Burke Alan, Borot Florence, Du Xing, Churchill Michael, Ding Jian, Grass Albert Mridul, DeSouza Philip, Ali Abdullah Mahmood, Mukherjee Siddhartha
Department of Medicine, Columbia University Irving Medical Center, Columbia University, New York, NY, 10032, USA.
Department of Molecular Pharmacology and Therapeutics, Columbia University Irving Medical Center, Columbia University, New York, NY, 10032, USA.
Oncogene. 2023 Jan;42(1):26-34. doi: 10.1038/s41388-022-02532-2. Epub 2022 Nov 10.
Many therapeutic bispecific T-cell engagers (BiTEs) are in clinical trials. A modular and efficient process to create BiTEs would accelerate their development and clinical applicability. In this study, we present the design, production, and functional activity of a novel bispecific format utilizing synthetic orthogonal heterodimers to form a multichain modular design. Further addition of an immunoglobulin hinge region allowed a stable covalent linkage between the heterodimers. As proof-of-concept, we utilized CD33 and CD3 binding scFvs to engage leukemia cells and T-cells respectively. We provide evidence that this novel bispecific T-cell engager (termed IgGlue-BiTE) could bind both CD3+ and CD33+ cells and facilitates robust T-cell mediated cytotoxicity on AML cells in vitro. In a mouse model of minimal residual disease, we showed that the novel IgGlue-BiTE greatly extended survival, and mice of this treatment group were free of leukemia in the bone marrow. These findings suggest that the IgGlue-BiTE allows for robust simultaneous engagement with both antigens of interest in a manner conducive to T cell cytotoxicity against AML. These results suggest a compelling modular system for bispecific antibodies, as the CD3- and CD33-binding domains can be readily swapped with domains binding to other cancer- or immune cell-specific antigens.
许多治疗性双特异性T细胞衔接器（BiTEs）正在进行临床试验。创建BiTEs的模块化高效流程将加速其开发和临床应用。在本研究中，我们展示了一种新型双特异性形式的设计、生产和功能活性，该形式利用合成正交异二聚体形成多链模块化设计。进一步添加免疫球蛋白铰链区可使异二聚体之间形成稳定的共价连接。作为概念验证，我们利用结合CD33和CD3的单链抗体片段（scFvs）分别与白血病细胞和T细胞结合。我们提供的证据表明，这种新型双特异性T细胞衔接器（称为IgGlue - BiTE）能够结合CD3 +和CD33 +细胞，并在体外促进T细胞介导的对急性髓系白血病（AML）细胞的强大细胞毒性。在微小残留病小鼠模型中，我们表明新型IgGlue - BiTE大大延长了生存期，并且该治疗组的小鼠骨髓中没有白血病。这些发现表明，IgGlue - BiTE能够以有利于T细胞对AML进行细胞毒性作用的方式，与两种目标抗原同时进行强大的结合。这些结果表明了一种引人注目的双特异性抗体模块化系统，因为结合CD3和CD33的结构域可以很容易地与结合其他癌症或免疫细胞特异性抗原的结构域进行交换。
