The authors have no acknowledgements.

I dati su tutti i pazienti trattati con cisplatino EBUS-guidata sono stati rivisti. Il comitato di revisione istituzionale presso l&#39;Università della Florida ha approvato questo studio (# IRB201400823). Tutti i dati sono stati raccolti prospetticamente e analizzati retrospettivamente. Utilizzare i seguenti criteri di iscrizione. 1. Selezione del paziente <ol><li> Selezionare i pazienti di età compresa tra i 18 ei 80 anni che hanno un non-piccolo cancro biopsia-confermato cellule del polmone (NSCLC) o carcinoma polmonare a piccole cellule (SCLC) e hanno una recidiva patologicamente confermato tra. Arruolare pazienti che hanno ricevuto almeno 50 Gy di radiazioni al ilare e strutture mediastiniche per 6 mesi o più. Includere i pazienti con recidiva limitata al ilare, mediastino, e le strutture peribronchiali (linfonodi, noduli, e le masse) accessibili attraverso EBUS e metastasi a distanza. </li><li> Presentare tutti questi pazienti ad una scheda di Oncologia Toracica tumore multidisciplinare e di ottenere il consenso su di procedere con l&#39;acisplatino tratumoral a causa della mancanza di altre opzioni terapeutiche localizzate. La decisione di procedere con EBUS-guidata ITC dovrebbe essere una raccomandazione congiunta del Consiglio di oncologia toracica tumore istituzionale. </li></ol> 2. endobronchiale a ultrasuoni (EBUS) di iniezione-guidata Cisplatino <ol><li> Utilizzare un EBUS convessa-sonda, che ha una visualizzazione in tempo reale integrato sonda a ultrasuoni su un broncoscopio flessibile e consente di ilare, mediastino, e le strutture peribronchiali per ITC. </li><li> Assicurarsi che i pazienti astenersi dal cibo per via orale e liquidi per almeno 6 ore prima della procedura. </li><li> Creare una soluzione cisplatino acquosa ad una concentrazione di 1 mg / ml con una dose massima di 40 mg per sessione, sulla base della letteratura precedentemente pubblicato 9, 10. Sciogliere la polvere liofilizzata cisplatino nel 0,9% soluzione di NaCl al momento dell&#39;uso. </li><li> Dare 10 mg di desametasone e 8 mg di ondansetron endovenosa all&#39;inining della procedura per prevenire la nausea. </li><li> Eseguire la procedura con sedazione cosciente con midazolam e fentanyl per via endovenosa (IV). Utilizzare anestesia topica sotto forma di 5 mL di 4% lidocaina nebulizzazione prima della procedura. </li><li> Inizia con una iniezione IV di 2 mg di midazolam e 50 mcg di fentanil. Rivalutare il paziente ogni 1-2 minuti e dare un ulteriore midazolam 1 mg e 25 mg in bolo di fentanyl per raggiungere e mantenere la sedazione cosciente (una coscienza minimamente depresso in modo tale che il paziente è in grado di mantenere costantemente e indipendentemente pervietà delle vie aeree, mantenere riflessi protettivi, e rimanere reattivo ai comandi verbali e la stimolazione fisica). Non superare le dosi massime ammissibili di fentanyl e midazolam, 200 mg e 10 mg, rispettivamente. </li><li> Utilizzare monitoraggio standard, tra cui un elettrocardiogramma continuo a 3 derivazioni monitoraggio della frequenza cardiaca, pulsossimetria continua, e le misurazioni frequenza respiratoria. Fornire Anest localehesia e O supplementare 2 come richiesto. </li><li> Far avanzare la portata attraverso le corde vocali nelle vie aeree. Identificare e localizzare la lesione bersaglio. </li><li> Accendere il processore EBUS e gonfiare il palloncino sulla punta della portata utilizzando 1-2 mL di soluzione fisiologica. Flettere la portata attraverso la parete delle vie aeree e localizzare la lesione. La posizione della lesione dipenderà risultati CT / PET, ma alcuni dei luoghi comuni includono l&#39;area sottocarenali, il diritto e le aree paratracheali a sinistra, e il diritto e le aree ilari sinistra. </li><li> Distribuire un G EBUS ago 22 che si trova in una guaina attraverso il canale di lavoro del campo di applicazione e bloccarla in posizione. NOTA: Anche se non ci sono le coordinate chirurgiche precise per il sito di iniezione, alcuni dei siti comunemente trattate includono il sottocarenali (mediale a livello della giunzione dei bronchi principali destro e sinistro), il paratracheale destra (lateralmente al bronco principale destro, appena sopra a destra dell&#39;orifizio lobo superiore), il paratracheale sinistra(All&#39;incrocio della trachea distale e la principale di sinistra tra l&#39;arco dell&#39;aorta e l&#39;arteria polmonare sinistra), l&#39;ilare destra (lateralmente al bronco intermedio prossimale, sotto il diritto di apertura lobo superiore), e l&#39;ilare sinistra (a la giunzione del lobo superiore di sinistra e la sinistra in basso bronchi del lobo). </li><li> Sotto guida ecografica in tempo reale, forare la parete tracheobronchiale e posizionare l&#39;ago nella lesione bersaglio. Rimuovere il mandrino entro l&#39;ago ed iniettare cisplatino nella lesione 10 mL (1 mg / ml di cisplatino) alla volta. Ritrarre l&#39;ago all&#39;interno del foglio. </li><li> Trattare ogni lesione con quattro forature per sessione in luoghi diversi per facilitare l&#39;iniezione del farmaco in tutto. Iniettare 10 mg di cisplatino per foratura. </li><li> Trattare uno a due lesioni per sessione. Se più di una lesione è trattata per sessione, iniettare ogni sito con 20 mg di cisplatino, con dose totale per sessione non superiore a 40 mg. </li><li> Rimuovere l&#39;ago dal workicanale ng del campo di applicazione. </li><li> Aspirare qualsiasi farmaco aggiuntivo dalle vie aeree distali e rimuovere il broncoscopio. </li><li> Lasciare i pazienti di recuperare come da protocollo ospedaliero e li scaricano lo stesso giorno. </li><li> Ripetere l&#39;iniezione cisplatino (passi 2,7-2,13) ​​una volta a settimana per un totale di 4 settimane (nei giorni 1, 8, 15, e 22). NOTA: una volta trattati, un particolare della lesione non dovrebbe mai essere preso in considerazione per il ritrattamento con ITC in incontri successivi. </li></ol> 3. post-iniezione <ol><li> Valutare la risposta da una tomografia torace follow-up computerizzata (TC) o una tomografia ad emissione di positroni (PET / TC) 8-12 settimane dopo l&#39;ultima sessione di trattamento. Definire la recidiva locale come la ricorrenza presso il sito di trattamento e la recidiva regionale la ricorrenza nel mediastino, ilo, o fossa sovraclaveare. </li></ol>

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The authors have nothing to disclose.

<html> Il nostro manoscritto si concentra sulla gestione di isolati recidive del mediastino e ilari di cancro al polmone per i pazienti in cui più radiazioni non è un&#39;opzione. L&#39;incidenza di IMHR è pari a circa il 9% 11. Diverse opzioni di trattamento sono stati provati in questi pazienti, tra cui la chemioterapia palliativa, più fasci esterni radioterapia o terapia di supporto e sorveglianza. Nel nostro istituto, in cui trattiamo i pazienti con cisplatino intratumorale guidata dalla b...

Il cancro al polmone è il tumore più diffuso in tutto il mondo in entrambi gli uomini e le donne e gli account per 1 a 5 di tutti i decessi per cancro. Ha un tasso di mortalità di 0,87 1, 2. Recidiva dopo il trattamento iniziale rappresenta la maggior parte dei decessi in pazienti con tumore polmonare. Le opzioni terapeutiche per il cancro del polmone ricorrenti sono limitati. Cancro al polmone ricorrente porta anche a una significativa compromissione della qualità di vita e di significativo il burnout caregiver e richiede ulteriori cure di supporto, ognuno dei quali diminuire la probabilità di accettazione e di tolleranza di ulteriori interventi 3. Quasi il 30% dei pazienti con NSCLC hanno recidiva loco-regionale dopo la radiazione 4. Le opzioni di trattamento per questi pazienti sono limitati e comprendono la chemioterapia sistemica, che ha un tasso di risposta obiettivo di solo il 10% 5, 6.Anche se le radiazioni al torace ripetizione è fattibile, è stata studiata principalmente per scopi palliative e sollievo dai sintomi e non è stato utilizzato per la remissione della malattia 3. Nel nostro istituto, tutti i pazienti con recidiva isolata mediastinica e ilare (IMHR) accessibili tramite broncoscopia sono stati trattati con una ecografia endobronchiale (EBUS)-guidata iniezione intratumorale transbronchiale di cisplatino (ITC). Il trattamento concomitante con chemioterapia sistemica e / o radioterapia a fasci esterni (EBRT) è stato ammesso come da discrezionalità gli oncologi trattamento. Questo manoscritto evidenzia la sicurezza, l&#39;efficacia, la fattibilità, e metodi dettagliate del protocollo utilizzato presso il nostro istituto.

<table border="0" cellpadding="0" cellspacing="0" width="261">
	<tbody>
		<tr height="20">
			<td colspan="2" height="20" style="height:20px;width:261px;">Bronchoscope</td>
		</tr>
		<tr height="20">
			<td colspan="2" height="20" style="height:20px;">22 Guage olympus EBUS needle</td>
		</tr>
		<tr height="20">
			<td colspan="2" height="20" style="height:20px;">40 g of cisplatin in 40 mls of normal saline</td>
		</tr>
	</tbody>
</table>

endobronchial ultrasound-guided intratumoral injection of cisplatin for the treatment of isolated mediastinal recurrence of lung cancer

Isolato ilari e la ricorrenza del mediastino (IMHR) dopo radioterapia esterna (EBRT), in pazienti con carcinoma polmonare è comune. Questi pazienti non hanno molte opzioni di trattamento e di solito sono offerti chemioterapia palliativa o migliore terapia di supporto. ecografia endobronchiale (EBUS)-guidata iniezione intratumorale di cisplatino (ITC) è un nuovo approccio per questi pazienti. La procedura viene eseguita in sedazione cosciente. La lesione si trova con una broncoscopia con EBUS, e un ago EBUS 22-gauge è avanzato attraverso il canale di lavoro del campo di applicazione e bloccato in posizione. Sotto guida ecografica, la parete dell&#39;albero tracheobronchiale è perforato e l&#39;ago viene spostato nella lesione target. Il mandrino ago viene poi rimosso e cisplatino (40 mg / 40 ml) viene iniettato nella lesione. Da uno a due siti vengono trattati per ogni sessione. I dettagli della procedura sono descritti nella sezione del protocollo di carta. Presso il nostro centro, 50 siti sono stati trattati in 36 pazienti(19 maschi, 17 femmine). L&#39;età media della nostra coorte era 61,9 ± 8,5 anni. Abbiamo eseguito analisi finali su 35 pazienti e 41 siti. 24/35 (69%) hanno avuto una risposta completa o parziale (responder), mentre 11/35 (31%) hanno avuto una stabilizzazione della malattia o progressiva (non-responder). Nel complesso, la sopravvivenza nel nostro gruppo è stato di 8 mesi (95% CI 6-11 mesi), con pazienti che hanno risposto hanno significativamente migliore sopravvivenza rispetto a quelli che non hanno.

La risposta è stata misurata mediante follow-up di imaging 8-12 settimane dopo la terapia. La risposta è stata classificata come la remissione completa (CR), remissione parziale (PR), malattia stabile (SD), malattia progressiva (PD), o in grado di valutare la risposta in base a RECIST 1.1 criteri 8. I pazienti con CR e PR sono stati considerati responder, e gli altri sono stati classificati come non-responder. Gli esiti secondari includevano la risposta sulla ba...

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Endobronchial Ultrasound-guided Intratumoral Injection of Cisplatin for the Treatment of Isolated Mediastinal Recurrence of Lung Cancer

The management of isolated recurrent lung cancer in a previously-irradiated field is challenging. Here, we describe an endobronchial ultrasound (EBUS)-guided cisplatin injection for the management of patients with localized lung cancer recurrence in a previously-radiated field.

Endobronchiale ecoguidata iniezione intratumorale di cisplatino per il trattamento di isolato mediastinica ricorrenza del cancro del polmone

Isolated hilar and mediastinal recurrence (IMHR) following external beam radiation therapy (EBRT) in patients with lung cancer is common. These patients ...

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10.3K Views.  University of Florida College of Medicine. The management of isolated recurrent lung cancer in a previously-irradiated field is challenging. Here, we describe an endobronchial ultrasound (EBUS)-guided cisplatin injection for the management of patients with localized lung cancer recurrence in a previously-radiated field. 

Video: Endobronchial Ultrasound-guided Intratumoral Injection of Cisplatin for the Treatment of Isolated Mediastinal Recurrence of Lung Cancer

Operating Procedures of the Electrochemotherapy for Treatment of Tumor in Dogs and Cats

Electrochemotherapy (ECT) is a local approach which is used for treating solid tumors of different histologies. Its mechanism is based on cell membrane permeabilization by means of &#34;electroporation&#34;. To achieve the &#34;electroporation&#34; of the cells, electric pulses are generated by a generator and delivered to the target tissue by the use of electrodes. Electroporation is a physical method which is used to introduce molecules, like cytostatic drugs, into the cells that could not pass the cell membrane on their own. In electrochemotherapy, currently, cisplatin and bleomycin are clinically used. Electrochemotherapy antitumor effectiveness is high, for example up to 100% complete response of canine mast cell tumors smaller than 2 cm3 was achieved. Additionally, electrochemotherapy can be used for the treatment of inoperable tumors. One of the important characteristics of electrochemotherapy is that it can be effective as a one-time treatment only. However, in the case of failure or partial tumor response it can be repeated several times with equal or improved effectiveness. Electrochemotherapy is already a standard treatment for cutaneous and subcutaneous tumors of various histologies in human and veterinary oncology. Furthermore, several clinical studies exploiting electrochemotherapy for deep-seated tumors are on-going.

Electrochemotherapy is the local treatment of solid tumors. The main mechanism is electroporation-mediated permeabilization of the tumor cells' membrane which enables increased internalization of cytostatics like bleomycin and cisplatin. Thus, antitumor effectiveness is potentiated at the site of the application of electric pulses to the tumors.

Procedure operative della elettrochemioterapia per il trattamento del tumore in cani e gatti

Electrochemotherapy (ECT) is a local approach which is used for treating solid tumors of different histologies. Its mechanism is based on cell membrane ...

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The Establishment of a Lung Colonization Assay for Circulating Tumor Cell Visualization in Lung Tissues

Metastasis is the major cause of cancer death. The role of circulating tumor cells (CTCs) in promoting cancer metastasis, in which lung colonization by CTCs critically contributes to early lung metastatic processes, has been vigorously investigated. As such, animal models are the only approach that captures the full systemic process of metastasis. Given that problems occur in previous experimental designs for examining the contributions of CTCs to blood vessel extravasation, we established an in vivo lung colonization assay in which a long-term-fluorescence cell-tracer, carboxyfluorescein succinimidyl ester (CFSE), was used to label suspended tumor cells and lung perfusion was performed to clear non-specifically trapped CTCs prior to lung removal, confocal imaging, and quantification. Polymeric fibronectin (polyFN) assembled on CTC surfaces has been found to mediate lung colonization in the final establishment of metastatic tumor tissues. Here, to specifically test the requirement of polyFN assembly on CTCs for lung colonization and extravasation, we performed short term lung colonization assays in which suspended Lewis lung carcinoma cells (LLCs) stably expressing FN-shRNA (shFN) or scramble-shRNA (shScr) and pre-labeled with 20 &#956;M of CFSE were intravenously inoculated into C57BL/6 mice. We successfully demonstrated that the abilities of shFN LLC cells to colonize the mouse lungs were significantly diminished in comparison to shScr LLC cells. Therefore, this short-term methodology may be widely applied to specifically demonstrate the ability of CTCs within the circulation to colonize the lungs.

An animal model is needed to decipher the role of circulating tumor cells (CTCs) in promoting lung colonization during cancer metastasis. Here, we established and successfully performed an in vivo assay to specifically test the requirement of polymeric fibronectin (polyFN) assembly on CTCs for lung colonization.

L'istituzione di un'analisi di colonizzazione del polmone per la visualizzazione di cellule del tumore nei tessuti polmonari di circolazione

Metastasis is the major cause of cancer death. The role of circulating tumor cells (CTCs) in promoting cancer metastasis, in which lung colonization by ...

Utilization of Ultrasound Guided Tissue-directed Cellular Implantation for the Establishment of Biologically Relevant Metastatic Tumor Xenografts

Preclinical testing of anticancer therapies relies on relevant xenograft models that mimic the innate tendencies of cancer. Advantages of standard subcutaneous flank models include procedural ease and the ability to monitor tumor progression and response without invasive imaging. Such models are often inconsistent in translational clinical trials and have limited biologically relevant characteristics with low proclivity to produce metastasis, as there is a lack of a native microenvironment. In comparison, orthotopic xenograft models at native tumor sites have been shown to mimic the tumor microenvironment and replicate important disease characteristics such as distant metastatic spread. These models often require tedious surgical procedures with prolonged anesthetic time and recovery periods. To address this, cancer researchers have recently utilized ultrasound-guided injection techniques to establish cancer xenograft models for preclinical experiments, which allows for rapid and reliable establishment of tissue-directed murine models. Ultrasound visualization also provides a noninvasive method for longitudinal assessment of tumor engraftment and growth. Here, we describe the method for ultrasound-guided injection of cancer cells, utilizing the adrenal gland for NB and renal sub capsule for ES. This minimally invasive approach overcomes tedious open surgery implantation of cancer cells in tissue-specific locations for growth and metastasis, and abates morbid recovery periods. We describe the utilization of both established cell lines and patient derived cell lines for orthotopic injection. Pre-made commercial kits are available for tumor dissociation and luciferase tagging of cells. Injection of cell suspension using image-guidance provides a minimally invasive and reproducible platform for the creation of preclinical models. This method is utilized to create reliable preclinical models for other cancers such as bladder, liver and pancreas exemplifying its untapped potential for numerous cancer models.

Here, we present a protocol to utilize ultrasound-guided injection of neuroblastoma (NB) and Ewing's sarcoma (ES) cells (established cell lines and patient-derived tumor cells) at biologically relevant sites to create reliable preclinical models for cancer research.

L'utilizzo degli ultrasuoni guidati l'impianto cellulare diretto del tessuto per l'istituzione degli xenotrapianti del tumore metastatico biologicamente rilevanti

Preclinical testing of anticancer therapies relies on relevant xenograft models that mimic the innate tendencies of cancer. Advantages of standard ...

Evaluation of the Feasibility, Safety, and Accuracy of an Intraoperative High-intensity Focused Ultrasound Device for Treating Liver Metastases

Today, the only potentially curative option in patients with colorectal liver metastases is surgery. However, liver resection is feasible in less than 20% of patients. Surgery has been widely used in association with radiofrequency, cryotherapy, or microwaves to expand the number of treatments performed with a curative intent. Nevertheless, several limitations have been documented when using these techniques (i.e., a traumatic puncture of the parenchyma, a limited size of lesions, and an inability to monitor the treatment in real-time).High-intensity focused ultrasound (HIFU) technology can achieve precise ablations of biological tissues without incisions or radiation. Current HIFU devices are based on an extracorporeal approach with limited access to the liver. We have developed a HIFU device designed for intraoperative use. The use of a toroidal transducer enables an ablation rate (10 cm3&#183;min-1) higher than any other treatment and is independent of perfusion.
The feasibility, safety, and accuracy of intraoperative HIFU ablation were evaluated during an ablate-and-resect prospective study. This clinical phase I and IIa study was performed in patients undergoing hepatectomy for liver metastases. The HIFU treatment was performed on healthy tissue scheduled for resection.Liver metastases measuring less than 20 mm will be targeted during phase IIb (currently ongoing). This set-up allows the real-time evaluation of HIFU ablation while protecting participating patients from any adverse effects related to this new technique.
Fifteen patients were included in phase I&#8211;IIa and 30 HIFU ablations were safely created within 40 s and with a precision of 1&#8211;2 mm. The average dimensions of the HIFU ablations were 27.5 x 21.0 mm2, corresponding to a volume of approximately 7.5 cm3. The aim of the ongoing phase IIb is to ablate metastases of less than 20 mm in diameter with a 5 mm margin.

Here, we present an ablate-and-resect prospective study to evaluate the feasibility, safety, and accuracy of intraoperative high-intensity focused ultrasound ablation in patients undergoing hepatectomy for liver metastases.

Valutazione di fattibilità, la sicurezza e la precisione di un'intraoperatoria ad alta intensità focalizzato dispositivo a ultrasuoni per il trattamento di metastasi epatiche

Today, the only potentially curative option in patients with colorectal liver metastases is surgery. However, liver resection is feasible in less than 20% ...

Elastic Staining on Paraffin-embedded Slides of pT3N0M0 Gastric Cancer Tissue

The elastic lamina, which is usually located in the sub-mesothelial layer, adjacent to the peritoneum mesothelial cells, is amphiphilic on hematoxylin and eosin (H&amp;E) staining but can be visualized through elastic staining. One of the important benefits of elastic staining is that it makes it easy to determine whether tumor cells have invaded beyond the elastic lamina. This helps in examining the extent of peritoneal surface invasion, thereby distinguishing the pT3 and pT4 stages in gastrointestinal cancer. In this study, we present a protocol to identify elastic fibers in the formalin-fixed, paraffin-embedded pT3N0M0 gastric cancer tissue sections. We prepare 5-&#181;m paraffin sections fixed on slides, and then all the slides are deparaffinated and rehydrated during the staining procedure. Subsequently, all the sections are oxidized by potassium permanganate, bleached by oxalic acid, stained by elastic staining, and counterstained by Van Gieson's. Finally, we examine the effect of staining; elastic lamina is stained blue-black and collagen fibers are stained red, while the cancer cells are stained in varying shades of yellow. We also describe in detail the methods used for determining the positional relationship between cancer cells and elastic lamina. This method is simple, low-cost, and widely applicable in the identification of peritoneal surface invasion in gastrointestinal cancer because of its outstanding selectivity for elastic fibers and authentic results.

Here, we present a protocol of elastic staining to identify elastic fibers in pT3N0M0 gastric cancer tissues on formalin-fixed, paraffin-embedded sections. Subsequently, we describe a method to determine whether tumor cells invade beyond the elastic lamina.

Colorazione elastica su vetrini di tessuto gastrico pT3N0M0 gastrico

The elastic lamina, which is usually located in the sub-mesothelial layer, adjacent to the peritoneum mesothelial cells, is amphiphilic on hematoxylin and ...

Ultrasound-Guided Orthotopic Implantation of Murine Pancreatic Ductal Adenocarcinoma

The recent success of immune checkpoint blockade in melanoma and lung adenocarcinoma has galvanized the field of immuno-oncology as well as revealed the limitations of current treatments, as the majority of patients do not respond to immunotherapy. Development of accurate preclinical models to quickly identify novel and effective therapeutic combinations are critical to address this unmet clinical need. Pancreatic ductal adenocarcinoma (PDA) is a canonical example of an immune checkpoint blockade resistant tumor with only 2% of patients responding to immunotherapy. The genetically engineered KrasG12D+/-;Trp53R172H+/-;Pdx-1 Cre (KPC) mouse model of PDA recapitulates human disease and is a valuable tool for assessing therapies for immunotherapy resistant in the preclinical setting, but time to tumor onset is highly variable. Surgical orthotopic tumor implantation models of PDA maintain the immunobiological hallmarks of the KPC tissue-specific tumor microenvironment (TME) but require a time-intensive procedure and introduce aberrant inflammation. Here, we use an ultrasound-guided orthotopic tumor implantation model (UG-OTIM) to non-invasively inject KPC-derived PDA cell lines directly into the mouse pancreas. UG-OTIM tumors grow in the endogenous tissue site, faithfully recapitulate histological features of the PDA TME, and reach enrollment-sized tumors for preclinical studies by four weeks after injection with minimal seeding on the peritoneal wall. The UG-OTIM system described here is a rapid and reproducible tumor model that may allow for high throughput analysis of novel therapeutic combinations in the murine PDA TME.

We describe a protocol for the ultrasound guided implantation of murine-derived pancreatic ductal adenocarcinoma cell lines directly into the native tumor site. This approach resulted in pancreatic tumors detectable by ultrasound scanning within 2-4 weeks of injection, and significantly reduced the proportion tumor cell seeding on the peritoneal wall as compared to surgical orthotopic implantation.

Impianto ortotopico a ultrasuoni dell'adenocarcinoma dugalese Murine

The recent success of immune checkpoint blockade in melanoma and lung adenocarcinoma has galvanized the field of immuno-oncology as well as revealed the ...

Perfusion and Inflation of the Mouse Lung for Tumor Histology

The ability to evaluate lung histology is critical for the fields of lung cancer research and cancer metastasis. It is equally important to perform necropsies rapidly and efficiently from studies without sacrificing the quality of the tissues procured. The goal of this protocol is to present a method to rapidly perfuse, inflate, and fix mouse lungs for downstream histological analysis. This method does not standardize lung inflation; thus, it does not require any special procedures or equipment and instead simply instills fixative directly through the trachea following perfusion through the heart. This allows for sufficient estimation of tumor size, histology, and scoring. This also allows for the collection of frozen tissue prior to lung tissue fixation. This method is limited in that it does not allow for later morphometric quantification of the lung; however, it is more than sufficient for lung tumor analysis from genetically engineered mouse models (GEMMs), syngeneic models, as well as xenograft tumor and metastasis studies.

The purpose of this method is to present a simple and efficient method for the perfusion, inflation, and fixation of mouse lungs for the examination of lung tumor pathology and evaluation of metastases to the lung.

Perfusione e inflazione del polmone del topo per l'istologia tumorale

The ability to evaluate lung histology is critical for the fields of lung cancer research and cancer metastasis. It is equally important to perform ...

Pathological Analysis of Lung Metastasis Following Lateral Tail-Vein Injection of Tumor Cells

Metastasis, the primary cause of morbidity and mortality for most cancer patients, can be challenging to model preclinically in mice. Few spontaneous metastasis models are available. Thus, the experimental metastasis model involving tail-vein injection of suitable cell lines is a mainstay of metastasis research. When cancer cells are injected into the lateral tail-vein, the lung is their preferred site of colonization. A potential limitation of this technique is the accurate quantification of the metastatic lung tumor burden. While some investigators count macrometastases of a pre-defined size and/or include micrometastases following sectioning of tissue, others determine the area of metastatic lesions relative to normal tissue area. Both of these quantification methods can be exceedingly difficult when the metastatic burden is high. Herein, we demonstrate an intravenous injection model of lung metastasis followed by an advanced method for quantifying metastatic tumor burden using image analysis software. This process allows for investigation of multiple end-point parameters, including average metastasis size, total number of metastases, and total metastasis area, to provide a comprehensive analysis. Furthermore, this method has been reviewed by a veterinary pathologist board-certified by the American College of Veterinary Pathologists (SEK) to ensure accuracy.

Intravenous injection of cancer cells is often used in metastasis research, but the metastatic tumor burden can be difficult to analyze. Herein, we demonstrate a tail-vein injection model of metastasis and include a novel approach to analyze the resulting metastatic lung tumor burden.

Analisi patologica delle metastasi polmonari a seguito di iniezione laterale della vena della coda di cellule tumorali

Metastasis, the primary cause of morbidity and mortality for most cancer patients, can be challenging to model preclinically in mice. Few spontaneous ...

Modeling Brain Metastases Through Intracranial Injection and Magnetic Resonance Imaging

Metastatic spread of cancer is an unfortunate consequence of disease progression, aggressive cancer subtypes, and/or late diagnosis. Brain metastases are particularly devastating, difficult to treat, and confer a poor prognosis. While the precise incidence of brain metastases in the United States remains hard to estimate, it is likely to increase as extracranial therapies continue to become more efficacious in treating cancer. Thus, it is necessary to identify and develop novel therapeutic approaches to treat metastasis at this site. To this end, intracranial injection of cancer cells has become a well-established method in which to model brain metastasis. Previously, the inability to directly measure tumor growth has been a technical hindrance to this model; however, increasing availability and quality of small animal imaging modalities, such as magnetic resonance imaging (MRI), are vastly improving the ability to monitor tumor growth over time and infer changes within the brain during the experimental period. Herein, intracranial injection of murine mammary tumor cells into immunocompetent mice followed by MRI is demonstrated. The presented injection approach utilizes isoflurane anesthesia and a stereotactic setup with a digitally controlled, automated drill and needle injection to enhance precision, and reduce technical error. MRI is measured over time using a 9.4 Tesla instrument in The Ohio State University James Comprehensive Cancer Center Small Animal Imaging Shared Resource. Tumor volume measurements are demonstrated at each time point through use of ImageJ. Overall, this intracranial injection approach allows for precise injection, day-to-day monitoring, and accurate tumor volume measurements, which combined greatly enhance the utility of this model system to test novel hypotheses on the drivers of brain metastases.

Intracranial brain metastasis modeling is complicated by an inability to monitor tumor size and response to treatment with precise and timely methods. The presented methodology couples intracranial tumor injection with magnetic resonance imaging analysis, which when combined, cultivates precise and consistent injections, enhanced animal monitoring, and accurate tumor volume measurements.

Modellazione delle metastasi cerebrali attraverso l'iniezione intracraniciale e la risonanza magnetica

Metastatic spread of cancer is an unfortunate consequence of disease progression, aggressive cancer subtypes, and/or late diagnosis. Brain metastases are ...

Modeling Brain Metastasis Via Tail-Vein Injection of Inflammatory Breast Cancer Cells

Metastatic spread to the brain is a common and devastating manifestation of many types of cancer. In the United States alone, about 200,000 patients are diagnosed with brain metastases each year. Significant progress has been made in improving survival outcomes for patients with primary breast cancer and systemic malignancies; however, the dismal prognosis for patients with clinical brain metastases highlights the urgent need to develop novel therapeutic agents and strategies against this deadly disease. The lack of suitable experimental models has been one of the major hurdles impeding advancement of our understanding of brain metastasis biology and treatment. Herein, we describe a xenograft mouse model of brain metastasis generated via tail-vein injection of an endogenously HER2-amplified cell line derived from inflammatory breast cancer (IBC), a rare and aggressive form of breast cancer. Cells were labeled with firefly luciferase and green fluorescence protein to monitor brain metastasis, and quantified metastatic burden by bioluminescence imaging, fluorescent stereomicroscopy, and histologic evaluation. Mice robustly and consistently develop brain metastases, allowing investigation of key mediators in the metastatic process and the development of preclinical testing of new treatment strategies.

We describe a xenograft mouse model of breast cancer brain metastasis generated via tail-vein injection of an endogenously HER2-amplified inflammatory breast cancer cell line.

Modellazione delle metastasi cerebrali tramite iniezione di vena della coda di cellule infiammatorie del cancro al seno

Metastatic spread to the brain is a common and devastating manifestation of many types of cancer. In the United States alone, about 200,000 patients are ...