Questa ricerca è stata sostenuta dalla Custodia Comprehensive Cancer Center (P30 CA43703).

1. Radiochirurgia stereotassica Cyberknife consultazione <ol><li> Descrivere il trattamento radiochirurgia Cyberknife. Radiochirurgia Cyberknife prevede l&#39;utilizzo di un acceleratore lineare montato su un braccio robotico industriale simile a macchine usate per rendere le automobili. Il braccio robotico sposta l&#39;acceleratore lineare in uno spazio tridimensionale intorno ad un paziente che si svilupperà attraverso &#39;fermate dei trattamenti multipli per la consegna del fascio. Cross-piano sono ottenuti raggi X durante il trattamento per verificare il paziente è in una posizione di corretto trattamento. Poiché fascio consegna del trattamento non è limitato ad un piano bidimensionale, questo sistema migliora ulteriormente la capacità di erogare dosi di radiazione concentrata a bersagli tumorali, riducendo al minimo la dose di radiazione alla pelle critica e strutture organi viscerali. </li><li> Discuti rischi radiochirurgia CyberKnife. Cyberknife radiochirurgia può provocare possibili concia e arrossamento della pelle, affaticamento, nausea poco frequenti o diarrhea, rare lesioni di organi viscerali, muscolo, nervo e lesione ossea, e il rischio molto ridotto di secondo cancro. </li></ol> 2. Radiochirurgia stereotassica Cyberknife Placement Fiducial <ol><li> Descrivere il posizionamento fiduciale. Le donne in trattamento per il persistente o ricorrente tumori ginecologici sottoposti operative o TC-guidata inserimento di almeno tre singoli 1,6 x 3 mm fiducial dei tessuti molli in oro (circa le dimensioni di un chicco di riso) all&#39;interno o in prossimità del bersaglio cancro. Fermagli chirurgici posti al momento dell&#39;intervento chirurgico prima di densità non sono sufficienti per essere utilizzata come fiducial radiochirurgici. Clip chirurgiche non interferiscono con radiochirurgico di mira a causa della differenza di densità. </li><li> Indicare il posizionamento fiducial rispetto al bersaglio radiochirurgico. Fiducial sono messi a varie profondità del tessuto intorno al bersaglio radiochirurgico e devono essere separati da due centimetri o più. Fiducial sono posizionati entro 4 a 6 centimetri del target. </li></ol> 3. Radiochirurgia stereotassica Cyberknife Pianificazione del trattamento <ol><li> Discuti pianificazione del trattamento radiochirurgico. Almeno una settimana dopo il posizionamento fiducial (vale a dire, in modo che la guarigione si è verificato e il movimento fiducial è ridotto al minimo), i pazienti sottoposti a TC-guidata simulazione del trattamento SBRT. Nel nostro programma, due poli localizzato evacuato vuoto-bag immobilizzazione pelvica viene utilizzato. </li><li> Indicare il posizionamento del paziente sul tavolo di trattamento vengono trattati. Donne in una posizione di trattamento supino sul tavolo radiochirurgia Cyberknife piatto. </li><li> Discuti di destinazione e immobilizzazione del paziente. Paziente può subire evacuate a vuoto-bag immobilizzazione per ridurre il movimento intrafraction durante la consegna della dose di radiazione. Una volta immobilizzato, le donne subiscono non contrastata contigua assiale CT imaging ad alta risoluzione (1 a 1 passo, 1,0 mm di spessore fetta, tensione 120 kVp, 450 mAs). </li><li> Discutere l&#39;uso di tecnologie avanzatedi imaging per la localizzazione di destinazione. Dopo CT imaging, le donne subiscono preferibilmente convenzionale 18 F-FDG PET / TC nella stessa posizione di trattamento SBRT. In alternativa, le donne possono subire convenzionale contrasto immagine a risonanza magnetica pelvica. </li><li> Descrivere l&#39;immagine co-registrazione per la pianificazione radiochirurgico trattamento. Immagini da TC ad alta risoluzione e 18 F-FDG PET / CT sono importati per la pianificazione inversa trattamento con radiazioni sul sistema 3.5.2 Pianificazione del trattamento Multiplan (Accuray). </li><li> Descrivere approccio di squadra alla pianificazione radiochirurgico trattamento. Per la pianificazione del trattamento, sia un radioterapista e oncologo di contorno ginecologiche cancro volumi bersaglio bersaglio clinico (CTV). Altre malattie a rischio il tessuto è sagomato ed è incluso in televisori a colori. Vicine strutture dei tessuti normali, come il piccolo intestino, il retto, vescica, fegato, reni, polmoni, bilaterali femore prossimale, la vagina, e le radici dei nervi sacrali vengono contornati dala radiazione oncologo. </li><li> Descrivere dose di prescrizione di radiazione. Una dose di radiazione di prescrizione 3 × 800 cGy = 2400 cGy (comunemente alla linea isodose 70%) è stato selezionato. In un calcolo equivalente biologico dose di 200 cGy, quasi 6.170 cGy vengono forniti con questa prescrizione, nell&#39;ipotesi di un rapporto &quot;α / β&quot; del 10 per tumore. La radiazione emessa da un acceleratore lineare SBRT viene collimato o utilizzando uno dei collimatori di tungsteno 12 circolare fissa (5 a 60 mm) o utilizzando un tungsteno-rame segmentato esagono collimatore IRIS. Organi 14 normali hanno una tolleranza di radiazione e l&#39;uso di dose-volume parametri istogramma per il vincolo del tessuto normale durante la radiochirurgia stereotassica ablative sono elencati nella Tabella 1. </li></ol> 4. Radiochirurgia stereotassica Cyberknife Trattamento di consegna <ol><li> Eseguire cross-piano di verifica delle immagini. Durante Cyberknife SBRT dei tessuti molli fiducial(O in alcuni casi rigidi punti di repere anatomici quali il cranio per le lesioni endocraniche e la spina dorsale per le lesioni della colonna vertebrale) sono tracciati da cross-plane radiografie e confermato di essere nella posizione di destinazione prevista dal sistema di destinazione localizzazione (TLS) sottosistema. </li><li> Verificare il posizionamento del target. Fiducial sono tracciati in tre e fino a sei gradi di libertà (ad esempio, x, y, z, beccheggio, rollio e imbardata). Immagini generate dal TLS sono automaticamente registrati e confrontati con radiografie digitalmente ricostruite (DRR) generati dal trattamento iniziale di pianificazione TC. </li><li> Verificare posizionamento bersaglio. Se i risultati di registrazione automatica indicano che fiducial hanno spostato oltre le tolleranze predefiniti in uno qualsiasi dei sei gradi di libertà (x, y, z, beccheggio, rollio e imbardata), il trattamento viene automaticamente in pausa. Riposizionamento del paziente si verifica allora. </li><li> Considerare il movimento di destinazione. Obiettivi che si muovono con il movimento respiratorio sono treated utilizzando il sottosistema Synchrony Respiratory Tracking (Synchrony). Con l&#39;utilizzo di una telecamera montata nella camera di trattamento, il sistema rileva continuamente modello respiratorio del paziente (cioè, ottenere dati di posizione esterni) con l&#39;uso di diodi emettitori di luce (LED) apposti su una maglia intorno torace del paziente. 10 insieme con le informazioni di localizzazione fiducial (dati di posizione interni) ottenuto da cross-piano raggi X, il sistema Synchrony costruisce un modello di correlazione con i dati esterni ei dati interni. Questo modello di correlazione permette al braccio robotico di seguire qualsiasi respiratorio movimento causato del bersaglio, mentre durante la consegna di qualsiasi fasci di trattamento. </li><li> Il trattamento può comportare 100-150 posizioni di trattamento consegnabili del braccio robotico. I trattamenti possono durare da 30 a 90 minuti. </li></ol> 5. Risultati rappresentativi Corpo la terapia stereotassica radiazioni (SBRT) possono coinvolgere molti fasci di radiazione individuali (<html> vettori blu) tutti convergenti su obiettivi strettamente associate singole o multiple radiazione clinici, come mostrato nella Figura 1A. Un rappresentante buon esito pianificazione radiochirurgico dà un risultato finale SBRT trattamento con alte dosi di radiazioni copertura del volume bersaglio e cancro bersaglio conformalità cancro. Figure 1B-D mostrano 131 travi stati utilizzati per trattare una ricaduta pelvico di un bersaglio chemorefractory cancro ovarico oltre 42 minuti. SBRT prescritto alla linea isodose 80% resa 100% di copertura clinico volume bersaglio con un indice conformalità di 1,94 per una dose totale di 2400 cGy in tre frazioni giornalieri 800 cGy. Gli istogrammi dose-volume per l&#39;obiettivo clinico (rosso) e per le strutture critiche del retto (marrone), vescica (giallo), piccolo intestino (luce blu) nervi sacrali (tan) e fianchi (arancione) sono illustrati nella Figura 2. <img alt="Figura 1" src="/files/ftp_upload/3793/3793fig1.jpg" /> Figura 1.tp :/ / www.jove.com/files/ftp_upload/3793/3793fig1large.jpg &quot;target =&quot; _blank &quot;&gt; Clicca qui per ingrandire la figura. <img alt="Figura 2" src="/files/ftp_upload/3793/3793fig2.jpg" /> Figura 2. Istogrammi dose-volume per l&#39;obiettivo clinico (rosso) e per le strutture critiche del retto (marrone), vescica (giallo), piccolo intestino (luce blu) nervi sacrali (tan) e fianchi (arancione).

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	<li>Sherouse, G., Bourland, J., Reynolds, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Virtual+simulation+in+the+clinical+setting:+some+practical+considerations.">Virtual simulation in the clinical setting: some practical considerations.</a> Int. J. Radiat. Oncol. Biol. Phys. 19, 1059-1065 (1990).</li><li>Dickesn, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Personalized+fixation+using+a+vacuum+consolidation+technique.">Personalized fixation using a vacuum consolidation technique.</a> Br. J. 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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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Biol. 54, 5359-5380 (2009).</li><li>Choi, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Image-guided+stereotactic+body+radiation+therapy+in+patients+with+isolated+para-aortic+lymph+node+metastases+from+uterine+cervical+and+corpus+cancer.">Image-guided stereotactic body radiation therapy in patients with isolated para-aortic lymph node metastases from uterine cervical and corpus cancer.</a> Int. J. Radiat. Oncol. Biol. Phys. 74, 147-153 (2009).</li></ol>

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Incoraggiare i risultati SBRT primi hanno alimentato la sperimentazione clinica di radiochirurgia per il trattamento di tumori ginecologici persistenti o ricorrenti. 7, 8, 15 Dati in discussione gli effetti radiobiologici e le modalità di morte cellulare risultante da SBRT. Piccoli studi clinici hanno dimostrato che dosi di radiazioni ablativi fornite da SBRT produrre mirata di aliquote di controllo della malattia superiore al 90%. A differenza della radioterapia convenzionale, è stato piuttosto impegnativo per combinare SBRT con le chemioterapie radiosensibilizzante e citotossici / citostatici. Meglio di mira il cancro tramite espansioni di trattamento un aumento di volume e di pianificazione attraverso l&#39;inserimento di 18 F-FDG PET / CT immagini è migliorato gli esiti clinici. Mentre è indispensabile esaminare altri metodi di consegna dose-escalation radiazioni ad alta precisione, rimane indeciso se SBRT in grado di fornire equivalente efficacia terapeutica più bassa e alta rateo di dose brachiterapia. Infatti, la brachiterapia è il più comunemente e validatodatata tecnica per ottenere aumento graduale della dose di radiazione negli obiettivi di tumori ginecologici. In quanto tale, sia l&#39;entusiasmo e la prudenza sono appropriate nella lettura dei dati SBRT disponibili per il trattamento dei tumori ginecologici.

<table border="1"><tbody><tr><td> Nome del dispositivo </td><td> Azienda </td><td> Numero di catalogo </td><td> Comments </td></tr><tr><td> Sistema Cyberknife </td><td> Accuray (Sunnyvale, CA) </td><td></td><td></td></tr></tbody></table>

stereotactic radiosurgery for gynecologic cancer

Corpo radioterapia stereotassica (SBRT) si distingue per necessita più rigida immobilizzazione del paziente, che rappresenta il movimento respiratorio, la pianificazione del trattamento intricato, a bordo delle immagini e riduzione del numero di dosi di radiazione al cancro ablativi si rivolge di solito refrattario alla chemioterapia e alla radioterapia convenzionale. Campi di trattamento Steep dose di radiazioni SBRT drop-off permessi stretta &#39;fascio matita&#39; essere utilizzato per il trattamento ablativo radiazioni condensato in 1 a 3 terapie. Trattare i medici devono capire che SBRT arriva in un pericolo maggiore di lesioni del tessuto normale e la possibilità di Miss tumore geografica. Entrambi devono essere affrontate con l&#39;immobilizzazione degli obiettivi del cancro e di alta precisione consegna del trattamento. Obiettivo di immobilizzazione Cancer è stato raggiunto attraverso l&#39;uso di calchi personalizzati indicizzati polistirolo, sacchi di fagioli evacuate, o corpo-fix stampi con il paziente indipendente compressione addominale. Movimento Intrafraction 1-3 di Targe cancroa causa della respirazione ts ora può essere ridotto mediante tecniche di respirazione dei pazienti rispondono attesa, 4 boccaglio paziente attivo di coordinamento respirazione, 5 respirazione-correlata tomografia computerizzata, 6 o guidata da immagini tracciabilità dei fiducial impiantati dentro e attorno a un tumore in movimento. 7-9 L&#39; Cyberknife sistema (Accuray [Sunnyvale, CA]) utilizza un acceleratore lineare radiazione montata su un braccio robotico industriale che segue con precisione il movimento del paziente respiratorio da una telecamera-tracciato insieme di diodi emettitori di luce (LED) impregnati su un giubbotto montato su un paziente. 10 sostanziali riduzioni dei margini di radioterapia può essere ottenuto mediante il tracciamento del movimento, in ultima analisi, rendendo più piccoli a volumi bersaglio di pianificazione che vengono irradiati con precisione submillimetrica. 11-13 Obiettivi di cancro trattati con SBRT sono irradiati da fasci convergenti, ben collimati. Dose di radiazione risultante a istogrammi bersaglio del cancro del volume hanno un pronome piùced radiazione &quot;spalla&quot; che indica alta percentuale di copertura prevista e una piccola alte dosi di radiazioni &quot;coda&quot;. Così, conformalità bersaglio maggiore va a scapito della dose ridotta uniformità nella destinazione cancro SBRT. Questo può avere implicazioni sia per il controllo del tumore successiva l&#39;obiettivo SBRT e la tolleranza tessuto normale degli organi a rischio. A causa della forte dose di falloff SBRT, la possibilità di malattia occulta sfuggire dose di radiazioni ablativa si verifica se gli obiettivi di cancro non sono pienamente riconosciuti e inadeguate dosi SBRT margini vengono applicate. Clinical volume bersaglio (CTV) di espansione da 0,5 cm, risultante in un volume più grande di pianificazione di destinazione (PTV), è associata a controllo di destinazione senza indebito aumento lesione dei tessuti normali. 7,8 Ulteriore riduzione della probabilità di mancato geografica può essere realizzato per incorporazione di 2 - [18 F] fluoro-2-deossi-D-glucosio (18F-FDG), tomografia ad emissione di positroni (PET) 8 Uso di 18 F-FDG PET / TC in SBRT Cure.pianificazione nt è solo l&#39;inizio di tentativi per scoprire nuove firme bersaglio molecolare di imaging per tumori ginecologici.

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

Stereotactic Radiosurgery for Gynecologic Cancer

Corpo radioterapia stereotassica (SBRT) comporta guidata dalle immagini, la radiazione ablativa consegnato al cancro obiettivi refrattario alla chemioterapia o alla radioterapia convenzionale. Il braccio robotico del sistema Cyberknife SBRT, si utilizza la localizzazione sofisticata target, offre ipofrazionata dosi di radiazioni in grado di sterilizzare gli obiettivi del cancro. Questo articolo prenderà in considerazione i nuovi ruoli di SBRT terapeutiche per i tumori ginecologici.

Radiochirurgia stereotassica for Gynecologic Cancer

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

stereotactic-radiosurgery-for-gynecologic-cancer

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Medicine

17.5K Views.  University Hospitals Case Medical Center and Case Western Reserve University School of Medicine. Corpo radioterapia stereotassica (SBRT) comporta guidata dalle immagini, la radiazione ablativa consegnato al cancro obiettivi refrattario alla chemioterapia o alla radioterapia convenzionale. Il braccio robotico del sistema Cyberknife SBRT, si utilizza la localizzazione sofisticata target, offre ipofrazionata dosi di radiazioni in grado di sterilizzare gli obiettivi del cancro. Questo articolo prenderà in considerazione i nuovi ruoli di SBRT terapeutiche per i tumori gi...

Video: 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.

Multispettrali in tempo reale immagini a fluorescenza per il rilevamento intraoperatoria del linfonodo sentinella in Oncologia Ginecologica

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 ...

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Medicina

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.

Stereotassica impianto intracranico e<em&gt; In vivo</em&gt; Imaging Bioluminescent di xenotrapianti di tumori in un sistema modello del mouse di 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 ...

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.

Luminescenza Imaging Cerenkov (CLI) per il monitoraggio Cancer Therapy

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.

Bioluminescente modello ortotopico di cancro al pancreas Progressione

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.

Profiling di microRNA regolati dagli estrogeni in cellule di carcinoma mammario

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.

Utilizzando Murine inducibile telomerasi alleli in Studi di Tissue Degeneration / Regeneration e il cancro

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.

Studiare cancro del pancreas staminali caratteristiche cellulari per lo sviluppo di nuove strategie di trattamento

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.

Polmone dinamico Tumore inseguimento per stereotassica ablativo corpo Radioterapia

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.

Lavorare con tessuti umani per 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 ...

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.

Modellazione Piazza precoci di cancro ovarico Diffusione in una cultura Organotipica della Cavità peritoneale umano

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