CXC ligand 13 orchestrates an immunoactive microenvironment and enhances immunotherapy response in head and neck squamous cell carcinoma.

Objectives: This study aims to systematically explore the role of chemokine CXC ligand 13 (CXCL13) in head and neck squamous cell carcinoma (HNSCC). Methods: The Genotype-Tissue Expression (GTEx) and The Cancer Genome Atlas (TCGA) databases provided the RNA-seq data for cancer and normal tissues, respectively. Gene set enrichment analysis was applied to search the cancer hallmarks associated with CXCL13 expression. TIMER2.0 was the main platform used to investigate the immune cell infiltration related to CXCL13. Immunohistochemistry was applied to explore the relationship between CXCL13 and patients' prognosis and the relationship between CXCL13 and tertiary lymphoid structures (TLSs). Results: The expression of CXCL13 was upregulated in most tumors, including HNSCC. The higher expression of CXCL13 was closely related to the positive prognosis of HNSCC. CXCL13 was mainly expressed in B cells and CD8 + T cells, revealing the relationship between its expression and immune activation in the tumor microenvironment. Furthermore, immunohistochemistry and multiple fluorescence staining analysis of HNSCC samples showed a powerful correlation between CXCL13 expression, TLSs formation, and positive prognosis. Finally, CXCL13 significantly increased the response to cancer immunotherapy. Conclusions: CXCL13 may function as a potential biomarker for predicting prognosis and immunotherapy response and associate with TLSs in HNSCC.

Yin Yang 1-Induced Long Noncoding RNA DUXAP9 Drives the Progression of Oral Squamous Cell Carcinoma by Blocking CDK1-Mediated EZH2 Degradation.

LncRNAs play a critical role in oral squamous cell carcinoma (OSCC) progression. However, the function and detailed molecular mechanism of most lncRNAs in OSCC are not fully understood. Here, a novel nuclear-localized lncRNA, DUXAP9 (DUXAP9), that is highly expressed in OSCC is identified. A high level of DUXAP9 is positively associated with lymph node metastasis, poor pathological differentiation, advanced clinical stage, worse overall survival, and worse disease-specific survival in OSCC patients. Overexpression of DUXAP9 significantly promotes OSCC cell proliferation, migration, invasion, and xenograft tumor growth and metastasis, and upregulates N-cadherin, Vimentin, Ki67, PCNA, and EZH2 expression and downregulates E-cadherin in vitro and in vivo, whereas knockdown of DUXAP9 remarkably suppresses OSCC cell proliferation, migration, invasion, and xenograft tumor growth in vitro and in vivo in an EZH2-dependent manner. Yin Yang 1 (YY1) is found to activate the transcriptional expression of DUXAP9 in OSCC. Furthermore, DUXAP9 physically interacts with EZH2 and inhibits EZH2 degradation via the suppression of EZH2 phosphorylation, thereby blocking EZH2 translocation from the nucleus to the cytoplasm. Thus, DUXAP9 can serve as a promising target for OSCC therapy.

Targeting cellular metabolism in head and neck cancer precision medicine era: A promising strategy to overcome therapy resistance.

Head and neck squamous cell carcinoma (HNSCC) is among the most prevalent cancer worldwide, with the most severe impact on quality of life of patients. Despite the development of multimodal therapeutic approaches, the clinical outcomes of HNSCC are still unsatisfactory, mainly caused by relatively low responsiveness to treatment and severe drug resistance. Metabolic reprogramming is currently considered to play a pivotal role in anticancer therapeutic resistance. This review aimed to define the specific metabolic programs and adaptations in HNSCC therapy resistance. An extensive literature review of HNSCC was conducted via the PubMed including metabolic reprogramming, chemo- or immune-therapy resistance. Glucose metabolism, fatty acid metabolism, and amino acid metabolism are closely related to the malignant biological characteristics of cancer, anti-tumor drug resistance, and adverse clinical results. For HNSCC, pyruvate, lactate and almost all lipid categories are related to the occurrence and maintenance of drug resistance, and targeting amino acid metabolism can prevent tumor development and enhance the response of drug-resistant tumors to anticancer therapy. This review will provide a better understanding of the altered metabolism in therapy resistance of HNSCC and promote the development of new therapeutic strategies against HNSCC, thereby contribute to a more efficacious precision medicine.

Unicystic ameloblastoma: A retrospective study on recurrent factors from a single institute database.

OBJECTIVE: Unicystic ameloblastomas are a variant of ameloblastoma with a definite recurrence rate because of the biological behaviours of the tumour. The risk factors associated with disease recurrence were analysed in this retrospective study. METHODS: A total of 132 patients with primary unicystic ameloblastoma reported in a tertiary hospital from 2005 to 2015 were analysed to identify the clinic-pathological and radiological factors associated with recurrence using univariate and multivariate Cox regression analyses. RESULTS: The mean volume was 30.54cm3  ± 12.55 cm3 , and this value differed significantly according to recurrence (p < 0.001). Root resorption and bone cortex/soft tissue invasion were also significantly associated with recurrence among unicystic ameloblastoma patients (p = 0.017 vs. p < 0.001, respectively). A new stage classification system was developed to predict disease recurrence of patients. The multivariate Cox regression analysis revealed that the new stage classification system was the only predictor of disease recurrence in unicystic ameloblastoma patients (p < 0.001), regardless of root resorption, position and site characteristics. CONCLUSIONS: Volume, root resorption and bone cortex/soft tissue invasion were risk factors for disease recurrence among unicystic ameloblastoma patients. The new stage classification was an independent predictor of disease recurrence in patients with unicystic ameloblastoma.

Application of mixed reality-based surgical navigation system in craniomaxillofacial trauma bone reconstruction.

OBJECTIVES: This study aimed to build a surgical navigation system based on mixed reality (MR) and optical positioning technique and evaluate its clinical applicability in craniomaxillofacial trauma bone reconstruction. Me-thods We first integrated the software and hardware platforms of the MR-based surgical navigation system and explored the system workflow. The systematic error, target registration error, and osteotomy application error of the system were then analyzed via 3D printed skull model experiment. The feasibility of the MR-based surgical navigation system in craniomaxillofacial trauma bone reconstruction was verified via zygomatico-maxillary complex (ZMC) reduction experiment of the skull model and preliminary clinical study. RESULTS: The system error of this MR-based surgical navigation system was 1.23 mm±0.52 mm, the target registration error was 2.83 mm±1.18 mm, and the osteotomy application error was 3.13 mm±1.66 mm. Virtual surgical planning and the reduction of the ZMC model were successfully conducted. In addition, with the guidance of the MR-based navigation system, the frontal bone defect was successfully reconstructed, and the clinical outcome was satisfactory. CONCLUSIONS: The MR-based surgical navigation system has its advantages in virtual reality fusion effect and dynamic navigation stability. It provides a new method for doctor-patient communications, education, preoperative planning, and intraoperative navigation in craniomaxillofacial surgery.

Cancer cells co-opt nociceptive nerves to thrive in nutrient-poor environments and upon nutrient-starvation therapies.

Although nutrient-starvation therapies can elicit strong anti-tumor effects in multiple carcinomas, it has been convincingly demonstrated that cancer cells exploit the tumor microenvironment to thrive in nutrient-poor environments. Here, we reveal that cancer cells can co-opt nociceptive nerves to thrive in nutrient-poor environments. Initially examining the low-glucose environment of oral mucosa carcinomas, we discovered that cancer cells employ ROS-triggered activation of c-Jun to secrete nerve growth factor (NGF), which conditions nociceptive nerves for calcitonin gene-related peptide (CGRP) production. The neurogenic CGRP subsequently induces cytoprotective autophagy in cancer cells through Rap1-mediated disruption of the mTOR-Raptor interaction. Both anti-glycolysis and anti-angiogenesis-based nutrient-starvation therapies aggravate the vicious cycle of cancer cells and nociceptive nerves and therapeutically benefit from blocking neurogenic CGRP with an FDA-approved antimigraine drug. Our study sheds light on the role of the nociceptive nerve as a microenvironmental accomplice of cancer progression in nutrient-poor environments and upon nutrient-starvation therapies.

A novel motionless calibration method for augmented reality surgery navigation system based on optical tracker.

Augmented reality (AR) surgery navigation systems display the pre-operation planned virtual model at the accurate position in the real surgical scene to assist the operation. Accurate calibration of the mapping relationship between the virtual coordinate and the real world is the key to the virtual-real fusion effect. Former calibration methods require the doctor user to conduct complex manual procedures before usage. This paper introduces a novel motionless virtual-real calibration method. The method only requires to take a mixed reality image containing both virtual and real marker balls using the built-in forward camera of the AR glasses. The mapping relationship between the virtual and real spaces is calculated by using the camera coordinate system as a transformation medium. The composition and working process of the AR navigation system is introduced, and then the mathematical principle of the calibration is designed. The feasibility of the proposed calibration scheme is verified with a verification experiment, and the average registration accuracy of the scheme is around 5.80mm, which is of same level of formerly reported methods. The proposed method is convenient and rapid to implement, and the calibration accuracy is not dependent on the user experience. Further, it can potentially realize the real-time update of the registration transformation matrix, which can improve the AR fusion accuracy when the AR glasses moves. This motionless calibration method has great potential to be applied in future clinical navigation research.

Classifying and standardizing panfacial trauma according to anatomic categories and Facial Injury Severity Scale: a 10-year retrospective study.

BACKGROUND: The purpose of this study was to identify the epidemiologic factors of panfacial fractures (PFs), and to evaluate the significance of anatomic PF categories and the Facial Injury Severity Scale (FISS) in classifying and standardizing panfacial injuries. METHODS: A retrospective review of all patients treated with PFs at our institution between June 2010 and April 2021 was performed. PF was defined as a concurrent fracture in at least 3 of 4 facial subunits (frontal, upper midface, lower midface, and mandible). Data regarding patient demographics, causes of injury, location of fractures, major concomitant injuries, and postinjury complications were collected, and the FISS score was collected from each patient. Statistical analysis was performed using IBM SPSS Statistics version 22.0. RESULTS: A total of 227 patients were enrolled. The most commonly fractured bones were the maxillary sinus wall (92.1%), mandible (82.8%), and zygomatic arch (75.3%), and the most common fracture sites in PFs were graphically presented. Four PF patterns were defined: FULM (n = 60), FUL (n = 39), ULM (n = 127), and FUM (n = 1). There was a significant association between PF patterns and sex (p = 0.018), the number of concomitant injuries (p = 0.014), and early surgical airway management (p = 0.003). Different PF patterns were significantly correlated with different types of concomitant injuries and complications. The FISS score showed a significant difference with PF patterns (p = 0.000) and sex (p = 0.007), and a FISS value of 11 or more is the appropriate cutoff for the prediction of multiple concomitant injuries and complications. CONCLUSIONS: Both the anatomic PF categories and FISS were significantly correlated with various concomitant injuries and complications. The combination of PF categories and FISS provided a better positive and negative prediction of concomitant injuries and complications for PF patients. Patients with FULM and FISS > 11 had an obviously higher proportion of the need for multiprofessional treatment.

Effect of the regulator of G-protein signaling 2 on the proliferation and invasion of oral squamous cell carcinoma cells and its molecular mechanism.

OBJECTIVES: This study aims to investigate the effect of the regulator of G-protein signaling 2 (RGS2) on the proliferation and invasion of oral squamous cell carcinoma (OSCC) cells and its potential molecular mechanism. Metho⁃ds The expression status and clinical significance of RGS2 in head and neck squamous cell carcinomas and matched adjacent normal tissues were evaluated using TCGA database. Three OSCC cell lines (i.e., SCC-9, Cal27, and Fadu) were overexpressed with RGS2, and the effect of RGS2 on cell proliferation and invasion was determined using the Transwell, clone formation, and cell counting kit (CCK)-8 assays. Moreover, the yeast two-hybrid scree-ning and co-immunoprecipitation (Co-IP) assays were conducted to detect the correlation of RGS2, four and a half LIM domains protein 1 (FHL1), and damage DNA-binding protein 1 (DDB1). RESULTS: The expression level of RGS2 in OSCC was significantly lower than that in matched adjacent normal tissues (P=0.023). The high RGS2 expression level was negatively correlated with lymphovascular invasion (P<0.001). After transfection with lentiv-RGS2, the expression of RGS2 was increased, and the invasion and proliferation abilities of OSCC cell lines were evidently inhibited. FHL1 could competitively bind with RGS2, which decreased the integration of DDB1 and RGS2, inhibited the ubiquitination process of RGS2, and maintained the stability of the RGS2 protein. CONCLUSIONS: RGS2 plays an important role in the inhibition of OSCC proliferation and invasion. The structure stability of RGS2 is competitively regulated by FHL1 and DDB1.

Dysregulation of FOXD2-AS1 promotes cell proliferation and migration and predicts poor prognosis in oral squamous cell carcinoma: a study based on TCGA data.

FOXD2 adjacent opposite strand RNA 1 (FOXD2-AS1) plays an important role in the pathogenesis of some cancers. However, its functional role in oral squamous cell carcinoma (OSCC) remains largely unknown. In this study, we conducted expressional and functional analyses of FOXD2-AS1 using data from the Cancer Genome Atlas (TCGA) and in vitro OSCC assays. FOXD2-AS1 dysregulation was remarkably associated with radiation therapy, anatomic location, high histologic grade, and lymphovascular invasion (P < 0.05). A nomogram based on FOXD2-AS1 expression was constructed for use as a diagnostic indicator for OSCC patients, and multivariate cox regression analysis showed that FOXD2-AS1 expression was an independent prognostic factor for OSCC patients. KEGG, gene set enrichment analysis, and immune infiltration evaluations indicated that FOXD2-AS1 was involved in tumor progression via epithelial-to-mesenchymal transition and cell cycle regulation and was negatively associated with mast cell, DCs, iDCs, and B cells. FOXD2-AS1 silencing suppressed the proliferation and migration of Cal27 cells. Our findings showed that an aberrantly high FOXD2-AS1 expression predicts poor prognosis in OSCC; FOXD2-AS1 may act as an oncogenic protein by regulating cell proliferation and migration and may suppress adaptive immunity by modulating the number and function of antigen-presenting cells.

Calcitonin gene-related peptide: A promising bridge between cancer development and cancer-associated pain in oral squamous cell carcinoma.

Nerves have been widely demonstrated to exert major effects in tumor-associated microenvironments. Due to the characteristic innervation of the oral cavity and the fact that cancer-associated pain is a distinct feature of oral squamous cell carcinoma (OSCC), the sensory nerves may dominate in the OSCC-nerve microenvironment. As the most abundant neuropeptide in the trigeminal ganglion, the calcitonin gene-related peptide (CGRP) exerts a dual effect on cancer development and cancer-associated pain in various types of cancer. The present review explored the potential molecular mechanisms of the roles of CGRP in cancer development and cancer-associated pain, suggesting that CGRP may be a promising therapeutic target for OSCC.

Nerve growth factor (NGF)-TrkA axis in head and neck squamous cell carcinoma triggers EMT and confers resistance to the EGFR inhibitor erlotinib.

Understanding the molecular mechanisms regulating tumor dissemination and therapeutic resistance is of central importance for effective cancer therapies. Here, we report that nerve growth factor (NGF) and its receptor TrkA facilitate epithelial-mesenchymal transition (EMT) and EGFR inhibitor resistance via STAT3 activation in head and neck squamous cell carcinoma (HNSCC). Both NGF and TrkA expression were elevated in HNSCC, indicating poor clinical outcomes. NGF was highly expressed in cancer cells and nerves in perineural niche, whereas TrkA expression was higher in cancer cells with perineural invasion. The NGF/TrkA axis could promote HNSCC cell dissemination and trigger EMT via STAT3 activation. Moreover, we discovered that the NGF/TrkA axis conferred resistance to the EGFR inhibitor erlotinib via EMT processes in HNSCC cells. Blocking TrkA signaling markedly reversed EMT and sensitized HNSCC cells to erlotinib in both in vitro and in vivo models. Overall, our results demonstrate novel evidence that the paracrine NGF/TrkA axis favors EMT and confers EGFR-targeted therapeutic resistance in HNSCC.

Recurrence factors in pediatric ameloblastoma: Clinical features and a new classification system.

BACKGROUND: Ameloblastomas of jaw in the pediatric population are a rare clinical entity and have not been well addressed in the literatures. The present retrospective study analyzed the risk factors associated with recurrence of pediatric ameloblastomas. METHODS: Cases of primary pediatric ameloblastomas seen in a tertiary hospital between 2005 and 2015 were analyzed to identify the clinical factors associated with recurrence. RESULTS: There were a total of 104 cases of primary pediatric ameloblastomas. The overall mean maximum tumor diameter was 4.11 ± 1.339 cm. The receiver operating characteristic curve and the Youden Index showed an optimal cutoff point of 4.95 cm to accurately predict recurrence. Bone cortex/soft tissue invasion were associated with tumor recurrence (P < .001). CONCLUSIONS: The maximum tumor diameter, root resorption, and bone cortex/soft tissue invasion were risk factors for recurrence of pediatric ameloblastomas. The new classification system may serve as a predictor of recurrence in pediatric ameloblastomas.

GDNF secreted by nerves enhances PD-L1 expression via JAK2-STAT1 signaling activation in HNSCC.

Programmed death ligand 1 (PD-L1) functions as a key immune inhibitory factor by binding with its receptor, programmed death 1 (PD-1), to induce immune cell dysfunction and escape of the immune system. However, the mechanisms of PD-L1 expression under growth factor stimulation are not well characterized. Here, we demonstrate a novel role for glial cell line-derived neurotrophic factor (GDNF) in upregulating PD-L1 expression in head and neck squamous cell carcinoma (HNSCC). The expression and correlation of PD-L1, GDNF and perineural invasion (PNI) status were evaluated by bioinformatics analysis of TCGA database and IHC assays from 145 HNSCC patients. PD-L1 expression was investigated by flow cytometry, Western blot and real-time PCR analyses in HNSCC cells after GNDF incubation. The cell signaling pathways activated by GDNF were analyzed with an antibody array and blocked by specific signaling inhibitors in cancer cell lines. PD-L1 expression was significantly higher in cancer cells that exhibited PNI in the HNSCC specimens, and elevated PD-L1 expression was significantly correlated with GDNF levels. GDNF not only enhanced cancer cell PNI in a co-culture of dorsal root ganglions and cancer cells but also had a potent role in inducing PD-L1 expression through the JAK2-STAT1 signaling pathway. Moreover, a JAK2 inhibitor attenuated GDNF-induced PD-L1 and enhanced tumor cell susceptibility to NK cell killing. Our findings provide clinically novel evidence that nerve-derived GDNF can increase PD-L1 levels in cancer cells around the perineural niche and that regulatory signaling is critical for cancer cell escape from immune surveillance in the nerve-cancer microenvironment.

CD73 is associated with poor prognosis in HNSCC.

CD73 is a cell surface immunosuppressive enzyme involved in tumor progression and metastasis. While patients whose cancer cells express elevated CD73 are typically associated with an unfavorable outcome, the clinical impact of CD73 expression in patients with Head and neck squamous cell carcinoma (HNSCC) remains unclear. In the present study, we investigated the prognostic significance of CD73 in HNSCC using gene and protein expression analyses. Our results demonstrate that high levels of CD73 are significantly associated with reduced overall survival in patients with HNSCC. We also investigated the functional role of CD73 in vitro and demonstrated that CD73 promotes HNSCC migration and invasion through adenosine A3R stimulation and the activation of EGF/EGFR signaling. Moreover, in vivo xenograft studies demonstrated that CD73 promotes tumorigenesis. In conclusion, our study highlights a role for CD73 as a poor prognostic marker of patient survival and also as a candidate therapeutic target in HNSCCs.

Osteosarcoma of head and neck: A retrospective study on prognostic factors from a single institute database.

BACKGROUND: Osteosarcoma is a common bone malignancy occurring infrequently in head and neck region, the NCDB database of osteosarcoma suggests that the survival and prognosis of the head and neck osteosarcomas lie midway among other sites of occurrence, poorest survival is pelvic region and best is upper extremity. The influence of other prognostic factors independently effecting survival and recurrence are not studied widely because of scarce data even in databases. More over; these database are underrepresent eastern population. The authors institute treated around 160 head and neck osteosarcomas (HNOS) from 2007 to 2013 which were evaluated retrospectively for prognostic factors effecting survival and recurrence in specific population. PATIENTS AND METHOD: The historical records of patients treated for head and neck osteosarcomas from 2007 to 2013 were charted. The clinical and pathological factors affecting the local recurrence (LR), overall survival (OS), disease free survival (DFS), metastasis (MT) were analyzed in univariate and multivariate cox regression model for survival. RESULTS: A total of 160 HNOS patients were treated in the given time period, and 137 patients with follow up were analyzed. The median period of follow up was 3.067±0.356years for the alive patients. In the multivariate cox regression model for OS; surgical margin (p=0.000) was most significant, histological grade was borderline (p=0.062). For LR: surgical margin (p=0.002), histological subtype (p=0.048) and histological grade (p=0.024). For MT; surgical margin (p=0.000) was the significant factor. CONCLUSION: Histological grade and unclear margins are the significant independent prognostic factors effecting disease outcome of HNOS.

Elevated RET expression enhances EGFR activation and mediates EGFR inhibitor resistance in head and neck squamous cell carcinoma.

BACKGROUND AND AIM: Co-activation of EGFR by alternative receptor tyrosine kinases (RTKs) might mediate resistance to EGFR inhibition in head and neck squamous cell carcinoma (HNSCC). Here we found a novel mechanism to improve the efficacy of EGFR inhibitor erlotinib on HNSCC. METHOD: Immunohistochemistry, western blot, cell migration and invasion assays, cell proliferation, cell cycle analysis and in vivo serial transplantation assays were used to evaluate the role of RET on HNSCC cells. RESULTS: The elevated levels of a rearranged during transfection (RET) are observed in HNSCC and that high levels of RET correlate with increased tumor size, advanced tumor stage and decreased overall survival rate. The HNSCC cell proliferation and invasion were inhibited by RET knockdown in vitro and in vivo. The inhibition of RET expression markedly reduced EGFR phosphorylation and downstream EGFR signaling. The inhibition of RET signaling significantly increased the sensitivity of HNSCC cells to the EGFR inhibitor erlotinib in both in vitro and in vivo models. CONCLUSION: Our results offer a preclinical proof-of-concept supporting a role for RET signaling inhibition in a targeted therapeutic approach to improve the efficacy of EGFR inhibition in HNSCC.