TY - JOUR
T1 - Nanoscale CAR Organization at the Immune Synapse Correlates with CAR-T Effector Functions
AU - Sajman, Julia
AU - Yakovian, Oren
AU - Unger Deshet, Naamit
AU - Almog, Shaked
AU - Horn, Galit
AU - Waks, Tova
AU - Globerson Levin, Anat
AU - Sherman, Eilon
N1 - Publisher Copyright:
© 2023 by the authors.
PY - 2023/9/12
Y1 - 2023/9/12
N2 - T cells expressing chimeric antigen receptors (CARs) are at the forefront of clinical treatment of cancers. Still, the nanoscale organization of CARs at the interface of CAR-Ts with target cells, which is essential for TCR-mediated T cell activation, remains poorly understood. Here, we studied the nanoscale organization of CARs targeting CD138 proteoglycans in such fixed and live interfaces, generated optimally for single-molecule localization microscopy. CARs showed significant self-association in nanoclusters that was enhanced in interfaces with on-target cells (SKOV-3, CAG, FaDu) relative to negative cells (OVCAR-3). CARs also segregated more efficiently from the abundant membrane phosphatase CD45 in CAR-T cells forming such interfaces. CAR clustering and segregation from CD45 correlated with the effector functions of Ca++ influx and target cell killing. Our results shed new light on the nanoscale organization of CARs on the surfaces of CAR-Ts engaging on- and off-target cells, and its potential significance for CAR-Ts’ efficacy and safety.
AB - T cells expressing chimeric antigen receptors (CARs) are at the forefront of clinical treatment of cancers. Still, the nanoscale organization of CARs at the interface of CAR-Ts with target cells, which is essential for TCR-mediated T cell activation, remains poorly understood. Here, we studied the nanoscale organization of CARs targeting CD138 proteoglycans in such fixed and live interfaces, generated optimally for single-molecule localization microscopy. CARs showed significant self-association in nanoclusters that was enhanced in interfaces with on-target cells (SKOV-3, CAG, FaDu) relative to negative cells (OVCAR-3). CARs also segregated more efficiently from the abundant membrane phosphatase CD45 in CAR-T cells forming such interfaces. CAR clustering and segregation from CD45 correlated with the effector functions of Ca++ influx and target cell killing. Our results shed new light on the nanoscale organization of CARs on the surfaces of CAR-Ts engaging on- and off-target cells, and its potential significance for CAR-Ts’ efficacy and safety.
KW - CAR-T
KW - CD138
KW - CD45
KW - adoptive cell immunotherapy
KW - cancer
KW - immune synapse
KW - nanoclusters
KW - super resolution microscopy
UR - http://www.scopus.com/inward/record.url?scp=85172791195&partnerID=8YFLogxK
U2 - 10.3390/cells12182261
DO - 10.3390/cells12182261
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C2 - 37759484
AN - SCOPUS:85172791195
SN - 2073-4409
VL - 12
JO - Cells
JF - Cells
IS - 18
M1 - 2261
ER -
