Michler Dennis, Seifert Oliver, Pfizenmaier Klaus, Kontermann Roland E
Institute of Cell Biology and Immunology, University of Stuttgart, Allmandring 31, 70569, Stuttgart, Germany.
Stuttgart Research Center Systems Biology (SRCSB), University of Stuttgart, Nobelstraße 15, 70569, Stuttgart, Germany.
Sci Rep. 2025 May 6;15(1):15801. doi: 10.1038/s41598-025-00476-7.
Fusion proteins combining TNF-related apoptosis inducing ligand (TRAIL) and antibody building blocks have emerged as a strategy for the targeted treatment of cancer cells. Using a single-chain derivative of homotrimeric TRAIL (scTRAIL), several targeted and non-targeted scTRAIL fusion proteins of varying geometries and valencies for TRAIL receptors and target antigens, all comprising an Fc region, were generated. These fusion proteins comprised either 1 or 2 scTRAIL units, i.e. are tri- or hexavalent for TRAIL receptors and in the targeted versions, 1 or 2 binding sites for EGFR. These fusion proteins were analyzed for cell binding and cell death induction using the EGFR-expressing colorectal cancer cell lines Colo205 and HCT116. In line with previous findings, all fusion proteins that were hexavalent for TRAIL receptors exhibited a strongly increased cell killing activity compared to the trivalent ones. Interestingly, the fusion proteins comprising one scTRAIL unit, did not benefit from targeting to EGFR. In contrast, the hexavalent scTRAIL fusion proteins further benefited from EGFR targeting, resulting in an approximately 6- to 30-fold increase in cell killing. In summary, this study shed further light on the influence of geometry and valency of TRAIL fusion proteins and confirmed IgG-scTRAIL fusion proteins as highly potent cell death inducers.
将肿瘤坏死因子相关凋亡诱导配体（TRAIL）与抗体构建模块相结合的融合蛋白已成为一种靶向治疗癌细胞的策略。利用同源三聚体TRAIL的单链衍生物（scTRAIL），制备了几种针对TRAIL受体和靶抗原具有不同几何形状和价态的靶向和非靶向scTRAIL融合蛋白，它们均包含一个Fc区域。这些融合蛋白包含1个或2个scTRAIL单元，即对TRAIL受体具有三价或六价，在靶向版本中，对表皮生长因子受体（EGFR）具有1个或2个结合位点。使用表达EGFR的结肠癌细胞系Colo205和HCT116对这些融合蛋白进行细胞结合和细胞死亡诱导分析。与先前的研究结果一致，所有对TRAIL受体具有六价的融合蛋白与三价融合蛋白相比，细胞杀伤活性显著增强。有趣的是，包含一个scTRAIL单元的融合蛋白并未从靶向EGFR中获益。相反，六价scTRAIL融合蛋白从EGFR靶向中进一步获益，导致细胞杀伤增加约6至30倍。总之，本研究进一步揭示了TRAIL融合蛋白的几何形状和价态的影响，并证实IgG-scTRAIL融合蛋白是高效的细胞死亡诱导剂。
