Engineering tumor constructs to study matrix-dependent angiogenic signaling of breast cancer cells.

Breast cancer is the leading cause of cancer deaths among females globally. The crosstalk between tumor microenvironment and neoplastic cells is the key for promoting tumor growth, stimulating tumor angiogenesis, and metastasis to distant organs. Thus, it is highly important to investigate tumor cell-matrix interactions to facilitate screening of different anti-cancer agents, individually or in combination. We, herein report, the development of an in vitro three-dimensional (3D) breast cancer model to investigate the effect of stromal crosslinking and consequent, stiffening on the angiogenic activity of cancer cells. Crosslinking of collagen gels was altered via non-enzymatic glycation and highly aggressive breast cancer cells, MDA-MB-231, were encapsulated in these gels. Cells encapsulated in glycated/stiffer matrices displayed an increased expression of pro-angiogenesis-related signals. Inhibition of mechanotransduction pathways on the angiogenic activity of aggressive tumor cells in stiff matrices was investigated using Y-27632, blebbistatin, and cytochalasin D. Rho-associated kinase (ROCK) inhibitor, Y-27632, diminished the pro-angiogenic signaling, thereby suggesting the potential dependence of breast cancer cells on the Rho/ROCK pathway in regulating tumor angiogenesis. Our findings highlight the potential of the developed model to be used as a tool to investigate matrix-associated tumor angiogenesis and screen different therapeutic agents towards inhibiting it.