Cellular Localization, Invasion, and Turnover Are Differently Influenced by Healthy and Tumor-Derived Extracellular Matrix

Abstract

The interplay between tumor cells and the microenvironment has been recognized as one of the hallmarks of cancer biology. To assess the role of extracellular matrix (ECM) in the modulation of tissue homeostasis and tumorigenesis, we developed a protocol for the purification of tissue-derived ECM using mucosae from healthy human colon, perilesional area, and colorectal carcinoma (CRC). Matched specimens were collected from the left colon of patients undergoing CRC resection surgery. ECMs were obtained from tissues that were decellularized with hypotonic solutions containing ionic and nonionic detergents, hypertonic solution, and endonuclease in the absence of denaturing agents. Mucosae-derived ECMs maintained distribution and localization of proteins and glycoproteins typical of the original tissues, and showed different three-dimensional (3D) structures among normal versus perilesional and tumor-derived stroma. The three types of ECM differentially regulated the localization and organization of seeded monocytes and cancer cells that were located and organized as in the original tissue. Specifically, healthy, perilesional, and CRC-derived ECMs sustained differentiation and polarization of cancer epithelial cells. In addition, healthy, but not perilesional and CRC-derived ECM constrained invasion of cancer cells. All three ECMs sustained turnover between cell proliferation and death up to 40 days of culture, although each ECM showed different ability in supporting cell proliferation, with tumor > perilesional > healthy-derived ECMs. Healthy-, perilesional-and CRC-derived ECM differently modulated cell homeostasis, spreading in the stroma and turnover between proliferation and death, and equally supported differentiation and polarization of cancer epithelial cells, thus highlighting the contribution of different ECMs modulating some features of tissue homeostasis and tumorigenesis. Moreover, these ECMs provide competent scaffolds useful to assess efficacy of antitumor drugs in a 3D setting that more closely recapitulates the native microenvironment. Further, ECM-based scaffolds may also be beneficial for future studies seeking prognostic and diagnostic stromal markers and targets for antineoplastic drugs.

Linearized texture of three-dimensional extracellular matrix is mandatory for bladder cancer cell invasion

“…Recent evidence has suggested that tumor-related extracellular environment may play a leading role, and it is not just a supporting actor over the initiation of the tumor; indeed, the dysregulation of the composition4 and stiffness21 of the ECM are associated with both the lack of asymmetric cellular division and of the differentiation of stem cells, and with the epithelial-mesenchymal transition of cancer stem cells as well, thus regulating tissue homeostasis and sustaining the onset and progression of cancer419. In agreement with these latter observations, we recently reported that an increased organization of ECM fibrils, but not biochemical composition, of the perilesional area of the colorectal carcinoma22 is associated with proliferation and infiltration of metastatic cancer cells23.…”

“…2b) and MIBC (Fig. 2c) were isolated according to our previously published protocol validated on the human colon23. The hematoxylin-eosin stained sections of the non-neoplastic urinary bladder showed the different cells layers: (i) mucosa, formed by transitional epithelium and lamina propria, and (ii) muscolaris propria, known as detrusor muscle, formed by three layers of smooth muscle.…”

“…ECMs were isolated according to a protocol recently published and validated on the human colon23. A minor modification to the original protocol was introduced, as components of solutions A and B were merged together thus resulting in a solution composed of 86.8% 0.1x PBS, 5 mM EDTA, 10% DMSO, 1% Triton X-100, 1% protease inhibitor cocktail and 1% pen/strep.…”

“…Cells and nuclei in the tissues and ECMs were evaluated by hematoxylin-eosin, cellular antigens (cytokeratin, vimentin and α-smooth muscle actin) and the presence of stromal component collagen IV by immunohistochemistry, as it has been previously reported2340. Tissues and ECMs were fixed in 10% buffered formalin for 24–48 h at room temperature and then embedded in paraffin.…”

In Vitro Modeling of Tissue-Specific 3D Microenvironments and Possibile Application to Pediatric Cancer Research

“…Synthetic matrices could also be regarded as the future of engineering tumor microenvironments, but the high number of biomaterials proposed in the literature need to be further optimized before being effectively applied in the tumor biology fields [128]. In effect, in the design of these matrices, several key factors should be taken into account, and, among them, landscape/ topography, fiber orientation, porosity (macro-, micro-, nanoscale conditioning), matrix viscoelasticity, matrix remodeling, and growth factors' and cytokines' binding sites.…”