A Novel 3D In Vitro Platform for Pre-Clinical Investigations in Drug Testing, Gene Therapy, and Immuno-oncology

Abstract

Tumors develop within complex cell-to-cell interactions, with accessory cells playing a relevant role starting in the early phases of cancer progression. This event occurs in a three-dimensional (3D) environment, which to date, has been difficult to reproduce in vitro due to its complexity. While bi-dimensional cultures have generated substantial data, there is a progressive awareness that 3D culture strategies may rapidly increase the understanding of tumor development and be used in anti-cancer compound screening and for predicting response to new drugs utilizing personalized approaches. However, simple systems capable of rapidly rebuilding cancer tissues ex-vivo in 3D are needed and could be used for a variety of applications. Therefore, we developed a flat, handheld and versatile 3D cell culture bioreactor that can be loaded with tumor and/or normal cells in combination which can be monitored using a variety of read-outs. This biocompatible device sustained 3D growth of tumor cell lines representative of various cancers, such as pancreatic and breast adenocarcinoma, sarcoma, and glioblastoma. The cells repopulated the thin matrix which was completely separated from the outer space by two gas-permeable membranes and was monitored in real-time using both microscopy and luminometry, even after transportation. The device was tested in 3D cytotoxicity assays to investigate the anti-cancer potential of chemotherapy, biologic agents, and cell-based therapy in co-cultures. The addition of luciferase in target cancer cells is suitable for comparative studies that may also involve parallel in vivo investigations. Notably, the system was challenged using primary tumor cells harvested from lung cancer patients as an innovative predictive functional assay for cancer responsiveness to checkpoint inhibitors, such as nivolumab. This bioreactor has several novel features in the 3D-culture field of research, representing a valid tool useful for cancer investigations, drug screenings, and other toxicology approaches.

Development of a 3D functional assay and identification of biomarkers, predictive for response of high-grade serous ovarian cancer (HGSOC) patients to poly-ADP ribose polymerase inhibitors (PARPis): targeted therapy

“…Research in oncology is now becoming more aware of the importance of 3D culture applications to understand how tumors develop, including their utility in compound screening for predicting drug response in patients [46,47]. Studying prolonged and long term effects of treatments is more feasible in a 3D setting due to the lower proliferation rates exhibited by cells in 3D cultures [48], in addition to lower levels of apoptosis, as well as various parameters relating to cellular motility, and cell morphology [49].…”

Exploitation of the chick embryo chorioallantoic membrane (CAM) as a platform for anti-metastatic drug testing

“…Recently, optimization of 3D primary cultures has provided biologically relevant information on tumor growth and response to various stimuli 32 . However, the complex interactions within the organism are hardly mimicked in 3D co-culture techniques 2 and complementary preclinical tools are needed. Up until now, many successful drug tests were performed on primary nodules of the CAM and confirm utilization of this model as a reliable preclinical model for testing novel therapeutics 17,26,33 .…”

“…Furthermore, costs related to establishment and husbandry of mouse models and ethical issues considerably limit their use. Recently, 3D organoids have emerged as novel robust tools to model tumor heterogeneity and perform drug screening assays 2 . Great efforts have been put into generating 3D co-cultures to simulate tumor microenvironment ex vivo.…”

Arming Mesenchymal Stromal/Stem Cells Against Cancer: Has the Time Come?

“…In addition, is possible to consistently isolate and modify MSCs from human adipose tissue by minimally invasive surgical procedures ( Foppiani et al., 2019 ; Starnoni et al., 2019 ). The wild-type gene coding for membrane-bound TRAIL, as well as modified cassettes expressing soluble ligand forms, have been used in MSC-based therapeutic strategies, demonstrating antitumor effects in vitro and in vivo in a wide variety of human solid neoplasms, including lung cancer, pancreatic cancer, glioblastoma, sarcoma, and hepatocarcinoma ( Loebinger et al., 2009 ; Sasportas LS et al., 2009 ; Grisendi et al., 2010 ; Grisendi et al., 2011 ; Yan et al., 2014 ; D’Souza et al., 2015 ; Grisendi et al., 2015 ; Guiho et al., 2016 ; Golinelli et al., 2018 ; Candini et al., 2019 ; Rossignoli et al., 2019 ; Spano et al., 2019 ).…”