Institute of Cell Biology and Immunology, University of Stuttgart, Allmandring 31, 70569, Stuttgart, Germany.
Department of Dermatology, Experimental Dermatology, TU-Dresden, 01307 Dresden, Germany.
Sci Rep. 2018 May 17;8(1):7808. doi: 10.1038/s41598-018-24450-8.
Single-chain formats of TNF-related apoptosis inducing ligand (scTRAIL) can serve as effector components of tumour-associated antigen-targeted as well as non-targeted fusion proteins, being characterized by high tumour cell-specific induction of apoptosis through death receptor activation. We studied the suitability of immunoglobulin G as a scaffold for oligovalent and bispecific TRAIL fusion proteins. Thus, we developed novel targeted hexa- and dodecavalent IgG-scTRAIL molecules by fusing scTRAIL to the C-terminus of either light (LC-scTRAIL) or heavy immunoglobulin chain (HC-scTRAIL), or to both ends (LC/HC-scTRAIL) of the anti-EGFR IgG antibody hu225. The binding specificity to EGFR and death receptors was retained in all IgG-scTRAIL formats and translated into high antigen-specific bioactivity on EGFR-positive Colo205, HCT116 and WM1366 tumour cell lines, with or without sensitization to apoptosis by bortezomib. In vivo, therapeutic potential was assessed for one of the targeted variants, HC-scTRAIL, compared to the non-targeted Fc-scTRAIL. Both molecules showed a significant reduction of tumour volume and synergism with a Smac mimetic in a Colo205 xenograft tumour model. The IgG-scTRAIL format allows directing a defined, highly bioactive form of TRAIL to a wide variety of tumour antigens, enabling customized solutions for a patient-specific targeted cancer therapy with a reduced risk of side effects.
单链格式的肿瘤坏死因子相关凋亡诱导配体（scTRAIL）可以作为肿瘤相关抗原靶向和非靶向融合蛋白的效应组件，其特征是通过死亡受体激活高度特异性诱导肿瘤细胞凋亡。我们研究了免疫球蛋白 G 作为多价和双特异性 TRAIL 融合蛋白的支架的适用性。因此，我们通过将 scTRAIL 融合到轻链（LC-scTRAIL）或重链（HC-scTRAIL）的 C 末端，或融合到抗 EGFR IgG 抗体 hu225 的两端（LC/HC-scTRAIL），开发了新型靶向六价和十二价 IgG-scTRAIL 分子。所有 IgG-scTRAIL 格式均保留了对 EGFR 和死亡受体的结合特异性，并转化为对 EGFR 阳性 Colo205、HCT116 和 WM1366 肿瘤细胞系的高抗原特异性生物活性，无论是否用硼替佐米敏化诱导凋亡。在体内，与非靶向 Fc-scTRAIL 相比，评估了一种靶向变体 HC-scTRAIL 的治疗潜力。这两种分子在 Colo205 异种移植肿瘤模型中均显著减少了肿瘤体积，并与 Smac 模拟物具有协同作用。IgG-scTRAIL 格式允许将定义明确、高度生物活性的 TRAIL 导向多种肿瘤抗原，为具有降低副作用风险的个体化靶向癌症治疗提供定制解决方案。
