Identification of drugs as single agents or in combination to prevent carcinoma dissemination in a microfluidic 3D environment

Abstract

Experiments were performed in a modified microfluidic platform recapitulating part of the in vivo tumor microenvironment by co-culturing carcinoma cell aggregates embedded in a three-dimensional (3D) collagen scaffold with human umbilical vein endothelial cells (HUVECs). HUVECs were seeded in one channel of the device to initiate vessel-like structures in vitro prior to introducing the aggregates. The lung adenocarcinoma cell line A549 and the bladder carcinoma cell line T24 were tested. Dose-response assays of four drugs known to interfere with Epithelial Mesenchymal Transition (EMT) signaling pathways were quantified using relative dispersion as a metric of EMT progression. The presence of HUVECs in one channel induces cell dispersal in A549 which then can be inhibited by each of the four drugs. Complete inhibition of T24 aggregate dispersal, however, is not achieved with any single agent, although partial inhibition was observed with 10 μM of the Src inhibitor, AZD-0530. Almost complete inhibition of T24 dispersal in monoculture was achieved only when the four drugs were added in combination, each at 10 μM concentration. Coculture of T24 with HUVECs forfeits the almost-complete inhibition. The enhanced dispersal observed in the presence of HUVECs is a consequence of secretion of growth factors, including HGF and FGF-2, by endothelial cells. This 3D microfluidic co-culture platform provides an in vivo-like surrogate for anti-invasive and anti-metastatic drug screening. It will be particularly useful for defining combination therapies for aggressive tumors such as invasive bladder carcinoma.

Warburg metabolism in tumor-conditioned macrophages promotes metastasis in human pancreatic ductal adenocarcinoma

“…3 consisting of a central gel channel flanked by two fluidic channels was utilized. Similar microfluidic platforms have been used for investigating angiogenesis, 13,14 EMT, 15,16 cancer cell-macrophage interactions, 17 intravasation 18 and extravasation. 19,20 For the present work, GFPexpressing HUVECs were seeded into one of the fluidic channels to form a uniform monolayer mimicking a blood vessel wall (Fig.…”

Janus kinases and Src family kinases in the regulation of EGF-induced vimentin expression in MDA-MB-468 breast cancer cells

“…The SFKs consist of nine members including: Src, Lck, Hck, Fyn, Blk, Lyn, Fgr, Yes, and Yrk (Boggon and Eck, 2004), while the JAK family has four members: JAK1, JAK2, JAK3 and TYK2 (Babon et al, 2014). In addition to being implicated in the progression of different epithelial tumors, including ovarian (Abubaker et al, 2014;Colomiere et al, 2009;Yue et al, 2012), lung (Zhang et al, 2007) and breast (Hedvat et al, 2009;Silva, 2004), selected members of each family (particularly Src and JAK2) are also associated with the regulation of the mesenchymal phenotype in various in vitro models of EMT (Bai et al, 2015;Balanis et al, 2013;Chua et al, 2012;Colomiere et al, 2009;Jo et al, 2009;Liu et al, 2014;Lo et al, 2007;Maschler et al, 2010;Nagathihalli and Merchant, 2012;Nam et al, 2002;Switzer et al, 2012;Yadav et al, 2011). The JAK and SFK signaling pathways therefore represent potential therapeutic targets in the treatment and prevention of invasive disease.…”

Heralding a new paradigm in 3D tumor modeling

“…Microfluidic platforms that enable the precise integration of various biophysical and biochemical factors that govern tumor-vasculature interactions have also been developed [129–131]. In one such study, Bai et al co-cultured a highly invasive bladder carcinoma cell line as aggregates embedded in a collagen scaffold in close proximity to an endothelial cell-lined channel mimicking a blood vessel to screen a panel of therapeutics (and combinations thereof) that can inhibit EMT [131].…”

“…In one such study, Bai et al co-cultured a highly invasive bladder carcinoma cell line as aggregates embedded in a collagen scaffold in close proximity to an endothelial cell-lined channel mimicking a blood vessel to screen a panel of therapeutics (and combinations thereof) that can inhibit EMT [131]. Using the degree of tumor aggregate dispersion as a measure of drug efficacy, it was found that the presence of endothelial cells enhanced the drug resistance of the bladder carcinoma cells.…”