Name: a preclinical murine model of hepatic metastases
Uploaded: 2014-09-27T13:00:00.000+00:00
Duration: 6 min 51 s
Description: Watch this Scientific Journal Video about A Preclinical Murine Model of Hepatic Metastases at JoVE.com

KF und KO sind Co ersten Autoren zusammen mit KS Diese Arbeit wurde zum Teil durch die AHPBA Fellowship (KS), NIH K23 CA148964-01 (LZ), Johns Hopkins School of Medicine Clinical Scientist Award (LZ), Viragh Stiftung und das Überspringen Viragh Bauchspeicheldrüsenkrebszentrum an der Johns Hopkins (EMJ und unterstützt LZ), die National Foundation Pankreas (LZ), die Lefkofsky Family Foundation (LZ), das NCI SPORE in Magen-Darm-Krebs P50 CA062924 (EMJ, LZ), der Lustgarten-Stiftung (LZ) und die Sol Goldman Bauchspeicheldrüsenkrebszentrum Zuschüsse (KS , BHE, LZ).

Alle Tierversuche entsprach den Richtlinien der Animal Care und Verwenden Ausschuss der Johns Hopkins University, und die Tiere wurden in Übereinstimmung mit den Richtlinien der Gesellschaft für Evaluierung und Akkreditierung von Laborbetreuung (AAALAC) gehalten. Das chirurgische Verfahren wird unter sterilen Bedingungen im Operationssaal, die ein Minimum von fünfzehn Luftwechsel pro Stunde unterzogen geführt. Alle Instrumente werden durch Autoklavieren vor der Prozedur und erneut mit 70% Ethanol in zwischen jeder Maus während des Verfahrens sterilisiert. Alle Mäuse nicht erwartet, dass das chirurgische Verfahren überleben werden unter CO 2 für 3 min, gefolgt durch Genickbruch noch unter Narkose eingeschläfert. 1. Zellpräparation <ol><li> Resuspendieren kultiviert Panc02 Zellen in phosphatgepufferter Salzlösung mit 2 x 10 7 Zellen / ml, und auf Eis halten. </li></ol> 2. Maus Vorbereitung <ol><li> Verwalten KetaminE (100 mg / kg) mit Xylazin (10 mg / kg) gemischt intraperitoneal 8-12 Wochen alte weibliche C57BL / 6 Mäuse in geteilten Dosen. </li><li> Überprüfen Sie, ob der Zeh Prise Rückzug Reflex wird abgeschafft, um sicherzustellen, dass die Maus vollständig betäubt wird. </li><li> Verwaltung zusätzlicher Dosen von Ketamin (100 mg / kg) mit Xylazin (10 mg / kg) nach Bedarf, um die Mäuse vollständig betäuben gemischt. </li><li> Anwenden Puralube Vet Salbe auf die Augen der Mäuse um das Trocknen zu verhindern, während die Mäuse unter Anästhesie. </li><li> Rasur das Operationsgebiet der Mäuse zu einer Kontamination der Wunde zu vermeiden. </li><li> Bereiten Sie die linke subkostalen Bereich der Schnittführung mit 70% Ethanol und dann Iod. </li><li> Legen Sie ein Operationstuch um den Bauch, um die Sterilität zu erhalten. </li></ol> 3. Laparotomie <ol><li> Sie links subkostalen Einschnitt im Einklang mit dem linken Ohr durch die Haut und durch das Peritoneum mit einem Skalpell. </li><li> Ausdruck der Milz durch den Einschnitt durch die Anwendung gleichzeitiger digitaler pruck entlang der kranialen und kaudalen Aspekte des Einschnitts. </li><li> Suchen Sie die Milz Blutgefäße am unteren Ende der Milz. </li><li> Teilen Sie die Milz, indem zwei Horizon mittlerer Größe Liga Clips in der Mitte der Milz vorsichtig, um eine Beschädigung der Milz-Blutgefäße an der Milz Pole. </li><li> Legen Sie den oberen Pol der Milz zurück in die Bauchhöhle, um Verunreinigungen zu vermeiden. </li></ol> 4. Tumorinjektion <ol><li> Auszuarbeiten 150 ul Phosphat-gepufferter Kochsalzlösung in einer 26 G x 8.5 &quot;Spritze. </li><li> Auszuarbeiten 100 ul Panc02 (2 x10 6 Zellen) in die gleiche Spritze Aufrechterhaltung der Spritze in einer aufrechten Position zu jedem Zeitpunkt während der Injektion. </li><li> Tauchen Sie die Nadel in einem 70% igen Ethanollösung. Die Nadel wird in 70% Ethanol eingetaucht, um Sterilität zu gewährleisten. </li><li> Injizieren die Zellen langsam in die freigelegte hemispleen wobei Sie die Spritze aufrecht zu allen Zeiten gehalten. Die Abschrägung der syringe zu allen Zeiten durch die Milz Kapsel beobachtet werden, um zu vermeiden Injektion der Zellen in der Milz. Eine Weißfärbung der Milz und Blutgefäße sollte bei Injektion beobachtet werden. </li><li> Erhöhen die Milz und gelten ein Medium Horizont Clip unter der Milz, die Milz Blutgefäße verschließen. </li><li> Wenden Sie einen kleinen Horizont Clip auf die meisten distalen Aspekt der Bauchspeicheldrüse und Milz Gefäße. </li><li> Ligieren die Bauchspeicheldrüse und Milz Schiffe aus der hemispleen durch Schneiden über dem Horizont Clips. </li><li> Platzieren Sie die Maus auf seiner &#39;Seite, und bewässern den Schnitt mit destilliertem Wasser. </li></ol> 5. Bauchdeckenverschluss und aus der Narkose <ol><li> Schließen Sie das Bauchfell mit einem 4-0 Lauf Stich. </li><li> Gelten zwei vor drei Hautklammern, um den Hautschnitt zu schließen. </li><li> Verwalten 0,1 mg / kg Buprenorphin oder 5-10 mg / kg Carprofen subkutan postoperative Schmerzen zu lindern. </li><li> Platzieren Sie die Maus auf einem Heizkissen für recOvery, bis Handy und die Maus zeigt, regelmäßige Atmung. Tiere werden nur in der Gesellschaft von anderen Tieren, nach völlig von der Operation erholt zurückgekehrt. </li></ol> 6. Necropsy Prüfung und Ernten der Leber <ol><li> Zwischen 30 bis 60 Tage nach der Operation, wird Mäusen beginnen, klinische Symptome der Metastasierung zeigen und Euthanasie verlangen. Anzeichen von Metastasen erfordern humane Sterbehilfe sind vergrößerte Bauch und die Entwicklung von Aszites. Euthanize Mäuse durch Inhalation von CO 2. </li><li> Nach Einatmen, führen Genickbruch. </li><li> Nachdem das Tier festgestellt worden, die auf nicht sein, einen Einschnitt durch das Peritoneum, um den Bauch mit einer Schere aus. </li><li> Mit einer Pinzette und Schere vorsichtig schneiden die Leber aus dem Bauch. </li><li> Legen Sie die Leber in OCT-Verbindung und Flash mit flüssigem Stickstoff einfrieren. Bewahren Sie die Leber bei -80 ° C. Alternativ kann das Liver kann in 16% neutral gepuffertem Formalin bei RT für 24 h und in Paraffin eingebetteten fixiert werden. </li></ol>

<ol>
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Die Autoren haben keine relevanten Interessenkonflikte offen zu legen.

Diese präklinischen Mausmodell für Bauchspeicheldrüsenkrebs Metastasierung in die Leber über eine hemispleen Injektionstechnik ist das erste Modell beschrieben, die viele der klinischen und immunologischen Bedingungen der Patienten mit metastatischer Erkrankung der Leber spiegelt. Dieses Modell bietet mehrere Vorteile gegenüber anderen Mausmodellen. Erstens, anders als transgene Modelle von Pankreaskrebs, bei dem nur 30% der Mäuse entwickeln Metastasen in der Leber und dem Zeitpunkt der Metastasenbildung ist sehr ...

Duktalen Adenokarzinom des Pankreas (PDA) ist die vierthäufigste Ursache von Krebs Todesfälle in den Vereinigten Staaten 1. Der fünfjährige Überlebensrate für Patienten mit metastasiertem PDA ist 2-12% 1. Obwohl Adjuvans und neoadjuvante Chemotherapie bei Bauchspeicheldrüsenkrebs ist heute effektiver als je zuvor, bleibt es immer noch einen verzweifelten Bedarf an neuen Therapien. Die Entwicklung neuartiger Therapeutika erfordert zuverlässige präklinischen Tiermodellen. Obwohl subkutanen Tumormodellen sind einfach zu bedienen, sind Nachteile, die Unfähigkeit dieser Tumoren, Metastasen zu bilden und imitieren die Tumorprogression und Mikroumgebungen in menschlichen Krebserkrankungen gesehen. Gentechnisch veränderte Modelle, wie der gentechnisch veränderte Maus enthalten, Kras und p53-Knock-in onkogene Mutationen (die KPC-Modell) 2 und die KPC-Modell mit einem YFP-Linie Tracer (die PKCY Modell) 3 ermöglichen die Untersuchung von natürlich vorkommenden Tumoren in immunocompetent Mäusen. Zusätzlich ist der Tumor-Mikroumgebung unverändert und ähnelt mehr der in menschlichen Tumorprogression angesehen. Leider ist die Zeitsteuerung der Tumorentwicklung Variable innerhalb dieser Modelle, was es schwierig experimentelle Therapien zu studieren. Zusätzlich Rückkreuzung dieser Mäuse erfordert eine erhebliche Menge an Zeit und finanziellen Engagement, das nicht machbar ist immer. Schließlich implantierten Xenograft PDA Mausmodelle erforderlich machen immungeschwächten Mäusen, die verändert wurden oder nicht vorhanden Tumormikroumgebungen. Als ein Ergebnis wird die Wirksamkeit von experimentellen Behandlungen in diesen Tiermodellen gezeigt werden oft in klinischen Studien am Menschen 4 nicht reproduzierbar. Diese Hemisplenektomie Mausmodell verwendet Panc02 Tumorzellen, die ein hoch tumorigenen, Methylcholanthren induzierten Pankreastumorzelllinie aus C57BL / 6 Mäusen, 5, 6 ableiten. Zusätzlich anderen murinen PDA Zelllinien aus der KPC mic abgeleitete kann in diesem Modell verwendet werden. Schulick et al. Zuvor entwickelte eine Technik Hemisplenektomie und schuf eine syngenen Mausmodell von Darmkrebs Metastasen 7. Diese Hemisplenektomie Modell (Abbildung 1) bietet konsistente und gleich Tumorimpfung bei immunkompetenten Mäusen Kredit sich auf die Untersuchung von neuen therapeutischen Mittel 7-10.

<table><tbody><tr><td>Cell culture media and components</td><td></td><td></td><td>will vary depending on cell line</td></tr><tr><td>Phosphate Buffered Saline</td><td>Gibco by Life Technologies</td><td>10010-023</td><td></td></tr><tr><td>15 ml centrifuge tubes</td><td>Corning</td><td>430052</td><td></td></tr><tr><td>Curved Operating Scissor</td><td>Roboz Surgical Store</td><td>RS-6835</td><td></td></tr><tr><td>Micro Dissecting Graefe Forceps</td><td>Roboz Surgical Store</td><td>RS-5130</td><td></td></tr><tr><td>Needle holder (regular)</td><td>World Precision Instruments, Inc</td><td></td><td></td></tr><tr><td>3-0 or 4-0 suture</td><td></td><td></td><td>courtesy of dept of surgery, Johns Hopkins</td></tr><tr><td>Weck Horizon Open Ligating Clip Applier, small, angled</td><td>Teleflex</td><td>137085</td><td></td></tr><tr><td>Weck Horizon Open Ligating Clip Applier, medium, angled</td><td>Teleflex</td><td>237115</td><td></td></tr><tr><td>Weck Horizon medium ligating clips</td><td>Teleflex</td><td>002200</td><td></td></tr><tr><td>Weck Horizon small ligating clips</td><td>Teleflex</td><td>001200</td><td></td></tr><tr><td>Syringe, 1 cc, tb syringe, 3/8&quot; slip tip</td><td>Becton Dickinson</td><td>309625</td><td></td></tr><tr><td>Syringe, 1 cc, tb syringe, 5/8&quot; slip tip</td><td>Becton Dickinson</td><td>309597</td><td></td></tr><tr><td>9 mm Autoclip Applier</td><td>Mikron</td><td>427630</td><td></td></tr><tr><td>9 mm Autoclip</td><td>Becton Dickinson</td><td>427631</td><td></td></tr><tr><td>Heating pad</td><td>Sunbeam</td><td>CAT93</td><td></td></tr><tr><td>70% Ethanol</td><td></td><td></td><td>sold by numerous vendors</td></tr><tr><td>Ketamine Hydrochloride&amp;nbsp;</td><td>Hospira</td><td>22395DD</td><td></td></tr><tr><td>Xylazine hydrochloride</td><td>Sigma</td><td>X1251</td><td></td></tr></tbody></table>

a preclinical murine model of hepatic metastases

Zahlreiche murine Modelle wurden entwickelt, um menschliche Karzinome zu studieren und zu fördern, das Verständnis der Krebsbehandlung und Entwicklung. Hier, ein präklinischer, murinen Pankreastumormodell von Lebermetastasen über eine Injektion von syngenen hemispleen murinen Pankreastumorzellen beschrieben. Dieses Modell imitiert viele der klinischen Bedingungen bei Patienten mit Metastasen in die Leber. Mäuse konsequent entwickeln Metastasen in der Leber so dass für die Untersuchung des metastatischen Prozess, experimentelle Therapie Testen und Tumorimmunologie Forschung.

Panc02 Tumorzellen in das hemispleen spritzt (Figur 1) bilden Lebermetastasen bei 100% der Mäuse, während die Lunge und Peritonealmetastasen werden nicht beobachtet, wenn das Verfahren vollständig durchgeführt wird. Diese Technik hat zu über einhundert Mäusen mit Panc02 Zellen in mehrere, unabhängige Experimente durchgeführt worden, und reproduzierbar, sterben alle unbehandelten Mäuse mit Lebermetastasen zwischen 30 und 60 Tagen nach der Hemisplenektomie Verfahren (Figur 2). Um...

Watch this Scientific Journal Video about A Preclinical Murine Model of Hepatic Metastases at JoVE.com

A Preclinical Murine Model of Hepatic Metastases

Eine präklinische, Mausmodell der Lebermetastasen über eine hemispleen Injektionstechnik durchgeführt.

Einer präklinischen Mausmodell der Lebermetastasen

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

a-preclinical-murine-model-of-hepatic-metastases

Research

JoVE Journal

Medicine

28.5K Aufrufe.  The Johns Hopkins University School of Medicine.  Eine präklinische, Mausmodell der Lebermetastasen über eine hemispleen Injektionstechnik durchgeführt. 

Serie: A Preclinical Murine Model of Hepatic Metastases

Murine Bioluminescent Hepatic Tumour Model

This video describes the establishment of liver metastases in a mouse model that can be subsequently analysed by bioluminescent imaging. Tumour cells are administered specifically to the liver to induce a localised liver tumour, via mobilisation of the spleen and splitting into two, leaving intact the vascular pedicle for each half of the spleen. Lewis lung carcinoma cells that constitutively express the firefly luciferase gene (luc1) are inoculated into one hemi-spleen which is then resected 10 minutes later. The other hemi-spleen is left intact and returned to the abdomen. Liver tumour growth can be monitored by bioluminescence imaging using the IVIS whole body imaging system. Quantitative imaging of tumour growth using IVIS provides precise quantitation of viable tumour cells. Tumour cell death and necrosis due to drug treatment is indicated early by a reduction in the bioluminescent signal. This mouse model allows for investigating the mechanisms underlying metastatic tumour-cell survival and growth and can be used for the evaluation of therapeutics of liver metastasis. 

This article describes a procedure for the induction of orthotopic bioluminescent liver tumours in mice, and subsequent analysis of tumour growth confined to the liver using live whole body luminescence imaging.

Murine Bioluminescent Leber Tumor-Modell

This video describes the establishment of liver metastases in a mouse model that can be subsequently analysed by bioluminescent imaging. Tumour cells are ...

Forschung

Medizin

Advanced Animal Model of Colorectal Metastasis in Liver: Imaging Techniques and Properties of Metastatic Clones

Patients with a limited number of hepatic metastases and slow rates of progression can be successfully treated with local treatment approaches1,2. However, little is known about the heterogeneity of liver metastases, and animal models capable of evaluating the development of individual metastatic colonies are needed. Here, we present an advanced model of hepatic metastases that provides the ability to quantitatively visualize the development of individual tumor clones in the liver and estimate their growth kinetics and colonization efficiency. We generated a panel of monoclonal derivatives of HCT116 human colorectal cancer cells stably labeled with luciferase and tdTomato and possessing different growth properties. With a splenic injection followed by a splenectomy, the majority of these clones are able to generate hepatic metastases, but with different frequencies of colonization and varying growth rates. Using the In Vivo&#160;Imaging System (IVIS), it is possible to visualize and quantify metastasis development with in vivo luminescent and ex vivo fluorescent imaging. In addition, Diffuse Luminescent Imaging Tomography (DLIT) provides a 3D distribution of liver metastases in vivo. Ex vivo fluorescent imaging of harvested livers provides quantitative measurements of individual hepatic metastatic colonies, allowing for the evaluation of the frequency of liver colonization and the growth kinetics of metastases. Since the model is similar to clinically observed liver metastases, it can serve as a modality for detecting genes associated with liver metastasis and for testing potential ablative or adjuvant treatments for liver metastatic disease.

The ability of metastatic clones to colonize distant sites depends on their proliferation capacity and/or their ability to survive in the host microenvironment without significant proliferation. Here, we present an animal model that allows quantitative visualization of both types of liver colonization by metastatic clones.

Erweiterte Tiermodell für Colorectal Metastase in der Leber: Imaging-Techniken und Eigenschaften des metastasierten Clones

Patients with a limited number of hepatic metastases and slow rates of progression can be successfully treated with local treatment approaches1,2. ...

Krebsforschung

Surgical Procedures and Methodology for a Preclinical Murine Model of De Novo Mammary Cancer Metastasis

A rate-limiting aspect of transgenic mouse models of mammary adenocarcinoma is that primary tumor burden in mammary tissue typically defines study end-points. Thus, studies focused on elucidating mechanisms of late-stage de novo metastasis are compromised, as are studies examining efficacy of anti-cancer therapies targeting mediators of metastasis in the adjuvant setting. Numerous murine mammary cancer models have been developed via targeted expression of dominant oncoproteins to mammary epithelial cells yielding models variably mimicking histopathologic and transcriptome-defined breast cancer subtypes common in women1. While much has been learned regarding the biology of mammary carcinogenesis with these models, their utility in identifying molecules regulating growth of late-stage metastasis are compromised as mice are typically euthanized at earlier time points due to significant primary tumor burden. Moreover, since a significant percentage of women diagnosed with breast cancer receive adjuvant therapy after surgical resection of primary tumors and prior to presence of detectable metastatic disease, preclinical models of de novo metastasis are urgently needed as platforms to evaluate new therapies aimed at targeting metastatic foci. To address these deficiencies, we developed a murine model of de novo mammary cancer metastasis, wherein primary mammary tumors are surgically resected, and metastatic foci subsequently develop over a 115 day post-surgical period. This long latency provides a tractable model to identify functionally significant regulators of metastatic progression in mice lacking primary tumor, as well as a model to evaluate preclinical therapeutic efficacy of agents aimed at blocking functionally significant molecules aiding metastatic tumor survival and growth.

Surgical Procedures and Methodology for a Preclinical Murine Model of De Novo Mammary Cancer Metastasis

Pre-clinical models evaluating adjuvant therapy targeting breast cancer metastasis are lacking. To address this, we developed a murine model of de novo pulmonary mammary adenocarcinoma metastasis, wherein therapies administered in the adjuvant setting (post surgical resection of primary tumors) can be evaluated for efficacy in impacting previously seeded pulmonary metastases.

Chirurgische Verfahren und Methodik für ein präklinisches Murine - Modell von<em&gt; De Novo</em&gt; Mamma Krebs Metastase

A rate-limiting aspect of transgenic mouse models of mammary adenocarcinoma is that primary tumor burden in mammary tissue typically defines study ...

A Portal Vein Injection Model to Study Liver Metastasis of Breast Cancer

Breast cancer is the leading cause of cancer-related mortality in women worldwide. Liver metastasis is involved in upwards of 30% of cases with breast cancer metastasis, and results in poor outcomes with median survival rates of only 4.8 - 15 months. Current rodent models of breast cancer metastasis, including primary tumor cell xenograft and spontaneous tumor models, rarely metastasize to the liver. Intracardiac and intrasplenic injection models do result in liver metastases, however these models can be confounded by concomitant secondary-site metastasis, or by compromised immunity due to removal of the spleen to avoid tumor growth at the injection site. To address the need for improved liver metastasis models, a murine portal vein injection method that delivers tumor cells firstly and directly to the liver was developed. This model delivers tumor cells to the liver without complications of concurrent metastases in other organs or removal of the spleen. The optimized portal vein protocol employs small injection volumes of 5 - 10 &#956;l, &#8805; 32 gauge needles, and hemostatic gauze at the injection site to control for blood loss. The portal vein injection approach in Balb/c female mice using three syngeneic mammary tumor lines of varying metastatic potential was tested; high-metastatic 4T1 cells, moderate-metastatic D2A1 cells, and low-metastatic D2.OR cells. Concentrations of &#8804; 10,000 cells/injection results in a latency of ~ 20 - 40 days for development of liver metastases with the higher metastatic 4T1 and D2A1 lines, and &#62; 55 days for the less aggressive D2.OR line. This model represents an important tool to study breast cancer metastasis to the liver, and may be applicable to other cancers that frequently metastasize to the liver including colorectal and pancreatic adenocarcinomas.

A surgical procedure was developed to deliver mammary tumor cells to the murine liver via portal vein injection. This model permits investigation of late stages of liver metastasis in a fully immune competent host, including tumor cell extravasation, seeding, survival, and metastatic outgrowth in the liver.

Ein Portal Veneninjektion Modell zur Untersuchung der Leber Metastasierung von Brustkrebs

Breast cancer is the leading cause of cancer-related mortality in women worldwide. Liver metastasis is involved in upwards of 30% of cases with breast ...

An Oncogenic Hepatocyte-Induced Orthotopic Mouse Model of Hepatocellular Cancer Arising in the Setting of Hepatic Inflammation and Fibrosis

The absence of a clinically relevant animal model addressing the typical immune characteristics of hepatocellular cancer (HCC) has significantly impeded elucidation of the underlying mechanisms and development of innovative immunotherapeutic strategies. To develop an ideal animal model recapitulating human HCC, immunocompetent male C57BL/6J mice first receive a carbon tetrachloride (CCl4) injection to induce liver fibrosis, then receive histologically-normal oncogenic hepatocytes from young male SV40 T antigen (TAg)-transgenic mice (MTD2) by intra-splenic (ISPL) inoculation. Androgen generated in recipient male mice at puberty initiates TAg expression under control of a liver-specific promoter. As a result, the transferred hepatocytes become cancer cells and form tumor masses in the setting of liver fibrosis/cirrhosis. This novel model mimics human HCC initiation and progression in the context of liver fibrosis/cirrhosis and reflects the most typical features of human HCC including immune dysfunction.

Here, we describe the development of a clinically relevant murine model of liver cancer recapitulating the typical immune features of hepatocellular cancer (HCC).

Ein onkogenes Hepatozyten-induziertes orthotopisches Mausmodell von hepatozellulärem Krebs, der bei der Einstellung von Leberentzündungen und Fibrose entsteht

The absence of a clinically relevant animal model addressing the typical immune characteristics of hepatocellular cancer (HCC) has significantly impeded ...

Murine Model of Metastatic Liver Tumors in the Setting of Ischemia Reperfusion Injury

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce multiple tissue and organ damage. Despite recent advances and therapeutic approaches, the overall morbidity has remained unsatisfactory especially in patients with underlying parenchymal abnormalities. In the context of aggressive cancer growth and metastasis, surgical I/R is suspected to be the promoter regulating tumor recurrence. This article aims to describe a clinically relevant murine model of liver I/R and colorectal liver metastasis. In doing so, we aim to assist other investigators in establishing and perfecting this model for their routine research practice to better understand the effects of liver I/R on promoting liver metastases.

We describe in detail a clinically relevant colorectal cancer liver metastases (CRLM) tumor model and the influence of liver ischemia reperfusion (I/R) in tumor growth and metastasis. This model can help to better understand the mechanisms underlying surgery-induced promotion of liver metastatic growth.

Murine Modell der metastasierenden Lebertumoren in der Einstellung der Ischämie Reperfusionsverletzung

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce ...

Portal Vein Injection of Colorectal Cancer Organoids to Study the Liver Metastasis Stroma

Hepatic metastasis of colorectal cancer (CRC) is a leading cause of cancer-related death. Cancer-associated fibroblasts (CAFs), a major component of the tumor microenvironment, play a crucial role in metastatic CRC progression and predict poor patient prognosis. However, there is a lack of satisfactory mouse models to study the crosstalk between metastatic cancer cells and CAFs. Here, we present a method to investigate how liver metastasis progression is regulated by the metastatic niche and possibly could be restrained by stroma-directed therapy. Portal vein injection of CRC organoids generated a desmoplastic reaction, which faithfully recapitulated the fibroblast-rich histology of human CRC liver metastases. This model was tissue-specific with a higher tumor burden in the liver when compared to an intra-splenic injection model, simplifying mouse survival analyses. By injecting luciferase-expressing tumor organoids, tumor growth kinetics could be monitored by in vivo imaging. Moreover, this preclinical model provides a useful platform to assess the efficacy of therapeutics targeting the tumor mesenchyme. We describe methods to examine whether adeno-associated virus-mediated delivery of a tumor-inhibiting stromal gene to hepatocytes could remodel the tumor microenvironment and improve mouse survival. This approach enables the development and assessment of novel therapeutic strategies to inhibit hepatic metastasis of CRC.

Portal vein injection of colorectal cancer (CRC) organoids generates stroma-rich liver metastasis. This mouse model of CRC hepatic metastasis represents a useful tool to study tumor-stroma interactions and develop novel stroma-directed therapeutics such as adeno-associated virus-mediated gene therapies.

Portalveneninjektion von Darmkrebs-Organoiden zur Untersuchung der Lebermetastasierung Stroma

Hepatic metastasis of colorectal cancer (CRC) is a leading cause of cancer-related death. Cancer-associated fibroblasts (CAFs), a major component of the ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

Our laboratory has developed a novel orthotopic implantation model of human prostate cancer (PCa). As PCa death is not due to the primary tumor, but rather the formation of distinct metastasis, the ability to effectively model this progression pre-clinically is of high value. In this model, cells are directly implanted into the ventral lobe of the prostate in Balb/c athymic mice, and allowed to progress for 4-6 weeks. At experiment termination, several distinct endpoints can be measured, such as size and molecular characterization of the primary tumor, the presence and quantification of circulating tumor cells in the blood and bone marrow, and formation of metastasis to the lung. In addition to a variety of endpoints, this model provides a picture of a cells ability to invade and escape the primary organ, enter and survive in the circulatory system, and implant and grow in a secondary site. This model has been used effectively to measure metastatic response to both changes in protein expression as well as to response to small molecule therapeutics, in a short turnaround time.

This orthotopic model of human prostate cancer allows for quantification of tumor size, circulating tumor cells, and formation of distinct metastasis to the lung. As cells must escape the primary organ, enter the blood stream, and implant into a secondary site, this model effectively recapitulates the scenario in humans.

Ein orthotopen Mausmodell der menschlichen Prostatakrebs-Metastasen

Our laboratory has developed a novel orthotopic implantation model of  human prostate cancer (PCa). As PCa death is not due to the primary tumor, but  ...

Non-Invasive PET/MR Imaging in an Orthotopic Mouse Model of Hepatocellular Carcinoma

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and evaluate novel therapeutic approaches. Non-invasive whole-body imaging using positron emission tomography (PET) provides critical insights into the in vivo characteristics of tissues at the molecular level in real-time. We present here a protocol for orthotopic HCC xenograft creation with and without hepatic artery ligation (HAL) to induce tumor hypoxia and the assessment of their tumor metabolism in vivo using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG) PET/magnetic resonance (MR) imaging. Tumor hypoxia could be readily visualized using the hypoxia marker [18F]FMISO, and it was found that the [18F]FMISO uptake was higher in HCC mice that underwent HAL than in the non-HAL group, whereas [18F]FDG could not distinguish tumor hypoxia between the two groups. HAL tumors also displayed a higher level of hypoxia-inducible factor (HIF)-1&#945; expression in response to hypoxia. Quantification of HAL tumors showed a 2.3-fold increase in [18F]FMISO uptake based on the standardized value uptake (SUV) approach.

Here, we present a protocol to create orthotopic hepatocellular carcinoma xenografts with and without hepatic artery ligation and perform non-invasive positron emission tomography (PET) imaging of tumor hypoxia using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG).

Nicht-invasive PET/MR-Bildgebung in einem orthotopen Mausmodell des hepatozellulären Karzinoms

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and ...

A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of cancer metastasis are still unclear. Animal models are essential for elucidating the mechanisms and for evaluating novel strategies for the treatment of metastatic cancers. Here, an in-depth description of a &#8220;patient-like&#8221; orthotopic syngeneic mouse model for exploring the mechanisms of metastasis of solid organ tumors is provided. The survival surgical implantation of BNL 1ME A.7R.1 mouse hepatocellular carcinoma cells directly into the liver (the organ of origin) of the inbred wild-type immune competent laboratory mouse strain, BALB/c is described. The success and reproducibility of this methodology recommends it for widespread use in elucidating the biological mechanisms of solid organ cancer metastasis.

The metastatic spread of cancer is the major cause of cancer-related deaths. We provide an in-depth description of our survival surgery methodology for establishing a &#8220;patient-like&#8221; orthotopic syngeneic mouse model system for studying the mechanisms of metastasis in solid organ tumors.

Ein &quot;Patient-Like&quot; Orthotope Syngene Maus-Modell der Leberzellkarzinom Metastasen

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of ...

Einer präklinischen Mausmodell der Lebermetastasen

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