Contribution à la compréhension de la dynamique de l’infiltrat immunitaire au cours de la carcinogénèse orale

Oral squamous cell carcinomas (OSCC), the 16th most common cancer worldwide, are associated with high morbidity and mortality, in particular due to frequent loco-regional/metastatic relapses (>50%) and second primary cancers (>30%). OSCC can develop from oral potentially malignant disorders (OMPD) with an estimated rate of malignant transformation of 3 to 50%, reflecting their heterogeneity. Despite recent therapeutic advances, the OSCC prognosis remains poor. To prevent the malignant transformation of OPMD (secondary prevention) as well as the development of relapse/second cancers (tertiary prevention), the immune microenvironment (IME) appears to be a promising therapeutic target provided its better characterization. Firstly, we sought to understand the dynamics of the IME during oral carcinogenesis by studying transcriptomic changes (laser microdissection and RNA-Sequencing) as well as immune infiltrates (immunohistochemistry) associated with histological changes (normal mucosa, hyperplasia, dysplasia, and invasive tumor) observed in a pre-clinical model of oral carcinogenesis: the 4-nitroquinoline (4- NQO) murine model. We report that most of the gene expression changes were observed in the stromal (not epithelial) compartment and were related to immune biological processes, involving the macrophagic population. This population was the most important quantitatively, especially at the stage of dysplasia. In 86 patients with OPMD, three M2 macrophage signatures were independently associated with better oral cancer-free survival. This first study has allowed us to better understand the dynamics of IME during oral carcinogenesis and provides a rationale for the development of immunoprevention strategies, in particular targeting the macrophage population for OPMD patients. Secondly, we used multiplex immunofluorescence (mIF) with two panels including markers for epithelial cells (PanCK), tumor infiltrating lymphocytes (TIL) (CD3, CD8) macrophages (CD68), inhibitory (PD-1, PD-L1, TIM-3, LAG-3, VISTA) or stimulatory (OX-40, ICOS) immune checkpoints (ICP) to characterize the IME in 44 treatment-naive OSCC patients, totaling 1246 mIF images. In addition to cell densities, spatial proximity and infiltration patterns were studied within four areas of interest: the tumor core, the invasion front, adjacent dysplasia, and adjacent non-tumor and non-dysplastic mucosa (also called adjacent normal mucosa). Finally, the mIF data were correlated with clinical and transcriptomic data. We found that OSCC are characterized by a remarkable intra-tumor’ and inter-patient’ infiltrates heterogeneity impacting on patient prognosis. Surprisingly, in the adjacent normal mucosa exclusively, a high density of PD-L1+/- macrophages and PD-1+ PD-L1+ T-cells was significantly associated with worse progression free survival. Spatial analyses showed that OSCC are characterized by remarkable intra-tumor’ and inter-patient’ infiltration patterns heterogeneity and by specific infiltration patterns called "PD-L1+ cells nests" whose presence in normal mucosa was significantly associated with worse prognosis. This study confirms the importance of the IME of OSCC and suggests that incorporation of its analysis including spatial analyses, especially in adjacent normal mucosa, may improve the prognostic evaluation of OSCC patients and the development of immunoprevention strategies.Les carcinomes épidermoïdes de la cavité orale (CECO), 16ième cancer le plus fréquent dans le monde, sont associé à une morbi-mortalité élevée, en particulier lié aux fréquentes rechutes loco-régionale/métastatique (>50%) et deuxième cancer primitifs (>30%). Les CECO peuvent se développer à partir de lésion orale à potentiel malin (LOPM) dont le taux de transformation maligne est estimé de 3 à 50%, témoignant de l(hétérogénéité de ces lésions. Malgré les récents progrès thérapeutique, le pronostic des CECO reste réservé. Afin de prévenir la transformation maligne des LOPM (prévention secondaire) ainsi que le développement de rechute/deuxième cancer (prévention tertiaire), le micro-environnement immunitaire (MEI) apparaît comme une cible thérapeutique prometteuse à condition de mieux le caractériser. Nous avons donc d’abord cherché à comprendre la dynamique du MEI durant la carcinogénèse orale en étudiant les changements transcriptomiques (microdissection laser et séquençage des ARN) ainsi que d’infiltrats immunitaires (immunohistochimie) associés aux changements histologiques (muqueuse normale, hyperplasie, dysplasie et tumeur invasive) observés dans un modèle pré-clinique de carcinogenèse orale : le modèle murin induit par le 4-nitroquinoléine (4- NQO). Nous rapportons que la plupart des changements d'expression génique ont été observés dans le compartiment stromal et sont liés à des processus biologiques immunitaires, impliquant en particulier la population macrophagique. Cette population était la plus importante quantitativement, en particulier au stade de la dysplasie. Chez 86 patients atteints de LOPM, trois signatures de macrophages M2 ont été indépendamment associées à une meilleure survie sans cancer oral. Cette première étude nous a permis de mieux comprendre la dynamique du MEI au cours de la carcinogenèse orale et apporte un rationnel pour le développement de stratégie d’immunoprévention, ciblant en particulier la population macrophagique, chez les patients porteurs de LOPM. En ce qui concerne la prévention tertiaire, nous avons utilisé l’immunofluorescence multiplex (mIF) avec deux panels incluant des marqueurs pour les cellules épithéliales (PanCK), des lymphocytes infiltrant la tumeur (TIL) (CD3, CD8), des macrophages (CD68), des points de contrôle immunitaire (ICP) inhibiteurs (PD-1, PD-L1, TIM-3, LAG-3, VISTA) ou stimulateurs (OX-40, ICOS) pour caractériser le MEI chez 44 patients atteints de CECO naïfs de traitement, totalisant ainsi 1246 images mIF. Outre les densités cellulaires, la proximité spatiale et les patterns d’infiltrations ont été étudiés au sein de quatre aires d’intérêt : le cœur tumoral, le front d'invasion, la dysplasie adjacente et la muqueuse adjacente non-tumorale et non-dysplasique (également appelée muqueuse normale adjacente). Enfin, les données mIF ont été corrélées avec les données cliniques et transcriptomiques. Nous avons mis en évidence trouvé que les CECO sont caractérisés par une remarquable hétérogénéité des densités d'infiltrats intra-tumorale et inter-patients ayant un impact sur le pronostic des patients. De manière surprenante, dans la zone adjacente non tumorale exclusivement, une forte densité de macrophages PD-L1+/- et de cellules T PD-1+ PD-L1+ était significativement associée à une plus mauvaise PFS. Les analyses spatiales ont montré que les CECO sont caractérisés par une hétérogénéité intra-tumorale et inter-patients remarquable des modèles d'infiltration et par des modèles d'infiltration spécifiques appelés "nids de cellules PD-L1+" dont la présence dans la muqueuse normale est significativement associée à un pronostic plus sombre. Cette étude confirme l'importance du MEI des CECO et suggère que l'incorporation de l'analyse du MEI incluant des analyses spatiales, en particulier dans la muqueuse normale adjacente, pourrait améliorer l'évaluation pronostique des patients atteints de CECO et également aider au développement de stratégies d’immunoprévention

Contribution à la compréhension de la dynamique de l’infiltrat immunitaire au cours de la carcinogénèse orale

Les carcinomes épidermoïdes de la cavité orale (CECO), 16ième cancer le plus fréquent dans le monde, sont associé à une morbi-mortalité élevée, en particulier lié aux fréquentes rechutes loco-régionale/métastatique (>50%) et deuxième cancer primitifs (>30%). Les CECO peuvent se développer à partir de lésion orale à potentiel malin (LOPM) dont le taux de transformation maligne est estimé de 3 à 50%, témoignant de l(hétérogénéité de ces lésions. Malgré les récents progrès thérapeutique, le pronostic des CECO reste réservé. Afin de prévenir la transformation maligne des LOPM (prévention secondaire) ainsi que le développement de rechute/deuxième cancer (prévention tertiaire), le micro-environnement immunitaire (MEI) apparaît comme une cible thérapeutique prometteuse à condition de mieux le caractériser. Nous avons donc d’abord cherché à comprendre la dynamique du MEI durant la carcinogénèse orale en étudiant les changements transcriptomiques (microdissection laser et séquençage des ARN) ainsi que d’infiltrats immunitaires (immunohistochimie) associés aux changements histologiques (muqueuse normale, hyperplasie, dysplasie et tumeur invasive) observés dans un modèle pré-clinique de carcinogenèse orale : le modèle murin induit par le 4-nitroquinoléine (4- NQO). Nous rapportons que la plupart des changements d'expression génique ont été observés dans le compartiment stromal et sont liés à des processus biologiques immunitaires, impliquant en particulier la population macrophagique. Cette population était la plus importante quantitativement, en particulier au stade de la dysplasie. Chez 86 patients atteints de LOPM, trois signatures de macrophages M2 ont été indépendamment associées à une meilleure survie sans cancer oral. Cette première étude nous a permis de mieux comprendre la dynamique du MEI au cours de la carcinogenèse orale et apporte un rationnel pour le développement de stratégie d’immunoprévention, ciblant en particulier la population macrophagique, chez les patients porteurs de LOPM. En ce qui concerne la prévention tertiaire, nous avons utilisé l’immunofluorescence multiplex (mIF) avec deux panels incluant des marqueurs pour les cellules épithéliales (PanCK), des lymphocytes infiltrant la tumeur (TIL) (CD3, CD8), des macrophages (CD68), des points de contrôle immunitaire (ICP) inhibiteurs (PD-1, PD-L1, TIM-3, LAG-3, VISTA) ou stimulateurs (OX-40, ICOS) pour caractériser le MEI chez 44 patients atteints de CECO naïfs de traitement, totalisant ainsi 1246 images mIF. Outre les densités cellulaires, la proximité spatiale et les patterns d’infiltrations ont été étudiés au sein de quatre aires d’intérêt : le cœur tumoral, le front d'invasion, la dysplasie adjacente et la muqueuse adjacente non-tumorale et non-dysplasique (également appelée muqueuse normale adjacente). Enfin, les données mIF ont été corrélées avec les données cliniques et transcriptomiques. Nous avons mis en évidence trouvé que les CECO sont caractérisés par une remarquable hétérogénéité des densités d'infiltrats intra-tumorale et inter-patients ayant un impact sur le pronostic des patients. De manière surprenante, dans la zone adjacente non tumorale exclusivement, une forte densité de macrophages PD-L1+/- et de cellules T PD-1+ PD-L1+ était significativement associée à une plus mauvaise PFS. Les analyses spatiales ont montré que les CECO sont caractérisés par une hétérogénéité intra-tumorale et inter-patients remarquable des modèles d'infiltration et par des modèles d'infiltration spécifiques appelés "nids de cellules PD-L1+" dont la présence dans la muqueuse normale est significativement associée à un pronostic plus sombre. Cette étude confirme l'importance du MEI des CECO et suggère que l'incorporation de l'analyse du MEI incluant des analyses spatiales, en particulier dans la muqueuse normale adjacente, pourrait améliorer l'évaluation pronostique des patients atteints de CECO et également aider au développement de stratégies d’immunoprévention.Oral squamous cell carcinomas (OSCC), the 16th most common cancer worldwide, are associated with high morbidity and mortality, in particular due to frequent loco-regional/metastatic relapses (>50%) and second primary cancers (>30%). OSCC can develop from oral potentially malignant disorders (OMPD) with an estimated rate of malignant transformation of 3 to 50%, reflecting their heterogeneity. Despite recent therapeutic advances, the OSCC prognosis remains poor. To prevent the malignant transformation of OPMD (secondary prevention) as well as the development of relapse/second cancers (tertiary prevention), the immune microenvironment (IME) appears to be a promising therapeutic target provided its better characterization. Firstly, we sought to understand the dynamics of the IME during oral carcinogenesis by studying transcriptomic changes (laser microdissection and RNA-Sequencing) as well as immune infiltrates (immunohistochemistry) associated with histological changes (normal mucosa, hyperplasia, dysplasia, and invasive tumor) observed in a pre-clinical model of oral carcinogenesis: the 4-nitroquinoline (4- NQO) murine model. We report that most of the gene expression changes were observed in the stromal (not epithelial) compartment and were related to immune biological processes, involving the macrophagic population. This population was the most important quantitatively, especially at the stage of dysplasia. In 86 patients with OPMD, three M2 macrophage signatures were independently associated with better oral cancer-free survival. This first study has allowed us to better understand the dynamics of IME during oral carcinogenesis and provides a rationale for the development of immunoprevention strategies, in particular targeting the macrophage population for OPMD patients. Secondly, we used multiplex immunofluorescence (mIF) with two panels including markers for epithelial cells (PanCK), tumor infiltrating lymphocytes (TIL) (CD3, CD8) macrophages (CD68), inhibitory (PD-1, PD-L1, TIM-3, LAG-3, VISTA) or stimulatory (OX-40, ICOS) immune checkpoints (ICP) to characterize the IME in 44 treatment-naive OSCC patients, totaling 1246 mIF images. In addition to cell densities, spatial proximity and infiltration patterns were studied within four areas of interest: the tumor core, the invasion front, adjacent dysplasia, and adjacent non-tumor and non-dysplastic mucosa (also called adjacent normal mucosa). Finally, the mIF data were correlated with clinical and transcriptomic data. We found that OSCC are characterized by a remarkable intra-tumor’ and inter-patient’ infiltrates heterogeneity impacting on patient prognosis. Surprisingly, in the adjacent normal mucosa exclusively, a high density of PD-L1+/- macrophages and PD-1+ PD-L1+ T-cells was significantly associated with worse progression free survival. Spatial analyses showed that OSCC are characterized by remarkable intra-tumor’ and inter-patient’ infiltration patterns heterogeneity and by specific infiltration patterns called "PD-L1+ cells nests" whose presence in normal mucosa was significantly associated with worse prognosis. This study confirms the importance of the IME of OSCC and suggests that incorporation of its analysis including spatial analyses, especially in adjacent normal mucosa, may improve the prognostic evaluation of OSCC patients and the development of immunoprevention strategies

Bilateral bifid condyles: A rare etiology of temporomandibular joint disorders.

Basckgroud: Bifid mandibular condyle (BMC) is a rare etiology of temporomandibular joint (TMJ) disorders characterized by a duplication of the head of the mandibular condyle. Case report: The authors report the case of a 20-year-old patient complaining of a painful and clicking TMJ and mandibular hypomobility, which had been progressing for several months. Radiological investigations (dental panoramic radiograph and X-ray CT scan) revealed right and left abnormalities of the TMJ due to bilateral BMC requiring surgical management. Conclusion: Despite a prevalence of 0.31% to 1.82% and the controversies surrounding its pathophysiology, maxillofacial surgeons should be aware of BMC to avoid misdiagnosis related to the clinical presentation (pain, clicking, hypomobility, or ankylosis) and provide adequate management

Functional and Aesthetic Factors Associated with Revision of Rhinoplasty

Background:. Surgical revision rate of rhinoplasty is from 5% to 15% in literature. The aims of our study were to define the rate and the predictive factors for surgical revision of rhinoplasty. Methods:. We have realized a single-center case/control study including 62 patients who underwent surgical revision among 732 patients who underwent closed rhinoplasty between 2005 and 2015. Data of each rhinoplasty were collected from medical records and photographs. Statistical analyses were used. Results:. The surgical revision rate was 8.6%. After multivariate analysis, 4 factors were statistically significant and independently associated with surgical revision: “preexisting respiratory functional disorder” [odds ratio OR = 3.30; 95% CI (1.47–7.76); P = 0.004], “wide nasal bone and side walls” [OR = 3.94; 95% CI (1.49–11.25); P = 0.007], “deviated nasal bone and side walls” [OR = 2.68; 95% CI [1.14–6.58]; P = 0.02] and the use of camouflage grafts [OR = 0.26; 95% CI [0.07–0.89]; P = 0.04]. Conclusions:. Closed rhinoplasties have similar revision rate to open techniques. Revision surgeries are justified by functional or aesthetic disorders. The interests of this study are to better inform patients and to adapt operative management. We provide here some recommendations with focus on the keys to successful rhinoplasty surgery

Challenges in lower face soft tissue reconstruction: The value of the historical bipedicled scalp flap procedure.

Dear Sir, Lower lip and chin reconstructions are challenging, since satisfactory aesthetic and function outcomes in the mouth region are difficult to achieve. Although various techniques for lower lip and chin reconstruction have been described, some cannot be applied in patients with disease recurrence, the failure of previous procedures, or a history of radiotherapy. [...

Endosteal blood supply of the mandible: anatomical study of nutrient vessels in the condylar neck accessory foramina.

In the mandible, the condylar neck vascularization is commonly described as mainly periosteal; while the endosteal contribution is still debated, with very limited anatomical studies. Previous works have shown the contribution of nutrient vessels through accessory foramina and their contribution in the blood supply of other parts of the mandible. Our aim was to study the condylar neck's blood supply from nutrient foramina. Six latex-injected heads were dissected and two hundred mandibular condyles were observed on dry mandibles searching for accessory bone foramina. Latex-injected dissections showed a direct condylar medular arterial supply through foramina. On dry mandibles, these foramina were most frequently observed in the pterygoid fovea in 91% of cases. However, two other accessory foramina areas were identified on the lateral and medial sides of the mandibular condylar process, confirming the vascular contribution of transverse facial and maxillary arteries. The maxillary artery indeed provided both endosteal and periosteal blood supply to the condylar neck, with three different branches: an intramedullary ascending artery (arising from the inferior alveolar artery), a direct nutrient branch and some pterygoid osteomuscular branches

A three-gene expression signature associated with positive surgical margins in tongue squamous cell carcinomas: Predicting surgical resectability from tumour biology?

International audienceObjectives: Achieving complete tumour resection is one of the main goals of surgery for head and neck squamous cell carcinoma (HNSCC) tumours. Whether biological characteristics of tumours contribute to the surgical resectability and the presence of positive surgical margins (SM) after resection of HNSCC is unclear. We aimed to address this issue.Materials and methods: We used data from The Cancer Genome Atlas (TCGA) to relate the SM status of 356 HNSCC tumours covering five major primary locations (tongue, larynx, tonsils, floor of mouth and buccal mucosa) with data from multiple omics approaches (transcriptomic, genomic and proteomic analyses).Results: We identified three differentially expressed genes whose expression was significantly associated with the presence of positive SM in tongue tumours (n = 144). The three genes (CCDC66, ZRANB2 and VCPKMT) displayed significantly higher mRNA levels in tongue tumours with positive SM compared to tumours with negative SM. The corresponding gene expression signature identified tongue tumours with a positive SM with high sensitivity and specificity (85% and 76%, respectively, Area Under the Curve (AUC) = 0.84). Tongue tumours with this signature were characterised by a high grade, elevated proliferation levels and a tumour stroma with fewer fibroblasts and endothelial cells.Conclusion: Positive SM were found to be strikingly associated with tumour biology in tongue tumours. These findings offer interesting perspectives for biomarker identification and precision surgery in these tumours

DIVA, a 3D virtual reality platform, improves undergraduate craniofacial trauma education

International audienceCraniofacial fractures management is challenging to teach due to the complex anatomy of the head, even when using three-dimensional CT-scan images. DIVA is a software allowing the straightforward visualization of CT-scans in a user-friendly three-dimensional virtual reality environment. Here, we assess DIVA as an educational tool for craniofacial trauma for undergraduate medical students. Three craniofacial trauma cases (jaw fracture, naso-orbital-ethmoid complex fracture and Le Fort 3 fracture) were submitted to 50 undergraduate medical students, who had to provide diagnoses and treatment plans. Each student then filled an 8-item questionnaire assessing satisfaction, potential benefit, ease of use and tolerance. Additionally, 4 postgraduate students were requested to explore these cases and to place 6 anatomical landmarks on both virtual reality renderings and usual slice-based three-dimensional CT-scan visualizations. High degrees of satisfaction (98%) without specific tolerance issues (86%) were reported. The potential benefit in a better understanding of craniofacial trauma using virtual reality was reported by almost all students (98%). Virtual reality allowed a reliable localization of key anatomical landmarks when compared with standard three-dimensional CT-scan visualization. Virtual reality interfaces such DIVA are beneficial to medical students for a better understanding of craniofacial trauma and allow a reliable rendering of craniofacial anatomy

Longitudinal assessment of PD-L1 expression and gene expression profiles in patients with head and neck cancer reveals temporal heterogeneity

International audienc