State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, People's Republic of China.
Department of Applied Biology, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, People's Republic of China.
J Membr Biol. 2020 Apr;253(2):139-152. doi: 10.1007/s00232-020-00109-2. Epub 2020 Jan 31.
Endosomal escape is a rate-limiting step in the cytosolic delivery of therapeutic drugs. Overcoming this barrier is crucial to achieve an effective biological based therapy. In this work, we evaluated the ability of a synthetic biomimetic peptide derived from the GALA to facilitate endosomal escape of protein drugs. Our results showed that the cytoplasmic distribution of GALA fusion proteins changed according to the hydrophobicity of GALA. One of the synthetic peptides, GALA3, significantly enhanced the endosomal escape efficiency of protein drugs. The cytosolic delivery capacity of GALA3 was significantly higher than that of several previously reported endosomal escape peptides, including hemagglutinin 2 (HA2). Moreover, when GALA3 was fused to BLF1-HBP, a ribosome-inactivating protein with cell-penetrating peptide HBP, the cytotoxicity of the fusion protein was significantly increased in various cell lines, including H460, HeLa, A549, and SMCC-7721. The growth inhibition effect of GALA3-BLF1-HBP was at least 20 times greater than that of BLF1-HBP alone in different tumor cell lines. GALA3 effectively promoted the endosomal escape of BLF1-HBP in a pH-dependent manner and greatly enhanced the apoptotic activity of BLF1-HBP. Taken together, our data show that by adjusting the hydrophobicity of GALA we obtained a more effective endosomal escape peptide. Therefore, GALA3-fusions can improve the efficiency of therapeutic protein drugs.
内体逃逸是治疗药物胞质递送的限速步骤。克服这一障碍对于实现有效的基于生物学的治疗至关重要。在这项工作中，我们评估了源自 GALA 的合成仿生肽促进蛋白药物内体逃逸的能力。我们的结果表明，GALA 融合蛋白的细胞质分布根据 GALA 的疏水性而变化。其中一种合成肽 GALA3 显著提高了蛋白药物的内体逃逸效率。GALA3 的胞质递送能力明显高于先前报道的几种内体逃逸肽，包括血凝素 2 (HA2)。此外，当 GALA3 与具有穿透肽 HBP 的核糖体失活蛋白 BLF1-HBP 融合时，融合蛋白在包括 H460、HeLa、A549 和 SMCC-7721 在内的各种细胞系中的细胞毒性显著增加。在不同的肿瘤细胞系中，GALA3-BLF1-HBP 的生长抑制作用至少比单独的 BLF1-HBP 高 20 倍。GALA3 以 pH 依赖性方式有效促进 BLF1-HBP 的内体逃逸，并大大增强了 BLF1-HBP 的凋亡活性。总之，我们的数据表明，通过调整 GALA 的疏水性，我们获得了一种更有效的内体逃逸肽。因此，GALA3 融合可以提高治疗性蛋白药物的效率。
