Role of integrin β1 as a biomarker of stemness in head and neck squamous cell carcinoma Publication date: September 2019 Source: Oral Oncology, Volume 96 Author(s): Jung Hwa Moon, Young Soo Rho, Sang Hyuk Lee, Bon Seok Koo, Hyun Joo Lee, Sung Im Do, Jae Hoon Cho, Young Gyu Eun, Min Woo Park, Hyang Ae Shin, Young Chang Lim Abstract Objectives Signaling between cancer stem cells (CSC) and their extracellular matrix has a crucial role in CSC progression and maintenance. However, mediators of this signaling pathway in head and neck squamous cell carcinoma (HNSCC) are largely unknown. Here, we explored whether integrin β1, which is one of the key regulators of the communication between cells and their microenvironment, affected the stemness of HNSCC cells. Materials and methods We examined self-renewal capacity, chemoresistance, and xenograft tumorigenicity after knockdown of integrin β1 in primary HNSCC cells. In addition, we studied the role of focal adhesion kinase (FAK), an intracellular downstream molecule of integrin signaling, in influencing stemness of HNSCC. The relevance of Notch1 and integrin β1 interactions in HNSCC cells was also examined. Finally, immunohistochemical analysis was carried out to test whether the coexpression of integrin β1 and Notch1 in the samples from HNSCC patients correlated with their survival. Results Targeting integrin β1 in HNSCC cells inhibited self-renewal, chemoresistance, and in vivo tumor-forming capacity. Treatment with an inhibitor of FAK decreased self-renewal capacities and expression of various putative stem cell markers (Oct4, Sox2, and Nanog) in a dose-dependent manner. Moreover, knockdown of integrin β1 decreased the expression of Notch1 and its target genes (Hey1 and Hes1). Notably, HNSCC patients demonstrating simultaneous expression of integrin β1 and Notch1 in their tissue samples had significantly worse survival rate. Conclusion Integrin β1/Notch1 axis has a significant role in the regulation of stemness in HNSCC.

Non-invasive screening of a microRNA-based dysregulation signature in oral cancer and oral potentially malignant disorders Publication date: September 2019 Source: Oral Oncology, Volume 96 Author(s): T. Yap, C. Seers, K. Koo, L. Cheng, L.J. Vella, A.F. Hill, E. Reynolds, A. Nastri, N. Cirillo, M. McCullough Abstract Introduction We have previously shown that oral swirls are a robust source of microRNA protected by extracellular vesicles, potentially useful to detect oral squamous cell carcinoma (OSCC)-associated molecular aberration. Objectives To study a developed dysregulation score and risk classification algorithm based upon a panel of OSCC-associated microRNA in oral swirls from individuals with OSCC and oral potentially malignant disorders (OPMDs). Materials and methods An OSCC-associated panel of 5 microRNAs (miR-24; miR-21; miR-99a; let-7c; miR-100;) was quantified by qPCR in 190 individuals with and without mucosal abnormalities, including OSCC (n = 53) and OPMDs (n = 74). Each sample was analyzed using a developed dysregulation score (dSCORE) and risk classification algorithm, allocating a LOW- or HIGH-RISK score. The influence of demographic, systemic, oral health and mucosal disease factors on the developed test was analyzed. Results MicroRNA for analysis can be predictably isolated from oral swirls sourced from individuals with a range of demographic, systemic and oral health findings. Utilizing the presence of HIGH-RISK identified OSCC patients with 86.8% sensitivity and 81.5% specificity. Older age and female gender were associated with higher dSCOREs and higher proportions of HIGH-RISK classification amongst individuals with no mucosal abnormalities. The dSCOREs for all subgroups of OPMDs were significantly different from the OSCC group. Conclusion This is the first comparison of microRNA sourced from oral swirls from individuals with OPMDs with individuals with and without OSCC. A HIGH-RISK dysregulation signature was found to be accurate in indicating the presence of OSCC and exampled to parallel malignant transformation.

Value of diffusion MR imaging in differentiation of recurrent head and neck malignancies from post treatment changes Publication date: September 2019 Source: Oral Oncology, Volume 96 Author(s): Ankush Jajodia, Deepa Aggarwal, Arvind K. Chaturvedi, Avinash Rao, Vivek Mahawar, Munish Gairola, Mudit Agarwal, Sumit Goyal, Venkata Pradeep Babu Koyyala, Sunil Pasricha, Rupal Tripathi Abstract Purpose Role of diffusion-weighted (DW) MR imaging in differentiating residual or recurrent neck malignancies from postoperative/post-radiation changes with histopathological correlation and comparison with PET-CT. Methods and materials Prospective observational study for a period of 1 year in 62 post-radiation/post-operative patients suspected to have residual/recurrent tumors of neck with lesion diameter more than 5 mm measured on MRI. Results Mean ADC for recurrent/residual tumors: 1.008 ± 0.220 × 10−3 mm2/s - significantly lower than mean ADC value for post-treatment changes of 1.69 ± 0.40 × 10−3 mm2/s (p < 0.0001). The overall diagnostic accuracy, positive predictive value (PPV) and negative predictive value (NPV) of the qualitative assessment for the use of DWI in differentiating tumors recurrence from post-treatment changes were 96.6%, 96% and 83.3%, respectively. Upon quantitative analysis of the DW imaging data, a threshold ADC value of 1.3 × 10−3 mm2/s used for differentiating between post-treatment changes and recurrent cancers showed the highest combined sensitivity of 94%, specificity of 83.3%, accuracy of 93.6%, positive predictive value of 95.9%, and negative predictive value of 83.3%. Conclusion DW MRI is a promising non-invasive MRI technique used to differentiate recurrent/residual head and neck malignancies from posttreatment changes based on ADC values. DWI offers advantage as it has a short scanning time and can be safely added to standard MRI protocol with minimum patient discomfort. Complementary use of DWI and PET/CT imaging may increase diagnostic confidence for differentiating recurrent disease from radiation therapy-induced changes after 6–12 months in posttreatment cases.

Immune profiles in primary squamous cell carcinoma of the head and neck Publication date: September 2019 Source: Oral Oncology, Volume 96 Author(s): Vassiliki Saloura, Evgeny Izumchenko, Zhixiang Zuo, Riyue Bao, Michael Korzinkin, Ivan Ozerov, Alex Zhavoronkov, David Sidransky, Atul Bedi, Mohammad O. Hoque, Hartmut Koeppen, Michaela K. Keck, Arun Khattri, Nyall London, Nikita Kotlov, Aiman Fatima, Theodore Vougiouklakis, Yusuke Nakamura, Mark Lingen, Nishant Agrawal Abstract Objectives In this study we describe the tumor microenvironment, the signaling pathways and genetic alterations associated with the presence or absence of CD8+ T-cell infiltration in primary squamous cell carcinoma of the head and neck (SCCHN) tumors. Materials and Methods Two SCCHN multi-analyte cohorts were utilized, the Cancer Genome Atlas (TCGA) and the Chicago Head and Neck Genomics (CHGC) cohort. A well-established chemokine signature classified SCCHN tumors into high and low CD8+ T-cell inflamed phenotypes (TCIP-H, TCIP-L respectively). Gene set enrichment and iPANDA analyses were conducted to dissect differences in signaling pathways, somatic mutations and copy number aberrations for TCIP-H versus TCIP-L tumors, stratified by HPV status. Results TCIP-H SCCHN tumors were enriched in multiple immune checkpoints irrespective of HPV-status. HPV-positive tumors were enriched in markers of T-regulatory cells (Tregs) and HPV-negative tumors in protumorigenic M2 macrophages. TCIP-L SCCHN tumors were enriched for the β-catenin/WNT and Hedgehog signaling pathways, had frequent mutations in NSD1, amplifications in EGFR and YAP1, as well as CDKN2A deletions. TCIP-H SCCHN tumors were associated with the MAPK/ERK, JAK/STAT and mTOR/AKT signaling pathways, and were enriched in CASP8, EP300, EPHA2, HRAS mutations, CD274, PDCD1LG2, JAK2 amplifications. Conclusions Our findings support that combinatorial immune checkpoint blockade and depletion strategies targeting Tregs in HPV-positive and M2 macrophages in HPV-negative tumors may lead to improved antitumor immune responses in patients with TCIP-H SCCHN. We highlight novel pathways and genetic events that may serve as candidate biomarkers and novel targeted therapies to enhance the efficacy of immunotherapy in SCCHN patients.

Beavertail modification of the radial forearm free flap in total oral glossectomy reconstruction: Technique and functional outcomes Publication date: September 2019 Source: Oral Oncology, Volume 96 Author(s): Peter T. Dziegielewski, Jana Rieger, Mohamed A. Shama, Daniel A. O'Connell, Jeffrey R. Harris, Hadi Seikaly Abstract Objective The total oral tongue (anterior 2/3 glossectomy) defect is seldom addressed in the literature. This is the first series to describe a consistent technique for its reconstruction. The aim of the study is to describe the use of the beavertail modified radial forearm free flap (BTRFFF) to reconstruct a total oral tongue defect and the functional and quality of life outcomes associated with it. Study design Retrospective review of prospectively collected data from 2000 to 2010. Methods All patients at the University of Alberta undergoing head and neck free flap surgery are enrolled in a prospective functional outcomes program. Pre-operatively and at set post-operative time points patients complete videofluoroscopic swallowing studies (VFSS), speech evaluations and quality of life questionnaires (EORTC H&N-35). Peri-operative outcomes were also measured. Results 17 consecutive patients were included. All were gastrostomy tube free at 12 months post-operatively and tolerating a full soft diet with aspiration scores of 0. Swallowing transit times increased by a mean of 0.4 s (p = 0.32). Speech intelligibility remained high with mean sentence intelligibility at 75% and single word intelligibility at 62%. Quality of life scores returned to baseline and remained satisfactory. Complications related to the BTRFFF were limited to scarring. Conclusions The BTRFFF provides a robust reconstructive option for the total oral tongue defect with excellent long term functional outcomes and quality of life.

Transoral 980-nm/1470-nm dual-wavelength fiber laser microsurgery for early-stage glottic carcinoma Publication date: September 2019 Source: Oral Oncology, Volume 96 Author(s): Faya Liang, Zhiwen Xiao, Renhui Chen, Pin Han, Peiliang Lin, Yuzhang Huang, Xiaoming Huang Abstract Objectives To investigate the effective and safety of transoral 980-nm/1470-nm dual-wavelength fiber laser microsurgery for early-stage glottic carcinoma by compared with CO2 laser surgery. Materials and Methods From September 2015 to July 2018, 44 patients with early glottic carcinoma underwent transoral microsurgery were divided into 980-nm/1470-nm dual-wavelength fiber laser surgery (Dual-wavelength fiber laser group) and CO2 laser surgery (CO2 laser group). The operative time, number of other hemostatic devices used, postoperative blood loss, surgical complications and postoperative length of hospital stay The time of mucosal epithelialization and Voice Handicap Index-10(VHI-10) in pre-operation, 1-month postoperation and 6-month postoperation in both two groupswere retrospectively analyzed. Results All the patients underwent successful operation and all the tumors received en-bloc resection with negative margins. The median operative time in Dual-wavelength laser group was faster than CO2 laser group (32.00 min vs 37.50 min, p = 0.014). There was no statistically significant difference between the two groups in the median postoperative hospital stay and the median time of mucosal epithelialization. No patient need feeding tubes place temporarily or permanently in both two groups. Tongue numbness, tear of the palatal arch, postoperative vocal cord adhesion, VHI-10 score in Pre-operation, 1-month postoperation and 6-month postoperation were similar in both two group. No recurrence was reported in both groups during follow-up. Conclusion Compared to the CO2 laser surgery, transoral 980-nm/1470-nm dual-wavelength fiber laser microsurgery is a safe and feasible procedure for early-stage glottic carcinoma. It can provide clearer surgical field without hemorrhage and make the operation simpler, smoother and faster.

Treatment for oral squamous cell carcinoma: Impact of surgeon volume on survival Publication date: September 2019 Source: Oral Oncology, Volume 96 Author(s): Timothy Liu, Michael David, Owen Ellis, Tsu-Hui (Hubert) Low, Carsten E. Palme, Jonathan Clark, Martin Batstone Abstract Background The volume-outcome relationship is a well-known phenomenon in surgical oncology. The aim of this study was to quantify the impact of surgeon volume on the treatment outcome of oral squamous cell carcinoma (OSCC) patients. Methods All new OSCC cases treated with curative intent between 2008 and 2013 were included. A heterogeneous set of predictor variables was collected, including patient, tumour and treatment factors. The outcomes of interest were recurrence-free survival (RFS), overall survival (OS) and disease-specific survival (DSS). To investigate the cut-off in surgeon volume, the number of OSCC resections was analysed in multiplies of 5 cases per annum according to DSS, using univariable regression analysis. Results 534 cases were recruited. Independently, the negative predictors for patient survival were age, perineural invasion, worsening tumour staging, and extracapsular spread. High-volume surgeon was determined to be most significant at 20 cases per annum and significantly associated with improved RFS (HR: 0.67), OS (HR: 0.44), and DSS (HR: 0.39). Conclusions Results from this study support the rationalisation of OSCC management at high-volume centres and in the hands of experienced surgeons for better patient survival. Head and neck surgeons should perform a minimum of 20 OSCC cases per year to maintain competency in OSCC ablation.

Exome sequencing of oral leukoplakia and oral squamous cell carcinoma implicates DNA damage repair gene defects in malignant transformation Publication date: September 2019 Source: Oral Oncology, Volume 96 Author(s): Camile S. Farah, Maryam Jessri, Nigel C. Bennett, Andrew J. Dalley, Kate D. Shearston, Simon A. Fox Abstract Objectives To map the genomic pathways of patients with oral leukoplakia (OLK) which transformed to cancer (progressive) and those which did not (non-progressive), and to compare their exomic profiles. Materials and methods Whole exome sequencing was performed on 42 sequential samples from five progressive and eight non-progressive patients. Association of genomic variant frequencies with progression or lesion severity were analysed by non-parametric tests (Kruskal-Wallis and Mann-Whitney-Wilcoxon) and multivariate sparse partial least squares discriminant analysis (sPLS-DA). Enrichment analysis was used to characterise the effect of mutations upon biological pathways. Confirmatory studies used qPCR and immunohistochemistry. Results Using sPLS-DA, the variant frequency of a small number of genes could be used to classify the samples based on lesion severity or progressive status. Enrichment analysis showed that DNA damage repair gene related pathways were highly impacted in lesions which progressed to cancer. Multivariate analysis of a set of 148 DNA damage repair genes could be used to classify progressive lesions using mutation frequency. BRCA1, BRCA2 and other double strand break (DSB) repair Fanconi anaemia (FA)/BRCA pathway genes were prominent contributors to this classification. Conclusion Patients with progressive and non-progressive OLK can be differentiated using the frequency of exomic variants, particularly in DNA damage repair pathway genes. To our knowledge, this is the first report of FA/BRCA (DSB) pathway involvement in malignant transformation of OLK to oral squamous cell carcinoma (OSCC).

Elective neck dissection for salvage laryngectomy: A systematic review and meta-analysis Publication date: September 2019 Source: Oral Oncology, Volume 96 Author(s): Chen Lin, Sidharth V. Puram, Mustafa G. Bulbul, Rosh K. Sethi, James W. Rocco, Matthew O. Old, Stephen Y. Kang Abstract Objective Elective neck dissection (END) for salvage laryngectomy remains controversial due to variability in reported occult nodal metastasis rates and postoperative complications. We performed a meta-analysis to examine the role of END for treatment of the clinically N0 (cN0) neck in the salvage setting. Methods A PubMed search, without limit on years searched, was conducted for English language articles. Additional sources were found by reviewing bibliographies of pertinent articles. Studies had to include END data for salvage laryngectomy for locally recurrent squamous cell carcinoma of the larynx with clinically negative regional metastasis. For patients who underwent END, pathological node status had to be reported. Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) recommendations were followed. Data were pooled using a random-effects model. Results Nineteen studies were included in the analysis. Within the END group, 31% were supraglottic, 61% were glottic, 6% were transglottic, and 1% were subglottic. The pooled rate of occult nodal metastasis was 14% (95% CI = 0.11–0.17) for all subsites. In subsite-specific analyses, occult nodal metastasis rates were 24% for supraglottic, 9% for glottic, and 17% for transglottic recurrences. Occult nodal metastasis was higher in recurrent T3/4 tumors (21%) compared to recurrent T1/2 tumors (9%) (relative risk (RR) = 2.17, 95% CI = 1.23–3.63, p = 0.003). The RR of postoperative complications with END compared to observation was 1.72 (95% CI = 0.96–3.10, p = 0.07). Conclusions The highest rates of occult nodal metastasis are associated with supraglottic recurrence and recurrent T3/T4 tumors. These data should be considered when deciding whether to perform END for salvage laryngectomy.

Clinical benefits from endoscopy screening of esophageal second primary tumor for head and neck cancer patients: Analysis of a hospital-based registry Publication date: September 2019 Source: Oral Oncology, Volume 96 Author(s): Chen-Shuan Chung, Wu-Chia Lo, Kuan-Chih Chen, Cheng-Lu Lin, Ming-Hsun Wen, Chen-Hsi Hsieh, Shih-Chiang Lin, Li-Jen Liao Abstract Objectives Esophageal second primary tumors (SPTs) in head and neck cancer (HNC) patients is not uncommon. The impact of image-enhanced endoscopy (IEE) screening for esophageal SPT on the outcome of HNC patients has not been well clarified. Methods and methods Patients with malignancies of the head and neck region and esophagus were recruited from a hospital-based cancer registry between July 2000–December 2016. IEE screening included magnifying endoscopy with narrow-band imaging and chromoendoscopy with Lugol's solution. Biopsied specimens with revised Vienna classification categories 1 and 2 were defined as group I, and those with categories 3 to 5 were defined as group II. The Kaplan-Meier estimate and Cox regression model were used for survival analysis. Results Totally 1577 HNC and 501 esophageal cancer patients were enrolled. The 5-year overall survival (OS) rates of stage I/II HNC, stage III/IV HNC and esophageal cancer patients were 58%, 29%, and 8%, respectively (p < 0.01). The 5-year OS rate of HNC patients with negative IEE results was higher than that of HNC patients without IEE screening, followed by IEE screening groups I, II and esophageal cancer patients (44% vs. 39% vs. 35% vs. 11% vs. 8%, respectively, p for trend <0.01). Among advanced HNC patients, those who received IEE screening had a trend of better prognosis than those without screening (5-year OS rate of 31% vs. 28%, p = 0.17). Conclusions IEE screening for esophageal SPTs is helpful in risk stratification and prognosis prediction for HNC patients. Routine IEE screening is recommended in HNC patients.