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Suijker, J.

Publications and source records attributed to Suijker, J..

2 recordsLinked to original sources
bioRxiv2026

A scalable tumor-vasculature-on-chip for CAR T cell trafficking and efficacy studies

Most cellular therapies, like CAR T cells, remain ineffective in solid tumors. This is primarily due to a complex tumor microenvironment (TME), which creates biochemically hostile and often immunosuppressive conditions that limit efficacy of immunotherapies. Besides, cellular therapy efficacy is still often established in traditional 2D cultures that fail to simulate relevant aspects of solid tumor biology. Recent advances in three-dimensional (3D) and organ-on-chip culture systems have provided more physiologically relevant models for immunotherapy testing. These microphysiological systems (MPS) not only offer a 3D environment that alters tumor cell sensitivity to therapy but also enable inclusion of TME components and assessment of processes such as extravasation and infiltration, key steps in CAR T cell activity in vivo. This study focuses on applying an advanced culture technique and further building on the use of a scalable on-chip platform, the OrganoPlate, to grow EpCAM-positive and EpCAM-negative tumor cells in co-culture with an endothelial vessel to study EpCAM-targeting CAR T cell migration and killing kinetics. The CAR T cells specifically targeted and killed EpCAM-positive HT-29 tumor cells while EpCAM-negative A375 tumor cells were not affected. In addition, target cell killing was dependent on the ratio between CAR T and tumor cells (E:T ratio) and was enhanced by addition of IL-2. Inflammatory cytokines like INF-{gamma}, TNF and IL-6 increased overtime in cultures containing CAR T cells. Morphometric analyses of the endothelial compartment showed E:T ratio dependent disruption of endothelial vessels. Additionally, this system was able to distinguish EpCAM ScFv-CD28-CD3z and EpCAM ScFv-TM-4-1BB-CD3z CAR T cells killing abilities and was used for studying the effect of immune checkpoint inhibitors and Temozolomide, a DNA targeting drug, on CAR T cell performance. Altogether, this work adds to the available advanced culture techniques for immunotherapy developers by describing a model that is modular, scalable, and suitable for phenotypic and functional characterization of CAR T cells.

bioengineering↗
bioRxiv2022

Lung Tumor Microphysiological System with 3D Endothelium to Evaluate Modulators of T-Cell Infiltration

Lung cancer is a leading cause of death worldwide, with only a fraction of patients responding to immunotherapy. The correlation between increased T-cell infiltration and positive patient outcomes has motivated the search for therapeutics promoting T-cell infiltration. While transwell and spheroid platforms have been employed, these models lack flow and endothelial barriers, and cannot faithfully model T-cell adhesion, extravasation and migration through 3D tissue. Presented here is a 3D chemotaxis assay, in a lung tumor on chip model with 3D endothelium (LToC-Endo), to address this need. The described assay consists of a vascular tubule cultured under rocking flow, through which T-cells are added; a collagenous stromal barrier, through which T-cells infiltrate; and a chemoattractant/tumor compartment. Here, activated T-cells extravasate and infiltrate in response to gradients of rhCXCL11 and rhCXCL12. Adopting a T-cell activation protocol with a rest period enables proliferative burst prior to introducing T-cells into chips, increases T-cell expression of CXCR3 and CXCR4 receptors, and enhances assay sensitivity. In addition, incorporating this rest recovers endothelial activation in response to rhCXCL12. As a final control, we show that blocking ICAM-1 interferes with T-cell adhesion and chemotaxis. This microphysiological system, which mimics in vivo stromal and vascular barriers, can be used to evaluate potentiation of immune chemotaxis into tumors while probing for vascular responses to potential therapeutics. Finally, we propose a translational strategy by which this assay could be linked to preclinical and clinical models to support human dose prediction, personalized medicine, and the reduction, refinement, and replacement of animal models.

bioengineering↗