Name: ex vivo treatment response of primary tumors and/or associated metastases for preclinical and clinical development of therapeutics
Uploaded: 2014-10-02T15:00:00
Description: Watch this Scientific Journal Video about Ex Vivo Treatment Response of Primary Tumors and/or Associated Metastases for Preclinical and Clinical Development of Therapeutics at JoVE.com

The authors would like to thank the MSKCC Tissue Procurement Service Team (TPS), specifically, Maria Corazon Mariano, Priscilla McNeil, Anas Idelbi, Daniel Navarrete and Katrina Allen, in all of their efforts in the successful pursuit of this project and funding from the following sources: 5 R21 CA158609-02 and the Conquer Cancer Foundation and the Breast Cancer Research Foundation. In addition, the authors would like to thank Eric Cottington PhD, Vice President of the Office of Research and Project Administration, the Office of Technology Development, Research Outreach and Compliance and RTM Information Systems Support, in the support of the submission of this manuscript.

REMARQUE: l&#39;obtention de tissus des patients a été autorisé par l&#39;examen institutionnel (CISR) -approved des échantillons biologiques et des protocoles cliniques (numéros de protocole 09-121 et 11-041, respectivement) au Memorial Sloan Kettering Cancer Center. 1. tissus achats <ol><li> Achat de patients tumeur primaire / métastases NOTE: A ce jour, ce protocole a été effectuée sur les types enlevés chirurgicalement pancréatiques, gastriques et tumeurs du sein, ain...

<ol>
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biologistes du cancer font face à des défis importants en essayant de développer des stratégies thérapeutiques efficaces. Test de médicaments en développement sur ​​des lignées cellulaires de cancer établis ne peut pas refléter avec précision en réponse vivo et in vivo sur des modèles PDX sont laborieux et très coûteux. Compte tenu de ce qui précède, l&#39;application de techniques ex vivo de tumeurs primaires du patient 14, 15 est maintenant positionn?...

Développement de traitements efficaces contre le cancer s&#39;est avéré être extrêmement difficile. lignes de cancer des cellules et des explants de tumeur - ainsi que les xénogreffes ont été utilisées dans la recherche sur le cancer pour plus d&#39;un demi siècle 1,2,3. A ce jour, l&#39;analyse moléculaire de la sensibilité aux médicaments et la résistance dans les deux lignées de cellules cancéreuses établies et des xénogreffes provenant de patients (PDX) est indispensable. Cependant, le test de composés sur des lignées cellulaires de cancer établies n&#39;est pas souvent prédictifs de l&#39;efficacité in vivo, et correspondant à des études in vivo chez les animaux, en particulier dans les modèles PDX, est très coûteux et prend du temps. Les limites de ces systèmes modèles, à savoir l&#39;incapacité d&#39;informer sur l&#39;influence du micro-environnement natif dans la progression tumorale et la réponse à des stratégies thérapeutiques, a conduit le domaine de la recherche afin de développer des méthodes supplémentaires pour compléter ces analyses. Récente, une attention accrue est accordée à une analyse ex vivo de son tour maladeou explants 4, 5 en raison de la plus grande compréhension que la réponse thérapeutique du cancer n&#39;est pas exclusif à la composition moléculaire inhérente des cellules cancéreuses mais est fortement influencée par le microenvironnement des cellules tumorales 6, 7 une fonction qui ne peut être récapitulé par des méthodes de culture traditionnelles et / ou PDX. analyse ex vivo dans le contexte ci-dessus (c.-à-., l&#39;influence de la tumeur adjacente entourant cellule microenvironnement) l&#39;évaluation des sections tumeur / métastases primaires viables implique, plutôt que ex vivo l&#39;analyse d&#39;isolats cellulaires 8, 9. Nous rapportons ici sur une technique ex vivo (c.-à-., Sections de précision tranches fraîches tissus) des deux tumeurs primaires de patients et métastases associées (par exemple, les ganglions lymphatiques) qui informe fidèlement réponse (IC50), les effets hors-cible et permet de moléculaire analyse des mécanismes de résistance et de rétroaction. En outre, une analyse corrélative de therapeUTIC sensibilité / résistance en fonction de biomarqueurs et profil d&#39;expression génique peut être réalisée dans un effort pour identifier les patients les plus susceptibles de répondre à la drogue expérimentale d&#39;intérêt (c.-à-., la réponse médicamenteuse élevée correspond à des patients en particulier le profil biologique). Application de la technique et l&#39;évaluation ex vivo dans un mode multi-paramètre est un mouvement vers la sélection des patients et l&#39;amélioration globale des résultats cliniques. Analyse ex vivo de la réponse au traitement pourrait devenir un outil standard dans le développement préclinique et clinique de traitements contre le cancer et est envisagée comme une étape vers une approche de médecine personnalisée dans les stratégies de développement thérapeutique.

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			<td height="43" style="height:43px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:96px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Vibratome Leica VT1000s</td>
			<td style="width:96px;">Leica</td>
			<td style="width:119px;">14047235613</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">UltraPure agarose</td>
			<td style="width:96px;">Invitrogen</td>
			<td style="width:119px;">16500500</td>
			<td style="width:167px;">Prepare 4% and 6% before use</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Injector blade</td>
			<td style="width:96px;">Ted Pella</td>
			<td style="width:119px;">121-4</td>
			<td style="width:167px;"></td>
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		<tr height="63">
			<td height="63" style="height:63px;width:216px;">MEM with Penicillin&#160; + Streptomycin</td>
			<td style="width:96px;">Media Core Facilities (MSKCC)</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">The media is prepared at Memorial Sloan Kettering Cancer Center&#160;</td>
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		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Scalpel no. 10</td>
			<td style="width:96px;">Thermo Scientific</td>
			<td style="width:119px;">31-200-32</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Disposable forceps</td>
			<td style="width:96px;">Cole-Parmer</td>
			<td style="width:119px;">84011182</td>
			<td style="width:167px;"></td>
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		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Embedding mold</td>
			<td style="width:96px;">Electron Microscopy Science</td>
			<td style="width:119px;">70181</td>
			<td style="width:167px;"></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">FBS (heat inactivated)</td>
			<td style="width:96px;">Gemini</td>
			<td style="width:119px;">100106</td>
			<td style="width:167px;"></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">24 well plates</td>
			<td style="width:96px;">Corning</td>
			<td style="width:119px;">3524</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Formalin (10%)</td>
			<td style="width:96px;">Sigma Diagnostics</td>
			<td style="width:119px;">SDHT501128&#160;</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">16% Formaldehyde solution</td>
			<td style="width:96px;">Thermo Scientific</td>
			<td style="width:119px;">28908</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Embedding microsettes</td>
			<td style="width:96px;">Simport</td>
			<td style="width:119px;">M503-2</td>
			<td style="width:167px;"></td>
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		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Ethanol (70%)</td>
			<td style="width:96px;">Fisher Scientific</td>
			<td style="width:119px;">A405P-4</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Waterbath</td>
			<td style="width:96px;">Fisher Scientific</td>
			<td style="width:119px;">15-462-2SQ</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Microwave</td>
			<td style="width:96px;">General Electric</td>
			<td style="width:119px;">ModelJES2051DNBB</td>
			<td style="width:167px;"></td>
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		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Adhesive (Ethyl Cyanoacrylate)</td>
			<td style="width:96px;">Sigma-Aldrich</td>
			<td style="width:119px;">E1505-5G</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">10mm dishes</td>
			<td style="width:96px;">BD Falcon</td>
			<td style="width:119px;">353003</td>
			<td style="width:167px;"></td>
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		<tr height="22">
			<td height="22" style="height:22px;width:216px;">15ml tubes</td>
			<td style="width:96px;">BD Falcon</td>
			<td style="width:119px;">352096</td>
			<td style="width:167px;"></td>
		</tr>
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ex vivo treatment response of primary tumors and/or associated metastases for preclinical and clinical development of therapeutics

The molecular analysis of established cancer cell lines has been the mainstay of cancer research for the past several decades. Cell culture provides both direct and rapid analysis of therapeutic sensitivity and resistance. However, recent evidence suggests that therapeutic response is not exclusive to the inherent molecular composition of cancer cells but rather is greatly influenced by the tumor cell microenvironment, a feature that cannot be recapitulated by traditional culturing methods. Even implementation of tumor xenografts, though providing a wealth of information on drug delivery/efficacy, cannot capture the tumor cell/microenvironment crosstalk (i.e., soluble factors) that occurs within human tumors and greatly impacts tumor response. To this extent, we have developed an ex vivo (fresh tissue sectioning) technique which allows for the direct assessment of treatment response for preclinical and clinical therapeutics development. This technique maintains tissue integrity and cellular architecture within the tumor cell/microenvironment context throughout treatment response providing a more precise means to assess drug efficacy.

Pour cette étude, la technique ex vivo a été utilisé dans une analyse de corrélation thérapeutique sensibilité / résistance d&#39;un inhibiteur de la protéine de choc thermique 90 (Hsp90i). Dans une évaluation préclinique de cette Hsp90i, la tumeur primaire de cancer du sein, un ER + carcinome canalaire invasif (IDC), et associé métastase des ganglions lymphatiques ont été analysés ex vivo de la réponse au traitement &#160; (Figure 1). Plusieurs 200 um...

Watch this Scientific Journal Video about Ex Vivo Treatment Response of Primary Tumors and/or Associated Metastases for Preclinical and Clinical Development of Therapeutics at JoVE.com

Ex Vivo Treatment Response of Primary Tumors and/or Associated Metastases for Preclinical and Clinical Development of Therapeutics

Lignées de cellules cancéreuses établies et les xénogreffes ont été le pilier de la recherche sur le cancer au cours des dernières décennies. Cependant, des données récentes suggèrent que la réponse thérapeutique est fortement influencée par le micro-environnement des cellules tumorales. Par conséquent, nous avons développé une analyse ex vivo des échantillons de tumeurs primaires à des fins de développement de médicaments.

La réponse au traitement ex vivo de tumeurs primaires et / ou métastases associées préclinique et développement clinique de la thérapeutique

The molecular analysis of established cancer cell lines has been the mainstay of cancer research for the past several decades. Cell culture provides both ...

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