B7-H3 Specific CAR T-Cell Three-Dimensional Killing Assay on Synovial Sarcoma Cell Lines
Session Number
MEDH 05
Advisor(s)
Dr. Seth Pollack, Northwestern University, Feinberg School of Medicine
Discipline
Medical and Health Sciences
Start Date
17-4-2024 10:45 AM
End Date
17-4-2024 11:00 AM
Abstract
The Pollack Lab focuses on treating sarcoma tumors by performing in-vitro two-dimensional and three-dimensional killing assays with chimeric antigen receptor (CAR) T-cell therapy, a form of immunotherapy. Specifically, CAR T-cells are made by isolating T-cells from a patient’s blood and genetically modifying them to specifically target the human B7-H3 gene on the surface of a tumor. B7-H3 is a tumor surface antigen that is broadly expressed in sarcoma tumors and responsible for tumor metastasis. The purpose of this experiment was to determine the efficacy of our CAR T-cells by first determining the density of the spheroids formed by each of our four synovial sarcoma cell lines, Fuji, SYO1, HSSY2, and Yamato. Depending on the type of spheroids formed, we can determine which synovial sarcoma cell lines most closely mimic a solid sarcoma tumor in a three-dimensional cell culture and seed them in either a two-dimensional or three-dimensional killing assay accordingly. To further establish CAR T-cell efficacy, we then measure the CAR T’s ability to kill sarcoma tumor spheroids in three-dimensional in-vitro killing assays with different ratios of effector cells to target cells (E: T) and concentrations of tumor cells.
B7-H3 Specific CAR T-Cell Three-Dimensional Killing Assay on Synovial Sarcoma Cell Lines
The Pollack Lab focuses on treating sarcoma tumors by performing in-vitro two-dimensional and three-dimensional killing assays with chimeric antigen receptor (CAR) T-cell therapy, a form of immunotherapy. Specifically, CAR T-cells are made by isolating T-cells from a patient’s blood and genetically modifying them to specifically target the human B7-H3 gene on the surface of a tumor. B7-H3 is a tumor surface antigen that is broadly expressed in sarcoma tumors and responsible for tumor metastasis. The purpose of this experiment was to determine the efficacy of our CAR T-cells by first determining the density of the spheroids formed by each of our four synovial sarcoma cell lines, Fuji, SYO1, HSSY2, and Yamato. Depending on the type of spheroids formed, we can determine which synovial sarcoma cell lines most closely mimic a solid sarcoma tumor in a three-dimensional cell culture and seed them in either a two-dimensional or three-dimensional killing assay accordingly. To further establish CAR T-cell efficacy, we then measure the CAR T’s ability to kill sarcoma tumor spheroids in three-dimensional in-vitro killing assays with different ratios of effector cells to target cells (E: T) and concentrations of tumor cells.