Tuesday, March 24, 2015

Phenotypic and MoA Analysis of Anti-Metastatic Molecules using 3D Tumoroid Invasion Assays, Kinetic Ligand Binding, and Cellular Microscopy Webinar Recap


Metastasis is the main cause of death in cancer patients and one of the most complex biological processes in human diseases. The development of therapies designed to forestall the metastatic activity of tumors has been met with multiple challenges, including the choice of an appropriate cell model. Tumors in vivo exist as a three-dimensional (3D) mass of multiple cell types, including cancer and stromal cells. Therefore, incorporating a 3D spheroid-type cellular structure that includes co-cultured cell types forming a tumoroid, provides a more predictive model than the use of individual cancer cells cultured on the bottom of a well in traditional two-dimensional (2D) format. A second hurdle has been accurate mechanism of action determination. Possessing knowledge of how invasion is interrupted provides a more complete picture before proceeding to clinical testing. A final hindrance which has been necessary to overcome is the proper capture and analysis of kinetic reader-based data and microscopic images during the tumor invasion process. Being able to monitor inhibitor binding, as well as tumor invasion through the matrix in a quantitative way is critical.

In the webinar presented last week, we demonstrated methods to perform live-cell, label-free, phenotypic analyses of 3D tumor invasion. Spheroid Microplates from Corning Life Sciences, coated with an ultra low attachment surface, were incorporated for tumoroid formation, and performance of the invasion process. Tumor invasion tracking and quantification was then performed via digital microscopy and cellular analysis using the Cytation 5 cell imaging multi-mode reader and Gen5 Data Analysis Software.

During the presentation, we also displayed the ability to determine inhibitor drug-target residence time using the Tag-lite® direct, homogeneous ligand binding assay from Cisbio. The compound set was then incorporated into the tumoroid invasion assay to determine their potential inhibitory effect on the invasion process. Finally, we illustrated how analysis of the involvement of matrix metalloproteinases during the invasion process could be assessed while simultaneously monitoring tumoroid invasion, using the EnSens Protease Activity Detection Technology from Enzium.

The combination of these techniques present accurate, yet easy-to-use methods to assess target-based and phenotypic effects of new, potential anti-metastatic drugs. We invite you to download and watch the recording of the webinar.

By: BioTek Instruments, Brad Larson, Principal Scientist

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