Optimizing CAR-T-cell therapy using 3D tumor models and real-time cell imaging

03:2523 months ago

Chimeric antigen receptor (CAR) T-cell therapy accounts for one of the most promising therapeutic advances in cancer immunotherapy. In this form of adoptive cell transfer, T-cells of a patient are engineered to express so-called ‘CARs’, in which the antigen-recognition capacity of antibodies is combined with T-cell activating domains. So far, CAR-T-cell therapy obtained its most impressive results in hematological malignancies resulting in the approval of five CAR-T cell products by the FDA for hematologic indications. However, CAR-T-cell therapy has not mirrored its success in solid tumors. The poor efficacy of CAR-T-cell therapy in solid tumors has, in part, been attributed to the lack of understanding in how CAR-T-cells function in a solid tumor microenvironment. Classical validation methods rely on the use of specificity and functionality assays in 2D models against adherent target cells or target cells in suspension. Yet, by using these models, observations made in vitro may differ greatly to an in vivo situation where tumors are engrafted in 3D structures. We developed a more relevant and translational 3D tumor model using eGFP+ target cells. This allows us to couple 3D tumor cell killing by CAR-T-cells to live-cell imaging, providing an efficient quantification of target cell death. As proof- of-concept, we used a 3D model of eGFP+ glioblastoma cells and CAR-T-cells targeting a pan-cancer antigen. This 3D glioblastoma model allowed us to show that classical scFv-based CAR-T-cell therapy of glioblastoma cells can be improved by nanoCAR-T-cells. Furthermore, combining nanoCAR-T-cell therapy with a genetic approach of nanobody-based anti-PD-L1 immune checkpoint blockade further increased the cytotoxicity of the nanoCAR-T-cell therapy.

Related

VitalTissue: scientific research can be more human(e)
Projects and initiatives
HealthInnovationIn vitro

VitalTissue: scientific research can be more human(e)

The goal of VitalTissue is to facilitate the availability of vital human residual tissue for all researchers in the Netherlands. This video shows how VitalTissue works. From a request from a researcher, the donation of the residual tissue by the patient and the transport to the lab. This process is the result of a feasibility study conducted with many stakeholders. The national tissue bank ETB-BISLIFE will implement VitalTissue in practice.
04:213 months ago
Helpathon #9 – Can you help Juan?
Meeting videos
HealthInnovationIn vitro

Helpathon #9 – Can you help Juan?

Juan is an experienced immunologist and scientific director of the cutting edge O2Flow facility for cytometry and cell sorting at the Amsterdam VU University Medical Center. Can you help Juan explore if and how he can transition his facility towards animal free antibodies? Are you using antibodies in your research do you want others to help you find animal free alternatives for your specific research let us know. More information can be found [here] (https://www.helpathonhotel.org/coming-up).
02:024 months ago
Development of 3D liver spheroids
Innovation examples
HealthToxicologyInnovationIn vitro

Development of 3D liver spheroids

Human-based in vitro models are increasingly being used in the hepatology field. And in addition to the obvious ethical arguments, they offer several advantages over the classical animal models. One of them is the ability to perform mechanistic research at the molecular level in a well-controlled setting and reduce species differences. These liver-based in vitro models can range from simple monolayer cultures of hepatocytes to the liver-on-chips systems in which all liver cells are cultured in a 3D configuration on a microfluidic platform. Liver-based in vitro models must be selected on a case-by-case basis and should fit the purpose of the research, which might go from fundamental to translational research.
01:045 months ago
Platform for in vitro airborne inhalation testing
Innovation examples
HealthToxicologyInnovationIn vitro

Platform for in vitro airborne inhalation testing

The air-liquid interface (ALI) technique uses lung cells cultured on a tiny polymer membrane in a cup. On one side of the membrane is a liquid containing the medium necessary for the cells to survive, while the other side is in contact with air. This is similar to the situation in the human lung. The compound to be tested is administered via an aerosol, vapor, or gas to mimic the situation in human lungs. By monitoring different parameters in the cell model before and after the compound is added, it is possible to measure the effects on lung cells. Depending on the test to be carried out, the lung cells can come from different regions in the respiratory tract and even from a variety of people, including individuals who smoke a lot or have specific diseases such as chronic obstructive pulmonary disease or asthma. In vitro ALI inhalation testing (https://doi.org/10.1021/acs.est.7b00493) adds value for e.g. pre-clinical trials and research in the pharmaceutical industry and testing (new) compounds for the chemical sector and beyond. The advantages of ALI inhalation testing are that it is a non-animal method, it reduces the use of in vivo experiments, pre-clinical testing with human-derived cell models is more realistic and limits clinical trial failures and it provides faster and more efficient testing of compound
04:135 months ago