Faculty Bibliography

The human homologue of the Drosophila segment polarity gene PTCH1, a tumor suppressor gene within the Sonic Hedgehog pathway has been implicated as the mutation responsible for nevoid basal cell carcinoma syndrome (NBCCS) as well as many other sporadic neoplasms. The calcifying epithelial odontogenic tumor (CEOT) is a rare and aggressive tumor of the jaws. The objective of this study was to investigate the role of the Sonic hedgehog pathway in the pathogenesis of the CEOT. We evaluated the protein distribution of PTCH and the transcription factors Gli1 and Gli2 within seven cases using immunohistochemistry. We also sought to confirm the findings by sequencing the PTCH1 gene from DNA extracted from the paraffin-embedded tissue of these cases. Seven cases of paraffin-embedded CEOT specimens were analyzed with immunohistochemistry. Immunoreactivity for Sonic hedgehog pathway proteins was evaluated using antibodies to the receptor PTCH as well as to the transcription factors Gli1 and Gli2. A keratocystic odontogenic tumor (KOT) from a 12year-old with NBCCS served as our positive control. Normal salivary gland tissue served as our negative control. PTCH gene sequencing was completed using PCR. Immunoreactivity to PTCH was seen in 6/7 cases, to Gli1 in 6/7 cases and to Gli2 in 6/7 cases. All three proteins were positive in the syndromic KOT and all proteins were negative in normal salivary tissue. Gene sequencing revealed five single-nucleotide polymorphisms (SNPs) of which two resulted in missense mutations. A missense mutation was also detected in the KOT. This study is the first to implicate the Sonic hedgehog pathway in the pathogenesis of the CEOT through sequencing. Similar to other odontogenic neoplasms gene mutations in PTCH1 are present in the CEOT

Endothelin-1 (ET-1) produced by various cancers is known to be responsible for inducing pain. While ET-1 binding to ETAR on peripheral nerves clearly mediates nociception, effects from binding to ETBR are less clear. The present study assessed the effects of ETBR activation and the role of endogenous opioid analgesia in carcinoma pain using an orthotopic cancer pain mouse model. mRNA expression analysis showed that ET-1 was nearly doubled while ETBR was significantly down-regulated in a human oral SCC cell line compared to normal oral keratinocytes (NOK). Squamous cell carcinoma (SCC) cell culture treated with an ETBR agonist (10(-4)M, 10(-5)M, and 10(-6) M BQ-3020) significantly increased the production of beta-endorphin without any effects on leu-enkephalin or dynorphin. Cancer inoculated in the hind paw of athymic mice with SCC induced significant pain, as indicated by reduction of paw withdrawal thresholds in response to mechanical stimulation, compared to sham-injected and NOK-injected groups. Intratumor administration of 3mg/kg BQ-3020 attenuated cancer pain by approximately 50% up to 3h post-injection compared to PBS-vehicle and contralateral injection, while intratumor ETBR antagonist BQ-788 treatment (100 and 300microg/kg and 3mg/kg) had no effects. Local naloxone methiodide (500microg/kg) or selective mu-opioid receptor antagonist (CTOP, 500microg/kg) injection reversed ETBR agonist-induced antinociception in cancer animals. We propose that these results demonstrate that peripheral ETBR agonism attenuates carcinoma pain by modulating beta-endorphins released from the SCC to act on peripheral opioid receptors found in the cancer microenvironment

Mediators involved in the generation of pain in patients with cancer are poorly understood. Using a combined molecular, pharmacologic, behavioral, and genetic approach, we have identified a novel mechanism of cancer-dependent allodynia induced by protease-activated receptor 2 (PAR2). Here we show that human head and neck carcinoma cells have increased levels of proteolytic activity compared to normal human cell controls. Supernatant from human carcinoma cells, but not controls, caused marked and prolonged mechanical allodynia in mice, when administered into the hindpaw. This nociceptive effect was abolished by serine protease inhibition, diminished by mast cell depletion and absent in PAR2-deficient mice. In addition, non-contact co-culture of trigeminal ganglion neurons with human head and neck carcinoma cells increased the proportion of neurons that exhibited PAR2-immunoreactivity. Our results point to a direct role for serine proteases and their receptor in the pathogenesis of cancer pain. This previously unrecognized cancer pain pathway has important therapeutic implications wherein serine protease inhibitors and PAR2 antagonists may be useful for the treatment of cancer pain

OBJECTIVE: To evaluate the early bone response to plateau root form dental implants with 4 different surface treatments. STUDY DESIGN: Surface treatments comprised (n = 12 each): as-machined (M), alumina-blasted/acid-etched (AB/AE), alumina-blasted/acid-etched + nanothickness bioceramic coating (Nano), and plasma-sprayed calcium phosphate (PSCaP). Implants were placed in the radius diaphyses of 12 beagle dogs, remaining in vivo for 3 and 5 weeks. After euthanasia, the implants were subjected to torque to interface fracture and subsequently nondecalcified for histomorphology. Statistical analysis was performed by a GLM analysis of variance model at 5% significance level. RESULTS: Torque to interface fracture was significantly greater for the PSCaP group than for other groups (P < .001). Histomorphologic analysis showed woven bone formation around all implant surfaces at 3 weeks, and its replacement by lamellar bone at 5 weeks. Time in vivo did not affect torque measures. CONCLUSION: The PSCaP surface increased the early bone biomechanical fixation of plateau root form implants.

The purpose of this study in rats was to identify whether a minimal dose of FK506 could enhance nerve regeneration along a 4-cm cross-chest saphenous nerve graft. Our center established a cross-chest nerve regeneration model previously using the contralateral C7 root transfer to the musculocutaneous nerve. Using this model, 10 adult male Sprague-Dawley rats were divided into two groups: group 1 (N = 5) consisted of animals that did not receive any further treatment, and group 2 (N = 5) consisted of animals that received a daily subcutaneous dose of 0.7 mg/kg FK506 for a period of 4 weeks. Evaluation methods of the study groups consisted of behavioral assessment, needle electromyography studies, and qualitative and quantitative morphometry. In the FK506 group, the middle of the graft and the musculocutaneous nerve contained larger axons and thicker myelin, bicep muscle weight recovered to an average of 68% of the normal (right) side, and overall behavioral results were better (P = 0.03175) than for untreated controls. Although the FK506 group achieved higher average myelinated fiber counts in all histologic sections, higher amplitude, and shorter latency results, there was no statistically significant difference between the two groups. Contralateral C7 transfer in the rat brachial plexus is a good experimental model to assess nerve regeneration and test treatments designed to enhance recovery in lesions with long nerve gaps (40 mm). FK506-treated animals demonstrated more advanced axonal regeneration, myelinated fiber maturation, and bicep muscle reinnervation. These results suggest a potential clinical use of low-dose FK506 in patients with severe nerve injuries

BACKGROUND: Half of the patients with head and neck squamous cell carcinoma (HNSCC) can be expected to fail therapy, indicating that more aggressive treatment is warranted for this group. We have developed a novel risk model that can become a basis for developing new treatment paradigms. Here we report on the performance of our model in a new multicenter cohort. DESIGN: Eligible patients from 3 institutions (Montefiore Medical Center, University of Manitoba, and New York University Medical Center) were identified and pathology slides from their resection specimens were reviewed by Margaret Brandwein-Gensler; risk category was assigned as previously published. Kaplan-Meier analysis was performed for disease progression and survival. Cox proportional hazards regression was performed, adjusted for potential confounders. A teaching module was also developed; attending pathologists were asked to score coded slides after a lecture and multiheaded microscope teaching session. Agreement was assessed by calculating Cohen unweighted kappa coefficients. RESULT: The validation cohort consisted of 305 patients, from the above institutions, with 311 primary HNSCC of the oral cavity, oropharynx, and larynx. The median follow-up period for all patients was 27 months. Risk category predicts time to disease progression (P=0.0005), locoregional recurrence (P=0.013), and overall survival (P=0.0000) by Kaplan-Meier analysis. High-risk status is significantly associated with decreased time to disease progression, adjusted for clinical confounders (P=0.015, hazard ratio 2.32, 95% confidence interval 1.18-4.58) compared with collapsed intermediate and low-risk groups. We also demonstrate substantial interrater agreement (kappa=0.64), and very good rater agreement when compared with the standard (kappa=0.87). CONCLUSIONS: We demonstrate significant predictive performance of the risk model in a new cohort of patients with primary HNSCC, adjusted for confounders. Our training experience also supports the feasibility of adapting the risk model in clinical practice

The purpose of this project was to study the effect of a subimmunosuppressive dose of FK506 (0.7 mg/kg per day) on nerve regeneration along a long nerve gap (4 cm), using the contralateral C7 nerve root transfer model for musculocutaneous nerve neurotization. Two types of tubes were applied to the nerve gap: a polycaprolactone tube and a collagen tube. Twenty adult male Sprague-Dawley rats were divided into 4 groups (n = 5). A polycaprolactone was used in groups 1 and 3 and a collagen tube in groups 2 and 4. Groups 1 and 2 were daily administered a subimmunosuppressive dose of FK506. Animals were euthanized on day 30. Evaluation consisted of behavioral assessment, needle electromyography studies, biceps muscle weight measurements, and qualitative and quantitative morphometry. Groups 1 and 2 showed higher mean values for fiber counts, axon diameters, myelin thickness and myelin area in C7, better functional evaluation results, and higher biceps weight left to right ratio than groups 3 and 4. There was no evidence of reinnervation potentials, and there were no axons detectable inside the tube lumen in any of the study groups. The present study demonstrated that there was nonsignificant improvement of the functional recovery, after systemic administration of a low dose of FK506. This was attributed to 3 factors: length of nerve gap; duration of follow up; and dose of FK506. However, FK506-treated animals tended to be in a more advanced stage of nerve regeneration compared with the control groups