciocutaneous free flap is a versatile donor site of sensate soft tissue for reconstruction and augmentation of the head and neck. The lateral arm flap can be quickly harvested without interference to the head and neck team and with minimal morbidity to the patient. For these reasons, this flap has become our soft-tissue flap of choice.

The cutaneous scapular free flap in head and neck reconstruction.

Successful surgical reconstruction of complicated soft-tissue defects of the head and neck region has been greatly enhanced by free-revascularized tissue transfers. The scapular free flap has become a favored reconstructive option in our department and has been reported previously for one-stage mandibular reconstruction. This flap can also be transferred as a cutaneous free flap. We present our clinical experience with the fasciocutaneous scapular free flap and review the anatomy, surgical technique, and utility of this versatile flap.

Primary adenocarcinoma of the temporal bone. A case with 40-year follow-up.

Adenomatous tumors of the temporal bone are rare neoplasms. This article records the 40-year course of a patient with adenocarcinoma of the temporal bone and reviews the literature pertinent to the biologic behavior, histologic appearance, prognosis, and treatment of this group of tumors.

Hemilaryngectomy for glottic carcinoma after radiation therapy failure.

OBJECTIVE: To determine the efficacy and safety of vertical hemilaryngectomy (VHL) for the treatment of early glottic carcinoma recurrent after radiation therapy (RT). DESIGN: Retrospective study. SETTING: Major referral center. PATIENTS: Forty patients were identified who underwent VHL for T1 or T2 glottic carcinoma between July 1975 and March 1991, and all were included in this study. Twenty-two patients had received full-course RT before VHL, and 18 patients underwent primary VHL. MAIN OUTCOME MEASURES: The local control rates were determined for T1 and T2 tumors in each group, along with actuarial survival rates and complications. RESULTS: Local control of tumor for VHL after RT failure was 85% for T1 tumors, 78% for T2 tumors, and 82% overall. Three of four of the local failures in this group occurred in patients who had contraindications to VHL. Total laryngectomy for treatment of local failures in this group increased the local control rate to 93% for T1 tumors, 89% for T2 tumors, and 91% overall. Local control rates for the primary VHL group were 90% for T1 tumors, 75% for T2 tumors, and 83% overall. Total laryngectomy for treatment of local recurrences increased local control to 87% for T2 tumors and 89% overall. Five-year actuarial survival was 85% for each group. Delayed tracheal decannulation occurred more frequently in the patients who had undergone RT. CONCLUSIONS: Our results support the oncologic safety and effectiveness of VHL for the surgical treatment of recurrent early glottic carcinomas after RT, with minimal increased morbidity.

The platysma myocutaneous flap. Indications and caveats.

The platysma myocutaneous flap has enjoyed limited popularity despite its versatility, dependability, and ease of harvesting. In this article, we describe 12 consecutive patients who underwent platysma flap reconstruction of various oral cavity and oropharyngeal defects. Complications included loss of one skin paddle, one pharyngocutaneous fistula, and one intraoral wound dehiscence. In all patients, bare bone was covered or appropriate spacing between the tongue and other structures was maintained to avoid ankyloglossia. We discuss specific indications and caveats.

Selective cricothyroid muscle reinnervation by muscle-nerve-muscle neurotization.

OBJECTIVE: To determine if selective reinnervation of the cricothyroid muscle could be achieved with muscle-nerve-muscle neurotization. DESIGN: Case series. SETTING: Tertiary referral center. PATIENTS: Three consecutive patients with high vagal lesions that resulted in unilateral laryngeal paralysis. INTERVENTIONS: Patients underwent laryngeal reinnervation with ansa hypoglossi to recurrent laryngeal nerve anastomosis. In addition, patients underwent selective cricothyroid muscle reinnervation by muscle-nerve-muscle neurotization technique. MAIN OUTCOME MEASURES: Objective and subjective improvement in voice quality and electromyographic evidence of selective reinnervation of the cricothyroid muscle. RESULTS: All patients recovered normal or near-normal speaking voice and had normal objective measures of voice quality. They also showed electromyographic evidence of cricothyroid muscle reinnervation. CONCLUSION: The muscle-nerve-muscle neurotization technique was successful in providing selective reinnervation of the cricothyroid muscle in our 3 patients.

Retargeting to EGFR enhances adenovirus infection efficiency of squamous cell carcinoma.

BACKGROUND: Adenovirus-mediated gene therapy has been used for squamous cell carcinoma of the head and neck (SCCHN), but the in vivo efficacy has been limited by a lack of tissue specificity and low infection efficiency. We are interested in improving cancer gene therapy strategies using targeted adenovirus vectors. OBJECTIVE: To determine if the infection efficiency of adenovirus-mediated gene transfer to SCCHN cells could be enhanced by retargeting to the epidermal growth factor receptor (EGFR), which is known to be overexpressed in these tumors. DESIGN: Epidermal growth factor receptor retargeting in SCCHN cells was accomplished with a bispecific antibody that recognized the knob domain of adenovirus as well as EGFR. Using this retargeting schema, we compared the infection efficiency and specificity of unmodified and EGFR-retargeted adenovirus. RESULTS: Squamous cell carcinoma of the head and neck cell lines were shown to be infected by adenovirus with low efficiency, which is likely because of the low level of adenovirus receptor expressed in the SCCHN cells. Epidermal growth factor receptor retargeting markedly enhanced transduction in both SCCHN cell lines and primary tumor tissue, as indicated by the elevated levels of reporter gene expression. Furthermore, retargeting enhanced infection of tumor tissue compared with normal tissue from the same patient. CONCLUSIONS: Epidermal growth factor receptor retargeting enhanced adenovirus infection of SCCHN cells and, in doing so, augments the potency of the vector. This modification makes the vector potentially more valuable in the clinical setting.

The oxidative decarboxylation of polyaminocarboxylic acidm-VI Reaction of diethylenetriaminepenta-acetic acid with cerium(IV) in sulphuric acid media.

The oxidation of diethylenetrianunepenta-acetic acid (DTPA) by Ce(IV) in sulphuric add was investigated spectrophotometrically by the stopped-flow technique. The rate of reaction is influenced by the acidity, but can be expressed by a simplified rate law At [H(2)SO(4)] below 0.75M the reaction proceeds stepwise as shown by formation of a 1:1 Ce(IV)-DTPA complex with measurable rate of formation and decay. At higher acid strengths, the formation of an intermediate is not evident. The rate is maximal in ~ 0.75M sulphuric add. The overall stoichiometry varies with time.