(T1530-03-17) Advancing Cancer Immunotherapy: Real-Time Immune Profiling and Cytokine Monitoring with an Integrated Biosensing Platform

Purpose: In the realm of cancer immunotherapy, understanding the role of immune cells and cytokines is crucial for efficacy and managing potential side effects1-2.  Current in vitro assessment methods have limitations, including being endpoint, time-intensive, and requiring large sample volumes. To address these challenges, we present an innovative solution: an integrated real-time 3D vascularized blood-brain barrier (BBB) chip 3-4. This platform aims to replicate the BBB environment infiltrated by immune T cells while enabling real-time monitoring of immune profiling and cytokine dynamics during immunotherapeutic interventions.

Methods: Our approach involves the development of an integrated real-time 3D vascularized BBB chip. This chip faithfully replicates the BBB environment and enables the infiltration of immune T cells. Crucially, it allows for the monitoring of immune profiling and real-time detection of cytokines during chimeric antigen receptor (CAR) T cell interactions with various cell types. The chip utilizes pillar microarrays with anti-cytokine capture antibodies (CAbs) immobilized as the primary layer to efficiently capture cytokines. Paired detection antibodies (DAbs) facilitate signal detection, enabling simultaneous detection of multiple cytokines within small sample sizes through parallel processing. The detection range spans from 1 to 10,000 pg/mL, requiring only a minimal sample volume of 4 μL.

Results: The integrated chip successfully monitors the dynamics of immune therapy-induced factor release without perturbing the cancer microenvironment. It allows for the evaluation of crucial aspects such as T cell and monocyte migration, cancer cell recognition, immune cell activation, and cytokine and neurotransmitter dynamics. The chip's innovative sensing mechanism enables real-time detection of cytokines within the biological milieu and offers a significant improvement over existing in vitro assessment methods in terms of sensitivity, sample volume requirement, and real-time monitoring capabilities

Conclusion: Our integrated real-time 3D vascularized BBB chip represents a significant advancement in the field of cancer immunotherapy. By providing a means to effectively monitor immune therapy-induced factor release without perturbing the cancer microenvironment, this platform empowers researchers to delve deeper into the intricacies of immunotherapy while enhancing its safety profile. This innovation holds promise for improving cancer treatment outcomes, ushering in a new era of precision and efficacy in cancer immunotherapy.

References: 1 Frankel, Timothy, Mirna Perusina Lanfranca, and Weiping Zou. "The role of tumor microenvironment in cancer immunotherapy." Tumor Immune Microenvironment in Cancer Progression and Cancer Therapy (2017): 51-64.
2Labanieh, Louai, Robbie G. Majzner, and Crystal L. Mackall. "Programming CAR-T cells to kill cancer." Nature biomedical engineering 2.6 (2018): 377-391.
3.Feng, Xiaojun, et al. "Microfluidic chip: next-generation platform for systems biology." Analytica Chimica Acta 650.1 (2009): 83-97.
4. Feng, Xiaojun, et al. "Microfluidic chip: next-generation platform for systems biology." Analytica Chimica Acta 650.1 (2009): 83-97.Friedrich, Juergen, et al. "Spheroid-based drug screen: considerations and practical approach." Nature protocols 4.3 (2009): 309-324.

Acknowledgements: This work was supported by the National Institutes of Health (NIH) MIRA R35GM133795 (P. Chen).


Figure1. CAR-T function in BBB chip. A. 3D capillary-like vascular structures in BBB chip. B. ZO-1 expression pattern in endothelial cells on brain barrier chip C. FITC-dextran penetration in the brain chamber D. The cellar attack of CAR-T on the chip. E. The HMGB1 levels in important cell components’. The ATP levels in important cell components. (The data are expressed as the mean ± standard deviation (SD), with a sample size of n = 3. Statistical significance is denoted by *** for P < 0.001. An unpaired t-test is utilized for comparisons between two groups, while one-way analysis of variance (ANOVA) is employed when assessing statistical significance among three or more groups.)