Abstract
Metastasis, the leading cause of death in cancer patients, requires the invasion of tumor cells through the stroma in response to migratory cues, in part 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 ECM proteins are more likely to have an effect on cancer invasion and metastasis. We evaluated the effect of four ECM proteins upregulated in breast tumor tissue in multiple human breast cancer cell lines in three assays. There was no linear relationship between cell adhesion to ECM proteins and ECM-driven 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. We find that ECM-driven 2D cell migration speed or persistence did not predict 3D invasion in response to the same cue. However, cell adhesion, and in particular cell elongation and shape irregularity, accurately predicted the magnitude of ECM-driven 2D migration and 3D invasion. Our models successfully predicted the effect of novel ECM proteins in a cell-line specific manner. 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 in tumor tissue and help prioritize strategies for targeting tumor-ECM interactions to treat metastasis.
Highlights
Metastasis, the dissemination of cells from the primary tumor to secondary organs in the body, is the leading cause of death in cancer
Our goal was to identify the relationship between extracellular matrix (ECM) responses in adhesion, 2D migration, and 3D invasion assays to develop strategies to predict the effect of novel ECM proteins on 3D cancer cell invasion, which is more relevant to the study of cancer metastasis. 3D invasion assays can be more complex, time consuming, and more challenging for follow up analysis, while 2D migration and adhesion assays are quicker, easy to analyze, and to use with other experimental approaches such as cell sorting, atomic force microscopy, or immunostaining
By evaluating the response of two triple-negative breast cancer cell lines to four ECM proteins known to be upregulated in metastatic breast cancers, we found that there is no linear relationship between metrics used to quantify these three assays
Summary
Metastasis, the dissemination of cells from the primary tumor to secondary organs in the body, is the leading cause of death in cancer. Metastasis involves the local invasion of tumor cells into the surrounding tissues, intravasation into the vasculature and lymphatics, and colonization of a distant site. Several chemical and biophysical cues have been shown to promote local invasion.[3] In particular, the extracellular matrix (ECM), which provides structure and support to our tissues, drives local invasion of tumor cells and metastasis, as well as colonization of secondary sites. The glycoprotein Fibronectin, which is produced by both tumor and stromal compartments in breast tumors,[4] can drive directional migration of breast cancer cells to drive metastasis.[5] The optimization of protocols to characterize the ECM of tumors has led to the identification of multiple ECM proteins abundant in tumor tissue that may be involved in promoting metastatic phenotypes.[4,6] The present study aims to develop a pipeline to assess which of these ECM proteins, alone or in combination, are more likely to affect invasion and metastasis, and are better targets as biomarkers or for drug development
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