Caractérisation d'un modèle cellulaire et animal orthotopique des cancers des VADS : du ciblage tumoral in vitro ou rôle de l'imagerie de fluorescence in vivo dans l'exérèse tumorale

Introduction: Targeted therapy of head and neck squamous cell carcinoma (HNSCC) requires the development of novel specific vectors that can deliver therapeutic molecules. These vectors could also be coupled to fluorophores to be used in near infrared fluorescence imaging-guided surgery.Objectives: The aim of our work is to test new targeted vectors of HNSCC and to study the role of the near infrared fluorescence imaging-guided surgery in HNSCC resection in a novel orthotopic animal model that we develop.Materials and Methods: The HNSCC cell line CAL33 is characterized in vitro and in vivo. Novel vectors that target one or more receptors of this cell line such as alpha v beta 3 integrin, EGFR and NRP1, are tested in vitro. Meanwhile, an orthotopic animal model of HNSCC is developed by implanting tumor fragments of CAL33 cells, in the oral cavity of nude mice. Surgical resection of orthotopic tumors is guided by the near infrared fluorescence imaging after systemic injection of RAFT-c[RGD]4 peptide coupled with a fluorophore. This peptide targets alpha v beta 3 integrin and is previously tested in vitro.Results: Our preliminary results show that bispecific vectors would present an increased binding to CAL33 cells in vitro. On the other hand, near infrared fluorescence imaging-guided surgery has a positive impact on the recurrence-free survival rate in our orthotopic model, by detecting fluorescent cancer foci that could remain unidentified if resection was performed exclusively under visual guidance. Our results show also that near infrared fluorescence imaging can also help to detect metastatic lymph nodes.Conclusion: Near-infrared fluorescence imaging-guided surgery improves the quality of tumor resection in our optimized orthotopic animal model of HNSCC. This preclinical stage is essential before testing this novel technique in humans.Introduction : La thérapie ciblée des cancers des VADS nécessite la mise au point de nouveaux vecteurs spécifiques. Ces vecteurs servent à acheminer des substances thérapeutiques, mais aussi ils peuvent être couplés à des fluorophores afin de les utiliser dans la chirurgie guidée par l'imagerie de fluorescence proche infrarouge.Objectifs : L'objectif de notre travail est de tester de nouveaux vecteurs des cancers des VADS et d'étudier l'apport de l'imagerie de fluorescence proche infrarouge dans la chirurgie des cancers des VADS chez un modèle animal orthotopique que nous mettons au point.Matériel et méthodes : La lignée cellulaire des cancers des VADS CAL33 est caractérisée in vitro et in vivo. De nouveaux vecteurs qui ciblent un ou plusieurs récepteurs des cellules CAL33 comme l'intégrine alpha v beta 3, l'EGFR et la NRP1, sont testés in vitro. Parallèlement, un modèle animal orthotopique des cancers des VADS est développé par implantation de fragments tumoraux des cellules CAL33, au niveau de la cavité buccale de la souris nude. La résection des tumeurs orthotopiques est guidée par l'imagerie de fluorescence proche infrarouge, après injection systémique du peptide RAFT-c[RGD]4 couplé à un fluorophore. Ce peptide cible l'intégrine alpha v beta 3 et est préalablement testé in vivo sur les cellules CAL33.Résultats : Nos résultats préliminaires montrent que certaines molécules bispécifiques présentent une liaison accrue in vitro aux cellules CAL33. Par ailleurs, la chirurgie guidée par l'imagerie de fluorescence proche infrarouge ciblant l'intégrine alpha v beta 3, présente un impact positif sur la survie sans rechute dans notre modèle orthotopique, à travers la détection de reliquats tumoraux qui pourraient passer inaperçus si l'exérèse tumorale avait été réalisée exclusivement d'une façon macroscopique. Elle permet aussi de détecter les adénopathies métastatiques.Conclusion : L'imagerie de fluorescence proche infrarouge améliore la qualité de l'exérèse tumorale dans notre modèle orthotopqiue optimisé des cancers des VADS. Cette étape préclinique est indispensable avant de tester cette technique chez l'être humain

Myeloid Sarcoma of the Uterine Cervix as Presentation of Acute Myeloid Leukaemia after Treatment with Low-Dose Radioiodine for Thyroid Cancer: A Case Report and Review of the Literature

The development of acute myeloid leukaemia after low-dose radioiodine therapy and its presentation as a myeloid sarcoma of the uterine cervix are both rare events. We report a case of acute myeloid leukaemia revealed by a myeloid sarcoma of the uterine cervix in a 48-year-old woman, 17 months after receiving a total dose of 100 mCi 131I for papillary thyroid cancer. A strict hematological follow-up of patients treated with any dose of 131I is recommended to accurately detect any hematological complications which might have been underestimated. Unusual presentations, such as chloroma of the uterine cervix, may reveal myeloid malignancy and should be kept in mind

Near-infrared fluorescence imaging-guided surgery improves recurrence-free survival rate in novel orthotopic animal model of head and neck squamous cell carcinoma

International audienceBackground. Appropriate animal models are required to test novel therapeutics for head and neck squamous cell carcinoma (HNSCC) such as near-infrared (NIR) imaging-guided surgery. Methods. We developed an optimized animal model of orthotopic HNSCC (in female athymic NMRI (Naval Medical Research Institute) nude mice) with a prolonged survival time. Resection of the orthotopic tumors was performed 30 days after implantation with or without the aid of a minia-turized clinical grade NIR optical imaging device, after systemic administration of a fluorescent RGD-based probe that targets a v b 3 integrin. Results. NIR optical imaging-guided surgery increased the recurrence-free survival rate by 50% through the detection of fluorescent cancer residues as small as 185 mm; these fragments could remain unidentified if resection was performed exclusively under unaided visual guidance. Conclusion. NIR optical imaging-guided surgery showed an improved HNSCC tumor resection quality in our optimized orthotopic animal model

Caractérisation d'un modèle cellulaire et animal orthotopique des cancers des VADS : du ciblage tumoral in vitro ou rôle de l'imagerie de fluorescence in vivo dans l'exérèse tumorale

Introduction: Targeted therapy of head and neck squamous cell carcinoma (HNSCC) requires the development of novel specific vectors that can deliver therapeutic molecules. These vectors could also be coupled to fluorophores to be used in near infrared fluorescence imaging-guided surgery.Objectives: The aim of our work is to test new targeted vectors of HNSCC and to study the role of the near infrared fluorescence imaging-guided surgery in HNSCC resection in a novel orthotopic animal model that we develop.Materials and Methods: The HNSCC cell line CAL33 is characterized in vitro and in vivo. Novel vectors that target one or more receptors of this cell line such as alpha v beta 3 integrin, EGFR and NRP1, are tested in vitro. Meanwhile, an orthotopic animal model of HNSCC is developed by implanting tumor fragments of CAL33 cells, in the oral cavity of nude mice. Surgical resection of orthotopic tumors is guided by the near infrared fluorescence imaging after systemic injection of RAFT-c[RGD]4 peptide coupled with a fluorophore. This peptide targets alpha v beta 3 integrin and is previously tested in vitro.Results: Our preliminary results show that bispecific vectors would present an increased binding to CAL33 cells in vitro. On the other hand, near infrared fluorescence imaging-guided surgery has a positive impact on the recurrence-free survival rate in our orthotopic model, by detecting fluorescent cancer foci that could remain unidentified if resection was performed exclusively under visual guidance. Our results show also that near infrared fluorescence imaging can also help to detect metastatic lymph nodes.Conclusion: Near-infrared fluorescence imaging-guided surgery improves the quality of tumor resection in our optimized orthotopic animal model of HNSCC. This preclinical stage is essential before testing this novel technique in humans.Introduction : La thérapie ciblée des cancers des VADS nécessite la mise au point de nouveaux vecteurs spécifiques. Ces vecteurs servent à acheminer des substances thérapeutiques, mais aussi ils peuvent être couplés à des fluorophores afin de les utiliser dans la chirurgie guidée par l'imagerie de fluorescence proche infrarouge.Objectifs : L'objectif de notre travail est de tester de nouveaux vecteurs des cancers des VADS et d'étudier l'apport de l'imagerie de fluorescence proche infrarouge dans la chirurgie des cancers des VADS chez un modèle animal orthotopique que nous mettons au point.Matériel et méthodes : La lignée cellulaire des cancers des VADS CAL33 est caractérisée in vitro et in vivo. De nouveaux vecteurs qui ciblent un ou plusieurs récepteurs des cellules CAL33 comme l'intégrine alpha v beta 3, l'EGFR et la NRP1, sont testés in vitro. Parallèlement, un modèle animal orthotopique des cancers des VADS est développé par implantation de fragments tumoraux des cellules CAL33, au niveau de la cavité buccale de la souris nude. La résection des tumeurs orthotopiques est guidée par l'imagerie de fluorescence proche infrarouge, après injection systémique du peptide RAFT-c[RGD]4 couplé à un fluorophore. Ce peptide cible l'intégrine alpha v beta 3 et est préalablement testé in vivo sur les cellules CAL33.Résultats : Nos résultats préliminaires montrent que certaines molécules bispécifiques présentent une liaison accrue in vitro aux cellules CAL33. Par ailleurs, la chirurgie guidée par l'imagerie de fluorescence proche infrarouge ciblant l'intégrine alpha v beta 3, présente un impact positif sur la survie sans rechute dans notre modèle orthotopique, à travers la détection de reliquats tumoraux qui pourraient passer inaperçus si l'exérèse tumorale avait été réalisée exclusivement d'une façon macroscopique. Elle permet aussi de détecter les adénopathies métastatiques.Conclusion : L'imagerie de fluorescence proche infrarouge améliore la qualité de l'exérèse tumorale dans notre modèle orthotopqiue optimisé des cancers des VADS. Cette étape préclinique est indispensable avant de tester cette technique chez l'être humain

Characterization of a cellular and an orthotopic animal model of head and neck cancer : from in vitro tumor targeting to in vivo fluorescence imaging-guided tumor resection

Introduction : La thérapie ciblée des cancers des VADS nécessite la mise au point de nouveaux vecteurs spécifiques. Ces vecteurs servent à acheminer des substances thérapeutiques, mais aussi ils peuvent être couplés à des fluorophores afin de les utiliser dans la chirurgie guidée par l'imagerie de fluorescence proche infrarouge.Objectifs : L'objectif de notre travail est de tester de nouveaux vecteurs des cancers des VADS et d'étudier l'apport de l'imagerie de fluorescence proche infrarouge dans la chirurgie des cancers des VADS chez un modèle animal orthotopique que nous mettons au point.Matériel et méthodes : La lignée cellulaire des cancers des VADS CAL33 est caractérisée in vitro et in vivo. De nouveaux vecteurs qui ciblent un ou plusieurs récepteurs des cellules CAL33 comme l'intégrine alpha v beta 3, l'EGFR et la NRP1, sont testés in vitro. Parallèlement, un modèle animal orthotopique des cancers des VADS est développé par implantation de fragments tumoraux des cellules CAL33, au niveau de la cavité buccale de la souris nude. La résection des tumeurs orthotopiques est guidée par l'imagerie de fluorescence proche infrarouge, après injection systémique du peptide RAFT-c[RGD]4 couplé à un fluorophore. Ce peptide cible l'intégrine alpha v beta 3 et est préalablement testé in vivo sur les cellules CAL33.Résultats : Nos résultats préliminaires montrent que certaines molécules bispécifiques présentent une liaison accrue in vitro aux cellules CAL33. Par ailleurs, la chirurgie guidée par l'imagerie de fluorescence proche infrarouge ciblant l'intégrine alpha v beta 3, présente un impact positif sur la survie sans rechute dans notre modèle orthotopique, à travers la détection de reliquats tumoraux qui pourraient passer inaperçus si l'exérèse tumorale avait été réalisée exclusivement d'une façon macroscopique. Elle permet aussi de détecter les adénopathies métastatiques.Conclusion : L'imagerie de fluorescence proche infrarouge améliore la qualité de l'exérèse tumorale dans notre modèle orthotopqiue optimisé des cancers des VADS. Cette étape préclinique est indispensable avant de tester cette technique chez l'être humain.Introduction: Targeted therapy of head and neck squamous cell carcinoma (HNSCC) requires the development of novel specific vectors that can deliver therapeutic molecules. These vectors could also be coupled to fluorophores to be used in near infrared fluorescence imaging-guided surgery.Objectives: The aim of our work is to test new targeted vectors of HNSCC and to study the role of the near infrared fluorescence imaging-guided surgery in HNSCC resection in a novel orthotopic animal model that we develop.Materials and Methods: The HNSCC cell line CAL33 is characterized in vitro and in vivo. Novel vectors that target one or more receptors of this cell line such as alpha v beta 3 integrin, EGFR and NRP1, are tested in vitro. Meanwhile, an orthotopic animal model of HNSCC is developed by implanting tumor fragments of CAL33 cells, in the oral cavity of nude mice. Surgical resection of orthotopic tumors is guided by the near infrared fluorescence imaging after systemic injection of RAFT-c[RGD]4 peptide coupled with a fluorophore. This peptide targets alpha v beta 3 integrin and is previously tested in vitro.Results: Our preliminary results show that bispecific vectors would present an increased binding to CAL33 cells in vitro. On the other hand, near infrared fluorescence imaging-guided surgery has a positive impact on the recurrence-free survival rate in our orthotopic model, by detecting fluorescent cancer foci that could remain unidentified if resection was performed exclusively under visual guidance. Our results show also that near infrared fluorescence imaging can also help to detect metastatic lymph nodes.Conclusion: Near-infrared fluorescence imaging-guided surgery improves the quality of tumor resection in our optimized orthotopic animal model of HNSCC. This preclinical stage is essential before testing this novel technique in humans

Stylohyoid and Posterior Digastric Measurement with Intramuscular EMG, Submental EMG and Swallowing Sound

International audienceThe stylohyoid and the posterior digastric muscles have essentially been measured through indirect imaging method because of the difficulty to measure them. They are small neck muscles, close to each other, that cannot easily be accessed independently. Yet, they showed promising results for a robust and safe indwelling detection of swallowing, both in terms of timing and recruitment. The possibility to thoroughly establish their relevance through their direct functional analysis would enable the development of an implantable active artificial larynx, that would protect the airway during swallowing detection. Therefore, we set up the first standardized procedure that allows their direct measurement through intramuscular electromyography (EMG) and that we report in this paper. We also used submental surface EMG and swallowing sound modalities to access the major time points of the swallowing process. Finally, various exercises, along with swallowing, were performed by the volunteers. 1 6 peoples were measured with our new procedure, and both the stylohyoid and the posterior digastric could be measured independently with no difficulty. Timings and tasks comparison are therefore ongoing

Stylohyoid and Posterior Digastric Potential Evaluation for a Swallowing Detection, With Intramuscular EMG

International audienceTotal laryngectomy consists of the removal of the larynx and requires a tracheostomy to allow breathing, where the trachea is sewn on the anterior throat. This permanently separates the airway from the esophagus and any attempt to set it back in place would require to emulate the protective mechanism of the larynx, to avoid any aspiration during swallowing. So, with the aim to allow for the feasibility of an implantable active artificial larynx, and based on our previous investigations 1-4, this paper focuses on the stylohyoid and the posterior digastric muscles to evaluate their potential for a real-time and implantable swallowing detection algorithm. Indeed, we have shown that they activate at the beginning of swallowing, predominantly for swallowing, and that they are easily accessible, with no further impairment required during any sensor implantation. So, we measured them with intramuscular electromyography (EMG), along with the surface EMG of the submental muscles, to provide a basis for comparison. The swallowing sound was also measured with an accelerometer, to measure the vibration it generates through the skin, and define a temporal limit for the detection. 4 swallowing tasks and 13 non-swallowing tasks were investigated, to assess the muscles in terms of detection performances and abilities to provide early detection. Sub-groups of participants with similar swallowing, based on the Euclidean distance, were also explored. Thus, we found that the stylohyoid and the posterior digastric outperform the submental muscles and previously reported results in the literature, with increased performances when combined. Besides, the tasks that are the most related to the oral preparatory stage of swallowing tend to provide similar activity than the swallowing tasks, and are responsible for the most drastic drop in performances. Finally, even though we found promising results in terms of earliness, a real-time swallowing detection would benefit both from a stratified approach and from the development of an algorithm that is inherently built around a temporal constraint

Stylohyoid and posterior digastric timing evaluation

International audienceIn the context of real-time and safe indwelling detection of swallowing, which would enable the conception of an implantable active artificial larynx, we evaluate the timing of the stylohyoid and the posterior digastric muscles. They were shown to activate at the beginning of swallowing, both from human and animal results, but only with an indirect imaging method on humans. Therefore, this paper analyses their onset, offset, and peak time through the first method, that we previously set up, which allowed their direct functional analysis with intramuscular electromyography, surface electromyography, and swallowing sound. We found that both muscle activate at the beginning of swallowing, are relatively stable, while the posterior digastric duration were the shortest

Toward a robust swallowing detection for an implantable active artificial larynx: a survey

International audienceTotal laryngectomy consists in the removal of the larynx and is intended as a curative treatment for laryngeal cancer, but it leaves the patient with no possibility to breathe, talk, and swallow normally anymore. A tracheostomy is created to restore breathing through the throat, but the aero-digestive tracts are permanently separated and the air no longer passes through the nasal tracts, which allowed filtration, warming, humidification, olfaction, and acceleration of the air for better tissue oxygenation. As for phonation restoration, various techniques allow the patient to talk again. The main one consists of a tracheo-esophageal valve prosthesis that makes the air passes from the esophagus to the pharynx, and makes the air vibrate to allow speech through articulation. Finally, swallowing is possible through the original tract as it is now isolated from the trachea. Yet, many methods exist to detect and assess a swallowing, but none is intended as a definitive restoration technique of the natural airway, which would permanently close the tracheostomy and avoid its adverse effects. In addition, these methods are non-invasive and lack detection accuracy. The feasibility of an effective early detection of swallowing would allow to further develop an implantable active artificial larynx and therefore restore the aero-digestive tracts. A previous attempt has been made on an artificial larynx implanted in 2012, but no active detection was included and the system was completely mechanic. This led to residues in the airway because of the imperfect sealing of the mechanism. An active swallowing detection coupled with indwelling measurements would thus likely add a significant reliability on such a system as it would allow to actively close an artificial larynx. So, after a brief explanation of the swallowing mechanism, this survey intends to first provide a detailed consideration of the anatomical region involved in swallowing, with a detection perspective. Second, the swallowing mechanism following total laryngectomy surgery is detailed. Third, the current non-invasive swallowing detection technique and their limitations are discussed. Finally, the previous points are explored with regard to the inherent requirements for the feasibility of an effective swallowing detection for an artificial larynx