Cette recherche a été financée par le Centre de cas Comprehensive Cancer (P30 CA43703).

1. Stéréotaxique radiochirurgie CyberKnife consultation <ol><li> Décrivent le traitement radiochirurgie Cyberknife. Cyberknife radiochirurgie implique l&#39;utilisation d&#39;un accélérateur linéaire monté sur un bras robotique industriel similaire à des machines utilisées pour fabriquer des automobiles. Bras robotique déplace l&#39;accélérateur linéaire dans l&#39;espace tridimensionnel autour d&#39;un patient à mesure qu&#39;elle progresse à travers &#39;butées des traitements multiples pour une livraison de faisceau. Croisée plan de rayons X sont obtenus pendant le traitement pour vérifier le patient est dans une position correcte de traitement. Depuis la livraison du faisceau de traitement ne se limite pas à un plan à deux dimensions, ce système améliore encore la capacité de délivrer des doses de rayonnement concentrés pour cibles cancéreuses tout en minimisant la dose de rayonnement pour la peau critique et des structures des organes viscéraux. </li><li> Discuter des risques radiochirurgie CyberKnife. Radiochirurgie CyberKnife peut se traduire par de tannage et de rougeur de la peau, la fatigue, des nausées fréquentes ou diarrhea, rare lésion des organes viscéraux, des muscles, des nerfs, et une lésion osseuse, et très faible risque de deuxième cancer. </li></ol> 2. Stéréotaxique radiochirurgie CyberKnife placement Fiducial <ol><li> Décrire le placement fiduciaire. Femmes traitées pour cancers gynécologiques, persistants ou récurrents subir coopérative ou CT-guidée de placement d&#39;au moins trois simples 1,6 x 3 mm repères or tissus mous (environ la taille d&#39;un grain de riz) à l&#39;intérieur ou près de la cible contre le cancer. Agrafes chirurgicales placés au moment de la chirurgie antérieur ne sont pas d&#39;une densité suffisante pour être utilisé comme repères radiochirurgicaux. Agrafes chirurgicales n&#39;interfèrent pas avec radiochirurgical ciblage en raison de la différence de densité. </li><li> Indiquer l&#39;emplacement des repères d&#39;alignement par rapport à la cible de radiochirurgie. Repères sont placés à différentes profondeurs de tissu autour de la cible de radiochirurgie et doivent être séparés par deux centimètres ou plus. Repères sont positionnés dans les 4 à 6 centimètres de l&#39;tArget. </li></ol> 3. Stéréotaxique de planification de traitement de radiochirurgie Cyberknife <ol><li> Discuter de la planification du traitement radiochirurgical. Au moins une semaine après le placement de repère (c.-à-, de sorte que la guérison s&#39;est produite et le mouvement fiduciaire est réduit au minimum), les patients subissent CT-guidée de simulation de traitement SBRT. Dans notre programme, à deux broches localisée évacués sous vide sac immobilisation pelvienne est utilisée. </li><li> Indiquez le positionnement du patient sur ​​la table de traitement. Les femmes sont traitées dans une position de traitement en décubitus dorsal sur la table Cyberknife radiochirurgical plat. </li><li> Discutez cible et immobilisation du patient. Le patient peut subir évacués sous vide sac d&#39;immobilisation à réduire le mouvement intrafraction lors de l&#39;accouchement dose de rayonnement. Une fois immobilisé, les femmes subissent non-opposition contiguë axiale CT imagerie à haute résolution (1 à 1 emplacement, épaisseur 1,0 mm tranche, tension 120 kV, 450 mAs). </li><li> Discuter de l&#39;utilisation des avancéesd&#39;imagerie pour la localisation de cibles. Après l&#39;imagerie CT, les femmes subissent de préférence classique 18 F-FDG PET / CT Imagerie en position de traitement SBRT même. Alternativement, les femmes peuvent subir classique contraste d&#39;imagerie par résonance magnétique pelvienne. </li><li> Décrire l&#39;image de co-inscription pour la planification du traitement radiochirurgical. Images de haute résolution CT et 18 F-FDG PET / CT sont importés pour la planification d&#39;une radiothérapie inverse sur le MultiPlan 3.5.2 Système de planification de traitement (Accuray). </li><li> Décrire l&#39;approche d&#39;équipe à la planification du traitement radiochirurgical. Pour la planification du traitement, à la fois un radio-oncologue et gynécologiques oncologue contour des volumes cibles cancéreuses cibles cliniques (TVC). Autres maladies des tissus à risque est profilée et est incluse dans les TVC. À proximité des structures tissulaires normales telles que l&#39;intestin grêle, du rectum, vessie, foie, reins, poumons, bilatéraux fémurs proximaux, le vagin, et les racines nerveuses sacrées sont profilées parle radio-oncologue. </li><li> Décrivez la prescription de dose de rayonnement. Une dose de prescription rayonnement de 3 × 800 = 2400 cGy cGy (communément à la ligne isodose 70%) a été sélectionné. Dans un 200 cGy équivalent de dose de calcul biologique, près de 6170 cGy sont livrés avec cette prescription en supposant un «α / β&quot; ratio de 10 pour une tumeur. Le rayonnement émis à partir d&#39;un accélérateur linéaire SBRT est collimaté soit en utilisant l&#39;un des 12 collimateurs de tungstène circulaire fixe (5 à 60 mm) ou en utilisant un alliage de tungstène-cuivre segmenté hexagone IRIS collimateur. 14 organes normaux ont une tolérance de rayonnement et d&#39;utiliser des paramètres histogramme dose-volume pour les tissus normaux au cours de la contrainte radiochirurgie stéréotaxique ablatif sont énumérés dans le tableau 1. </li></ol> 4. Stéréotaxique de livraison Cyberknife traitement de radiochirurgie <ol><li> Effectuez la vérification d&#39;image croix-plan. Pendant Cyberknife SBRT, les tissus mous repères(Ou, dans certains cas rigides repères anatomiques tels que le crâne pour les lésions intracrâniennes et la colonne vertébrale pour les lésions du rachis) sont suivis par la Croix-plan d&#39;imagerie radiographique et a confirmé être à l&#39;emplacement cible prévue par le système cible localisation (TLS) sous-système. </li><li> Vérifier le positionnement cible. Repères sont suivis dans trois et jusqu&#39;à six degrés de liberté (c.-à-x, y, z, tangage, roulis et lacet). Images générées par les TLS sont automatiquement enregistrées et comparées avec des radiographies reconstruites numériquement (DRR) générés à partir du traitement initial de planification CT scan. </li><li> Vérifier le positionnement cible. Si les résultats de l&#39;enregistrement automatique indiquent que repères ont changé au-delà des tolérances prédéfinies dans l&#39;une quelconque des six degrés de liberté (x, y, z, tangage, roulis et lacet), le traitement est automatiquement mise en pause. Repositionnement du patient se produit alors. </li><li> Considérez mouvement de la cible. Les cibles qui se déplacent avec les mouvements respiratoires sont treated utilisant le sous-système de suivi Synchrony respiratoire (Synchrony). Avec l&#39;utilisation d&#39;une caméra montée dans la chambre de traitement, le système surveille en permanence la fréquence respiratoire du patient (par exemple, obtenir des données de position externes) avec l&#39;utilisation de diodes électroluminescentes (LED) apposés sur une veste autour du thorax du patient. 10 Ensemble les informations de localisation repère (données de position internes) obtenu à partir transversale plan de rayons X, le système Synchrony construit un modèle de corrélation avec les données externes et les données internes. Ce modèle de corrélation permet au bras robotique de suivre tout mouvement respiratoire induite par de la cible tandis que lors de la livraison de tout et de tous les faisceaux de traitement. </li><li> Le traitement peut entraîner 100-150 postes de traitement livrables du bras robotique. Les traitements peuvent durer de 30 à 90 minutes. </li></ol> 5. Les résultats représentatifs Radiothérapie stéréotaxique corps (SBRT) peut impliquer de nombreux faisceaux de rayonnement individuelles (<html> vecteurs bleus) qui convergent toutes sur un seul ou plusieurs objectifs étroitement liés rayonnement cliniques, comme le montre la figure 1A. Un représentant de bons résultats planification radiochirurgical donne un traitement SBRT livrable avec la dose de rayonnement couverture élevée du volume cible et le cancer conformalité cible le cancer. Figures 1B-D montrent 131 faisceaux ont été utilisés pour traiter une rechute pelvienne d&#39;un cancer de l&#39;ovaire cible chemorefractory plus de 42 minutes. SBRT prescrit à la ligne isodose 80% rendu 100% de couverture du volume cible clinique avec un indice de 1,94 conformalité pour une dose totale de 2400 cGy en fractions quotidiennes trois cGy 800. Les histogrammes dose-volume pour la cible clinique (rouge) et pour les structures critiques du rectum (brun), de la vessie (jaune), l&#39;intestin grêle (lumière bleue) sacrés nerfs (tan) et les hanches (orange) sont représentés dans la figure 2. <img alt="Figure 1" src="/files/ftp_upload/3793/3793fig1.jpg" /> Figure 1.tp :/ / www.jove.com/files/ftp_upload/3793/3793fig1large.jpg &quot;target =&quot; _blank &quot;&gt; Cliquez ici pour agrandir la figure. <img alt="Figure 2" src="/files/ftp_upload/3793/3793fig2.jpg" /> Figure 2. Histogrammes dose-volume pour la cible clinique (rouge) et pour les structures critiques du rectum (brun), de la vessie (jaune), l&#39;intestin grêle (lumière bleue) sacrés nerfs (tan) et les hanches (orange).

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Radiol. 54, 257-258 (1981).</li><li>Wunderink, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+of+respiratory+liver+tumor+motion+by+abdominal+compression+in+stereotactic+body+frame,+analyzed+by+tracking+fiducial+markers+implanted+in+liver.">Reduction of respiratory liver tumor motion by abdominal compression in stereotactic body frame, analyzed by tracking fiducial markers implanted in liver.</a> Int. J. Radiat. Oncol. Biol. Phys. 71, 907-915 (2008).</li><li>Mageras, G., Yorke, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deep+inspiration+breath+hold+and+respiratory+gating+strategies+for+reducing+organ+motion+in+radiation+treatments.">Deep inspiration breath hold and respiratory gating strategies for reducing organ motion in radiation treatments.</a> Semin. Radiat. 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Phys. 30, 88-97 (2003).</li><li>Kunos, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyberknife+radiosurgery+for+squamous+cell+carcinoma+of+the+vulva+after+prior+pelvic+radiation+therapy.">Cyberknife radiosurgery for squamous cell carcinoma of the vulva after prior pelvic radiation therapy.</a> Technol. Cancer Res. Treat. 7, 375-380 (2008).</li><li>Kunos, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stereotactic+body+radiosurgery+for+pelvic+relapse+of+gynecologic+malignancies.">Stereotactic body radiosurgery for pelvic relapse of gynecologic malignancies.</a> Technol. Cancer Res. Treat. 8, 393-400 (2009).</li><li>Kilby, W., Dooley, J., Kuduvalli, G., Sayeh, S., Maurer, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Cyberknife+robotic+radiosurgery+system+in+2010.">The Cyberknife robotic radiosurgery system in 2010.</a> Technol. Cancer Res. Treat. 9, 431-438 (2010).</li><li>Ozhasoglu, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synchrony+&#x2013;+Cyberknife+Respiratory+Compensation+Technology.">Synchrony &#x2013; Cyberknife Respiratory Compensation Technology.</a> Med. 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Nous (CAK, JMB, RD) n&#39;ont rien à révéler.

Encourager les résultats SBRT début ont alimenté la recherche clinique de la radiochirurgie pour le traitement de persistants ou récurrents, les cancers gynécologiques. 7, 8, 15 données en question les effets radiobiologiques et le mode de la mort cellulaire résultant de SBRT. Les petites études cliniques ont montré que des doses de rayonnement ablatifs fournis par SBRT produire ciblées taux de contrôle des maladies de plus de 90%. Contrairement à rayonnement classique, il a été difficile de combiner avec des chimiothérapies SBRT radiosensibilisation et cytotoxique / cytostatique. Mieux cibler le cancer grâce à l&#39;expansion de planification accrus volume de traitement et par l&#39;inclusion de 18 F-FDG PET / CT images a amélioré les résultats cliniques. S&#39;il est impératif de se pencher sur d&#39;autres méthodes de prestation de la dose de rayonnement escalade avec une grande précision, il reste indécis quant SBRT peut fournir une efficacité thérapeutique équivalente aussi bas et haut débit de dose de curiethérapie. En effet, la brachythérapie est le plus communément et valien date du technique pour obtenir d&#39;escalade de dose de rayonnement dans les objectifs de cancer gynécologique. En tant que tel, à la fois l&#39;enthousiasme et la prudence sont appropriés à la lecture des données SBRT disponibles pour le traitement des cancers gynécologiques.

<table border="1"><tbody><tr><td> Nom de l&#39;appareil </td><td> Entreprise </td><td> Numéro de catalogue </td><td> Commentaires </td></tr><tr><td> Cyberknife système </td><td> Accuray (Sunnyvale, CA) </td><td></td><td></td></tr></tbody></table>

stereotactic radiosurgery for gynecologic cancer

La radiothérapie stéréotaxique corps (SBRT) se distingue en nécessitant une immobilisation rigide plus patient, la comptabilisation des mouvements respiratoires, la planification du traitement complexe, à bord d&#39;imagerie, et le nombre réduit de doses de rayonnement ablatives au cancer vise habituellement réfractaires à la chimiothérapie et la radiothérapie conventionnelle. Domaines de traitement &quot;Pencil Beam&quot; Steep dose de rayonnement SBRT abandon au large de permis étroit pour être utilisé pour le traitement ablatif rayonnement condensé en 1 à 3 traitements. Les médecins traitants doivent être conscients que SBRT arrive à un plus grand risque de blessure des tissus normaux et la chance de miss tumeur géographique. Les deux doivent être abordés par l&#39;immobilisation des cibles du cancer et par l&#39;administration du traitement de haute précision. Immobilisation cible le cancer a été réalisé grâce à l&#39;utilisation des moulages en mousse de polystyrène indexés sur mesure, sacs de fèves évacuées, ou le corps-fix moules avec le patient indépendant compression abdominale. Mouvement Intrafraction 1-3 de targe le canceren raison de la respiration ct maintenant peut être réduit par le patient en réponse techniques l&#39;apnée, 4 embouchure pour patient respiratoire coordination active, 5 respiration en corrélation tomodensitométrie, 6 ou guidée par l&#39;image de suivi de repères implantés à l&#39;intérieur et autour d&#39;une tumeur mobile. 7-9 Le Cyberknife système (Accuray [Sunnyvale]) utilise un accélérateur de rayonnement linéaire monté sur un bras robotique industriel qui suit précisément le mouvement respiratoire du patient par une caméra-ensemble de chenilles diodes électroluminescentes (DEL) imprégnées sur un gilet muni d&#39;un patient. 10 Des réductions substantielles dans les marges de radiothérapie peut être réalisée par le suivi de mouvement, en fin de compte rendre un plus petits volumes cibles de planification qui sont irradiés avec une précision sub-millimétrique. 11-13 Cibles du cancer traités par SBRT sont irradiés par des faisceaux convergents, bien collimatés. Dose de rayonnement résultant de cancer histogrammes volumes cibles ont une plus pronomced rayonnement &quot;l&#39;épaule&quot;, indiquant une forte couverture cible de pourcentage et un petit haut-dose de rayonnement &quot;de la queue.&quot; Ainsi, conformalité cible accrue se fait au détriment de l&#39;uniformité des doses a diminué dans la cible du cancer SBRT. Cela peut avoir des implications à la fois pour le contrôle tumoral après dans la cible SBRT et la tolérance des tissus normaux d&#39;organes à risque. En raison de la forte dose falloff SBRT, la possibilité d&#39;échapper à la maladie occulte la dose de rayonnement ablatif se produit lorsque les objectifs du cancer ne sont pas pleinement reconnus et l&#39;insuffisance des marges de dose SBRT sont appliquées. Volume cible clinique (CTV) l&#39;expansion de 0,5 cm, ce qui entraîne un volume cible plus large de planification (PTV), est associée à la cible de maîtrise accrue sans lésion tissulaire excessive normale. 7,8 réduction supplémentaire de la probabilité de manquer géographique peut être réalisée par incorporation de 2 - [18 F] fluoro-2-désoxy-D-glucose (18 F-FDG) par émission de positons (TEP) 8 Utilisation de 18 F-FDG PET / CT dans SBRT treatme.planification nt n&#39;est que le début des tentatives pour découvrir de nouvelles signatures de cibles d&#39;imagerie moléculaire pour les cancers gynécologiques.

Watch this Scientific Journal Video about Stereotactic Radiosurgery for Gynecologic Cancer at JoVE.com

Stereotactic Radiosurgery for Gynecologic Cancer

La radiothérapie stéréotaxique corps (SBRT) implique guidée par l&#39;image, le rayonnement ablatif livré à un cancer cible réfractaire à la chimiothérapie ou à la radiothérapie conventionnelle. Le robot-armé Cyberknife SBRT système, en utilisant la localisation de cibles sophistiquée, offre des doses de rayonnement hypofractionnée capables de stérilisation cibles cancéreuses. Cet article examinera de nouveaux rôles thérapeutiques de SBRT pour les cancers gynécologiques.

Radiochirurgie stéréotaxique pour Gynecologic Cancer

Stereotactic body radiotherapy (SBRT) distinguishes itself by necessitating more rigid patient immobilization, accounting for respiratory motion, ...

stereotactic-radiosurgery-for-gynecologic-cancer

Research

JoVE Journal

Medicine

17.5K vues.  University Hospitals Case Medical Center and Case Western Reserve University School of Medicine. La radiothérapie stéréotaxique corps (SBRT) implique guidée par l'image, le rayonnement ablatif livré à un cancer cible réfractaire à la chimiothérapie ou à la radiothérapie conventionnelle. Le robot-armé Cyberknife SBRT système, en utilisant la localisation de cibles sophistiquée, offre des doses de rayonnement hypofractionnée capables de stérilisation cibles cancéreuses. Cet article examinera de nouveaux rôles thérapeutiques de SBRT pour les cance...

Vidéo: Stereotactic Radiosurgery for Gynecologic Cancer

Multispectral Real-time Fluorescence Imaging for Intraoperative Detection of the Sentinel Lymph Node in Gynecologic Oncology

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases.1-3 Surgical treatment most often combines radical excision of the tumor with a full lymphadenectomy in the drainage area of the tumor. However, removal of lymph nodes is associated with increased morbidity due to infection, wound breakdown and lymphedema.4,5 As an alternative, the sentinel lymph node procedure (SLN) was developed several decades ago to detect the first draining lymph node from the tumor.6 In case of lymphogenic dissemination, the SLN is the first lymph node that is affected (Figure 1). Hence, if the SLN does not contain metastases, downstream lymph nodes will also be free from tumor metastases and need not to be removed. The SLN procedure is part of the treatment for many tumor types, like breast cancer and melanoma, but also for cancer of the vulva and cervix.7 The current standard methodology for SLN-detection is by peritumoral injection of radiocolloid one day prior to surgery, and a colored dye intraoperatively. Disadvantages of the procedure in cervical and vulvar cancer are multiple injections in the genital area, leading to increased psychological distress for the patient, and the use of radioactive colloid.
Multispectral fluorescence imaging is an emerging imaging modality that can be applied intraoperatively without the need for injection of radiocolloid. For intraoperative fluorescence imaging, two components are needed: a fluorescent agent and a quantitative optical system for intraoperative imaging. As a fluorophore we have used indocyanine green (ICG). ICG has been used for many decades to assess cardiac function, cerebral perfusion and liver perfusion.8 It is an inert drug with a safe pharmaco-biological profile. When excited at around 750 nm, it emits light in the near-infrared spectrum around 800 nm. A custom-made multispectral fluorescence imaging camera system was used.9.
The aim of this video article is to demonstrate the detection of the SLN using intraoperative fluorescence imaging in patients with cervical and vulvar cancer. Fluorescence imaging is used in conjunction with the standard procedure, consisting of radiocolloid and a blue dye. In the future, intraoperative fluorescence imaging might replace the current method and is also easily transferable to other indications like breast cancer and melanoma.

Fluorescence imaging is a promising innovative modality for image-guided surgery in surgical oncology. In this video we describe the technical procedure for detection of the sentinel lymph node using fluorescence imaging as showcased in gynecologic oncologicy. A multispectral fluorescence camera system, together with the fluorescent agent indocyanine green, is applied.

Multispectrale en temps réel d&#39;imagerie par fluorescence pour la détection peropératoire du ganglion sentinelle en oncologie gynécologique

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases.1-3 Surgical treatment most often combines radical ...

Recherche

Médecine

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality treatment consisting of surgical resection, post-operative radiation therapy and temozolomide chemotherapy 1. New therapeutic approaches are clearly needed to improve patient survival and quality of life. The development of more effective treatment strategies would be aided by animal models of GBM that recapitulate human disease yet allow serial imaging to monitor tumor growth and treatment response. In this paper, we describe our technique for the precise stereotactic implantation of bio-imageable GBM cancer cells into the brains of nude mice resulting in tumor xenografts that recapitulate key clinical features of GBM 2. This method yields tumors that are reproducible and are located in precise anatomic locations while allowing in vivo bioluminescent imaging to serially monitor intracranial xenograft growth and response to treatments 3-5. This method is also well-tolerated by the animals with low perioperative morbidity and mortality.

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

We describe an integrated method for the precise, stereotactic implantation of human glioblastoma multiforme cells into the brains of nude mice and subsequent serial in vivo imaging to monitor growth and response to treatment of the resultant xenografts.

Implantation stéréotaxique intracrânienne et<em&gt; In vivo</em&gt; Imagerie bioluminescente de xénogreffes tumorales dans un système de modèle de souris de glioblastome multiforme

Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality ...

Cerenkov Luminescence Imaging (CLI) for Cancer Therapy Monitoring

In molecular imaging, positron emission tomography (PET) and optical imaging (OI) are two of the most important and thus most widely used modalities1-3. PET is characterized by its excellent sensitivity and quantification ability while OI is notable for non-radiation, relative low cost, short scanning time, high throughput, and wide availability to basic researchers. However, both modalities have their shortcomings as well. PET suffers from poor spatial resolution and high cost, while OI is mostly limited to preclinical applications because of its limited tissue penetration along with prominent scattering optical signals through the thickness of living tissues.
 Recently a bridge between PET and OI has emerged with the discovery of Cerenkov Luminescence Imaging (CLI)4-6. CLI is a new imaging modality that harnesses Cerenkov Radiation (CR) to image radionuclides with OI instruments. Russian Nobel laureate Alekseyevich Cerenkov and his colleagues originally discovered CR in 1934. It is a form of electromagnetic radiation emitted when a charged particle travels at a superluminal speed in a dielectric medium7,8. The charged particle, whether positron or electron, perturbs the electromagnetic field of the medium by displacing the electrons in its atoms. After passing of the disruption photons are emitted as the displaced electrons return to the ground state. For instance, one 18F decay was estimated to produce an average of 3 photons in water5. 
 Since its emergence, CLI has been investigated for its use in a variety of preclinical applications including in vivo tumor imaging, reporter gene imaging, radiotracer development, multimodality imaging, among others4,5,9,10,11. The most important reason why CLI has enjoyed much success so far is that this new technology takes advantage of the low cost and wide availability of OI to image radionuclides, which used to be imaged only by more expensive and less available nuclear imaging modalities such as PET.
 Here, we present the method of using CLI to monitor cancer drug therapy. Our group has recently investigated this new application and validated its feasibility by a proof-of-concept study12. We demonstrated that CLI and PET exhibited excellent correlations across different tumor xenografts and imaging probes. This is consistent with the overarching principle of CR that CLI essentially visualizes the same radionuclides as PET. We selected Bevacizumab (Avastin; Genentech/Roche) as our therapeutic agent because it is a well-known angiogenesis inhibitor13,14. Maturation of this technology in the near future can be envisioned to have a significant impact on preclinical drug development, screening, as well as therapy monitoring of patients receiving treatments.

Cerenkov Luminescence Imaging CLI for Cancer Therapy Monitoring

Use of Cerenkov Luminescence Imaging (CLI) for monitoring preclinical cancer treatment is described here. This method takes advantage of Cerenkov Radiation (CR) and optical imaging (OI) to visualize radiolabeled probes and thus provides an alternative to PET in preclinical therapeutic monitoring and drug screening.

Luminescence Cerenkov Imaging (CLI) pour la surveillance de la thérapie du cancer

In  molecular imaging, positron emission tomography (PET) and optical imaging (OI)  are two of the most important and thus most widely used modalities1-3. ...

Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%. New therapeutic options are critically needed and depend on improved understanding of pancreatic cancer biology. To better understand the interaction of cancer cells with the pancreatic microenvironment, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression. Luciferase-tagged pancreatic cancer cells are resuspended in Matrigel and delivered into the pancreatic tail during laparotomy. Matrigel solidifies at body temperature to prevent leakage of cancer cells during injection. Primary tumor growth and metastasis to distant organs are monitored following injection of the luciferase substrate luciferin, using in vivo imaging of bioluminescence emission from the cancer cells. In vivo imaging also may be used to track primary tumor recurrence after resection. This orthotopic model is suited to both syngeneic and xenograft models and may be used in pre-clinical trials to investigate the impact of novel anti-cancer therapeutics on the growth of the primary pancreatic tumor and metastasis.

Improved understanding of pancreatic cancer biology is critically needed to enable the development of better therapeutic options to treat pancreatic cancer. To address this need, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression using in vivo bioluminescence imaging.

Modèle Orthotopique bioluminescente la progression du cancer du pancréas

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%.  New therapeutic options are critically needed and depend on improved ...

Profiling of Estrogen-regulated MicroRNAs in Breast Cancer Cells

Estrogen plays vital roles in mammary gland development and breast cancer progression. It mediates its function by binding to and activating the estrogen receptors (ERs), ER&#945;, and ER&#946;. ER&#945; is frequently upregulated in breast cancer and drives the proliferation of breast cancer cells. The ERs function as transcription factors and regulate gene expression. Whereas ER&#945;&#39;s regulation of protein-coding genes is well established, its regulation of noncoding microRNA (miRNA) is less explored. miRNAs play a major role in the post-transcriptional regulation of genes, inhibiting their translation or degrading their mRNA. miRNAs can function as oncogenes or tumor suppressors&#160;and are also promising biomarkers. Among the miRNA assays available, microarray and quantitative real-time polymerase chain reaction (qPCR) have been extensively used to detect and quantify miRNA levels. To identify miRNAs regulated by estrogen signaling in breast cancer, their expression in ER&#945;-positive breast cancer cell lines were compared before and after estrogen-activation using both the &#181;Paraflo-microfluidic microarrays and Dual Labeled Probes-low density arrays. Results were validated using specific qPCR assays, applying both Cyanine dye-based and Dual Labeled Probes-based chemistry. Furthermore, a time-point assay was used to identify regulations over time. Advantages of the miRNA assay approach used in this study is that it enables a fast screening of mature miRNA regulations in numerous samples, even with limited sample amounts. The layout, including the specific conditions for cell culture and estrogen treatment, biological and technical replicates, and large-scale screening followed by in-depth confirmations using separate techniques, ensures a robust detection of miRNA regulations,&#160;and eliminates false positives and other artifacts. However, mutated or unknown miRNAs, or regulations at the primary and precursor transcript level, will not be detected. The method presented here represents a thorough investigation of estrogen-mediated miRNA regulation.

Molecular signaling through both estrogen and microRNAs are critical in breast cancer development and growth. Estrogen activates the estrogen receptors, which are transcription factors. Many transcription factors can regulate the expression of microRNAs, and estrogen-regulated microRNAs can be profiled using different large-scale techniques.

Profilage de microARN œstrogène réglementés dans les cellules du cancer du sein

Estrogen plays vital roles in mammary gland development and breast cancer progression. It mediates its function by binding to and activating the estrogen ...

Utilizing Murine Inducible Telomerase Alleles in the Studies of Tissue Degeneration/Regeneration and Cancer

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that cause tissue degeneration during ageing. Cancer, with incidence rates greatly increasing with age, is characterized by short telomere lengths and high telomerase activity. To study the roles of telomere dysfunction and telomerase reactivation in ageing and cancer, the protocol shows how to generate two murine inducible telomerase knock-in alleles 4-Hydroxytamoxifen (4-OHT)-inducible TERT-Estrogen Receptor (mTERT-ER) and Lox-Stopper-LoxTERT (LSL-mTERT). The protocol describes the procedures to induce telomere dysfunction and reactivate telomerase activity in mTERT-ER and LSL-mTERT mice in vivo. The representative data show that reactivation of telomerase activity can ameliorate the tissue degenerative phenotypes induced by telomere dysfunction. In order to determine the impact of telomerase reactivation on tumorigenesis, we generated prostate tumor model G4 PB-Cre4 PtenL/L p53L/L LSL-mTERTL/L and thymic T-cell lymphoma model G4 Atm-/- mTERTER/ER. The representative data show that telomerase reactivation in the backdrop of genomic instability induced by telomere dysfunction can greatly enhance tumorigenesis. The protocol also describes the procedures used to isolate neural stem cells (NSCs) from mTERT-ER and LSL-mTERT mice and reactivate telomerase activity in NSCs in vitro. The representative data show that reactivation of telomerase can enhance the self-renewal capability and neurogenesis in vitro. Finally, the protocol describes the procedures for performing telomere FISH (Fluorescence In Situ Hybridization) on both mouse FFPE (Formalin Fixed and Paraffin Embedded) brain tissues and metaphase chromosomes of cultured cells.

Telomere and telomerase play essential roles in ageing and tumorigenesis. The goal of this protocol is to show how to generate two murine inducible telomerase knock-in alleles and how to utilize them in the studies of tissue degeneration/regeneration and cancer.

Utilisant murin inductible télomérase allèles dans les études de la dégénérescence des tissus / Régénération et cancer

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor initiation, metastasis and resistance to radio- and chemotherapy. Here we describe a specific methodology for culturing primary human pancreatic CSCs as tumor spheres in anchorage-independent conditions. Cells are grown in serum-free, non-adherent conditions in order to enrich for CSCs while their more differentiated progenies do not survive and proliferate during the initial phase following seeding of single cells. This assay can be used to estimate the percentage of CSCs present in a population of tumor cells. Both size (which can range from 35 to 250 micrometers) and number of tumor spheres formed represents CSC activity harbored in either bulk populations of cultured cancer cells or freshly harvested and digested tumors 1,2. Using this assay, we recently found that metformin selectively ablates pancreatic CSCs; a finding that was subsequently further corroborated by demonstrating diminished expression of pluripotency-associated genes/surface markers and reduced in vivo tumorigenicity of metformin-treated cells. As the final step for preclinical development we treated mice bearing established tumors with metformin and found significantly prolonged survival. Clinical studies testing the use of metformin in patients with PDAC are currently underway (e.g., NCT01210911, NCT01167738, and NCT01488552). Mechanistically, we found that metformin induces a fatal energy crisis in CSCs by enhancing reactive oxygen species (ROS) production and reducing mitochondrial transmembrane potential. In contrast, non-CSCs were not eliminated by metformin treatment, but rather underwent reversible cell cycle arrest. Therefore, our study serves as a successful example for the potential of in vitro sphere formation as a screening tool to identify compounds that potentially target CSCs, but this technique will require further in vitro and in vivo validation to eliminate false discoveries.

Pancreatic cancer stem cells (CSCs) can be expanded in vitro using the anchorage-independent sphere culture technique, which represents a powerful tool to study CSC biology and can serve as the first step to develop novel CSC-targeting therapies. Here the methodology for expanding, analyzing and targeting of pancreatic CSCs is provided.

Etudier le cancer du pancréas Stem Cell Caractéristiques pour le développement de nouvelles stratégies de traitement

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

Dynamic Lung Tumor Tracking for Stereotactic Ablative Body Radiation Therapy

Physicians considering stereotactic ablative body radiation therapy (SBRT) for the treatment of extracranial cancer targets must be aware of the sizeable risks for normal tissue injury and the hazards of physical tumor miss. A first-of-its-kind SBRT platform achieves high-precision ablative radiation treatment through a combination of versatile real-time imaging solutions and sophisticated tumor tracking capabilities. It uses dual-diagnostic kV x-ray units for stereoscopic open-loop feedback of cancer target intrafraction movement occurring as a consequence of respiratory motions and heartbeat. Image-guided feedback drives a gimbaled radiation accelerator (maximum 15 x 15 cm field size) capable of real-time &#177;4 cm pan-and-tilt action. Robot-driven &#177;60&#176; pivots of an integrated &#177;185&#176; rotational gantry allow for coplanar and non-coplanar accelerator beam set-up angles, ultimately permitting unique treatment degrees of freedom. State-of-the-art software aids real-time six dimensional positioning, ensuring irradiation of cancer targets with sub-millimeter accuracy (0.4 mm at isocenter). Use of these features enables treating physicians to steer radiation dose to cancer tumor targets while simultaneously reducing radiation dose to normal tissues. By adding respiration correlated computed tomography (CT) and 2-[18F] fluoro-2-deoxy-&#7429;-glucose (18F-FDG) positron emission tomography (PET) images into the planning system for enhanced tumor target contouring, the likelihood of physical tumor miss becomes substantially less1. In this article, we describe new radiation plans for the treatment of moving lung tumors.

Stereotactic ablative body radiation therapy (SBRT) involves the precise delivery of high-dose radiation to cancer tumor targets. A novel SBRT platform offers a first-of-its-kind gimbaled radiation accelerator mounted within a pivoting O-ring gantry capable of dynamic image-guided tumor tracking. Here, we describe dynamic tumor tracking for lung targets.

Dynamique de suivi de la tumeur du poumon pour stéréotaxique ablatif radiothérapie du corps

Physicians considering stereotactic ablative body radiation therapy (SBRT) for the treatment of extracranial cancer targets must be aware of the sizeable ...

Working with Human Tissues for Translational Cancer Research

Medical research for human benefit is greatly impeded by the necessity for human tissues and subjects. However, upon obtaining consent for human specimens, precious samples must be handled with the greatest care in order to ensure integrity of organs, tissues, and cells to the highest degree. Unfortunately, tissue processing by definition requires extraction of tissues from the host, a change which can cause great cellular stress and have major repercussions on subsequent analyses. These stresses could result in the specimen being no longer representative of the site from which it was retrieved. Therefore, a strict protocol must be adhered to while processing these specimens to ensure representativeness. The desired assay(s) must also be taken into consideration in order to ensure that an optimal technique is used for sample processing. Outlined here is a protocol for tissue retrieval, processing and various analyses which may be performed on processed tissue in order to maximize downstream production from limited tissue samples.

Translational cancer research is dependent on extraction of human tissues. Much work has gone into optimizing extraction methods for ex vivo analysis. Here, we describe tissue processing methods allowing for maximal data output from limited samples.

Travailler avec les tissus humains pour la recherche translationnelle sur le cancer

Medical research for human benefit is greatly impeded by the necessity for human tissues and subjects. However, upon obtaining consent for human ...

Modeling the Early Steps of Ovarian Cancer Dissemination in an Organotypic Culture of the Human Peritoneal Cavity

The pattern of ovarian cancer metastasis is markedly different from that of most other epithelial tumors, because it rarely spreads hematogenously. Instead, ovarian cancer cells exfoliated from the primary tumor are carried by peritoneal fluid to metastatic sites within the peritoneal cavity. These sites, most notably the abdominal peritoneum and omentum, are organs covered by a mesothelium-lined surface. To investigate the processes of ovarian cancer dissemination, we assembled a complex three-dimensional culture system that reconstructs the lining of the peritoneal cavity in vitro. Primary human fibroblasts and mesothelial cells were isolated from human omentum. The fibroblasts were then mixed with extracellular matrix and covered with a layer of the primary human mesothelial cells to mimic the peritoneal and omental surfaces encountered by metastasizing ovarian cancer cells. The resulting organotypic model is, as shown, used to examine the early steps of ovarian cancer dissemination, including cancer cell adhesion, invasion, and proliferation. This model has been used in a number of studies to investigate the role of the microenvironment (cellular and acellular) in early ovarian cancer dissemination. It has also been successfully adapted to high throughput screening and used to identify and test inhibitors of ovarian cancer metastasis.

Here, we present a protocol to construct a three-dimensional in vitro model of the lining of the peritoneal cavity, composed of primary human mesothelial cells and fibroblasts layered with extracellular matrix, as a tool to investigate ovarian cancer cell adhesion, invasion, and proliferation.

Modélisation des étapes précoces de cancer de l&#39;ovaire Diffusion dans une culture organotypique de la cavité péritonéale humain

The pattern of ovarian cancer metastasis is markedly different from that of most other epithelial tumors, because it rarely spreads hematogenously. ...