A Reproducible Bioprinted 3D Tumor Model Serves as a Preselection Tool for CAR T Cell Therapy Optimization

Abstract

Chimeric antigen receptor (CAR) T cell performance against solid tumors in mouse models and clinical trials is often less effective than predicted by CAR construct selection in two-dimensional (2D) cocultures. Three-dimensional (3D) solid tumor architecture is likely to be crucial for CAR T cell efficacy. We used a three-dimensional (3D) bioprinting approach for large-scale generation of highly reproducible 3D human tumor models for the test case, neuroblastoma, and compared these to 2D cocultures for evaluation of CAR T cells targeting the L1 cell adhesion molecule, L1CAM. CAR T cells infiltrated the model, and both CAR T and tumor cells were viable for long-term experiments and could be isolated as single-cell suspensions for whole-cell assays quantifying CAR T cell activation, effector function and tumor cell cytotoxicity. L1CAM-specific CAR T cell activation by neuroblastoma cells was stronger in the 3D model than in 2D cocultures, but neuroblastoma cell lysis was lower. The bioprinted 3D neuroblastoma model is highly reproducible and allows detection and quantification of CAR T cell tumor infiltration, representing a superior in vitro analysis tool for preclinical CAR T cell characterization likely to better select CAR T cells for in vivo performance than 2D cocultures.

Bioprinted Cancer Model of Neuroblastoma in a Renal Microenvironment as an Efficiently Applicable Drug Testing Platform

“…In a model composed of the neuroblastoma cell line SH-SY5Y in co-culture with mesenchymal stromal cells and human primary umbilical vein endothelial cells, the neuroblastoma cells formed Homer Wright-like rosettes and maintained their proliferative capacities [ 34 ]. Another bioprinted tumor model consisting of SK-N-BE(2) cells was used to investigate the infiltration of chimeric antigen receptor (CAR) T cells into tumor tissues [ 35 ]. In further studies, neuroblastoma cell lines have been used to develop neural tissue for studying neurodegenerative diseases [ 32 , 33 ].…”

“…As described above, the study of Grundwald et al [ 35 ] used a bioprinted neuroblastoma model consisting of SK-N-BE(2) cells to investigate tumor tissue penetration by CAR T cells. Our model, which consists of not only cancerous cells, but also normal fibroblasts, can now be used to investigate the specificity of the treatment for the destruction of the tumor.…”

“…The aim of the present study was to develop a bioprinted 3D model that mimics a tumor in a microenvironment exclusively composed of human cells. As neuroblastoma cells have been shown to be well suited for bioprinting approaches [ 30 , 31 , 32 , 33 , 34 , 35 ], this tumor type was chosen as an example. To the best of our knowledge, few studies exist that have embedded the tumor cells in an environment of normal cells to test the efficiency and specificity of cytostatic or other anticancer drugs.…”

Targeting High-Risk Neuroblastoma Patient-Derived Xenografts with Oncolytic Virotherapy

“…The use of 3D technology has been more widespread in the adult oncology literature [12,41] and has had only limited reports in pediatric oncology. Pediatric cancer researchers have printed SH-SY5Y neuroblastoma cells to create a neural network [42] and SK-N-BE neuroblastoma cells to study CAR-T cell therapy [43], but to our knowledge, no one has yet reported bioprinting neuroblastoma PDX cells. In the current investigations, we successfully created bioprinted microtumors of the COA6 and COA129 PDXs.…”

The next wave of cellular immunotherapies in pancreatic cancer

“… 146 In addition, bioprinted (neuroblastoma) organoids could be used to preselect CAR T cell constructs. 147 There is no standard cytotoxicity assay to test the efficacy of CARs with PDOs. One study established a luciferase-based endpoint assay and a live microscopy assay for continuous and cell-resolved analysis to monitor NK CAR-mediated cytotoxicity against colorectal cancer organoids.…”