Improving dual targeting selectivity in T-cell engagers via synapse-gated and affinity-tuned trispecific antibody design.

IF 7.3 2区 医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL
mAbs Pub Date : 2025-12-01 Epub Date: 2025-10-08 DOI:10.1080/19420862.2025.2570748
Peng Zhao, John Schardt, Chi-I Chiang, Pooja Shah, Gee Sung Eun, Jan Martinek, Matthew Cyr, Yoshimi Johnson, Bismark Amofah, Xiaoying Ye, Samuel Edwards, Xiaoru Chen, Mark Penney, Wenhai Liu, Chunning Yang, Keith Rickert, Amber Lee, Sterling Payne, Hanzhi Zhang, Garrett Kelly, Chunlei Wang, Allison Gerber, Kathy Mulgrew, Rajat Varma, Jonathan Boyd, Xiuling Li, John D Bagert, Even Walseng, Yariv Mazor
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引用次数: 0

Abstract

T-cell engagers (TCEs) represent a powerful drug modality for redirecting a patient's own T cells to recognize and eradicate cancer cells. Although TCEs have been effective in treating hematological cancers, their broad application for solid tumors has been more challenging due to the absence of tumor-specific antigens. This often leads to on-target, off-tumor toxicities and a low therapeutic index (TI). Strategies for dual-antigen targeting of double-positive cancer cells over single-positive normal tissue may improve the TI of TCEs. In this study, we report the development and characterization of a conditional dual tumor-associated antigen (TAA)-targeting trispecific antibody (TriMab) TCE composed of a non-active anchoring arm (i.e. anti-TAA1), deficient in mediating an active immunological synapse, and an affinity-tuned active arm (i.e. anti-TAA2), paired with an anti-CD3 domain to drive AND-gated targeting and elimination of dual-TAA tumors while sparing single-TAA healthy cells. Using an anti-receptor tyrosine kinase-like orphan receptor 1 (ROR1) mAb as a proof-of-concept anchoring arm and an array of affinity-modulated variants of the anti-epidermal growth factor receptor (EGFR) GA201 mAb as active arms, we show in vitro conditional engagement and elimination of double-positive human NCI-H358 non-small cell lung cancer cells over single-positive, non-target NCI-H358.ROR1.KO cells by affinity-modulated TriMab TCEs. In vivo, the TriMab TCE exhibits selective targeting and eradication of ROR1/EGFR double-positive tumors in a mouse xenograft model. We further demonstrate the generality of the anchoring arm in TriMab using anti-HER2 mAbs targeting different binding epitopes and discuss the interplay of factors regulating immunological synapse formation. Lastly, we demonstrate that the TriMab modality exhibits a favorable developability profile and mAb-like pharmacokinetic properties in human neonatal Fc receptor transgenic mice. Overall, this work presents a generalizable approach to utilizing the TriMab modality by leveraging avidity effects and molecular geometry to achieve conditional AND-gated dual TAA-targeting with a significantly improved TI.

通过突触门控和亲和调节三特异性抗体设计改善t细胞接合体的双重靶向选择性。
T细胞接合剂(TCEs)代表了一种强大的药物模式,可以重新引导患者自身的T细胞识别和根除癌细胞。虽然tce在治疗血液学癌症方面是有效的,但由于缺乏肿瘤特异性抗原,其在实体肿瘤中的广泛应用更具挑战性。这通常导致靶向、非肿瘤毒性和低治疗指数(TI)。双抗原靶向双阳性癌细胞在单阳性正常组织上的策略可能改善tce的TI。在这项研究中,我们报道了一种条件双肿瘤相关抗原(TAA)靶向三特异性抗体(TriMab) TCE的开发和特性,该抗体由一个非活性锚定臂(即抗taa1)组成,缺乏介导活性免疫突触,以及一个亲和力调节的活性臂(即抗taa2),与一个抗cd3结构域配对,驱动and门控靶向和消除双TAA肿瘤,同时保留单TAA健康细胞。使用抗酪氨酸激酶样孤儿受体1 (ROR1)单抗作为概念验证的锚定臂,使用一系列亲和调节的抗表皮生长因子受体(EGFR) GA201单抗作为活性臂,我们在体外条件地结合和消除了双阳性的人NCI-H358非小细胞肺癌细胞,而不是单阳性的非靶标NCI-H358。亲和调节的TriMab tce对KO细胞的影响。在体内,TriMab TCE在小鼠异种移植模型中表现出选择性靶向和根除ROR1/EGFR双阳性肿瘤。我们利用靶向不同结合表位的抗her2单抗进一步证明了TriMab中锚定臂的普遍性,并讨论了调节免疫突触形成的因素之间的相互作用。最后,我们证明了TriMab模式在人类新生Fc受体转基因小鼠中具有良好的可发展性和单克隆抗体样药代动力学特性。总的来说,这项工作提出了一种利用TriMab模式的通用方法,通过利用贪婪效应和分子几何来实现条件和门控双taa靶向,并显着改善TI。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
mAbs
mAbs 工程技术-仪器仪表
CiteScore
10.70
自引率
11.30%
发文量
77
审稿时长
6-12 weeks
期刊介绍: mAbs is a multi-disciplinary journal dedicated to the art and science of antibody research and development. The journal has a strong scientific and medical focus, but also strives to serve a broader readership. The articles are thus of interest to scientists, clinical researchers, and physicians, as well as the wider mAb community, including our readers involved in technology transfer, legal issues, investment, strategic planning and the regulation of therapeutics.
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