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Paloma, a junior at Harvard University studying Integrative Biology altered the architecture of CAR T Cells to produce more effective results in cancer patients

Cancer immunotherapy harnesses our bodies inherent ability to combat infections for the purpose of eliminating cancer. Chimeric Antigen Receptor (CAR) T Cell therapy involves genetically engineering a receptor into a patient’s T cells that detects antigens, such as CD19, on the surface of tumor cells and eliminates them.My time in the BSRP program was incredibly fulfilling and rewarding. Conducting cutting-edge research with the support of my mentors and instructors gave me the clarity to continue pursuing research as a career path. Engaging with my peers who have similar passions and interests as myself was the highlight of my summer research experience. I built a network of incredible colleagues and made friends along the way. Within the receptor design is a costimulatory domain that provides the necessary signaling to promote cytotoxicity and proliferation.These domains often utilize NFkB signaling to activate the T cell with some belonging to the Tumor Necrosis Factor Receptor (TNFR) family. Although the current generation of CARs have been successful in some patients, other patients’ T cells lose function prematurely. This issue has prompted us to assess NFkB activation and cytotoxicity in 8 other costimulatory domains from the TNFR family to potentially identify better CARs architectures. First, we cloned a plasmid with a CD19 CAR backbone and a costimulatory domain insert of interest. Then, we lentivirally transduced the CARs into NFkB Reporter Jurkat cells containing BFP and co cultured them with CD19+ Nalm6 cells. Using flow cytometry, we found that LTBR, TNFR2, and CD30 domains exhibited higher NFkB activation when exposed to different concentrations of CD19+ cells. To test for cytotoxicity, we conducted a similar experiment using primary human T cells and introduced K562 CD19- cells into the coculture. Using flow cytometry to quantify the remaining CD19+ cells, we found that BAFFR and CD27 CARs were more cytotoxic than the original CARs. This approach suggests that alternative domains could produce higher functioning CARs.

Project: Enhancing CAR T Cell Therapy Through Testing Alternative Costimulatory Domains

Mentors: Nelson Knudsen and Alex Rojas, Tumor Immunotherapy Discovery Engine (TIDE) program