Evaluation of Extracellular Matrix Composition to Improve Breast Cancer Modeling

Abstract

The development of resistance to therapy is a significant obstacle to effective therapeutic regimens. Evaluating the effects of oncology drugs in the laboratory setting is limited by the lack of translational models that accurately recapitulate cell-microenvironment interactions present in tumors. Acquisition of resistance to therapy is facilitated, in part, by the composition of the tumor extracellular matrix (ECM), with the primary current in vitro model using collagen I (COL I). Here we seek to identify the prevalence of COL I-enhanced expression in the triple-negative breast cancer (TNBC) subtype. Furthermore, we identify if methods of response to therapy are altered depending on matrix composition. We demonstrated that collagen content varies in patient tumor samples across subtypes, with COL I expression dramatically increased in typically less aggressive estrogen receptor (ER)-positive(ER + )/progesterone receptor (PGR)-positive (PGR + ) cancers irrespective of patient age or race. These findings are of significance considering how frequently COL I is implicated in tumor progression. In vitro analyses of ER + and ER-negative (ER -) cell lines were used to determine the effects of ECM content (collagen I, collagen IV, fibronectin, and laminin) on proliferation, cellular phenotype, and survival. Neither ER + nor ERcells demonstrated significant increases in proliferation when cultured on these ECM substrates. ERcells cultured on these substrates were sensitized to both chemotherapy and targeted therapy. In addition, MDA-MB-231 cells expressed different morphologies, binding affinities, and stiffness across these substrates. We also demonstrated that ECM composition significantly alters transcription of senescence-associated pathways across ER + and ERcell lines. Together, these results suggest that complex matrix composites should be incorporated into in vitro tumor models, especially for the drug-resistant TNBC subtype.

Substrate Type and Concentration Differently Affect Colon Cancer Cells Ultrastructural Morphology, EMT Markers, and Matrix Degrading Enzymes

“…Overproduction of collagens, increased crosslinking, and alignment of collagen fibers induce an increased peri-tumoral stiffness which favors cancer cell invasion and tumor malignancy [ 29 , 36 , 49 , 50 ]. Type I collagen new deposition is frequently related to tumor progression, but it has been considered as one of the ECM components also favoring chemoresistance [ 31 , 37 , 38 , 51 ]. The possible relation between collagen and drug resistance in CRC cells prompted us to evaluate the behavior and fine morphological characteristics of two cell types: LoVo-S cells, sensitive to doxorubicin, and LoVo-R, resistant to the same anticancer drug.…”

“…The first interesting event described next to the tumor mass includes a desmoplastic deposition of collagen, which might correspond to a physical defense of the host [ 66 ]. However, in human CRC, the highly expressed type I collagen and collagen fiber alignment also act as promoters of aggressiveness by inducing EMT, tumor progression, and poor patient disease-free survival [ 25 , 32 , 37 , 51 , 67 ]. Notably, the dense and stiff collagen type I fibers stimulate pro-tumorigenic signaling cascades in cancer, such as focal adhesion kinase (FAK), Src kinases family (SFKs), and extracellular regulated kinase (ERK)1/2 [ 67 ].…”

“…It is noteworthy that HPSE, a well-known component of exosomes/microvesicles, can favor both the production and docking of exosomes, thus playing a critical role in both tumor progression and chemoresistance [ 61 ]. Strong interactions between chemoresistance and peri-tumoral ECM composition or physical array alterations have been recently suggested: remodeling of ECM and prolonged chemotherapy can favor treatment resistance and ECM-cell communications [ 31 , 37 , 38 , 51 , 70 , 71 , 72 , 73 ]. In this study, we demonstrated that the CRC LoVo-S cells are affected by highly concentrated collagen, but once they become doxorubicin-resistant (LoVo-R cells), they maintain their intrinsic aggressiveness and are independent of the type of ECM substrate (Matrigel or collagen).…”

Differential Protein Citrullination in Human ER– and ER+ Tumor and Adjacent Healthy Breast Tissue

“…ECM is a major structural component for the tumor microenvironment, and its remodeling promotes tumorigenesis, metastasis, and drug resistance . The matrix composition plays a key role in the proliferation and progression of both ER+ and ER– cancer cells . Hormone receptor status and selective estrogen modulator treatment of tumors such as tamoxifen impact the occurrence of recurrence or secondary malignancies.…”

“… 23 The matrix composition plays a key role in the proliferation and progression of both ER+ and ER– cancer cells. 24 Hormone receptor status and selective estrogen modulator treatment of tumors such as tamoxifen impact the occurrence of recurrence or secondary malignancies. While having breast cancer likely increases the risk for recurrent breast cancer, more studies are needed to address the hypothesis that the ER status of tumors may be a risk factor for subsequent cancers in breast cancer.…”