The efficacy of cetuximab in a tissue-engineered three-dimensional in vitro model of colorectal cancer

Abstract

The preclinical development process of chemotherapeutic drugs is often carried out in two-dimensional monolayer cultures. However, a considerable amount of evidence demonstrates that two-dimensional cell culture does not accurately reflect the three-dimensional in vivo tumour microenvironment, specifically with regard to gene expression profiles, oxygen and nutrient gradients and pharmacokinetics. With this objective in mind, we have developed and established a physiologically relevant three-dimensional in vitro model of colorectal cancer based on the removal of interstitial fluid from collagen type I hydrogels. We employed the RAFT™ (Real Architecture For 3D Tissue) system for producing three-dimensional cultures to create a controlled reproducible, multiwell testing platform. Using the HT29 and HCT116 cell lines to model epidermal growth factor receptor expressing colorectal cancers, we characterized three-dimensional cell growth and morphology in addition to the anti-proliferative effects of the anti–epidermal growth factor receptor chemotherapeutic agent cetuximab in comparison to two-dimensional monolayer cultures. Cells proliferated well for 14 days in three-dimensional culture and formed well-defined cellular aggregates within the concentrated collagen matrix. Epidermal growth factor receptor expression levels revealed a twofold and threefold increase in three-dimensional cultures for both HT29 and HCT116 cells in comparison to two-dimensional monolayers, respectively (p < 0.05; p < 0.01). Cetuximab efficacy was significantly lower in HT29 three-dimensional cultures in comparison to two-dimensional monolayers, whereas HCT116 cells in both two-dimension and three-dimension were non-responsive to treatment in agreement with their KRAS mutant status. In summary, these results confirm the use of a three-dimensional in vitro cancer model as a suitable drug-screening platform for in vitro pharmacological testing.

Three‐dimensional cell culture models for anticancer drug screening: Worth the effort?

“…It also influences different types of cell behavior such as cell migration, differentiation, and communication (Antoni et al, 2015;Edmondson et al, 2014;Ravi et al, 2015). The gene expression profiles of cells grown in 3D are often more similar to expression patterns found in native tissues or primary tumor samples (Antoni et al, 2015;Edmondson et al, 2014;Magdeldin et al, 2014;Ravi et al, 2015Yoshii et al, 2011. Multiple studies compared protein or gene expression levels of liver monolayer cultures with liver tissues or spheroid cultures and suggest that expression patterns of spheroids more closely resemble those of native liver tissue (Olsavsky et al, 2007;Ramaiahgari et al, 2014;Takahashi et al, 2015;Wiśniewski et al, 2016).…”

A Bispecific Inhibitor of the EGFR/ADAM17 Axis Decreases Cell Proliferation and Migration of EGFR-Dependent Cancer Cells

“…As expected, the efficacy of the fusion protein E01-GS-TPD decreased in the KRAS mutant cell lines A549 (Figure 5c) and HCT116. The dependence of EGFR downstream molecular players such as KRAS in the responsiveness to EGFR-targeted therapies is widely reported [23], and lies behind the prescription rationale for similarly working drugs in the clinic, such as the anti-EGFR antibody cetuximab, which is only indicated for wild type KRAS cancers [3]. Similarly, no effects in growth inhibition were observed in the non-small cell lung cancer cell line HCC827, which harbors a mutation in EGFR rendering the receptor constitutively active, regardless of ligand binding [2].…”

Initial validation of equilibrium contrast imaging for extracellular volume quantification using a three‐dimensional engineered tissue model

“…At higher cell fractions, failure of preparations to solidify is likely the result of an inadequate supporting collagen matrix. Future 3D engineered tissue models may incorporate higher amounts of collagen in a more complex matrix able to support higher, more physiological cell fractions. Thus, the relationship between ECV (m) and ECV (EQ) over a wider range, and the effect of varying extracellular collagen quantity may be investigated.…”