Ability of Cytokines to Improve CAR T-Cell Efficacy

Molly working at lab bench

Students

2018
College

Faculty

Research Project Manager

Project Summary

This summer I worked in the lab of Dr. Carl June under the mentorship of John Scholler in the Smilow Center for Translational Research. My project focused on improving chimeric antigen receptor (CAR) T-cell immunotherapy for solid tumors. CAR T-cell therapy uses a patient’s own immune cells to treat their cancer by genetically modifying them to express a receptor that specifically recognizes and kills their tumor. This has been shown to be a successful method of treatment for hematological cancers, such as Acute Lymphoblastic Leukemia (ALL) and Diffuse Large B-cell Lymphoma (DLBCL). However, solid tumors are more difficult to treat since CAR T-cells have limited efficacy in their suppressive tumor microenvironment. In order to combat this, our lab hypothesized that co-expression of a CAR and particular combinations of cytokines could improve the efficacy of CAR T-cells. Cytokines are small proteins that are important in cell signaling and help to regulate cell responses. In addition to improving efficacy, they may help to enhance the functional persistence of CAR T-cells such that they survive and function better over time.

My goal this summer was to produce various CAR + cytokine combinations and determine which T-cells expressing these constructs demonstrated the best ability to expand and survive following antigen stimulation. I successfully designed and cloned different CAR + cytokine(s) DNA constructs, packaged them into virus, then infected T-cells with this virus such that they express the specified CAR and cytokines/receptors. Two different CARs were compared, each with four conditions: CAR alone, CAR-IL21, CAR-IL15R-IL15, and CAR + IL21-IL15R-IL15. Expression was analyzed via multi-parameter flow cytometry and all cell surface construct components were detected. I proceeded to monitor the cells during their initial expansion after CD3xCD28 activation before freezing them off as they rested down. Cells are thawed as needed for future use in various in vitro experiments. I am currently performing multiple rounds of antigen stimulation experiments with these therapeutic T-cells to determine which combination(s) of CAR + cytokine(s) perform best through analysis of cell survival, growth, and cellular phenotype.

Through the research I conducted this summer, I have gained a more holistic understanding of immune-therapeutic development and the process by which different DNA constructs can genetically modify T cells, resulting in varying tumor eradication capabilities. Furthermore, I have learned how to be an independent researcher, think critically, and find ways to overcome roadblocks encountered throughout my experiments. This independent research project served as a culmination of the skills and knowledge I have gained during my time so far in the June Lab and I am eager to determine what direction this project will take next based on our findings.

Finally, this project has contributed to my educational experience by confirming my passion for research and decision to pursue a PhD in immunology. It has been a true privilege to be a part of cutting edge research and to be able to work with such knowledgeable and professional mentors. I’d like to thank CURF and the June Lab for giving me this opportunity.