Identification of neural crest and melanoma cancer cell invasion and migration genes using high-throughput screening and deep attention networks.

Abstract

Background: Cell migration and invasion are well-coordinated in development and disease but remain poorly understood. We previously showed that the neural crest (NC) cell migratory wavefront shares a 45-gene panel with other cell invasion phenomena. To rapidly and systematically identify critical genes, we performed a high-throughput siRNA screen and statistical and deep learning analyses to determine changes in NC- versus non-NC-derived human cell line behaviors.

Results: We find 14 out of 45 genes significantly reduced c8161 melanoma cell migration; four of the 14 genes altered leader cell motility (BMP4, ITGB1, KCNE3, and RASGRP1). Deep learning identified marked disruptions in cell-neighbor interactions after BMP4 or RASGRP1 knockdown in c8161 cells. Recombinant proteins added to the culture media revealed five out of the 11 known secreted molecules stimulated c8161 cell migration. BMP4 knockdown severely reduced c8161 in vivo invasion in a chick embryo transplant model. Addition of BMP4 protein to the culture media of BMP4-siRNA-treated c8161 cells rescued cell migratory ability.

Conclusion: High-throughput screening and deep learning distilled a 45-gene panel to a small subset of genes critical to melanoma and warrant deeper in vivo functional analysis for their role and potential synergies in driving NC cell migration and invasion.