Department of Bionano Engineering and Department of Bionanotechnology, Center for Bionano Intelligence Education and Research, Hanyang University, Ansan 15588, Republic of Korea.
Department of Microbiology and Immunology, College of Medicine, Inje University, Busan 47392, Republic of Korea.
ACS Appl Mater Interfaces. 2023 Jul 12;15(27):32201-32214. doi: 10.1021/acsami.3c03989. Epub 2023 Jun 29.
Genetically engineered fusion polypeptides have been investigated to introduce unique bio-functionality and improve some therapeutic activity for anti-angiogenesis. We report herein that stimuli-responsive, vascular endothelial growth factor receptor 1 (VEGFR1) targeting fusion polypeptides composed of a VEGFR1 (fms-like tyrosine kinase-1 (Flt1)) antagonist, an anti-Flt1 peptide, and a thermally responsive elastin-based polypeptide (EBP) were rationally designed at the genetic level, biosynthesized, and purified by inverse transition cycling to develop potential anti-angiogenic fusion polypeptides to treat neovascular diseases. A series of hydrophilic EBPs with different block lengths were fused with an anti-Flt1 peptide, forming anti-Flt1-EBPs, and the effect of EBP block length on their physicochemical properties was examined. While the anti-Flt1 peptide decreased phase-transition temperatures of anti-Flt1-EBPs, compared with EBP blocks, anti-Flt1-EBPs were soluble under physiological conditions. The anti-Flt1-EBPs dose dependently inhibited the binding of VEGFR1 against vascular endothelial growth factor (VEGF) as well as tube-like network formation of human umbilical vein endothelial cells under VEGF-triggered angiogenesis in vitro because of the specific binding between anti-Flt1-EBPs and VEGFR1. Furthermore, the anti-Flt1-EBPs suppressed laser-induced choroidal neovascularization in a wet age-related macular degeneration mouse model in vivo. Our results indicate that anti-Flt1-EBPs as VEGFR1-targeting fusion polypeptides have great potential for efficacious anti-angiogenesis to treat retinal-, corneal-, and choroidal neovascularization.
基因工程融合多肽已被研究用于引入独特的生物功能并提高抗血管生成的某些治疗活性。我们在此报告，具有刺激响应性的血管内皮生长因子受体 1（VEGFR1）靶向融合多肽由 VEGFR1（fms 样酪氨酸激酶 1（Flt1））拮抗剂、抗 Flt1 肽和热响应弹性蛋白基多肽（EBP）组成，在遗传水平上进行了合理设计、生物合成，并通过逆转变环进行了纯化，以开发潜在的抗血管生成融合多肽来治疗新生血管疾病。一系列具有不同嵌段长度的亲水性 EBPs 与抗 Flt1 肽融合，形成抗 Flt1-EBPs，并研究了 EBP 嵌段长度对其理化性质的影响。虽然抗 Flt1 肽降低了抗 Flt1-EBPs 的相变温度，与 EBP 嵌段相比，但抗 Flt1-EBPs 在生理条件下是可溶的。抗 Flt1-EBPs 呈剂量依赖性抑制 VEGFR1 与血管内皮生长因子（VEGF）的结合，以及人脐静脉内皮细胞在体外 VEGF 触发的血管生成下管状网络形成，因为抗 Flt1-EBPs 与 VEGFR1 之间存在特异性结合。此外，抗 Flt1-EBPs 在体内湿性年龄相关性黄斑变性小鼠模型中抑制激光诱导的脉络膜新生血管化。我们的结果表明，作为 VEGFR1 靶向融合多肽的抗 Flt1-EBPs 具有有效的抗血管生成潜力，可用于治疗视网膜、角膜和脉络膜新生血管化。
