Nasopharyngeal cancer (NPC) is a malignant tumor. In a recent publication, we described the presence and distribution of CD8+ T cells in NPC and used the information to identify ‘inflamed’, ‘immune-excluded’, and ‘desert’ immune phenotypes, where ‘inflamed’ and ‘immune-excluded’ NPCs were correlated with CD8 T cell infiltration and survival. Arguably, more detailed and, in particular, spatially resolved data are required for patient stratification and for the identification of new treatment targets. In this study, we investigate the phenotype of CD45+ leukocytes in the previously analyzed NPC samples by applying multiplexed tissue analysis to assess the spatial distribution of cell types and to quantify selected biomarkers. A total of 47 specified regions-of-interest (ROIs) were generated based on CD45, CD8, and PanCK morphological staining. Using the GeoMx® Digital Spatial Profiler (DSP), 49 target proteins were digitally quantified from the selected ROIs of a tissue microarray consisting of 30 unique NPC biopsies. Protein targets associated with B cells (CD20), NK cells (CD56), macrophages (CD68), and regulatory T cells (PD-1, FOXP3) were most differentially expressed in CD45+ segments within ‘immune-rich cancer cell islet’ regions of the tumor (cf. ‘surrounding stromal leukocyte’ regions). In contrast, markers associated with suppressive populations of myeloid cells (CD163, B7-H3, VISTA) and T cells (CD4, LAG3, Tim-3) were expressed at a higher level in CD45+ segments in the ‘surrounding stromal leukocyte’ regions (cf. ‘immune-rich cancer cell islet’ regions). When comparing the three phenotypes, the ‘inflamed’ profile (cf. ‘immune-excluded’ and ‘desert’) exhibited higher expression of markers associated with B cells, NK cells, macrophages, and myeloid cells. Myeloid markers were highly expressed in the ‘immune-excluded’ phenotype. Granulocyte markers and immune-regulatory markers were higher in the ‘desert‘ profile (cf. ‘inflamed’ and ‘immune-excluded’). In conclusion, this study describes the spatial heterogeneity of the immune microenvironment in NPC and highlights immune-related biomarkers in immune phenotypes, which may aid in the stratification of patients for therapeutic purposes.

Purpose Sinonasal malignancies (SNM) represent a rare and complex group of cancers that includes a wide range of histopathological subtypes. Data from population-based cohorts are scarce but warranted as a basis for randomized controlled treatment trials (RCTs). Our aim was to assess overall and histology subset-specific outcomes for SNM patients treated at a tertiary referral centre. Methods A retrospective, population-based, consecutive cohort of patients with SNMs diagnosed from 2001 through 2019 was examined. Outcome was analysed in relation to age, gender, site, stage, histopathology, and treatment. Results Two-hundred and twenty-six patients were identified, whereof 61% presented with stage IV disease. 80% completed treatment with curative intent, which comprised surgery with neoadjuvant (29%) or adjuvant (37%) radiotherapy, monotherapy with surgery (22%), definitive chemoradiotherapy (7%), or radiotherapy (5%). Median follow-up was 106 months. The 5- and 10-year overall survival rates were 57% and 35%, respectively. Median overall survival was 76 months (esthesioneuroblastoma: 147 months; adenocarcinoma: 117; salivary carcinoma: 88; mucosal melanoma: 69; squamous cell carcinoma: 51, undifferentiated carcinoma: 42; neuroendocrine carcinoma: 9; and NUT-carcinoma 5). The 5- and 10-year disease-free survival rates were 63% and 54%, respectively, and disease-specific survival 83% and 66%. Increasing age, stage IVB, melanoma histopathology, and treatment with definitive chemoradiotherapy emerged as significant independent prognostic risk factors for disease-specific mortality ( p ≤ 0.001). Conclusion The results indicate a seemingly good outcome in comparison to previous reports, particularly for mucosal melanoma, adenocarcinoma, and undifferentiated carcinoma. The study provides additional background for future RCTs focusing on histology subset-specific treatment for SNM.

Ectopic thyroid is a rare condition most often found at the base of the tongue, lingual thyroid (LT). The majority of patients are asymptomatic. Recently, transoral robotic surgery (TORS) has emerged as an option for definitive treatment. Here, we present a 20-year-old patient with symptomatic LT, treated with TORS-assisted surgery without adverse events and with depletion of symptoms. We advocate TORS as a very promising means to be considered when encountering the rare condition of symptomatic LT.

Background: Nasopharyngeal cancer (NPC) is frequently associated with the Epstein-Barr virus (EBV). Accordingly, attempts have been made to utilise the presence of non-human EBV-antigen on NPC cells therapeutically by immunological interventions. In this context, data are lacking with regard to levels of EBV antigen within the tumour as well as to intralesional presence and distribution of CD8+ T-cells and antigen-presenting dendritic cells (DCs) and their expression of targets that may favour cross-presentation of antigen (e.g., the pattern recognition receptor CD207). Arguably, such data may reflect specific “cancer immune phenotypes”, which in turn may be linked to clinical features of disease including survival. Furthermore, pattern recognition receptors may be targeted by specific adjuvants; γ-PGA-Phe nanoparticles may represent one such possibility. Methods: Fresh NPC biopsies were processed by multi-colour flow-cytometry focusing on DC subsets and their expression of the C-lectin receptor CD207. Formalin-fixated and paraffin-embedded NPC samples were retrieved and analysed for EBV-DNA quantitates and by immunohistochemistry focusing on CD207+ DCs and CD8+ T-cells. Specific cancer immune phenotypes were explored, based on presence and distribution of lymphocytes, and quantitates of intralesional CD207+ DCs and CD8+ T-cells were assessed with digital imaging. The association between the biological parameters and the clinical features of disease was examined. In a rat model, effects of topical exposure of γ-PGA-Phe nanoparticles on middle ear mucosa were examined. Results: EBV-DNA was present in NPC lesions (range: 0.0005-94617 copies/cell). The presence of CD8+ T-cells was particularly high in EBV-DNA-rich tumours. Subsets of DCs, i.e. CD123+ pDCs, CD1c+ mDCs, CD141+ mDCs, and CD1c-CD141-mDCs, were observed and a high frequency of CD207 expression was seen among CD1c+ mDCs compared to the other subsets. In NPC, CD207+ cells, likely representing CD1c+ mDCs, were particularly frequent in cancer cell areas. Specific “cancer immune phenotypes” were detected: “inflamed” (61.7%), “excluded” (29.8%), and “deserted” (8.5%) and CD8 ratios aggregated as “inflamed” > “excluded” > “deserted”. An EBV-DNA load of more than 70 copies/cell was associated with prolonged disease-free survival for EBV-DNA positive patients (p=0.046). Between immune phenotypes, a statistically significant difference in disease-free survival was observed between the “inflamed” and “excluded” subtypes (p=0.0090). γ-PGA-Phe nanoparticles produced a type-1 response characterized by generation of pro-inflammatory cytokines (IL-1α, IL-1β, IL-6, MIP-1α, and TNF-α) and inflammatory histopathological changes. Conclusions: Specific intralesional DC subpopulations are present in NPC and the C-lectin receptor CD207, involved in antigen cross-presentation, is frequently expressed. CD207+ DCs are constantly encountered among cancer cells in NPC. Lymphocyte-based cancer immune phenotypes can be demonstrated in NPC and this observation is confirmed by CD8+ T-cell quantification. The cancer immune phenotypes and intralesional EBV-DNA load predicts survival in NPC. γ-PGA-Phe NPs, antigen carriers with adjuvant properties, induces a local mucosal response compatible with a type-1 response. All these findings are relevant for future antigen-specific active immunotherapy for NPC.

Nasopharyngeal cancer (NPC) is associated with the Epstein-Barr virus (EBV). The clinical presentation and prognosis of NPC is well described, but not in relation to intralesional EBV-DNA load. In a retrospective design, 48 patients with NPC were examined. Patient history was re-evaluated, and diagnostic biopsies were re-examined. Furthermore, intralesional EBV-DNA was quantitated and HPV status determined. Cancer stage, disease-free survival (DFS), and overall survival (OS) were assessed. Of the 48 patients, 36 (75%) patients featured lesions that were positive for EBER (Epstein–Barr virus-encoded small RNA) and 40 (83%) were positive for EBV-DNA. Seven patients (15%) were HPV positive. The levels of EBV-DNA ranged from 0.0005 to 94617 copies/cell. An EBV-DNA load of more than 70 copies/cell was associated with a prolonged DFS for EBV-DNA positive patients treated with curative intent (p = 0.046). In conclusion, the EBV-DNA load in NPC lesions appears to vary greatly. For patients with EBV-DNA positive NPC treated with curative intent, an EBV-DNA load of more than 70 copies/cell is associated with a better outcome in terms of 7-year DFS.

Background: Transoral robotic surgery (TORS) assisted base of the tongue (BOT) resection has been suggested as part of the work-up for head and neck squamous cell cancer with unknown primary (HNSCC-CUP). Success rates vary with regard to identification of primary BOT lesions, and cases with likely such lesions appear to be included in previous reports.Objectives: To analyse the possible benefits of a superficial TORS-assisted BOT resection in thoroughly investigated HNSCC-CUP.Material and methods: Retrospectively, 13 patients subjected to superficial TORS-assisted BOT resections due to HNSCC-CUP, where previous thorough work-ups including PET-scans had been performed and primary lesions had still not been identified, were reviewed.Results: Nodal status, according to the TNM-8 classification, was N1, N2, N2a, N2b, N2c, and N3, respectively, for 7, 1, 1, 4, 0, and 0 patients. In 38% of the cases, T1 BOT cancers were identified using superficial TORS-assisted BOT resections and treatments adjusted.Conclusion: Addition of a superficial TORS-assisted BOT resection to the work-up of HNSCC-CUP frequently identifies primary lesions and alters the treatment for this group of patients even after thorough work-up.

Background: The five Nordic countries with a population of 27 M people form a rather homogenous region in terms of health care. The management of head and neck cancer is centralized to the 21 university hospitals in these countries. Our aim was to gain an overview of the volume and role of transoral robotic surgery (TORS) and to evaluate the need to centralize it in this area as the field is rapidly developing. Materials and Methods: A structured questionnaire was sent to all 10 Departments of Otorhinolaryngology-Head and Neck Surgery in the Nordic countries having an active programme for TORS in December 2017. Results: The total cumulative number of performed robotic surgeries at these 10 Nordic centers was 528 and varied between 5 and 240 per center. The median annual number of robotic surgeries was 38 (range, 5-60). The observed number of annually operated cases remained fairly low (<25) at most of the centers. Conclusions: The present results showing a limited volume of performed surgeries call for considerations to further centralize TORS in the Nordic countries.

Nasopharyngeal cancer (NPC) is associated with Epstein-Barr virus (EBV) and EBV antigen may be utilized for therapeutic purposes, including targeting of dendritic cells (DCs). Although DCs may be present in NPC, the information is limited and not up-to-date with current knowledge on DC subsets. In the present study, biopsies from untreated NPC were obtained and subjected to multicolor flow-cytometry focusing on DC subtype markers: CD123 for plasmacytoid DCs (pDCs); and CD1c and CD141 for myeloid DCs (mDCs). Furthermore, subset-specific expression of the C-lectin receptor (CLR) CD207 (also termed langerin) was assessed. pDCs and mDCs were detected in the NPC lesions, contributing to a frequency mean average of 0.78% of CD45+ leukocytes in situ. Different subpopulations, previously not described in NPC, were observed, including: CD123+ pDCs; CD1c+ mDCs; CD141+ mDCs; and CD1c-CD141- mDCs. A high frequency of CD1c+ mDCs expressing CD207 was observed, compared with other subsets. In conclusion, different DC subsets are present in NPC lesions. The CLR CD207, a selective endocytic marker on CD1c+ mDCs, may be targeted for therapeutic purposes to facilitate cross-presentation of antigens and aid cell-mediated antitumor effects.

Choristomas, or foregut duplication cysts, are benign embryonic tumors characterized by normal tissue appearing in abnormal sites. If they are lined with respiratory epithelium they are termed bronchogenic cysts. When present, these rare lesions are usually intra-thoracic. We here present a case of a 4-year-old boy diagnosed with a bronchogenic cyst in the tongue.

CONCLUSION:Amphiphilic biodegradable nanoparticles (NPs) composed of poly(γ-glutamic acid) conjugated with L-phenylalanine ethylester (γ-PGA-Phe NPs) applied on the rat middle ear mucosa produce an inflammatory type 1 response. The observation is of relevance for the use of γ-PGA-Phe NPs as a concomitant antigen delivery system and adjuvant measure in the context of vaccinations.OBJECTIVES:To examine effects of topical mucosal administration of γ-PGA-Phe NPs as a potentially combined antigen delivery system and adjuvant.METHODS:γ-PGA-Phe NPs were administered on rat middle ear mucosa in a sham-controlled design and the response was monitored, focusing on soluble markers in mucosal surface liquids and on overall histopathology.RESULTS:γ-PGA-Phe NPs produced a dose- and time-dependent inflammatory response characterized by generation of proinflammatory cytokines (IL-1α, IL-1β, IL-6, MIP-1α, and TNF-α) and associated histopathological changes.