3D Tissue-Engineered Tumor Model for Ewing’s Sarcoma That Incorporates Bone-like ECM and Mineralization

Abstract

The tumor microenvironment harbors essential components required for cancer progression including biochemical signals and mechanical cues. To study the effects of microenvironmental elements on Ewing’s sarcoma (ES) pathogenesis, we tissue-engineered an acellular three-dimensional (3D) bone tumor niche from electrospun poly­(ε-caprolactone) (PCL) scaffolds that incorporate bone-like architecture, extracellular matrix (ECM), and mineralization. PCL-ECM constructs were generated by decellularizing PCL scaffolds harboring cultures of osteogenic human mesenchymal stem cells. The PCL-ECM constructs simulated in vivo-like tumor architecture and increased the proliferation of ES cells compared to PCL scaffolds alone. Compared to monolayer controls, 3D environments facilitated the downregulation of the canonical insulin-like growth factor 1 receptor (IGF-1R) signal cascade through mechanistic target of rapamycin (mTOR), both of which are targets of recent clinical trials. In addition to the downregulation of canonical IGF-1R signaling, 3D environments promoted a reduction in the clathrin-dependent nuclear localization and transcriptional activity of IGF-1R. In vitro drug testing revealed that 3D environments generated cell phenotypes that were resistant to mTOR inhibition and chemotherapy. Our versatile PCL-ECM constructs allow for the investigation of the roles of various microenvironmental elements in ES tumor growth, cancer cell morphology, and induction of resistant cell phenotypes.

In vitro three-dimensional cell cultures for bone sarcomas

“…Similarly, Santoro et al emphasized the role of IGF1/IGF-1R pathway and biomechanical TME stimulation in drug resistance by using similar 3D models [113] . These data are in agreement with the recent study published by Molina et al [114] . Indeed, these authors demonstrated that 3D TME favoured the downregulation of IGF-1R via mTOR pathway, which was accompanied by a reduction of the clathrin-dependent nuclear localisation and transcription activity of IGF-1R [114] .…”

“…These data are in agreement with the recent study published by Molina et al [114] . Indeed, these authors demonstrated that 3D TME favoured the downregulation of IGF-1R via mTOR pathway, which was accompanied by a reduction of the clathrin-dependent nuclear localisation and transcription activity of IGF-1R [114] . TC-71 3D culture was exposed to different shear stresses close to those observed in bone microenvironment in a flow perfusion bioreactor.…”

The Role of IGF/IGF-IR-Signaling and Extracellular Matrix Effectors in Bone Sarcoma Pathogenesis

“…Thus, in vitro 3D environments enhance the canonical IGF-IR signal cascade's attenuation through mechanistic target of rapamycin (mTOR). Notably, 3D environments facilitated a decrease in the clathrin-dependent nuclear localization and transcriptional activity of IGF-IR [159]. Therefore, modulating the matrix network could contribute to cancer therapies directed at the IGF-signaling pathway.…”

Going with the Flow: Modeling the Tumor Microenvironment Using Microfluidic Technology

“…While biomaterials can be natural or artificially made [ 89 ], most 3D tumor models use natural biomaterials purified from animals or plants that can be enzymatically digested by tumor and/or stromal cells. The selection of the biomaterial to be used as a scaffold for 3D tumor modeling is far from trivial as some are biologically active (e.g., ECM components driving cancer growth [ 89 ]) and can offer specific advantages, such as sustaining drug release or generating bone-like structures [ 90 , 91 ]. Commonly used natural biomaterials in 3D tumor models include collagen, gelatin, Matrigel, hydrogel, chitosan, alginate, silk, poly-ε-caprolactone (PCL), and hyaluronic acid [ 89 , 92 , 93 ].…”

“…Beyond that, some studies have also reported engineering biomaterials to have specific functions. For example, gelatin hydrogel microspheres (GM) have been used as a source to continually release drugs [ 90 , 94 ]; PCL scaffolds have been engineered to have bone-like architecture and mineralization [ 91 , 95 ].…”