Balancing activation and co-stimulation of CAR tunes signaling dynamics and enhances therapeutic potency
Balancing activation and co-stimulation of CAR tunes signaling dynamics and enhances therapeutic potency
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DOI：
10.1016/j.ymthe.2022.08.018
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发表时间：
2023-01-04
影响因子：
12.4
通讯作者：
Sun, Jie
中科院分区：
文献类型：
--
作者：
Duan, Yanting;Chen, Jiangqing;Sun, Jie
具有CD 28和CD 3 z信号传导结构域的靶向CD 19的嵌合抗原受体（汽车）已被美国FDA批准用于治疗B细胞恶性肿瘤。CD 3 z中基于免疫受体酪氨酸的激活基序（ITAM）的突变产生了含有1XX CAR的单ITAM，其在白血病小鼠模型中显示出上级抗肿瘤活性。在这里，我们研究了1XX设计是否可以增强对实体瘤的治疗效力。我们构建了CD 19和AXL特异性1XX汽车并将其体外和体内功能与野生型（WT）对应物进行了比较。1XX汽车在胰腺和黑素瘤小鼠模型中均显示出更好的抗肿瘤功效。详细的分析显示，1XX CAR-T细胞在体内持续时间更长，并且具有更高百分比的中央记忆细胞。使用基于荧光共振能量转移（FRET）的生物传感器，我们发现1XX中ITAM数量的减少导致类似的70 kDa zeta链相关蛋白（ZAP 70）激活，而1XX诱导更高的Ca 2+升高和更快的细胞外信号调节激酶（Erk）激活比WT CAR。因此，我们的结果证实了1XX在不同实体瘤模型中对两种靶点的优越性，并阐明了CAR信号转导的潜在分子机制，为1XX汽车在实体瘤中的临床应用铺平了道路。
CD19-targeting chimeric antigen receptors (CARs) with CD28 and CD3zsignaling domains have been approved by the US FDA for treating B cell malignancies. Mutation of immunoreceptor tyrosine-based activation motifs (ITAMs) in CD3z generated a single-ITAM containing 1XX CAR, which displayed superior antitumor activity in a leukemia mouse model. Here, we investigated whether the 1XX design could enhance therapeutic potency against solid tumors. We constructed both CD19- and AXL-specific 1XX CARs and compared their in vitro and in vivo functions with their wild-type (WT) counterparts. 1XX CARs showed better antitumor efficacy in both pancreatic and melanoma mouse models. Detailed analysis revealed that 1XX CAR-T cells persisted longer in vivo and had a higher percentage of central memory cells. With fluorescence resonance energy transfer (FRET)-based biosensors, we found that decreased ITAM numbers in 1XX resulted in similar 70-kDa zeta chain-associated protein (ZAP70) activation, while 1XX induced higher Ca2+ elevation and faster extracellular signal-regulated kinase (Erk) activation than WT CAR. Thus, our results confirmed the superiority of 1XX against two targets in different solid tumor models and shed light on the underlying molecular mechanism of CAR signaling, paving the way for the clinical applications of 1XX CARs against solid tumors.