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.

HINWEIS: Patientengewebebeschaffung wurde durch institutionell Review Board (IRB) zugelassenen -zugelassene Biospecimen und Klinische Protokolle (Protokoll-Nummern 09-121 und 11-041, beziehungsweise) am Memorial Sloan Kettering Cancer Center. 1. Gewebebeschaffung <ol><li> Beschaffung von Patienten Primärtumor / Metastasen HINWEIS: Bis heute hat dieses Protokoll auf chirurgisch entfernt Pankreas, Magen-und Brusttumorarten, sowie, Lymphom Metastasen durchgeführt. <ol><li> Richten Si...

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Krebs Biologen vor großen Herausforderungen, wenn man versucht, wirksame therapeutische Strategien zu entwickeln. Testen von Medikamenten in der Entwicklung auf etablierte Krebszelllinien nicht genau in vivo Reaktion und in vivo Versuchen an PDX Modelle sind arbeitsintensiv und sehr teuer. Angesichts des Vorstehenden wird die Anwendung der Ex-vivo-Techniken der Primärtumoren Patienten 14, 15 wird nun neben der molekularen Analyse von etablierten Tumorzelllinien und von einem Patie...

Entwicklung wirksamer Krebstherapeutika hat sich als äußerst schwierig. Krebszelllinien und Tumor Explantate - ebenso wie Heterotransplantate wurden in der Krebsforschung seit über einem halben Jahrhundert 1,2,3 verwendet. Bis heute ist die molekulare Analyse von Arzneimittelempfindlichkeit und Widerstand in beiden etablierten Krebszelllinien und Patienten abgeleitete Xenografts (PDX) unverzichtbar. Jedoch ist der Test-Verbindungen in etablierte Krebszelllinien oft nicht prädiktiv für die Wirksamkeit in vivo, und die entsprechenden in vivo-Untersuchungen an Tieren, insbesondere in PDX Modelle, ist sehr teuer und zeitaufwendig. Die Grenzen dieser Modellsysteme, nämlich der Unfähigkeit, über den Einfluss der nativen Mikroumgebung der Tumorprogression und der Reaktion auf therapeutische Strategien zu informieren, hat das Forschungsfeld, um zusätzliche Methoden, um diese Analysen zu entwickeln Kompliment geführt. Der letzten, ist erhöhte Aufmerksamkeit in Richtung Ex-vivo-Analyse von Patienten TUM bezahltoder Explantaten 4, 5 aufgrund der besseren Verständnis, dass Krebs therapeutische Antwort nicht exklusiv ist, um die inhärente molekulare Zusammensetzung von Krebszellen, sondern wird stark durch die Tumorzelle Mikro 6, 7 eine Funktion, die nicht durch herkömmliche Kultivierungsverfahren und rekapitulieren beeinflusst / oder PDX. Ex-vivo-Analyse in der obigen Kontext (dh., Einfluss der benachbarten Umgebung von Tumorzellen Mikroumgebung) impliziert Bewertung von lebensfähigen primären Tumor / Metastasen Abschnitte, statt ex vivo Analyse zellulärer Isolate 8, 9. Wir berichten hier über einen Ex-vivo-Technik (dh., Präzise geschnittenen frischen Gewebeschnitten) der beiden Patienten Primärtumoren und Metastasen assoziiert (dh Lymphknoten), die treu informiert über Antwort (IC50), Off-Target-Effekte und ermöglicht für die molekulare Analyse von Widerstand und Feedback-Mechanismen. Zusätzlich wird ein Korrelat Analyse therapeUTIC Empfindlichkeit / Resistenz gegen Biomarker und Genexpressionsprofil kann in dem Bemühen, Patienten eher zu dem experimentellen Medikament von Interesse reagieren zu identifizieren durchgeführt werden (dh., hohe Medikamentenantwort passt Patienten mit bestimmten biologischen Profil). Anwendung der Ex-vivo-Technik und Bewertung in einer Multi-Parameter-Mode ist Bewegung in Richtung Patientenauswahl und die allgemeine Verbesserung der klinischen Ergebnisse. Ex-vivo-Behandlung Response-Analyse konnte ein Standardwerkzeug in der präklinischen und klinischen Entwicklung von Krebstherapeutika zu werden und wird als ein Schritt hin zu einer personalisierten Medizin Ansatz in der therapeutischen Entwicklungsstrategien in Betracht gezogen.

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			<td style="width:96px;">Company</td>
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		<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>
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		<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>
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		<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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			<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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			<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>
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			<td height="21" style="height:21px;width:216px;">Disposable forceps</td>
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			<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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			<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>
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		<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>
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		<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>
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		<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>
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		<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>
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			<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>
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			<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>
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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.

Für diese Studie wurde die Ex-vivo-Technik in einem korrelativen Analyse der therapeutischen Empfindlichkeit / Widerstand eines Hitzeschockprotein 90-Inhibitor (Hsp90i) verwendet. In einer präklinischen Bewertung dieses Hsp90i, den Primärtumor Brustkrebs, einem ER + invasive duktale Karzinom (IDC) und die zugehörigen Lymphknoten-Metastasen wurden ex vivo zur Behandlung Reaktion analysiert &#160; (Abbildung 1). Mehrere 200 um serielle Schnitte wurden mit Vehikel all...

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

Ex-vivo-Ansprechen auf die Behandlung von Primärtumoren und / oder assoziierten Metastasen für die präklinische und klinische Entwicklung von Therapeutika

Gegründet Krebszelllinien und Heterotransplantate waren die Hauptstütze der Krebsforschung in den letzten Jahrzehnten. Jedoch schlägt letzten Beweis, dass therapeutische Reaktion wird durch die Tumorzelle Mikro beeinflusst. Daher haben wir eine ex vivo-Analyse des Primärtumorproben für die Arzneimittelentwicklung Zwecke entwickelt.

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

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Percutaneous thermal ablation is an emerging treatment option for many tumors of the abdomen not amenable to conventional treatments. During a thermal ...

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Monitoring Tumor Metastases and Osteolytic Lesions with Bioluminescence and Micro CT Imaging

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The use of patient-derived xenografts for modeling cancers has provided important insight into cancer biology and drug responsiveness. However, they are ...