A Comparison of the Diagnostic Accuracy and Reliability of Subjective Grading and Computer-Aided Assessment of Intranodal Vascularity in Differentiating Metastatic and Reactive Cervical Lymphadenopathy.

PURPOSE: Ultrasound is a well-established imaging modality in the assessment of malignant cervical lymphadenopathy. With the use of Doppler ultrasound, intranodal vascularity can be evaluated. However, the major limitation of ultrasound is operator dependency. Therefore, this study aimed to evaluate and compare the diagnostic accuracy and reliability of the subjective grading and computer-aided approach in assessing intranodal vascularity for the differentiation of benign and malignant lymph nodes. MATERIALS AND METHODS: The present study retrospectively assessed 99 power Doppler ultrasound images of cervical lymph nodes and evaluated the degree of intranodal vascularity using qualitative subjective grading (QSG) and quantitative computer-aided (QCA) methods. The diagnostic accuracy of the two methods in distinguishing metastatic and reactive nodes and their inter- and intra-rater reliability in assessing intranodal vascularity were evaluated and compared. RESULTS: The results showed that the QCA method was more accurate than the QSG method with a significantly higher sensitivity (67.8 % and 61.9 %, respectively, p < 0.05) and specificity (73.3 % and 57.3 %, respectively, p < 0.05). Using the intranodal vascularity index as determined by the QCA approach, the optimum cut-off to differentiate metastatic and reactive cervical lymph nodes was 32 %. The QCA method showed higher inter- and intra-rater reliability than the QSG method. CONCLUSION: In the assessment of the degree of intranodal vascularity, the QCA method was more accurate and reliable than the QSG method in distinguishing metastatic and reactive lymph nodes.

Phase I clinical study of vascular targeting fluorescent cationic liposomes in head and neck cancer.

The aim of this first-time-in-human non-randomized dose-escalating prospective phase I clinical trial was to analyze safety of two doses of fluorescent rhodamine-labeled cationic liposomes (LDF01) in head and neck squamous cell carcinoma (HNSCC). Patients had resectable UICC stadium I-IV A HNSCCs. LDF01 was administered before tumor resection under general anesthesia as an intravenous infusion with effective lipid doses of 0.5 or 2 mg/kg b.w., respectively. In addition to clinical monitoring for safety assessment, tumor biopsies were taken during the surgical procedure for fluorescence histological analysis. Eight patients were assigned to the two dose groups. During safety follow-up no clinically relevant adverse events occurred. Fluorescence histology revealed some evidence of favorable selectivity of LDF01 for tumor microvessels in the high-dose group. LDF01 is safe applied as infusion at both tested dose levels. Furthermore, LDF01 can be detected in the vicinity of tumor cells and could be assigned to the microvessel target in individual HNSSC cases. Detailed analysis of targeting properties of LDF01 has to be performed in upcoming clinical phase II trials.

Role of narrow-band imaging and high-definition television in the surveillance of head and neck squamous cell cancer after chemo- and/or radiotherapy.

Narrow-band imaging (NBI) is an endoscopic technique enhancing mucosal vasculature and better identifying superficial carcinomas due to their neo-angiogenic pattern. NBI accuracy is increased by combination with a high-definition television (HDTV) camera. The aim of this report was to evaluate the diagnostic improvement of NBI +/- HDTV in the evaluation of head and neck squamous cell cancer (HNSCC) previously treated by chemo-radiotherapy (CHT-RT) or RT. A total of 390 patients affected by HNSCC were prospectively evaluated by NBI and white light (WL) endoscopy +/- HDTV between April 2007 and April 2009 at a single academic institution. Among them, we focused on 59 (15%) patients who received CHT-RT or RT as part of their treatment. Of 59 patients, 13 (22%) showed adjunctive preoperative NBI findings when compared to the standard WL examination. These findings were always confirmed by intraoperative HDTV NBI, while only eight (62%) were visible with HDTV WL. Of 13 lesions, 12 received histopathologic confirmation (from carcinoma in situ to invasive carcinoma). The sensitivity of flexible NBI, HDTV WL, and HDTV NBI was 100, 66 and 100%, respectively. The specificity was 98, 100, and 98%. The positive predictive value was 92, 100, and 92%. The negative predictive value was 100, 94, and 100%. The accuracy was 98, 91, and 98%. NBI +/- HDTV after CHT-RT or RT was of value in detecting tumor persistence (n = 2), early recurrences (n = 6), and metachronous tumors (n = 4). By contrast, only 1 of 59 (2%) patients was found to be false positive.

Proangiogenic effects of ionizing irradiation on squamous cell carcinoma of the hypopharynx.

OBJECTIVE: There is experimental evidence that ionizing irradiation affects a proangiogenic response. However, the relevance of this effect on tumour growth in vivo is not in detail investigated yet. The present objectives were to examine the influence of ionizing radiation on the expression of the vascular endothelial growth factor (VEGF) and its receptors (flt-1 and flk-1), the microvessel density and the tumour proliferation, in head and neck squamous cell carcinoma (HNSCC). METHODS: We used a HNSCC-cell line, derived from a hypopharyngeal tumour, for subcutaneous injection in 16 athymic nude mice. After reaching an average diameter of 12-14 mm the xenografts were randomised and 8 out of the 16 animals (therapy group) were irradiated with a single fraction of 6 Gy while the control group remained without any intervention. The irradiated and the respective control tumours were prepared after 7 (T7) and 70 days (T70) for immunohistochemical analysis. The expression of VEGF, its receptors flk-1 and flt-1, the vessel density (CD31) and the proliferation rate (Ki67) were quantified. RESULTS: At the point of time T7 we observed a reduction of the tumour growth rate, of the proliferative activity and of the VEGF- as well as of the VEGF-R-expression. At the point of time T70 we found increased values for proliferation, microvessel density, VEGF- and flk-1 expression in the therapy group compared to the therapy group at T7 as well as to the control group at T70. CONCLUSION: These changes might suggest a long-term proangiogenic effect of irradiation, which might result in growth promotion of the remaining tumour after the end of therapy.

A Novel External Carotid Arterial Sheath System for Intra-arterial Infusion Chemotherapy of Head and Neck Cancer.

PURPOSE: The purpose of this study was to describe a novel system for treating advanced head and neck cancer consisting of an external carotid arterial sheath (ECAS) and a microcatheter to inject drugs retrogradely into multiple feeding arteries through the superficial temporal artery (STA). MATERIALS AND METHODS: Four consecutive patients with head and neck cancer that had more than one feeding artery were enrolled in this study. The ECAS was made of polyurethane and surface-coated with heparin resin to prevent thrombus formation, allowing it to remain in place for a prolonged period of time. The ECAS was inserted through the STA, and its tip was placed between the maxillary artery and facial artery. The tumor-feeding arteries were selected using a hooked-shaped microcatheter through the ECAS. RESULTS: A total of 13 target arteries were selected in the four patients. The microcatheter inserted via the ECAS was used to catheterize ten arteries (five lingual arteries and five facial arteries). The remaining three lingual arteries were directly selected by the catheter without ECAS. All of the target arteries were able to be catheterized superselectively. The technical success rate was 100%. Vascular occlusion, which might have been caused by the ECAS, was observed in one patient. No neurologic toxicities occurred. CONCLUSION: This ECAS system is a new approach for retrograde superselective intra-arterial chemotherapy that covers the entire tumor with anticancer drugs. It has the potential to increase the effectiveness of therapy for advanced head and neck cancer. LEVEL OF EVIDENCE: Level 4, Case Series.

Tumoural perfusion as measured by dynamic computed tomography in head and neck carcinoma.

PURPOSE: To investigate the intra- and interobserver variability, as well as the intra- and interpatient variability of CT-determined tumour perfusion in head and neck tumours, and to evaluate the preliminary value of this parameter as predictive factor of local failure after treatment by definitive radiotherapy. MATERIALS AND METHODS: In 41 patients the perfusion of a primary head and neck squamous cell carcinoma was estimated using dynamic CT. A 40-ml intravenous bolus of a low-osmolar non-ionic contrast agent was rapidly injected over 5 s (8 ml/s), while a dynamic acquisition of image data was obtained during the first pass at the level of the largest axial tumour surface. A time-density curve was constructed for the primary tumour and the carotid artery. The perfusion in the selected tumour region of interest was calculated by dividing the slope of the tumour-time density curve by the maximal value in arterial density. Tumour volume was calculated on the CT-images and correlated with perfusion rate. RESULTS: The mean perfusion rate was 86.4 ml/min per 100 g (median, 80.6; SD, 43.05; range, 31.7-239.8 ml/min per 100 g). No systematic difference was found between the measurements performed by two independent observers. The intratumoural COV was 0.22, the intertumoural COV 0.37. No correlation was found with tumour volume. Ten out of 20 patients with a perfusion rate < 80 ml/min per 100 g were not locally controlled, while nine out of 21 patients with a value > 80 ml/min per 100 g did show a local failure (P = 0.19). CONCLUSIONS: CT-determined perfusion measurements of head and neck tumours are feasible. No correlation with tumour volume and a sufficiently large COV were found to consider this parameter as a possible prognostic factor for outcome after radiotherapy. More patients need to be investigated to test the hypothesis that tumours with a low CT determined perfusion rate have a higher risk of local failure.

Pediatric head and neck lesions: assessment of vascularity by MR digital subtraction angiography.

BACKGROUND AND PURPOSE: Pediatric head and neck lesions can be difficult to characterize on clinical grounds alone. We investigated the use of dynamic MR digital subtraction angiography as a noninvasive adjunct for the assessment of the vascularity of these abnormalities. METHODS: Twelve patients (age range, 2 days to 16 years) with known or suspected vascular abnormalities were studied. Routine MR imaging, time-of-flight MR angiography, and MR digital subtraction angiography were performed in all patients. The dynamic sequence was acquired in two planes at one frame per second by using a thick section (6-10 cm) selective radio-frequency spoiled fast gradient-echo sequence and an IV administered bolus of contrast material. The images were subtracted from a preliminary mask sequence and viewed as a video-inverted cine loop. RESULTS: In all cases, MR digital subtraction angiography was successfully performed. The technique showed the following: 1) slow flow lesions (two choroidal angiomas, eyelid hemangioma, and scalp venous malformation); 2) high flow lesions that were not always suspected by clinical examination alone (parotid hemangioma, scalp, occipital, and eyelid arteriovenous malformations plus a palatal teratoma); 3) a hypovascular tumor for which a biopsy could be safely performed (Burkitt lymphoma); and 4) a hypervascular tumor of the palate (cystic teratoma). CONCLUSION: Our early experience suggests that MR digital subtraction angiography can be reliably performed in children of all ages without complication. The technique provided a noninvasive assessment of the vascularity of each lesion that could not always have been predicted on the basis of clinical examination or routine MR imaging alone.

Preoperative permanent balloon occlusion of internal carotid artery in patients with advanced head and neck squamous cell carcinoma.

OBJECTIVE/HYPOTHESIS: To determine the value of preoperative balloon occlusion in predicting the safety of carotid artery resection in advanced recurrent head and neck squamous cell carcinoma. STUDY DESIGN: Retrospective chart review of all cases undergoing planned carotid artery resection for recurrent disease at a major university hospital. METHODS: If the carotid artery was encased, a nonemergent carotid artery balloon test occlusion was performed for 30 minutes. If the patient tolerated this, he or she underwent permanent carotid artery occlusion. RESULTS: Twenty-three patients were prospectively evaluated for resection. Three underwent emergent carotid artery ligation. Twenty others underwent nonemergent carotid artery test occlusion. Of these, 5 patients failed preoperative carotid artery balloon occlusion and 15 patients successfully underwent permanent carotid balloon occlusion. Although eight of these patients died of recurrent disease in less than 1 year, seven patients survived more than 1 year with two patients surviving more than 2 years. CONCLUSIONS: Preoperative carotid balloon occlusion predicted patients who could tolerate permanent occlusion. All patients eventually developed recurrent disease, but in 14 of the 15 patients, no hemorrhages occurred.

Evaluation of paragangliomas presenting as a cervical mass on color-coded Doppler sonography.

OBJECTIVES: To assess the potential of color Doppler sonography to identify cervical mass lesions as paragangliomas. STUDY DESIGN: Prospective evaluation. METHODS: Fifteen patients with 18 paragangliomas (13 carotid body tumors, 3 vagal and 2 temporal) presenting as a mass in the neck were investigated using color Doppler sonography. RESULTS: All paragangliomas presented in B-mode sonography as solid, well-defined, and hypoechoic tumors. Color Doppler imaging revealed hypervascularity in 15 (82%) tumors. No flow signal was detected in 3 carotid body tumors at standard velocity encoding (30 cm/s). Only carotid body tumors could be assessed in their full extent. Temporal and vagal paragangliomas were only partially visible. Carotid body tumors showed a splaying of the carotid bifurcation with displacement of the external carotid anteriorly and both the internal carotid and the internal jugular vein posteriorly. Anterior displacement of both carotid arteries and posterior displacement of the internal jugular vein was found in the 3 vagal paragangliomas. The caudal tumor extension of the 2 temporal paragangliomas was recognized within the expanded lumen of the internal jugular vein. According to the direction of tumor growth and vascular supply, the intratumoral flow signal was predominantly directed upward in carotid body tumors and downward in vagal and temporal paragangliomas. CONCLUSIONS: Based on the appearance in the B-mode, the hypervascularity, the relationship toward the carotid arteries and the internal jugular vein, and the intratumoral flow direction, color Doppler sonography was able to establish the diagnosis and type of a paraganglioma.

Vascular endothelial growth factor receptor 2 (VEGFR2) expression in squamous cell carcinomas of the head and neck.

OBJECTIVES: Vascular endothelial growth factor receptor 2 (VEGFR2; Flk-1 [fetal liver kinase]/KDR [kinase insert domain containing receptor]) has been identified as a high affinity receptor for vascular endothelial growth factor (VEGF) on vascular endothelium. Head and neck squamous cell carcinomas (HNSCC) have already been shown to produce substantial amounts of VEGF. VEGFR2 is supposed to play a major role in tumor-neoangiogenesis. METHODS: We investigated 24 tumor specimens and 4 HNSCC cultured tumor cell lines for the incidence and distribution of VEGFR2 by immunohistochemistry using monoclonal antibodies (mAbs) and RT-PCR. RESULTS: Analysis of frozen sections by immunohistochemistry showed that in 90% of tumor specimens VEGFR2-positive cells were found which were associated with vascular endothelium. VEGFR2 was also expressed on tumor cells and vessels, which was confirmed by double immunolabeling of tumor cells with an a-cytokeratin mAb. Furthermore, 2 (JPPA, SCC9) of 4 HNSCC cultured tumor cell lines revealed positive VEGFR2 immunoreactivity. Synthesis of VEGFR2 mRNA on all 4 HNSCC cultured tumor cell lines (JPPA, SCC9, SCC25, and LFFR) and in 6 tumor specimens was confirmed by RT-PCR. In conclusion, our results showed that VEGFR2 is expressed in HNSCCs on tumor cells. VEGFR2 expression is associated with the beginning of vasculogenesis represented by accumulation of VEGFR2-positive cells budding into new vessels ("hot spots"). The focal expression pattern of VEGFR2 on tumor cells suggests an autocrine loop for VEGF in tumor cell growth.

Protein phosphatases 1 and 2A maintain endothelial cells in a resting state, limiting the motility that is needed for the morphogenic process of angiogenesis.

Angiogenesis that is induced by cancers, including those of the head and neck, requires endothelial cells to shift from a nonmotile resting state to an increased level of motility. Using a human microvascular endothelial cell line, this study shows the importance of the serine/threonine protein phosphatases 1 (PP1) and 2A (PP2A) in restricting endothelial cell motility. Treatment of endothelial cells with increasing concentrations of the PP1 and PP2A inhibitor okadaic acid resulted in cell rounding and increased motility, which was accompanied by cytoskeletal disorganization involving a loss of filamentous beta-tubulin and F-actin. These effects occurred at okadaic acid levels that selectively inhibit PP2A and became more prominent with higher levels that inhibit both PP2A and PP1. This study shows the importance of PP1 and PP2A in maintaining cytoskeletal organization, thereby limiting endothelial cell motility, and suggests that pharmacologic approaches to enhance PP1 and PP2A activities may be useful in preventing key events of the angiogenic process.

Tumor grade, microvessel density, and activities of malate dehydrogenase, lactate dehydrogenase, and hexokinase in squamous cell carcinoma.

Squamous cell carcinomas were evaluated with respect to tumor differentiation (through use of hematoxylin and eosin stain), microvessel density (through use of CD-34 immunocytochemical stain), and magnitudes of malate dehydrogenase (MDH), hexokinase, and lactate dehydrogenase (LDH) enzyme activities. Direct correlations were found between tumor grade, MDH activity, and microvessel density. Direct correlations were also found between hexokinase activity and MDH activity and microvessel density. Inverse correlations were found between LDH activity and both tumor grade and MDH activity. These results suggest that the high rate of glucose utilization (indicated by hexokinase activity) found in more poorly differentiated tumors has a higher component of aerobic oxidative metabolism (indicated by MDH activity) and a relatively lower contribution from anaerobic metabolism (indicated by LDH activity) than do the rates found in more differentiated tumors. It is also suggested that as the glycolytic rate increases, more pyruvate goes into the Krebs cycle than into lactate. The availability of glucose-derived pyruvate for oxidative metabolism would mean less of a dependency on glutamine as a carbon source in squamous cell carcinoma.

Estimation of relative blood volume in head and neck squamous cell carcinomas.

PURPOSE: MR based first-pass method can be utilized to obtain hemodynamic information in the head and neck region. The purpose of this study was to estimate the regional relative blood volume (rBV) in head and neck tumors, which is useful for tumor staging and tumor biopsy. METHODS: Eighteen patients with head and neck tumors (17 squamous cell carcinomas, 1 hemangiopericytoma) were studied on a 1.5-T system. Conventional T1-weighted MR images and T2-weighted images and sequential T2*-weighted images were obtained. During repetitive image sequence acquisition, a bolus (0.2 mmol/kg) of gadopentetate dimeglumine was mechanically injected. Image processing of the dynamic raw data was performed on a pixel-by-pixel basis. RESULTS: Regional relative blood volume maps of the head and neck were successfully reconstructed in all (18/18) patients. The regional relative blood volume values within the tumor area of squamous cell carcinoma were 7.0 +/- 2.8, normalized on muscle, whereas the rBV of a single hemangiopericytoma was 11.6. The difference of rBV values of tumor and muscle was highly significant at statistical evaluation (p < 0.001). CONCLUSIONS: Relative blood volume imaging of head and neck tumors is valid using MR-based first-pass method. This method provides hemodynamic information which is not available from conventional MR imaging and is promising for further characterization of head and neck tumors

[Embolization by direct puncture of hypervascularized ORL tumors]. / Embolisation par ponction directe des tumeurs hypervasculaires de la sphère ORL.

Direct intratumoral embolization was used to treat five patients with vascularized tumors of the head and neck: 2 nasopharyngeal angiofibroma, 2 glomus jugulare tumors and 1 hemangiopericytoma. Embolization was performed by direct puncture and intratumoral injection of a plastic mixture under angiographic control. In 4 patients, embolization was performed preoperatively. Devascularization was induced in at least 90% of the volume of these tumors, thus facilitating subsequent surgical removal of the tumor. In 1 patient, embolization was performed with palliative intent. Good regression of symptoms was obtained.

Using intravital microscopy to observe bevacizumab-mediated anti-angiogenesis in human head and neck squamous cell carcinoma xenografts.

CONCLUSION: The study showed the value of using intravital microscopy (IVM) analysis for the study of neoangiogenesis. It demonstrated that the model and the analytical methodology could be used to evaluate in detail the effects of treatment strategies for solid tumours. OBJECTIVES: Neoangiogenesis is a key component of tumour progression, invasion and metastasis. In clinical trials monoclonal antibodies specific for vascular endothelial growth factor - VEGF (bevacizumab) - have been shown to significantly affect tumour progression when given in combination with standard chemotherapy, and also to improve the overall survival of patients. For squamous cell carcinoma of the head and neck (HNSCC), we still await definitive evidence of the effect of such treatment. The present study was designed to investigate the anti-angiogenesis effect of beviacizumab in green fluorescent protein (GFP)-labelled HNSCC xenografts using IVM technology. METHODS: We performed IVM and used image analysis for quantification of angiogenesis and of effects of bevacizumab on cell viability, combined with histochemical and immunohistochemical analysis to standardize the digital analysis of changes in tumour vascularization and cell viability. RESULTS: We found significant effects of bevacizumab on angiogenesis and cancer cell survival in HNSCC. Repeated injections of bevacizumab were found to provide the greatest effects.

[Angiogenesis: a new therapeutic target of thoracic and laryngopharyngeal carcinoma]. / L'angiogenèse: une nouvelle cible thérapeutique des cancers thoraciques et ORL.

Angiogenesis or new blood vessel formation is a complex and fundamental event in the process of tumor growth and metastatic dissemination. Actually, most of antiangiogenic agents target the VEGF considered like the most potent proangiogenic factor. These molecules directly inhibit VEGF or the kinase activity of its receptor (VEGFR) and represent a significant therapeutic progress in several solid tumors types. First clinical studies of antiangiogenic agents in thoracic and laryngopharyngeal carcinomas have shown promise mainly in combination with other therapies (chemotherapy, other targeted therapies or radiotherapy). Besides common antiangiogenic therapies-induced adverse events, risks of bleeding caused by tumor necrosis mainly in squamous cell lung carcinomas have been observed during early clinical trials. Assessment of surrogate markers of target inhibition could allow a better selection of patients able to benefit from antiangiogenic treatments eventually combined with chemotherapy or molecules targeting others metabolic pathways.

Whole-tumor perfusion CT parameters and glucose metabolism measurements in head and neck squamous cell carcinomas: a pilot study using combined positron-emission tomography/CT imaging.

BACKGROUND AND PURPOSE: Previous (separately performed) perfusion CT (PCT) and PET studies have been inconclusive regarding the correlation of functional tumor characteristics. The purpose of this study was to perform dual assessment of head and neck squamous cell carcinomas (SCCAs) to examine the relationship between perfusion measurements derived from PCT and glucose standardized uptake values (SUV). MATERIALS AND METHODS: We prospectively evaluated 15 primary and recurrent SCCAs using combined positron-emission tomography (PET) and CT of the head and neck. SUV(mean), SUV(max), blood flow (BF), blood volume (BV), mean transit time (MTT), and permeability (PS) values were calculated with use of manually drawn regions of interest (ROIs) over the lesions and the healthy muscle tissue. Parametric comparison test, correlation coefficients, and regression analysis were performed. RESULTS: The mean (+/- SD) SUV(mean), SUV(max), BF, BV, MTT, and PS values in the tumor tissue were 6.26 (+/- 1.48), 15.25 (+/- 3.81), 91.50 (+/- 24.69), 5.08 (+/- 1.17), 7.51 (+/- 2.24), and 23.08 (+/- 8.77), respectively. All PET/CT and PCT parameters of muscle versus tumor tissue were statistically different (.0001 < P < .001). There were significant correlations between BF and SUV(max) as well as SUV(mean) (r = 0.57; P = .02 and r = 0.63; P = .011, respectively) in the tumors. Significant correlation was also found between PS and SUV(mean) (r = 0.53; P = .04) in the tumors. Regression analysis showed: SUV(max) = 0.09 x BF + 7.2 (R(2) = 0.33; P = .02), SUV(mean) = 0.05 x BF + 2.22 (R(2) = 0.45; P = .011), and SUV(mean) = 0.05 x PS + 5.36 (R(2) = 0.35; P = .04). The tumor to nontumor (muscle) SUV(mean) and SUV(max) ratio was 9.45 (+/- 3.55) and 17.58 (+/- 4.32), respectively. BF-ratio SUV(mean) and BF-ratio SUV(max) showed significant correlations (r = 0.64; P = .01 and r = 0.53; P = .04, respectively). Regression analysis showed ratio SUV(mean) = 0.14 x BF-3.48 (R(2) = 0.42; P = .01) and ratio SUV(max) = 0.14 x BF + 4.51 (R(2) = 0.29; P = .04). CONCLUSION: Tissue perfusion-metabolic coupling is evident in head and neck SCCAs and may provide additional diagnostic information in patients undergoing PET/CT studies.

Molecular angiogenic signaling in angiofibromas after embolization: implications for therapy.

OBJECTIVES: To examine (1) the molecular angiogenic relationship between endothelial and stromal cells of angiofibromas and how this may elucidate the pathogenesis of angiofibromas and (2) the effects of embolization on the expression of angiotrophic factors and proapoptotic and antiapoptotic factors within the tumor. DESIGN: The expression of mesenchymal and endothelial stem/progenitor cell-associated proteins (MECAPs) such as proangiogenic cytokine vascular endothelial growth factor (VEGF), VEGF receptors (VEGFR1, VEGFR2, and VEGFR3), angiopoietin receptors (Tie-1 and Tie-2), and stem cell subset marker CD133 was assessed by immunohistological staining in 7 embolized angiofibroma specimens. Expression of proapoptotic Bax, antiapoptotic Bcl-2 and Bcl-xL, nuclear proliferation protein MiB-1, and hypoxia-inducible factor 1alpha (Hif-1alpha) in peri-ischemic areas of the embolized angiofibromas was also assessed. SETTING: A single pediatric institution. PATIENTS: Seven patients (identified from medical records, January 1, 2001, through December 31, 2005) who were diagnosed as having juvenile angiofibroma and who underwent surgical treatment. Archival tissues were retrieved for immunostaining. MAIN OUTCOME MEASURES: The immunostaining results were evaluated by microscopy and the staining intensities were also recorded. RESULTS: All angiofibroma specimens expressed the stem cell subset marker CD133 and MECAPs except VEGFR3 (a few cases). In the only case tested, we found evidence of VEGF-induced angiogenic signaling as the expression of phosphorylated VEGFR2 (Tyr951). Endothelial cells expressed VEGFR1 and VEGFR2 and angiopoietin receptors Tie-1 and Tie-2 but not VEGF. In contrast, VEGF was expressed within stromal cells. Viable tumor adjacent to the ischemic areas demonstrated increased staining intensities to VEGFR2, Tie-1, Tie-2 (all cases), and VEGFR3 (2 cases) and increased nuclear proliferation (5%-20%). All cases expressed proapoptotic and antiapoptotic factors, and the expression of Hif-1alpha was unaffected by ischemia. CONCLUSIONS: Stromal cells appear to be similar to mesenchymal stem cells with endothelial differentiation potential in umbilical cord blood cells. Stromal cells support endothelial growth by providing VEGF as a paracrine factor. Under ischemic stress, the embolized tumor tissues show upregulation of angiogenic receptors, retention of Hif-1alpha, and increased nuclear proliferation rates. Specific angiogenesis blockers may represent a novel treatment strategy for angiofibromas.

Modulation of tumor cell growth in vivo by extracellular matrix metalloprotease inducer.

OBJECTIVE: To investigate if loss of extracellular matrix metalloprotease inducer (EMMPRIN) will inhibit the growth of head and neck squamous cell carcinoma (HNSCC) tumor cell lines in vivo. Tumor cell-derived EMMPRIN is highly overexpressed in HNSCC and is thought to be induced by surrounding fibroblasts to stimulate matrix metalloproteases, which modulate tumor cell invasion, growth, and angiogenesis. DESIGN: In vivo study using FaDu tumor xenografts. SETTING: Academic research facility. SUBJECTS: Severe combined immunodeficiency (SCID) mice. INTERVENTIONS: The HNSCC cell line FaDu was transfected with EMMPRIN (FaDu/E), control vector (FaDu), or plasmid-expressing small-interfering RNA against EMMPRIN (FaDu/siE). Tumor cells combined with fibroblast cells were xenografted onto the flank of SCID mice. Tumors were measured biweekly over 4 weeks, at which time the mice were killed, and tumor samples were analyzed for proliferation (Ki-67 immunohistochemical analysis), vascularization (factor VIII staining), and apoptosis (TUNEL [terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling] assay). MAIN OUTCOME MEASURE: Growth of head and neck cancer cell lines genetically engineered to express variable levels of EMMPRIN. RESULTS: Tumor growth positively correlated and animal survival negatively correlated with increasing EMMPRIN expression. FaDu/E tumor growth was significantly larger at 4 weeks compared with FaDu tumors (P = .006). Similarly, the control vector-transfected FaDu tumors were significantly larger than FaDu/siE (P < .001). Immunohistochemical analysis demonstrated increased Ki-67 in EMMPRIN-transfected cells, without a significant change in the rate of apoptosis between groups. Vascular density and tumor formation rate also increased significantly with EMMPRIN expression. CONCLUSION: This study suggests that anti-EMMPRIN-targeted therapy may prove to be a novel treatment option in HNSCC.

[Selective embolization and surgical resection for head and neck glomus tumours--clinical outcome analysis]. / Präoperative Embolisation mit chirurgischer Resektion von Glomustumoren im Kopf-Hals-Bereich: Analyse der klinischen Ergebnisse.

The effectiveness and safety of angiographic embolization was investigated as a preliminary step prior to surgical excision of glomus tumours in the head and neck region. Embolization was performed in 54 patients presenting with a total of 58 chemodectomas, jugular (n=30), tympanicum (n=24) and caroticum (n=4) between the years 1988 and 2006. Embolization was considered successful if complete occlusion of all tumor-feeding vessels was achieved. The procedure was performed using polyvinylalcohol particles and microcoils and lasted for a median duration of 159 minutes. Complete tumor embolization was achieved in 72 % of patients. In 23%, it was partly successful and in 4% it was unsuccessful. 16% of patients experienced minor events during the procedure including hypotension, bradycardia, and vertigo. Following embolization, almost all patients (98%) had their tumour completely excised. Although the majority experienced minor postoperative complications (69%), one patient developed meningitis. There were no reported deaths. Angiographic embolization of glomus tumours in the head and neck before definitive excision can be safe and effective, resulting in an improved surgical outcome and tumour resectability.