CD28 Costimulation Augments CAR Signaling in NK Cells via the LCK/CD3ζ/ZAP70 Signaling Axis.

Acharya, Sunil; Basar, Rafet; Daher, May; Rafei, Hind; Li, Ping; Uprety, Nadima; Ensley, Emily; Shanley, Mayra; Kumar, Bijender; Banerjee, Pinaki P; Melo Garcia, Luciana; Lin, Paul; Mohanty, Vakul; Kim, Kun H; Jiang, Xianli; Pan, Yuchen; Li, Ye; Liu, Bin; Nunez Cortes, Ana K; Zhang, Chenyu; Fathi, Mohsen; Rezvan, Ali; Montalvo, Melisa J; Cha, Sophia L; Reyes-Silva, Francia; Shrestha, Rejeena; Guo, Xingliang; Kundu, Kiran; Biederstädt, Alexander; Muniz-Feliciano, Luis; Deyter, Gary M; Kaplan, Mecit; Jiang, Xin R; Liu, Enli; Jain, Antrix; Roszik, Janos; Fowlkes, Natalie W; Solis Soto, Luisa M; Raso, Maria G; Khoury, Joseph D; Lin, Pei; Vega, Francisco; Varadarajan, Navin; Chen, Ken; Marin, David; Shpall, Elizabeth J; Rezvani, Katayoun     «

Acharya, Sunil; Basar, Rafet; Daher, May; Rafei, Hind; Li, Ping; Uprety, Nadima; Ensley, Emily; Shanley, Mayra; Kumar, Bijender; Banerjee, Pinaki P; Melo Garcia, Luciana; Lin, Paul; Mohanty, Vakul; Kim, Kun H; Jiang, Xianli; Pan, Yuchen; Li, Ye; Liu, Bin; Nunez Cortes, Ana K; Zhang, Chenyu; Fathi, Mohsen; Rezvan, Ali; Montalvo, Melisa J; Cha, Sophia L; Reyes-Silva, Francia; Shrestha, Rejeena; Guo, Xingliang; Kundu, Kiran; Biederstädt, Alexander; Muniz-Feliciano, Luis; Deyter, Gary M; Kaplan, Mec...     »

Multiple factors in the design of a chimeric antigen receptor (CAR) influence CAR T-cell activity, with costimulatory signals being a key component. Yet, the impact of costimulatory domains on the downstream signaling and subsequent functionality of CAR-engineered natural killer (NK) cells remains largely unexplored. Here, we evaluated the impact of various costimulatory domains on CAR-NK cell activity, using a CD70-targeting CAR. We found that CD28, a costimulatory molecule not inherently present in mature NK cells, significantly enhanced the antitumor efficacy and long-term cytotoxicity of CAR-NK cells both in vitro and in multiple xenograft models of hematologic and solid tumors. Mechanistically, we showed that CD28 linked to CD3ζ creates a platform that recruits critical kinases, such as lymphocyte-specific protein tyrosine kinase (LCK) and zeta-chain-associated protein kinase 70 (ZAP70), initiating a signaling cascade that enhances CAR-NK cell function. Our study provides insights into how CD28 costimulation enhances CAR-NK cell function and supports its incorporation in NK-based CARs for cancer immunotherapy. Significance: We demonstrated that incorporation of the T-cell-centric costimulatory molecule CD28, which is normally absent in mature natural killer (NK) cells, into the chimeric antigen receptor (CAR) construct recruits key kinases including lymphocyte-specific protein tyrosine kinase and zeta-chain-associated protein kinase 70 and results in enhanced CAR-NK cell persistence and sustained antitumor cytotoxicity.      «

Multiple factors in the design of a chimeric antigen receptor (CAR) influence CAR T-cell activity, with costimulatory signals being a key component. Yet, the impact of costimulatory domains on the downstream signaling and subsequent functionality of CAR-engineered natural killer (NK) cells remains largely unexplored. Here, we evaluated the impact of various costimulatory domains on CAR-NK cell activity, using a CD70-targeting CAR. We found that CD28, a costimulatory molecule not inherently prese...     »