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.

MERK: Pasient vev anskaffelser ble autorisert gjennom institusjonelt review board (IRB) -godkjent Biospecimen og kliniske protokoller (protokoll tall 09-121 og 11-041, henholdsvis) ved Memorial Sloan Kettering Cancer Center. 1. Tissue Procurement <ol><li> Anskaffelse av pasientens primærtumor / metastase MERK: Til dags dato har denne protokollen blitt utført på kirurgisk fjernet bukspyttkjertelen, mage og bryst krefttyper, samt, lymfom metastaser. <ol><li> Rett kirurgisk team for...

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Kreft biologer står overfor betydelige utfordringer når du forsøker å utvikle effektive terapeutiske strategier. Testing narkotika i utvikling på etablerte kreftcellelinjer kan ikke nøyaktig reflektere in vivo respons og in vivo eksperimenter på PDX modeller er arbeidskrevende og svært kostbart. Gitt de ovennevnte, tumorer ved anvendelse av ex vivo teknikker for primær pasienten 14, 15 er nå plassert ved siden av den molekylære analyser av etablerte cancercellelinjer og p...

Utvikle effektive kreftbehandling har vist seg å være svært utfordrende. Kreftcellelinjer og kreft eksplantater - samt xenografts har blitt brukt i kreftforskning i over et halvt 1,2,3-tallet. Til dags dato, er den molekylære analyser av stoffet følsomhet og resistens i både etablerte kreftcellelinjer og pasient avledet xenografts (PDX) uunnværlig. Imidlertid er testing av forbindelsene i etablerte kreftcellelinjer ofte ikke prediktiv for in vivo effekt, og tilsvarende in vivo-studier i dyr, spesielt i PDX-modeller, er svært kostbart og tidkrevende. Begrensningene av disse modellsystemer, nemlig den manglende evnen til å informere på påvirkning av den opprinnelige mikromiljøet i tumorprogresjon og respons på terapeutiske strategier har ført til forskning området på å utvikle andre metoder for å komplimentere disse analysene. Av nyere, er forsterket oppmerksomhet mot ex vivo analyse av pasient tumeller eksplantater 4, 5 på grunn av den større forståelse at kreft terapeutisk respons er ikke eksklusivt for den iboende molekylær sammensetning av kreftceller, men heller er sterkt påvirket av tumorcellemikro 6, 7 en funksjon som ikke kan rekapitulert av tradisjonelle dyrkningsmetoder og / eller PDX. Ex vivo-analyse i ovennevnte sammenheng (f.eks. innflytelse av tilstøtende omgir tumorcelle-mikromiljøet) innebærer vurdering av levedyktige primære tumor / metastase seksjoner, i stedet for ex vivo analyse av cellulære isolater 8, 9. Vi rapporterer her på en ex vivo teknikk (ie., Presisjon-skiver fersk vevssnitt) av både pasient primære svulster og tilknyttede metastaser (dvs. lymfeknuter) som trofast informerer på respons (IC50), off-target effekter og gir mulighet for molekylær analyse av motstand og tilbakekoblingsmekanismer. I tillegg er en samsvarende analyse av therapeUtic følsomhet / resistens mot biomarkør og genuttrykk profilen kan utføres i et forsøk på å identifisere pasienter mer sannsynlig å svare på den eksperimentelle narkotika av interesse (ie., høye medikamentrespons kamper pasient med særlig biologisk profil). Anvendelse av den ex vivo teknikk og vurdering i en multi-parameter mote er bevegelse mot pasientens utvalg og generell forbedring av de kliniske resultater. Ex vivo behandlingsrespons analyse kunne bli en standard verktøy i preklinisk og klinisk utvikling av kreftbehandling, og er tenkt som et skritt mot et personlig medisin tilnærming i terapeutiske utviklingsstrategier.

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			<td style="width:119px;">Catalog Number</td>
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			<td height="21" style="height:21px;width:216px;">Vibratome Leica VT1000s</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: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>
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			<td style="width:119px;">31-200-32</td>
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			<td height="21" style="height:21px;width:216px;">Disposable forceps</td>
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			<td height="21" style="height:21px;width:216px;">FBS (heat inactivated)</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>
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			<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>
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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.

For denne studien ble den ex vivo teknikk som brukes i en samsvarende analyse av terapeutisk sensitivitet / motstand av et varmesjokkprotein 90 inhibitor (Hsp90i). I en preklinisk vurdering av denne Hsp90i, brystkreft primærtumor, en ER + invasiv duktalt karsinom (IDC), og tilhørende lymfeknute metastaser ble analysert ex vivo for behandlingsrespons &#160; (Figur 1). Multiple 200 mikrometer seriesnitt ble behandlet med kjøretøy bare og økende doser (0,25, 0,50, 1,...

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

Etablerte kreftcellelinjer og xenografts har vært bærebjelken i kreftforskningen de siste tiårene. Imidlertid tyder nyere bevis på at terapeutisk respons er sterkt påvirket av svulsten celle mikromiljøet. Derfor har vi utviklet en ex vivo analyse av primære vevsprøver for narkotikautviklingsformål.

Ex Vivo Treatment Response av primære svulster og / eller Associated metastaser for preklinisk og klinisk utvikling av legemiddel

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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