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Document Type
Poster
Publication Date
Fall 10-1-2026
Abstract
Melanoma is commonly treated with the combination of targeted therapies and immune checkpoint inhibitors, but treatment resistance or relapse remains a major challenge. P-selectin glycoprotein ligand-1 (PSGL-1) is a cell-surface glycoprotein expressed on immune cells that can suppress T-cell activity within the tumor microenvironment. Silencing PSGL-1 expression may therefore help improve T-cell antitumor function. This project aimed to produce functional adeno-associated virus serotype 6 (AAV6) vectors carrying short hairpin RNA (shRNA) constructs targeting PSGL-1 and to evaluate their ability to reduce PSGL-1 expression in T cells. AAV6 vectors containing U6 or CD3 based shRNA constructs were produced in HEK293T cells using PEI transfection. Viral preparations were then harvested, quantified by qPCR, and used to transduce TK1 T cells. Transduction efficiency, cell viability, and PSGL-1 expression were evaluated by flow cytometry. Measurable viral titers were obtained for all constructs, and AAV6 efficiently transduced TK1 cells as indicated by mCherry expression. Cell viability was generally maintained following transduction. U6 based shRNA constructs produced greater reductions in PSGL-1 expression than the CD3-based constructs, while increasing viral dose did not clearly improve knockdown efficiency. These preliminary results suggesting that AAV6-mediated delivery of U6 shRNA constructs may be a promising strategy for reducing PSGL-1 expression in T cells. Because these results are based on a single experiment, additional replicates are needed to be done to confirm data reproducibility and optimize transduction conditions before evaluating functional effects on T-cell activity.
Department
Department of Pharmacology and Physiology, Drexel University College of Medicine, Philadelphia, PA
Project Mentor
Dr. Gabriele Romano
Recommended Citation
Le, Uyen '27; Vanlal, Eli; and Romano, Gabriele, "Production and Characterization of AAV6 Vectors for PSGL-1 Knockdown in T Cells" (2026). Annual Student Research Poster Session. 263.
https://scholarship.depauw.edu/srfposters/263
Funding and Acknowledgements
Thank you Eli Vanlal and Dr. Romano for the support and lab materials. Thank you Congressionally Directed Medical Research Program, American Cancer Society, WW Smith Charitable Trust, Sidney Kimmel Comprehensive Cancer Center and Dept. of Pharmacology and Physiology - DUCOM for fundings.