Investigation of Cancer Cell Migration and Proliferation on Synthetic Extracellular Matrix Peptide Hydrogels.

Zbigniev Balion,Airina Mazetyte-Godiene,Emilija Sipailaite,Ieva Seskeviciute,Jaywant Phopase,Aiste Jekabsone,Gabija Stasyte,Agne Vailionyte,Rasa Bernotiene

doi:10.3389/fbioe.2020.00773

Abstract

Chemical and mechanical properties of a tumor microenvironment are essential players in cancer progression, and it is important to precisely control the extracellular conditions while designing cancer in vitro models. The study investigates synthetic hydrogel matrices from multi-arm polyethylene glycol (PEG) functionalized with collagen-like peptide (CLP) CG(PKG)4(POG)4(DOG)4 alone and conjugated with either cell adhesion peptide RGD (mimicking fibronectin) or IKVAV (mimicking laminin). Human glioblastoma HROG36, rat C6 glioma cells, and A375 human melanoma cells were grown on the hydrogels and monitored for migration, proliferation, projected cell area, cell shape index, size and number, distribution of focal contacts in individual cells, and focal adhesion number. PEG-CLP-RGD induced migration of both glioma cell lines and also stimulated proliferation (assessed as metabolic activity) of HROG36 cells. Migration of C6 cells were also stimulated by PEG-CLP-IKVAV. These responses strongly correlated with the changes in adhesion and morphology parameters of individual cells – projected cell area, cell shape index, and focal contact number. Melanoma A375 cell proliferation was increased by PEG-CLP-RGD, and this was accompanied by a decrease in cell shape index. However, neither RGD nor IKVAV conjugated to PEG-CLP stimulated migratory capacity of A375 cells. Taken together, the study presents synthetic scaffolds with extracellular matrix (ECM)-mimicking peptides that allow for the exploration of the effect of ECM signaling to cancer cells.

Highlights

The design of a realistic cell culture microenvironment is an important aspect in the development of cancer in vitro models
We examine extracellular matrix (ECM)-mimicking hydrogels made of self-assembling collagen-like peptide attached to eight-armed polyethylene glycol (PEG-CLP; Islam et al, 2016) functionalized with fibronectin active site motif RGD (PEG-CLP-RGD) and laminin motif IKVAV (PEG-CLP-IKVAV) as matrices for cancer in vitro modeling
The quantitative conjugation of CLP, CLP-RGD, and CLP-IKVAV peptides with 8-arm-PEG maleimide was confirmed by the disappearance of the maleimide peak at δ 6.5–7.0 ppm after conjugation of peptides to PEG template

Summary

Introduction

The design of a realistic cell culture microenvironment is an important aspect in the development of cancer in vitro models. Besides soluble and extracellular vesicle-encapsulated factors, there are two key players instructing cancer cells to migrate and proliferate: cell– cell interaction and cell–extracellular matrix (ECM) interaction For studies into the latter, it is necessary to develop a substrate that: (i) would mimic a natural cancer cell environment, (ii) would have precisely controlled composition of signaling elements, and (iii) would support a standardized and easy-tomonitor and analyze cell culture. The cell migration directly depends on the focal adhesion size (Kim and Wirtz, 2013) These interactions between cells and ECM proteins control differentiation, shape, movement, cell phenotype, and viability (Colognato and Yurchenco, 2000; Smith et al, 2018). Here lies the challenge of controlled design, scaling, and standardization of such ECM mimetics, because the production of the proteins is expensive and contains the risk of relatively high batch-to-batch variations and biocontamination

Methods

Results

Conclusion