Moore Allyson E, Nault Hayley, Cummings Derek, Bojovic Bonnie, Serniuck Nick, Baker Christopher L, Aarts Craig, Venugopal Chitra, Singh Sheila K, Hammill Joanne A, Bramson Jonathan L
Centre for Discovery in Cancer Research, McMaster University, Hamilton, ON L8S 4K1, Canada.
McMaster Immunology Research Centre, McMaster University, Hamilton, ON L8S 4K1, Canada.
iScience. 2025 Mar 7;28(4):112142. doi: 10.1016/j.isci.2025.112142. eCollection 2025 Apr 18.
We describe a series of DAP12-associated receptors that can be used to achieve multi-targeting within a small genetic payload. Empirical evaluation of scaffold/binder combinations is required to define the optimal synthetic receptor configuration. When two DAP12-associated synthetic receptors were expressed in T cells from a single vector, the surface levels of individual receptors was reduced when compared to T cells engineered with vectors that express a single receptor. The reduction in receptor expression had a pronounced effect on early, but not late, signaling events and primarily affected cytokine production. The functional deficiency was overcome by increasing synthetic receptor levels demonstrating that there is no fundamental issue related to co-expression of multiple DAP12-associated synthetic receptors in a single T cell. Our data show that T cells can be engineered with multiple recombinant DAP12-based receptors to yield multi-target specific T cells, however, thoughtful design and optimization are necessary.
我们描述了一系列与DAP12相关的受体，可用于在小的遗传载荷内实现多靶点作用。需要对支架/结合物组合进行实证评估，以确定最佳的合成受体构型。当两个与DAP12相关的合成受体从单个载体在T细胞中表达时，与用表达单个受体的载体工程改造的T细胞相比，单个受体的表面水平降低。受体表达的降低对早期而非晚期信号事件有显著影响，并且主要影响细胞因子的产生。通过增加合成受体水平克服了功能缺陷，这表明在单个T细胞中共表达多个与DAP12相关的合成受体不存在根本问题。我们的数据表明，可以用多个基于重组DAP12的受体对T细胞进行工程改造，以产生多靶点特异性T细胞，然而，精心的设计和优化是必要的。
