Computer-assisted quantification of motile and invasive capabilities of cancer cells

Abstract

High-throughput analysis of cancer cell dissemination and its control by extrinsic and intrinsic cellular factors is hampered by the lack of adequate and efficient analytical tools for quantifying cell motility. Oncology research would greatly benefit from such a methodology that allows to rapidly determine the motile behaviour of cancer cells under different environmental conditions, including inside three-dimensional matrices. We combined automated microscopy imaging of two- and three-dimensional cell cultures with computational image analysis into a single assay platform for studying cell dissemination in high-throughput. We have validated this new approach for medulloblastoma, a metastatic paediatric brain tumour, in combination with the activation of growth factor signalling pathways with established pro-migratory functions. The platform enabled the detection of primary tumour and patient-derived xenograft cell sensitivity to growth factor-dependent motility and dissemination and identified tumour subgroup-specific responses to selected growth factors of excellent diagnostic value.

Crosstalk between SHH and FGFR Signaling Pathways Controls Tissue Invasion in Medulloblastoma

“…Since bFGF is a strong promoter of invasion in DAOY cells [15], we tested whether SAG treatment would affect bFGF-induced collagen I invasion. We performed spheroid invasion assays in the absence of stimulation and with SAG or bFGF alone or with a combination of both.…”

“…Images were acquired on an Axio Observer 2 mot plus fluorescence microscope using a 5× objective (Zeiss, Munich, Germany). Cell invasion is determined as the average of the distance invaded by the cells from the center of the spheroid, which was determined using automated cell dissemination counter (aCDc) with our cell dissemination counter software aSDIcs [15].…”

Synthetic Activators of Cell Migration Designed by Constructive Machine Learning

“…The candidate molecules suggested by the constructive machine learning approach were prioritized based on their pharmacophore similarity to the fine‐tuning set, and two of the de novo designs were synthesized. These compounds induced pronounced medulloblastoma cell migration in a phenotypic spheroid invasion assay (SIA) . RNAi mediated knockdown of CXCR4 abrogated the migration‐inducing effects, indicating the relevance of this chemokine receptor for the observed phenotypic activity.…”

Identification of Synthetic Activators of Cancer Cell Migration by Hybrid Deep Learning

“…Spheroid invasion assay (SIA) : The effect of compounds on cell invasion and the phenotype of the medulloblastoma tumor cell line were determined. DAOY cells were tested by SIA, as described . DAOY cells stably expressing lifeact (LA) enhanced green fluorescent protein (EGFP) produced by lentiviral transduction with pLenti‐LA‐EGFP were used for SIA.…”