Matrix Remodeling and Hyaluronan Production by Myofibroblasts and Cancer-Associated Fibroblasts in 3D Collagen Matrices

Abstract

The tumor microenvironment is a key modulator in cancer progression and has become a novel target in cancer therapy. An increase in hyaluronan (HA) accumulation and metabolism can be found in advancing tumor progression and are often associated with aggressive malignancy, drug resistance and poor prognosis. Wound-healing related myofibroblasts or activated cancer-associated fibroblasts (CAF) are assumed to be the major sources of HA. Both cell types are capable to synthesize new matrix components as well as reorganize the extracellular matrix. However, to which extent myofibroblasts and CAF perform these actions are still unclear. In this work, we investigated the matrix remodeling and HA production potential in normal human dermal fibroblasts (NHFB) and CAF in the absence and presence of transforming growth factor beta -1 (TGF-β1), with TGF-β1 being a major factor of regulating fibroblast differentiation. Three-dimensional (3D) collagen matrix was utilized to mimic the extracellular matrix of the tumor microenvironment. We found that CAF appeared to response insensitively towards TGF-β1 in terms of cell proliferation and matrix remodeling when compared to NHFB. In regards of HA production, we found that both cell types were capable to produce matrix bound HA, rather than a soluble counterpart, in response to TGF-β1. However, activated CAF demonstrated higher HA production when compared to myofibroblasts. The average molecular weight of produced HA was found in the range of 480 kDa for both cells. By analyzing gene expression of HA metabolizing enzymes, namely hyaluronan synthase (HAS1-3) and hyaluronidase (HYAL1-3) isoforms, we found expression of specific isoforms in dependence of TGF-β1 present in both cells. In addition, HAS2 and HYAL1 are highly expressed in CAF, which might contribute to a higher production and degradation of HA in CAF matrix. Overall, our results suggested a distinct behavior of NHFB and CAF in 3D collagen matrices in the presence of TGF-β1 in terms of matrix remodeling and HA production pointing to a specific impact on tumor modulation.

Stratified 3D Microtumors as Organotypic Testing Platforms for Screening Pancreatic Cancer Therapies

“…On this focus, increasing evidence suggests that stromal cells are the main players in fibrotic ECM production. [ 28–30 ] In order to evaluate the capacity of 3D STAMS to recapitulate the desmoplastic reaction and ECM deposition found in human pancreatic tumors, collagen and glycosaminoglycans (GAGs) were quantified. As demonstrated in Figure , both 3D co‐culture models showed an increased collagen secretion than their monoculture counterparts, highlighting the role of CAFs in fibrotic stroma production.…”

“…Such includes the accumulation of hyaluronic acid (HA), a large linear glycosaminoglycan composed of D‐glucuronic and N ‐acetyl‐D‐glucosamine that is reportedly to be overexpressed in PDAC. [ 29 ] Similar to collagen, GAGs quantification showed higher GAGs content in co‐culture models than monoculture systems. Moreover, GAGs deposition is slightly higher in STAMS than in Random microtissues (Figure 5B day 14).…”

Hydrogel Models with Stiffness Gradients for Interrogating Pancreatic Cancer Cell Fate

“…Activated CAFs promote tumor growth and metastasis through multiple mechanisms, such as ECM remodeling, secretion of cytokines, and immunosuppression [ 12 ]. Activation of CAFs also contributes to the stiffening of TME by matrix remodeling, including increased secretion of enzymes, excess deposition of ECM (e.g., HA) [ 14 , 15 , 16 ], and force-mediated matrix remodeling [ 17 , 18 ]. In particular, secretion of matrix metalloproteinase (MMP) by CAFs contributes to the matrix remodeling and cell invasion by forming “tracks” in the ECM, hence facilitating cancer cell migration [ 17 ].…”

Differential Expression Profiles of Cell-to-Matrix-Related Molecules in Adrenal Cortical Tumors: Diagnostic and Prognostic Implications

“…More intriguingly, HAS-1 expression in ACC tumor cells was associated with a lower mitotic rate, a lower prevalence of atypical mitotic figures and a better outcome. The prognostic significance of HAS-1 in cancers is equivocal and has been studied either in cancer cells or in cancer stroma, including cancer-associated fibroblasts [ 52 , 53 , 54 ]. However, in agreement with our findings, a negative prognostic role of HAS-1 downregulation in cancer cells has been described in lung cancer [ 55 ] and melanoma [ 56 ].…”