The transcription factor FOXM1 regulates the balance between proliferation and aberrant differentiation in head and neck squamous cell carcinoma

Sustained expression of FOXM1 is a hallmark of nearly all human cancers including squamous cell carcinomas of the head and neck (HNSCC). HNSCCs partially preserve the epithelial differentiation program, which recapitulates fetal and adult traits of the tissue of tumor origin but is deregulated by genetic alterations and tumor-supporting pathways. Using shRNA-mediated knockdown, we demonstrate a minimal impact of FOXM1 on proliferation and migration of HNSCC cell lines under standard cell culture conditions. However, FOXM1 knockdown in three-dimensional (3D) culture and xenograft tumor models resulted in reduced proliferation, decreased invasion, and a more differentiated-like phenotype, indicating a context-dependent modulation of FOXM1 activity in HNSCC cells. By ectopic overexpression of FOXM1 in HNSCC cell lines, we demonstrate a reduced expression of cutaneous-type keratin K1 and involucrin as a marker of squamous differentiation, supporting the role of FOXM1 in modulation of aberrant differentiation in HNSCC. Thus, our data provide a strong rationale for targeting FOXM1 in HNSCC. © 2019 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd

Head and neck cancer - prediction and modeling of regional progression

Head and neck squamous cell carcinoma (HNSCC), the sixth most common cancer worldwide, is responsible for over 500'000 new cases annually. Despite advances in surgical techniques and radiation therapy protocols, along with the emergence of targeted treatments and immunothérapies, the survival of HNSCC patients has not progressed in the last 20 years1. The presence of lymph node metastasis (LNM) is the most accurate predictor of cancer-related outcome in HNSCC patients2-4. Therefore, correct assessment of the neck is critical for risk stratification and treatment planning. Clinically negative neck (cNO) is defined as the absence of LNM based on pre-treatment clinical and radiological examinations. Currently, the recommended approach to treat cNO patients is to perform an elective neck dissection. As the prevalence of OLNM in only 20-30%, this means that at least 70% of neck dissections are useless. Sentinel node biopsy (SLNB) is a less invasive alternative to elective neck dissection, but SLNB is technically challenging and still requires an opening of the neck. In this work, we developed a différent approach, which enables the prédiction of occult LNM (OLNM) solely based on primary tumor features, thereby reserving neck dissection for high-risk patients. We showed that the lymphatic endothelial marker PROX1 and the pan-endothelial marker CD31, which are both expressed at the level of the primary tumor, were significantly associated with OLNM. We used a machine learning approach to combine CD31 and PROX1 with classical clinicopathological markers. This model reached 95% negative prédictive value while conserving a reasonably good positive prédictive value (PPV). While OLNM is a significant problem regarding the secondary effects of surgery, treatment failures account for a majority of HNSCC related deaths. To better understand how tumors relapse after surgery we had first to create a surgical mouse model. This model is based on the orthotopic implantation of mEERL5 and mEERL95 cells6 which initiate primary tumors in the neck, recapitulate the histology of poorly differentiated aggressive HNSCC and give rise to post-surgical récurrences. This model is the first immune-competent orthotopic mouse model of HNSCC disease progression suitable for pre-clinical studies. Post-surgical local, régional and distant relapses are initiated by cancer cell clones that escaped from the surgical field and subsequently promoted secondary tumor growth. To understand the clonal dynamics of tumor progression, mEERL, and mEERL95 cells were transduced with a highly complex genetic barcoding library. Following implantation of cells in the floor of the mouth of C57BL/6Rj mice, we micro-surgically resected primary tumors and followed mice until they developed local récurrence. Barcodes retrieved by PCR from genomic DNA were analyzed by next-generation sequencing (NGS). Barcoding revealed that HNSCC tumor progression is governed by a successive clonal shift aiong with an exponential enrichment of malignant clones following a stepwise advancement from the primary tumor and nodal métastasés to récurrent tumors, récurrent nodal and distant métastasés. The development of a prédictive model of OLNM now calls for a prospective randomized trial comparing RF model risk stratification versus elective neck dissection, whereas our insights on clonal évolution shall motivate novel approaches for the development of targeted therapies directed at advanced HNSCC cases. -- Les carcinomes épidermoïdes de la tête et du cou représentent la sixième cause de cancer dans le monde. Malgré de nombreuses avancées chirurgicales, radio- et immuno- thérapeutiques , la survie des patients ne s'est pas améliorées depuis près de 20 ans1. La présence de métastases ganglionnaires est le prédicteur le plus précis du pronostic de la maladie2-4. Les patients avec un cou cliniquement négatif (cNO), défini par l'absence de métastases ganglionnaires lors de l'examen initial, sont actuellement traité électivement par un évidement ganglionnaire, en raison du risque de métastases occultes, qui peut atteindre jusqu'à 30% des patients. En contrepartie, cela signifie que 70% des évidements cervicaux n'ont pas de réelle utilité. La biopsie du ganglion sentinelle est une technique alternative de pointe à la dissection ganglionnaire élective mais celle-ci demeure toujours invasive et requiert une collaboration étroite entre de nombreux spécialistes. Dans le présent travail nous avons montré que des marqueurs au niveau de la tumeur primaire, PROX1 et CD31, exprimés par les cellules endothéliales, étaient significativement associés à la présence de métastases ganglionnaires occultes. Nous avons utilisé une approche d'apprentissage automatique pour combiner ces marqueurs à des paramètres clinico- pathologiques classiques. Le modèle prédictif final a atteint une valeur prédictive négative de 95% tout en conservant une bonne valeur prédictive positive. Bien que les métastases ganglionnaires occultes soit un problème significatif, les échecs de traitements représentent la cause majeure de décès liés à cette maladie. Afin de mieux comprendre comment les tumeurs récidivent après la chirurgie, nous avons créé un nouveau modèle chirurgical murin. Ce dernier est basé sur l'implantation des cellules mEERL5 ou mEERL956 qui développent de manière reproductible des tumeurs primaires dans le cou, des récurrences locales, des métastases, et récapitulent bien l'histologie de carcinomes épidermoïdes agressifs et peu différenciés. Les récidives post-chirurgicales sont initiées par des clones cellulaires cancéreux qui ont échappés à la résection chirurgicale et permettent ainsi le développement de tumeurs secondaires. Pour comprendre la dynamique clonale des récurrences et métastases, nous avons transduit les cellules mEERL et mEERL95 avec une librairie hautement complexe de code-barres génétiques. Après l'implantation des cellules, nous avons réséqué micro-chirurgicalement les tumeurs primaires et suivi les animaux jusqu'à ce qu'ils développent des récurrences locales et des métastases. Les codes-barres ont été récoltés par PCR depuis de l'ADN génomique de cinq tissus différents, avant la déconvolution finale des codes- barres par des techniques de séquençage de nouvelle génération. Ceci a permis le suivi des clones à une échelle unicellulaire. En résumé, ceci nous a permis de mettre en valeur que la progression tumorale des carcinomes épidermoïdes de la tête et du cou est gouvernée par une transition clonale successive accompagnée d'un enrichissement exponentiel de clones malins, de la tumeur primaire à la récurrence locale, puis à aux récidives régionales et à distance. Le développement d'un modèle prédictif de métastases ganglionnaires occultes devra être suivi par un essai prospectif randomisé comparant une approche de stratification du risque à la dissection ganglionnaire élective. Quant à nos avancées concernant la description de l'évolution clonale, ces dernières devraient motiver de nouvelles approches pour le développement de thérapies ciblées des carcinomes épidermoïdes de la tête de du cou de stades avancés

Rosai-Dorfman disease presenting as stridor and hoarseness in a young female patient

Rosai-Dorfman disease is a non-Langherans cell histiocytosis typically revealed by lymphadenopathy. Extranodal involvement occurs in 43% and most commonly involves the head and neck, skin, and bones. Few reports have described laryngeal lesions. We report the case of a Rosai-Dorfman disease in a 27-year-old female, presenting as an obstructing transglottic mass. We provide the results of the MRI and PET-scanner examination. As the treatment relies on surgical excision and the diagnosis depends on pathological examination, we also detail the analysis results that followed the surgical resection. This report highlights the necessity to consider Rosai-Dorfman disease as a potential diagnosis in case of a laryngeal submucosal mass

Rosai-Dorfman disease presenting as stridor and hoarseness in a young female patient

Rosai-Dorfman disease is a non-Langherans cell histiocytosis typically revealed by lymphadenopathy. Extranodal involvement occurs in 43% and most commonly involves the head and neck, skin, and bones. Few reports have described laryngeal lesions. We report the case of a Rosai-Dorfman disease in a 27-year-old female, presenting as an obstructing transglottic mass. We provide the results of the MRI and PET-scanner examination. As the treatment relies on surgical excision and the diagnosis depends on pathological examination, we also detail the analysis results that followed the surgical resection. This report highlights the necessity to consider Rosai-Dorfman disease as a potential diagnosis in case of a laryngeal submucosal mass

Prediction of Occult Lymph Node Metastasis in Head and Neck Cancer with CD31 Vessel Quantification.

The management of occult lymph node metastasis (LNM) in head and neck squamous cell carcinoma has been a matter of controversy for decades. The vascular density within the tumor microenvironment, as an indicator of ongoing angiogenesis, could constitute an attractive predictor of LNM. The use of the panvascular endothelial antibody CD31 as a marker of occult LNM has never been reported. The aim of this study was to assess the predictive value of CD31 microvascular density for the detection of occult LNM in squamous cell carcinoma of the oral cavity and oropharynx. Case series with chart review. Tertiary university hospital. Intra- and peritumoral microvascular density values were determined in 56 cases of squamous cell carcinoma of the oral cavity (n = 50) and oropharynx (n = 6) with clinically negative necks using the CD31 marker. Statistical associations of CD31 microvascular densities with clinicopathologic data were then established. Peritumoral CD31 microvascular density was significantly associated with occult LNM in multivariate analysis ( P < .01). Recursive partitioning analysis for this parameter found a cutoff of 19.33, which identified occult LNM with a sensitivity of 91%, a specificity of 65%, a positive predictive value of 40%, a negative predictive value of 97%, and an overall diagnostic accuracy of 71%. Peritumoral CD31 microvascular density in primary squamous cell carcinoma of the oral cavity and oropharynx allows accurate prediction of occult LNM

CXCL9:SPP1 macrophage polarity identifies a network of cellular programs that control human cancers.

Tumor microenvironments (TMEs) influence cancer progression but are complex and often differ between patients. Considering that microenvironment variations may reveal rules governing intratumoral cellular programs and disease outcome, we focused on tumor-to-tumor variation to examine 52 head and neck squamous cell carcinomas. We found that macrophage polarity-defined by CXCL9 and SPP1 (CS) expression but not by conventional M1 and M2 markers-had a noticeably strong prognostic association. CS macrophage polarity also identified a highly coordinated network of either pro- or antitumor variables, which involved each tumor-associated cell type and was spatially organized. We extended these findings to other cancer indications. Overall, these results suggest that, despite their complexity, TMEs coordinate coherent responses that control human cancers and for which CS macrophage polarity is a relevant yet simple variable