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.

BEMÆRK: Patient vævsudtagning blev godkendt gennem institutionelt review board (IRB) -godkendte Biospecimen og kliniske protokoller (protokol numrene 09-121 og 11-041, henholdsvis) på Memorial Sloan Kettering Cancer Center. 1. vævsudtagning <ol><li> Indkøb af patient primær tumor / metastaser BEMÆRK: Til dato er denne protokol blevet udført på kirurgisk fjernet pancreas, mave og bryst tumortyper, samt, lymfom metastaser. <ol><li> Ret det kirurgiske team til at levere de pr?...

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Kræft biologer står over for betydelige udfordringer, når de forsøger at udvikle effektive terapeutiske strategier. Test narkotika i udviklingen på etablerede cancercellelinjer kan ikke præcist afspejler in vivo respons og in vivo forsøg på PDX modeller er arbejdskrævende og meget dyrt. I betragtning af ovenstående, er anvendelsen af ex vivo-teknikker til primær patient tumorer 14, 15 er nu placeret ved siden af den molekylære analyse af etablerede kræft cellelinjer og ...

Udvikling af effektive kræftlægemidler har vist sig at være ekstremt udfordrende. Cancercellelinjer og tumor eksplantater - samt xenotransplantater er blevet anvendt i kræftforskning i over et halvt 1,2,3 århundrede. Til dato er uundværlig den molekylære analyse af narkotika følsomhed og modstand i både etablerede cancer cellelinjer og patient-afledte xenografter (PDX). Men afprøvning af forbindelser i etablerede cancercellelinier er ikke ofte prædiktive for effektivitet in vivo, og den tilsvarende in vivo undersøgelser i dyr, især i PDX modeller er meget dyrt og tidskrævende. Begrænsningerne af disse modelsystemer, nemlig manglende evne til at informere om indflydelsen af ​​det native mikromiljø i tumorudvikling og respons på terapeutiske strategier har ført forskningsområde at udvikle yderligere metoder til at komplementere disse analyser. Af de seneste, forstærkes opmærksomhed mod ex vivo analyse af patientens tumeller eksplantater 4, 5 som følge af større forståelse for, at kræft terapeutisk respons er ikke eksklusivt til iboende molekylære sammensætning af cancerceller, men snarere er stærkt præget af tumorcellens mikromiljø 6, 7 en funktion, der ikke kan blive gentaget ved traditionelle dyrkningsmetoder og / eller PDX. Ex vivo analyse i ovennævnte sammenhæng (dvs.., påvirkning af tilstødende omgivende tumor celle mikromiljø) indebærer vurdering af levedygtige primære tumor / metastase sektioner, snarere end ex vivo analyse af cellulære isolater 8, 9. Vi rapporterer her om en ex vivo teknik (dvs.., Præcision-skiver friske vævssnit) af både patientens primære tumorer og tilhørende metastaser (dvs. lymfeknuder), der trofast informerer om respons (IC50), off-target effekter og muliggør molekylær Analyse af resistens og feedbackmekanismer. Derudover en korrelativ analyse af therapeutic følsomhed / resistens mod biomarkør og genekspression profil kan udføres i et forsøg på at identificere patienter mere tilbøjelige til at reagere på det eksperimentelle stof af interesse (dvs.. høj lægemiddelrespons matcher patient med særlige biologiske profil). Anvendelse af ex vivo teknik og vurdering i en multi-parameter mode er bevægelse i retning af patientens valg og en generel forbedring af de kliniske resultater. Ex vivo-behandling respons analyse kunne blive et standard værktøj i præklinisk og klinisk udvikling af kræftmedicin og er planlagt som et skridt i retning af en skræddersyet medicin tilgang i terapeutiske udviklingsstrategier.

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			<td style="width:119px;">16500500</td>
			<td style="width:167px;">Prepare 4% and 6% before use</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;">Waterbath</td>
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			<td style="width:119px;">15-462-2SQ</td>
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			<td height="42" style="height:42px;width:216px;">Microwave</td>
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			<td style="width:119px;">352096</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.

Til denne undersøgelse blev ex vivo teknik, der anvendes i en korrelativ analyse af terapeutisk følsomhed / resistens hos en varmechokprotein 90 inhibitor (Hsp90i). I en præklinisk vurdering af denne Hsp90i, brystkræft primær tumor, en ER + invasiv duktalt carcinom (IDC) og tilhørende lymfeknudemetastaser blev analyseret ex vivo til behandling respons &#160; (Figur 1). Multiple 200 um serielle sektioner blev behandlet med vehikel og stigende 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

Ex vivo-behandling Reaktion fra primære tumorer og / eller associerede Metastaser til præklinisk og klinisk udvikling af Therapeutics

Etablerede cancer cellelinjer og xenografter har været grundpillen i kræftforskning i de sidste mange årtier. Seneste oplysninger tyder dog på, at terapeutiske reaktion i høj grad er påvirket af tumorcellens mikromiljø. Derfor har vi udviklet en ex vivo analyse af primær tumor prøver for lægemiddeludvikling formål.

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

Research

JoVE Journal

Medicine

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

Thermal Ablation for the Treatment of Abdominal Tumors

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

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

Following intracardiac delivery of MDA-MB-231-luc-D3H2LN cells to Nu/Nu mice, systemic metastases developed in the injected animals. Bioluminescence ...

A High-throughput Automated Platform for the Development of Manufacturing Cell Lines for Protein Therapeutics

The fast-growing biopharmaceutical industry demands speedy development of highly efficient and reliable production systems to meet the increasing ...

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

A Novel Høj opløsning<em&gt; In vivo</em&gt; Imaging teknik til at undersøge den dynamiske respons af Intrakranielle Structures til tumorvækst og Therapeutics

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a ...

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Adaptation of Semiautomated Circulating Tumor Cell CTC Assays for Clinical and Preclinical Research Applications

Tilpasning af halvautomatiseret Cirkulerende Tumor Cell (CTC) analyser til kliniske og prækliniske forskning applikationer

The majority of cancer-related deaths occur subsequent to the development of metastatic disease. This highly lethal disease stage is associated with the ...

A Preclinical Murine Model of Hepatic Metastases

Et præklinisk musemodel af levermetastaser

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

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Isoleret Nedsat Perfusion som en behandling for levermetastaser af Uveal melanom

Isolated hepatic perfusion (IHP) is a procedure where the liver is surgically isolated and perfused with a high concentration of the chemotherapeutic ...

Using Micro-computed Tomography for the Assessment of Tumor Development and Follow-up of Response to Treatment in a Mouse Model of Lung Cancer

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Lung cancer is the most lethal cancer in the world. Intensive research is ongoing worldwide to identify new therapies for lung cancer. Several mouse ...

Generation of Microtumors Using 3D Human Biogel Culture System and Patient-derived Glioblastoma Cells for Kinomic Profiling and Drug Response Testing

Generation af Microtumors Brug 3D Menneskelig Biogel Kultur System og Patient-afledte glioblastomaceller for Kinomic Profilering and Drug respons Testing

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