| Link | https://uottawa.scholarsportal.info/ottawa/index.php/uojm-jmuo/issue/view/642/377 |
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| Author | Search for: Huang, J.1; Search for: Li, B.1ORCID identifier: https://orcid.org/0000-0002-1560-8866; Search for: Charlebois, C.1; Search for: Baumann, E.1; Search for: Bloemberg, D.1; Search for: Nguyen, T.1; Search for: Zafer, A.1; Search for: Liu, Z.1; Search for: Liu, Q.1; Search for: Stanimirovic, D. B.1; Search for: McComb, S.1ORCID identifier: https://orcid.org/0000-0002-4616-7864; Search for: Jezierski, A1 |
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| Affiliation | - National Research Council Canada. Human Health Therapeutics
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| Format | Text, Abstract |
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| Conference | Brain Health Research Day 2022, June 3, 2022, Ottawa, ON, Canada |
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| Abstract | Human blood brain barrier (BBB) models derived from induced pluripotent stem cell (iPSC) have become an important tool for discovery and preclinical evaluation of CNS targeting cell and gene -based therapies. Chimeric antigen receptor (CAR)-T is a revolutionary form of gene-mod-ified cell-based immunotherapy with potential for targeting solid tumors, such as glioblastomas. Crossing the BBB is an important step in the systemic application of CAR-T therapy for the treatment of glioblastomas and other CNS malignancies. In addition, CAR-T therapies are known to trigger CNS side-effects, including brain swelling due to BBB disruption. In this study, we used iPSC-derived brain endothelial cell (iBEC) transwell co-culture model to assess BBB extravasation of CAR-T based immunotherapies targeting U87MG human glioblastoma (GBM) cells overexpressing EGFRvIII (U87vIII). Two types of anti-EGFRvIII targeting CAR-T cells (CAR-F263 and CAR-F269) and control Mock T cells applied on the luminal side, triggered a decrease in transendothelial electrical resistance (TEER) and an increase in BBB permeability. CAR-T cell extravasation and U87vIII cytotoxicity were assessed from the abluminal compartment using flow cytometry and IncuCyte real-time viability imaging, respectively. A significant decrease in U87vIII cell viability was observed over 48hrs, with the most robust GBM cytotoxicity response observed for CAR-F263. CAR-F269 and Mock T cells showed a similar cytotoxic profile but were collectively approximately 4-fold less efficient at killing the U87vIII cells than CAR-F263, despite similar transmigration rates. Visualization of CAR-T cell extravasation across the BBB was further confirmed using iBEC-on-CHIP models. The BBB assay was able to discriminate the cytotoxic efficacies of the two different EGFR-CARs and to provide a measure of potential alterations to BBB integrity. Collectively, we illustrate how BBB models in vitro can be a valuable tool in deciphering the mechanisms of CAR-T–induced BBB disruption, accompanying toxicity and effector function on post-barrier target cells. |
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| Publication date | 2022-08 |
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| Publisher | University of Ottawa |
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| In | |
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| Other version | |
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| Language | English |
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| Peer reviewed | Yes |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | 75e4eab4-78a0-4316-9024-8b305cd4f93e |
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| Record created | 2024-09-13 |
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| Record modified | 2024-09-13 |
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