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: colheita de tecidos do paciente, autorizado por conselho de revisão institucional (IRB) -aprovado Biospecimen e Protocolos Clínicos (números de protocolo 09-121 e 11-041, respectivamente) no Memorial Sloan Kettering Cancer Center. Procurement 1 Tissue <ol><li> Aquisição de paciente primário tumor / metástase NOTA: Até à data, este protocolo foi realizado em pancreáticos, gástricos e tipos de tumor da mama removidos cirurgicamente, bem como, as metástases de linfoma...

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Biólogos câncer enfrentam desafios significativos ao tentar desenvolver estratégias terapêuticas eficazes. Testar medicamentos em desenvolvimento em linhas celulares de cancro estabelecidas não pode refletir com precisão em resposta vivo e in vivo em modelos PDX são trabalhoso e muito caro. Dado o acima, a aplicação de técnicas ex vivo de tumores primários do paciente 14, 15 está agora posicionada ao lado da análise molecular das linhas celulares de cancro e xenoenxerto...

O desenvolvimento de cancro terapêutica eficazes provou ser extremamente desafiador. Linhas de câncer de células tumorais e explantes - bem como enxertos foram usados ​​em pesquisa sobre o câncer há mais de meio século 1,2,3. Até à data, a análise molecular de sensibilidade à droga e resistência em ambas as linhas celulares de cancro e xenoenxertos estabelecidas derivadas de pacientes (PDX) é indispensável. No entanto, o teste de compostos em linhas celulares de cancro estabelecidas muitas vezes não é de previsão de eficácia in vivo, e estudos in vivo correspondente em animais, especialmente em modelos de PDX, é muito cara e demorada. As limitações destes sistemas modelo, ou seja, a incapacidade para informar sobre a influência do microambiente nativo em progressão do tumor e a resposta de estratégias terapêuticas, levou o campo de investigação para o desenvolvimento de métodos adicionais para complementar estas análises. De recente, maior atenção está sendo pago para ex vivo de análise de tum pacienteou explantes 4, 5, devido à maior compreensão de que a resposta terapêutica do câncer não é exclusivo para a composição molecular inerente das células cancerosas mas é muito influenciada pelo microambiente celular tumor 6, 7 um recurso que não pode ser reproduzido por meio de métodos e de cultivo tradicionais / ou PDX. análise ex vivo, no contexto acima (ie., influência do microambiente adjacente célula tumoral circundante) implica a avaliação de seções tumor / metástase de primário viáveis, ao invés de ex vivo de análise de isolados celulares 8, 9. Registramos aqui uma técnica ex vivo (ie., Seções cortado com precisão frescas de tecidos) de ambos os tumores primários de pacientes e metástases associadas (ou seja, os gânglios linfáticos), que informa fielmente na resposta (IC50), efeitos fora do alvo e permite molecular análise dos mecanismos de resistência e de feedback. Além disso, uma análise correlativa de therapeUTIC sensibilidade / resistência contra biomarcador e perfil de expressão do gene pode ser realizada em um esforço para identificar os pacientes com maior probabilidade de responder à droga experimental de interesse (ou seja., resposta de droga elevada corresponde paciente com particular perfil biológico). Aplicação da técnica ex vivo e avaliação de uma forma multi-parâmetro é movimento para a seleção dos pacientes e melhoria global dos resultados clínicos. Análise ex vivo da resposta ao tratamento pode se tornar uma ferramenta padrão no desenvolvimento pré-clínico e clínico da terapêutica do câncer e é visto como um passo em direção a uma abordagem da medicina personalizada em estratégias de desenvolvimento de terapias.

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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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			<td height="21" style="height:21px;width:216px;">Injector blade</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>
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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>
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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>
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			<td height="21" style="height:21px;width:216px;">Embedding microsettes</td>
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			<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>
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			<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>
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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.

Para este estudo, a técnica ex vivo foi utilizado na análise de um correlativo de terapêutica sensibilidade / resistência de um inibidor da proteína de choque térmico 90 (Hsp90i). Em uma avaliação pré-clínica deste Hsp90i, o tumor primário de câncer de mama, um ER + ductal carcinoma invasivo (IDC), e associado metástases em linfonodos foram analisados ​​ex vivo para a resposta ao tratamento &#160; (Figura 1). Vários 200 mM cortes seriados foram tratad...

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 resposta ao tratamento de tumores primários e / ou Associated Metástases para pré-clínica e clínica de Desenvolvimento Therapeutics

Linhas celulares de cancro estabelecidos e xenotransplantes têm sido o esteio da pesquisa do câncer para as últimas décadas. No entanto, evidências recentes sugerem que a resposta terapêutica é muito influenciada pelo microambiente de células tumorais. Portanto, foi desenvolvida uma análise ex vivo de amostras de tumor primário para fins de desenvolvimento de drogas.

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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