Three-dimensional prostate tumor model based on a hyaluronic acid-alginate hydrogel for evaluation of anti-cancer drug efficacy

Abstract In vitro cell-based assays are widely applied to evaluate anti-cancer drug efficacy. However, the conventional approaches are mostly based on two-dimensional (2D) culture systems, making it difficult to recapitulate the in vivo tumor scenario because of spatial limitations. Here, we develop an in vitro three-dimensional (3D) prostate tumor model based on a hyaluronic acid (HA)-alginate hybrid hydrogel to bridge the gap between in vitro and in vivo anticancer drug evaluations. In situ encapsulation of PCa cells was achieved by mixing HA and alginate aqueous solutions in the presence of cells and then crosslinking with calcium ions. Unlike in 2D culture, cells were found to aggregate into spheroids in a 3D matrix. The expression of epithelial to mesenchyme transition (EMT) biomarkers was found to be largely enhanced, indicating an increased invasion and metastasis potential in the hydrogel matrix. A significant up-regulation of proangiogenic growth factors (IL-8, VEGF) and matrix metalloproteinases (MMPs) was observed in 3D-cultured PCa cells. The results of anti-cancer drug evaluation suggested a higher drug tolerance within the 3D tumor model compared to conventional 2D-cultured cells. Finally, we found that the drug effect within the in vitro 3D cancer model based on HA-alginate matrix exhibited better predictability for in vivo drug efficacy.

[1]  S. Van Vlierberghe,et al.  Biopolymer-based hydrogels as scaffolds for tissue engineering applications: a review. , 2011, Biomacromolecules.

[2]  Glenn D Prestwich,et al.  Modular extracellular matrices: solutions for the puzzle. , 2008, Methods.

[3]  F. Lin,et al.  An alginate-based platform for cancer stem cell research. , 2016, Acta biomaterialia.

[4]  JONG BIN Kim,et al.  Three-dimensional tissue culture models in cancer biology. , 2005, Seminars in cancer biology.

[5]  L. Windus,et al.  In vivo biomarker expression patterns are preserved in 3D cultures of Prostate Cancer. , 2012, Experimental cell research.

[6]  N. B. Viana,et al.  Extracellular matrix secreted by reactive stroma is a main inducer of pro-tumorigenic features on LNCaP prostate cancer cells. , 2012, Cancer letters.

[7]  Jianmiao Liu,et al.  Agarose multi-wells for tumour spheroid formation and anti-cancer drug test , 2016 .

[8]  Shufang Zhang,et al.  Engineering a scaffold-free 3D tumor model for in vitro drug penetration studies. , 2010, Biomaterials.

[9]  Michael J. Wilson,et al.  Expression of matrix metalloproteinase‐2 and ‐9 and their inhibitors, tissue inhibitor of metalloproteinase‐1 and ‐2, in primary cultures of human prostatic stromal and epithelial cells * † , 2002, Journal of cellular physiology.

[10]  Donald C. Chang,et al.  Hyaluronan stimulates transformation of androgen-independent prostate cancer. , 2006, Carcinogenesis.

[11]  A Tampieri,et al.  HA/alginate hybrid composites prepared through bio-inspired nucleation. , 2005, Acta biomaterialia.

[12]  M. Boulton.  Cell Culture , 2010, Cell.

[13]  P. Stattin,et al.  Prostate cancer increases hyaluronan in surrounding nonmalignant stroma, and this response is associated with tumor growth and an unfavorable outcome. , 2011, The American journal of pathology.

[14]  A. Oloumi,et al.  Identification of genes differentially expressed in V79 cells grown as multicell spheroids , 2002, International journal of radiation biology.

[15]  Dietmar W. Hutmacher,et al.  Bioengineered 3D platform to explore cell-ECM interactions and drug resistance of epithelial ovarian cancer cells. , 2010, Biomaterials.

[16]  R E Durand,et al.  Detection of hypoxic cells in a murine tumor with the use of the comet assay. , 1992, Journal of the National Cancer Institute.

[17]  Samy Lamouille,et al.  Molecular mechanisms of epithelial–mesenchymal transition , 2014, Nature Reviews Molecular Cell Biology.

[18]  Jason A. Burdick,et al.  Hyaluronic Acid Hydrogels for Biomedical Applications , 2011, Advanced materials.

[19]  Christian G Elowsky,et al.  Spontaneous metastasis of prostate cancer is promoted by excess hyaluronan synthesis and processing. , 2009, The American journal of pathology.

[20]  Stephen J. Florczyk,et al.  3D Porous Chitosan–Alginate Scaffolds: A New Matrix for Studying Prostate Cancer Cell–Lymphocyte Interactions In Vitro , 2012, Advanced healthcare materials.

[21]  Jinmin Zhao,et al.  In vitro ovarian cancer model based on three-dimensional agarose hydrogel , 2014, Journal of tissue engineering.

[22]  K. Książek,et al.  Resveratrol inhibits ovarian cancer cell adhesion to peritoneal mesothelium in vitro by modulating the production of α5β1 integrins and hyaluronic acid. , 2014, Gynecologic oncology.

[23]  S. Sahoo,et al.  Characterization of porous PLGA/PLA microparticles as a scaffold for three dimensional growth of breast cancer cells. , 2005, Biomacromolecules.

[24]  David J Mooney,et al.  Cancer cell angiogenic capability is regulated by 3D culture and integrin engagement , 2009, Proceedings of the National Academy of Sciences.

[25]  J. Hubbell,et al.  Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering , 2005, Nature Biotechnology.

[26]  Jyrki Lötjönen,et al.  A Comprehensive Panel of Three-Dimensional Models for Studies of Prostate Cancer Growth, Invasion and Drug Responses , 2010, PloS one.

[27]  Kenneth M. Yamada,et al.  Modeling Tissue Morphogenesis and Cancer in 3D , 2007, Cell.

[28]  Martin Fussenegger,et al.  Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types. , 2003, Biotechnology and bioengineering.

[29]  Tae Woo Kim,et al.  A chitosan hydrogel-based cancer drug delivery system exhibits synergistic antitumor effects by combining with a vaccinia viral vaccine. , 2008, International journal of pharmaceutics.

[30]  Tobias Schmelzle,et al.  Engineering tumors with 3D scaffolds , 2007, Nature Methods.

[31]  Shuichi Takayama,et al.  High-throughput 3D spheroid culture and drug testing using a 384 hanging drop array. , 2011, The Analyst.

[32]  Hitoshi Shiku,et al.  Alginate gel microwell arrays using electrodeposition for three-dimensional cell culture. , 2013, Lab on a Chip.

[33]  Melvin E. Andersen,et al.  Organotypic liver culture models: Meeting current challenges in toxicity testing , 2012, Critical reviews in toxicology.

[34]  D. Lauffenburger,et al.  Migration of tumor cells in 3D matrices is governed by matrix stiffness along with cell-matrix adhesion and proteolysis. , 2006, Proceedings of the National Academy of Sciences of the United States of America.