An Interplay Between Reaction-Diffusion and Cell-Matrix Adhesion Regulates Multiscale Invasion in Early Breast Carcinomatosis

Abstract

The progression of cancer in the breast involves multiple reciprocal interactions between malignantly transformed epithelia, surrounding untransformed but affected stromal cells, and the extracellular matrix (ECM) that is remodeled during the process. A quantitative understanding of the relative contribution of such interactions to phenotypes associated with cancer cells can be arrived at through the construction of increasingly complex experimental and computational models. Herein, we introduce a multiscale three-dimensional (3D) organo- and pathotypic experimental assay that approximates, to an unprecedented extent, the histopathological complexity of a tumor disseminating into its surrounding stromal milieu via both bulk and solitary motility dynamics. End point and time-lapse microscopic observations of this assay allow us to study the earliest steps of cancer invasion as well as the dynamical interactions between the epithelial and stromal compartments. We then simulate our experimental observations using the modeling environment Compucell3D that is based on the Glazier–Graner–Hogeweg model. The computational model, which comprises adhesion between cancer cells and the matrices, cell proliferation and apoptosis, and matrix remodeling through reaction–diffusion–based morphogen dynamics, is first trained to phenocopy controls run with the experimental model, wherein one or the other matrices have been removed. The trained computational model successfully predicts phenotypes of the experimental counterparts that are subjected to pharmacological treatments (inhibition of N-linked glycosylation and matrix metalloproteinase activity) and scaffold modulation (alteration of collagen density). Further parametric exploration-based simulations suggest that specific permissive regimes of cell–cell and cell–matrix adhesions, operating in the context of a reaction–diffusion–regulated ECM dynamics, promote multiscale invasion of breast cancer cells and determine the extent to which the latter migrate through their surrounding stroma.

Heterogeneity in 2,6-Linked Sialic Acids Potentiates Invasion of Breast Cancer Epithelia

“… 35 Our model, calibrated to diverse matrix microenvironments and pharmacological perturbations, had earlier been shown to simulate single-cell and collective-cell invasion (individually and in combination, known as multiscale invasion) through consecutive barriers of lrBM-like collagen-like fibrillar environments, similar to the 3D invasion assay used above. 34 The cellular constituents of our model were digital medium and high 2,6-Sial cells (mimicking medium and high 2,6-Sial cells) with the former differing from the latter in exhibiting greater ECM adhesion. The ECM constituents of the model were digital lrBM and digital collagen (mimicking BM and Type 1 collagen) with the only difference being their nonfibrillar and fibrillar structure, respectively.…”

“…A 3D invasion assay was performed as described previously by our group. 34 Briefly, cancer clusters were made using 30 000 cells in a polyHEMA-coated (Sigma, P3932), 96-well plate, defined medium 48 supplemented with 4% rBM. After 48 h, clusters were collected and embedded in polymerizing rat tail collagen in a chambered cover glass.…”

“…Two different methods of analysis were used; the first one, “Area of minimal enclosing circle”, detailed in the previous publication 34 calculates the smallest possible circle, which encloses all cells of a certain cell type and can be considered as a quantification of invasion corresponding to that cell type in that initial configuration. The collected simulation images can be binarized for digital high 2,6-Sial cells (orange), digital medium 2,6-Sial cells (red), or both (orange and red; Figure 6 F) depending on the analysis.…”

Nanomotors Sense Local Physicochemical Heterogeneities in Tumor Microenvironments**

“…The fitting has two parameters which represent characteristic force ( ) and characteristic length scale ( ) for different cell lines. We used four different CC lines with variable extents of invasiveness and metastatic ability: MDA‐MB‐231, a triple negative cancer cell line that shows an invasive “stellate”‐like growth phenotype, when cultured on rBM; MCF‐7, HCC70 and T4−2 represent three breast cancer cell lines that show a milder “mass”‐like morphologies on rBM matrices [47, 48] (See SI: Table S1 for a description of the cell lines used in this study. Also see SI: Figure S4 for adhesion force calculation in other cell lines).…”

Nanorobots Sense Local Physiochemical Heterogeneities of Tumor Matrisome

“…A higher field would be necessary to overcome the adhesion and move further near the cell membrane. To quantify the adhesion data in more general terms, we fit an exponential function of the form: cancer cell lines that show a milder 'mass'-like morphologies on rBM matrices 54,55 (See SI: Table S1 for a description of the cell lines used in this study). Control experiments with untransformed breast epithelial cells HMLE and S1, the latter being an isogenic non-transformed counterpart of the malignant T42 mentioned above 56 did not show any adhesion for field strengths of 50 Gauss (see movie M2b and M2c).…”