Cell shape, and not 2D migration, predicts ECM-driven 3D cell invasion in breast cancer

Abstract

Metastasis, the leading cause of death in cancer patients, requires the invasion of tumor cells through the stroma in response to migratory cues, such as those provided by the extracellular matrix (ECM). Recent advances in proteomics have led to the identification of hundreds of ECM proteins which are more abundant in tumors relative to healthy tissue. Our goal was to develop a pipeline to easily predict which of these ECM proteins is more likely to have an effect on cancer invasion and metastasis. We evaluated the effect of 4 ECM proteins upregulated in breast tumor tissue in multiple human breast cancer cell lines in 3 assays. We found there was no linear relationship between the 11 cell shape parameters we quantified when cells adhere to ECM proteins and 2D cell migration speed, persistence or 3D invasion. We then used classifiers and partial-least squares regression analysis to identify which metrics best predicted ECM-driven 2D migration and 3D invasion responses. ECM-driven 2D cell migration speed or persistence did not correlate with or predict 3D invasion in response to that same cue. However, cell adhesion, and in particular cell elongation and irregularity accurately predicted the magnitude of ECM-driven 2D migration and 3D invasion in all cell lines. Testing predictions revealed that our models are good at predicting the effect of novel ECM proteins within a given cell line, but that ECM responses are cell-line specific. Overall, our studies identify the cell morphological features that determine 3D invasion responses to individual ECM proteins. This platform will help provide insight into the functional role of ECM proteins abundant tumor tissue and help prioritize strategies for targeting tumor-ECM interactions to treat metastasis. to M.J.O.]; Tufts University [Start-up funds from the School of Engineering to M.J.O.] and funds from NSF REU to A.W.Conflict-of-interest: None. Insight BoxMetastasis, the dissemination of tumor cells, is driven by the interaction of invading tumor cells with their local environment, in particular with the ECM, which provides structure and support to our tissues. This study presents an integrated approach to predict the effect of individual ECM proteins on 3D invasion and metastasis based on simple adhesion assays which quantify cell shape. Machine learning classification and partial-least squares regression models reveal that ECM-driven 2D cell migration metrics are not predictive of 3D invasion, and that cell shape of cells adhered to ECM can predict that protein's effect on 3D invasion. These data provide a pipeline for predicting the effect of ECM proteins on breast cancer cell invasion and metastasis.

Constricted migration is associated with stable 3D genome structure differences in cancer cells

“…However, like A375 cells, a higher fraction of MDA-MB-231 cells that underwent constricted migration for 10 rounds (Bottom-5) showed elongated cell bodies and further elongation of their nuclei compared with Control or Bottom-12 (Fig EV2C -E). These observations coincide with previous reports that elongated cell and nuclear shape correlated with higher migration propensity in MDA-MB-231 cells (Baskaran et al, 2020). As in A375 cells, the increased migratory efficiency of MDA-MB-231 Bottom-5 cells was not related to a change in overall Lamin A/C or H3K9me3 protein levels (Fig EV2H-I).…”

How does the extracellular matrix affect the rigidity of an embedded spheroid?

“…A key in vitro model system for studying the combined effects of cell-cell and cell-ECM interactions on tissue behavior is a tissue spheroid embedded in a collagen network. It has been shown that tensile forces and stiffer collagen fibers in the ECM facilitate tumor invasion [16,17], and that as various ECM proteins are modified, cell shapes may be a good predictor of invasion potential [18].…”