Asaf Rotem,1 Levi Garraway,2 Mei-Ju Su,2 Anindita Basu,3 Aviv Regev,3 Kevin Struhl4
1Dana Farber Cancer Institute (United States) 2Dana-Farber/Harvard Cancer Ctr. (United States) 3Broad Institute of MIT and Harvard (United States) 4Harvard Medical School (United States)
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Three-dimensional growth conditions reflect the natural environment of cancer cells and are crucial to be performed at drug screens. We developed a 3D assay for cellular transformation that involves growth in low attachment (GILA) conditions and is strongly correlated with the 50-year old benchmark assay-soft agar. Using GILA, we performed high-throughput screens for drugs and genes that selectively inhibit or increase transformation, but not proliferation.
This phenotypic approach is complementary to our genetic approach that utilizes single-cell RNA-sequencing of a patient sample to identify putative oncogenes that confer sensitivity to drugs designed to specifically inhibit the identified oncoprotein.
Currently, we are dealing with a big challenge in our field- the limited number of cells that might be extracted from a biopsy. Small patient-derived samples are hard to test in the traditional multiwell plate and it will be helpful to minimize the culture area and the experimental system. We managed to design a suitable microfluidic device for limited number of cells and perform the assay using image analysis.
We aim to test drugs on tumor cells, outside of the patient body- and recommend on the ideal treatment that is tailored to the individual. This device will help to minimize biopsy-sampling volumes and minimize interventions in the patient’s tumor.
Asaf Rotem,Levi Garraway,Mei-Ju Su,Anindita Basu,Aviv Regev, andKevin Struhl
"Miniaturizing 3D assay for high-throughput drug and genetic screens for small patient-derived tumor samples (Conference Presentation)", Proc. SPIE 10046, Visualizing and Quantifying Drug Distribution in Tissue, 100460H (19 April 2017); https://doi.org/10.1117/12.2256299
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Asaf Rotem, Levi Garraway, Mei-Ju Su, Anindita Basu, Aviv Regev, Kevin Struhl, "Miniaturizing 3D assay for high-throughput drug and genetic screens for small patient-derived tumor samples (Conference Presentation)," Proc. SPIE 10046, Visualizing and Quantifying Drug Distribution in Tissue, 100460H (19 April 2017); https://doi.org/10.1117/12.2256299