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.

NOTA: approvvigionamento dei tessuti del paziente è stato autorizzato attraverso la revisione bordo istituzionalmente (IRB) -approvato Biospecimen e protocolli clinici (numeri di protocollo 09-121 e 11-041, rispettivamente) al Memorial Sloan Kettering Cancer Center. Procurement 1 Tissue <ol><li> Approvvigionamento di paziente tumore primario / metastasi NOTA: Fino ad oggi, questo protocollo è stata eseguita su pancreatiche, gastriche e tipi di tumore al seno chirurgicamente rimoss...

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Biologi cancro devono affrontare sfide significative nel tentativo di sviluppare efficaci strategie terapeutiche. Test farmaci in sviluppo su linee cellulari di cancro stabilite non può riflettere con precisione in risposta vivo e in vivo su modelli PDX sono laborioso e molto costoso. Considerato quanto sopra, l&#39;applicazione di tecniche ex vivo di paziente primaria tumori 14, 15 è ora posizionato accanto alla analisi molecolare di consolidate linee cellulari tumorali ...

Lo sviluppo di terapie efficaci cancro ha dimostrato di essere estremamente impegnativo. Linee cellulari tumorali e espianti tumorali - così come xenotrapianti sono stati utilizzati nella ricerca sul cancro per oltre mezzo secolo 1,2,3. Ad oggi, l&#39;analisi molecolare della sensibilità di droga e resistenza in entrambe le linee di cellule di cancro affermati e xenotrapianti di derivazione paziente (PDX) è indispensabile. Tuttavia, i test di composti in linee cellulari di cancro stabiliti spesso non è predittivo di efficacia in vivo, e corrispondenti studi in vivo negli animali, soprattutto nei modelli PDX, è molto costoso e richiede tempo. I limiti di questi sistemi modello, vale a dire l&#39;impossibilità di informare l&#39;influenza del microambiente nativo nella progressione tumorale e la risposta alle strategie terapeutiche, ha portato il campo di ricerca per sviluppare metodi aggiuntivi per complimentarmi con queste analisi. Di recente, accresciuta attenzione viene rivolta verso ex vivo analisi di tum pazienteo espianti 4, 5 a causa della maggiore comprensione che il cancro risposta terapeutica non è esclusiva alla composizione molecolare intrinseca delle cellule tumorali, ma piuttosto è fortemente influenzato dal microambiente delle cellule tumorali 6, 7, una caratteristica che non può essere riassunta con metodi di coltura tradizionali e / o PDX. ex vivo analisi nel contesto di cui sopra (ad esempio., influenza adiacente tumorale circostante microambiente cellulare) implica la valutazione delle sezioni tumore / metastasi primarie vitali, piuttosto che ex vivo analisi degli isolati cellulari 8, 9. Riportiamo qui su una tecnica ex vivo (ad esempio., Sezioni di precisione-fette fresche di tessuto) di entrambi i tumori primari del paziente e metastasi associate (ad esempio, linfonodi) che segnala fedelmente alla risposta (IC50), gli effetti off-target e consente molecolare analisi dei meccanismi di resistenza e di feedback. Inoltre, l&#39;analisi correlativa di therapeUTIC sensibilità / resistenza contro biomarker e profilo di espressione genica può essere eseguita in uno sforzo per identificare i pazienti più probabilità di rispondere al farmaco sperimentale di interesse (ad esempio., la risposta alle alte farmaco corrisponde paziente con particolare profilo biologico). Applicazione della tecnica ex vivo e la valutazione in modo multi-parametro è movimento verso la selezione dei pazienti e il miglioramento complessivo dei risultati clinici. Trattamento ex vivo analisi di risposta potrebbe diventare uno strumento standard per lo sviluppo preclinico e clinico di terapie del cancro ed è immaginato come un passo verso un approccio di medicina personalizzata nelle strategie di sviluppo terapeutiche.

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			<td style="width:96px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</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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			<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>
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			<td height="63" style="height:63px;width:216px;">MEM with Penicillin&#160; + Streptomycin</td>
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			<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>
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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>
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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>
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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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			<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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			<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.

Per questo studio, la tecnica ex vivo è stato utilizzato in un&#39;analisi correlativa terapeutico sensibilità / resistenza di un inibitore della heat shock protein 90 (Hsp90i). In una valutazione preclinica di questo Hsp90i, il cancro al seno tumore primario, un ER + invasivo carcinoma duttale (IDC), e associata metastasi linfonodali sono stati analizzati ex-vivo per la risposta al trattamento &#160; (Figura 1). Più di 200 micron sezioni seriali sono stati trattati...

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

Fondata linee cellulari tumorali e xenotrapianti sono stati il ​​cardine della ricerca sul cancro negli ultimi decenni. Tuttavia, recenti evidenze suggeriscono che la risposta terapeutica è fortemente influenzata dal microambiente delle cellule tumorali. Di conseguenza, abbiamo sviluppato una analisi ex vivo di campioni tumorali primarie per scopi di sviluppo della droga.

Ex Vivo risposta al trattamento di tumori primari e / o associati per metastasi preclinici e sviluppo clinico di 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 ...

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