Andrea Pavesi1,2, Anthony T. Tan3, Sarene Koh4, Adeline Chia3, Marta Colombo5, Emanuele Antonecchia5, Carlo Miccolis5, Erica Ceccarello3, Giulia Adriani2, Manuela T. Raimondi5, Roger D. Kamm2,6 and Antonio Bertoletti3,4
1 Institute of Molecular and Cell Biology, Agency for Science, Technology, and Research, Singapore.
2 BioSystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, Singapore.
3 Emerging Infectious Disease Program, Duke-NUS Graduate Medical School, Singapore.
4 Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research, Singapore.
5 Department of Chemistry, Materials and Chemical Engineering “Giulio Natta,” Politecnico di Milano, Milan, Italy.
6 MechanoBiology Laboratory, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Authorship note: A. Pavesi and A.T. Tan contributed equally to this work
Published in JCI Insight on 15 June 2017. (doi:10.1172/jci.insight.89762)
The tumor microenvironment imposes physical and functional constraints on the antitumor efficacy of adoptive T cell immunotherapy. Preclinical testing of different T cell preparations can help in the selection of efficient immune therapies, but in vivo models are expensive and cumbersome to develop, while classical in vitro 2D models cannot recapitulate the spatiotemporal dynamics experienced by T cells targeting cancer. Here, we describe an easily customizable 3D model, in which the tumor microenvironment conditions are modulated and the functionality of different T cell preparations is tested. We incorporate human cancer hepatocytes as a single cell or as tumor cell aggregates in a 3D collagen gel region of a microfluidic device. Human T cells engineered to express tumor-specific T cell receptors (TCR–T cells) are then added in adjacent channels. The TCR–T cells’ ability to migrate and kill the tumor target and the profile of soluble factors were investigated under conditions of varying oxygen levels and in the presence of inflammatory cytokines. We show that only the 3D model detects the effect that oxygen levels and the inflammatory environment impose on engineered TCR–T cell function, and we also used the 3D microdevice to analyze the TCR–T cell efficacy in an immunosuppressive scenario. Hence, we show that our microdevice platform enables us to decipher the factors that can alter T cell function in 3D and can serve as a preclinical assay to tailor the most efficient immunotherapy configuration for a specific therapeutic goal.
Evaluation of TCR-engineered T cell function using a 3D microfluidic device. Factors influencing the functionality of TCR-engineered T cells were assessed using the microdevice, and the information obtained will then be used to improve the in vivo efficiency of the engineered T cells. (A–C) The workflow of the presented work is explained. (D and E) 3D rendering of the devices in disperse cell configuration and aggregate configuration. (F) The predominantly gravity-driven migration of engineered T cells along the z axis, as occurs in a 2D well-based assay, compared with the directional chemotaxis in a 3D microdevice.
For more information on Andrea PAVESI's lab, please click here.