A fusogenic dengue virus-derived peptide enhances antitumor efficacy of an antibody-ribonuclease fusion protein targeting the EGF receptor
A fusogenic dengue virus-derived peptide enhances antitumor efficacy of an antibody-ribonuclease fusion protein targeting the EGF receptor
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DOI：
10.1093/protein/gzu040
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发表时间：
2014-10-01
影响因子：
2.4
通讯作者：
Krauss, Juergen
中科院分区：
文献类型：
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作者：
Kiesgen, Stefan;Liebers, Nora;Krauss, Juergen
由于表皮生长因子受体（EGFR）在多种实体瘤中频繁过表达，其是上皮癌中治疗干预的公认靶点。为了靶向头颈癌中的EGFR，我们已经产生了核糖核酸酶（RNase）融合蛋白，其包含人源化抗EGFR抗体单链Fv片段（scFv）和Ranpirnase，一种来自蛙的RNase。豹蛙酶通过柔性甘氨酸-丝氨酸接头（G（4）S）与scFv的N-末端融合（3）导致融合蛋白的细胞毒性非常差。由于已经报道内体积累和溶酶体降解会降低核糖核酸酶或基于毒素的免疫剂的抗肿瘤功效，我们探索了来自登革病毒的融合肽，该融合肽已经报道参与病毒的内体逃逸。该肽作为豹蛙酶和scFv部分之间的接头引入。修饰的免疫核糖核酸酶对EGFR表达的头颈部细胞系表现出异常高的细胞毒性，而不影响特异性。这些结果表明，内体截留需要考虑豹蛙酶为基础的免疫剂，并可能克服使用定制的转导结构域从病毒蛋白。
Due to its frequent overexpression in a variety of solid tumors the epidermal growth factor receptor (EGFR) is a well-established target for therapeutic interventions in epithelial cancers. In order to target EGFR in head and neck cancer, we have generated a ribonuclease (RNase) fusion protein comprising a humanized anti-EGFR antibody single-chain Fv fragment (scFv) and Ranpirnase, an RNase from Rana pipiens. Fusion of Ranpirnase to the N-terminus of the scFv via a flexible glycine-serine linker (G(4)S)(3) resulted in very poor cytotoxicity of the fusion protein. As endosomal accumulation and lysosomal degradation have been reported to diminish the antitumor efficacy of ribonuclease or toxin-based immunoagents, we explored a fusion peptide from dengue virus that has been reported to be involved in the endosomal escape of the virus. This peptide was introduced as a linker between Ranpirnase and the scFv moiety. The modified immunoRNase exhibited exceptionally high cytotoxicity toward EGFR-expressing head and neck cell lines without affecting specificity. These results indicate that endosomal entrapment needs to be considered for Ranpirnase-based immunoagents and might be overcome by the use of tailored transduction domains from viral proteins.