Role of Collagen Fiber Morphology on Ovarian Cancer Cell Migration Using Image-Based Models of the Extracellular Matrix

Abstract

Remodeling of the extracellular matrix (ECM) is an important part in the development and progression of many epithelial cancers. However, the biological significance of collagen alterations in ovarian cancer has not been well established. Here we investigated the role of collagen fiber morphology on cancer cell migration using tissue engineered scaffolds based on high-resolution Second-Harmonic Generation (SHG) images of ovarian tumors. The collagen-based scaffolds are fabricated by multiphoton excited (MPE) polymerization, which is a freeform 3D method affording submicron resolution feature sizes (~0.5 µm). This capability allows the replication of the collagen fiber architecture, where we constructed models representing normal stroma, high-risk tissue, benign tumors, and high-grade tumors. These were seeded with normal and ovarian cancer cell lines to investigate the separate roles of the cell type and matrix morphology on migration dynamics. The primary finding is that key cell–matrix interactions such as motility, cell spreading, f-actin alignment, focal adhesion, and cadherin expression are mainly determined by the collagen fiber morphology to a larger extent than the initial cell type. Moreover, we found these aspects were all enhanced for cells on the highly aligned, high-grade tumor model. Conversely, the weakest corresponding responses were observed on the more random mesh-like normal stromal matrix, with the partially aligned benign tumor and high-risk models demonstrating intermediate behavior. These results are all consistent with a contact guidance mechanism. These models cannot be synthesized by other conventional fabrication methods, and we suggest this approach will enable a variety of studies in cancer biology.

Extracellular vesicle molecular signatures characterize metastatic dynamicity in ovarian cancer

“…The extracellular matrix (ECM) is a key component of the TME, and undergoes remodeling during many of the stages of metastasis establishment (63)(64)(65). This remodeling plays a key role particularly in the development and progression of many epithelial cancers including ovarian cancer (33,34,(66)(67)(68). Our 10-gene signature, which overlaps with the signature elucidated by Cheon et al, was thus focused on collagen remodeling genes that are implicated in invasion and metastases in cancer (34,67,68).…”

“…This remodeling plays a key role particularly in the development and progression of many epithelial cancers including ovarian cancer (33,34,(66)(67)(68). Our 10-gene signature, which overlaps with the signature elucidated by Cheon et al, was thus focused on collagen remodeling genes that are implicated in invasion and metastases in cancer (34,67,68). Through bioinformatic analysis using published datasets such as that of Cheon et al (28), a ten gene signature that was overexpressed in ovarian cancer (28,69) and was focused on collagen remodeling (Table 1) was selected.…”

Multiscale anisotropy analysis of second-harmonic generation collagen imaging of mouse skin

“…To this end, previous evidence suggests that collagen fiber spacing, density, and orientation play important roles in regulating tissue morphology, cell shape, and integrin-mediated signaling that collectively helps control cell behavior. 10 , 26 , 76 – 79 For example, changes in collagen architecture have been shown to modulate integrin binding and promote both normal and pathological processes such as wound healing, angiogenesis, and tumor growth and metastasis. 10 , 26 , 76 – 79 However, while considerable insight is available as to how collagen binding integrins can control gene expression, 24 , 25 , 29 much less is known regarding whether collagen-binding integrins play roles in regulating the orientation of collagen fibers.…”

Engineered Collagen Matrices

“…A rapidly developing area in the fabrication of engineered collagen matrices focuses upon replicating the complex geometrical and architectural features of collagen. While the majority of synthetic collagen-mimetic approaches have focused on its biochemical attributes, there is mounting evidence supporting the importance of collagen architecture in guiding cell function [ 112 , 134 , 143 , 154 ]. Many elements of the nano- and micro- structure of collagen are important for guiding proper cell responses, as well as for providing desired mechanical characteristics.…”

“…Approaches to yield aligned collagen fiber scaffolds include the use of microfluidic platforms [ 156 ] and the application of anisotropic strain during collagen gelation [ 157 ]. Techniques such as multi-photon excited fabrication take the architectural replication a step further and allow for direct recapitulation of exact, tissue-specific fiber features based upon scans of native tissue [ 154 ]. However, some of these platforms do not permit cell encapsulation at the time of scaffold fabrication, and have thus been limited to studying these fibrillar interactions in 2D [ 156 ].…”