Graduation Year


Date of Submission


Document Type

Open Access Senior Thesis

Degree Name

Bachelor of Arts



Second Department

W.M. Keck Science Department

Reader 1

Dr. Michael Barish

Reader 2

Dr. Melissa Coleman

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Terms of Use for work posted in Scholarship@Claremont.

Rights Information

© 2022 Arianna Livi


Chimeric Antigen Receptor T (CAR-T) cells have demonstrated anti-tumor activity against aggressive and invasive cancers such as glioblastoma (GBM); however, clinical response rates remain low in clinical trial studies. Tumor heterogeneity and tumor microenvironment conditions pose significant challenges for treatment of GBM, thus continuous optimization of CAR-T cell therapies and identification of novel, widely expressed, and highly specific GBM antigens are vital to better patient outcomes. A newly developed CAR-T cell construct incorporating chlorotoxin (CLTX) as the targeting domain exhibited broad GBM-targeting capabilities and elicited potent cytotoxic effects during preclinical studies and is currently being tested in a phase I clinical trial (NCT04214392) for recurrent GBM. Since much is still unknown about the binding mechanism and cytotoxic action of CLTX-CAR-T cells, characterization of the immunological synapse is important to be able to quantify and visualize structural, functional and signaling cascading components of CAR-T cell cytotoxicity. The purpose of this research project was to investigate the morphology of the immunological synapse between the novel CLTX-CAR-T cells and GBM tumor cells using key structural components such as receptor aggregation, actin accumulation, cytotoxic granule convergence and MTOC polarization to indicate effective cytotoxic activity and potentially predict clinical performance. Through confocal microscopy, visualization of the CLTX-CAR-T cell immunological synapse provided evidence of significant cytotoxic activity and matrix metaoproteinase-2 (MMP2) receptor localization at CLTX-CAR-T cells contact sites during tumor cell killing. These findings further our understanding of the CLTX-CAR-T cell binding complex and enhance the potential for optimizing the clinical efficacy of CLTX-CAR-T cells, thereby contributing to better treatment outcomes for patients with glioblastoma.