Khramtsov Y V, Bunin E S, Ulasov A V, Lupanova T N, Georgiev G P, Sobolev A S
Institute of Gene Biology, RAS, Moscow, Russia.
Moscow State University, Moscow, Russia.
Dokl Biochem Biophys. 2025 Feb;520(1):148-151. doi: 10.1134/S1607672924601252. Epub 2025 Jan 31.
In the previously created modular nanotransporter (MNT) capable of delivering a monobody to Keap1 into the cytosol, the endosomolytic module, translocation domain of diphtheria toxin (DTox), was replaced by the endosomolytic peptide GALA3. It was found that this substitution more than doubles the lifetime of MNT in the blood. Using confocal microscopy, it was shown that MNT with GALA3 was internalized into AML12 cells mainly due to binding to the epidermal growth factor receptor, and is also able to exit from endosomes into the cytosol. Using cellular thermal shift assay, it was shown that MNT with GALA3 and MNT with DTox are equally effective in disrupting the formation of the Nrf2 complex with Keap1, which led to similar protection of AML12 cells from the action of hydrogen peroxide. The obtained results allow not only optimizing the systemic use of MNT, but can also serve as a basis for creating agents aimed at treating diseases associated with oxidative stress.
在先前创建的能够将抗肌动蛋白单克隆抗体递送至Keap1并进入细胞质的模块化纳米转运体（MNT）中，溶酶体溶解模块（白喉毒素（DTox）的转位结构域）被溶酶体溶解肽GALA3所取代。研究发现，这种替换使MNT在血液中的寿命延长了一倍多。利用共聚焦显微镜观察发现，带有GALA3的MNT主要通过与表皮生长因子受体结合而被内化进入AML12细胞，并且还能够从内体进入细胞质。利用细胞热迁移分析表明，带有GALA3的MNT和带有DTox的MNT在破坏Nrf2与Keap1复合物形成方面同样有效，这导致AML12细胞对过氧化氢的作用具有相似的保护作用。所获得的结果不仅有助于优化MNT的全身应用，还可为开发旨在治疗与氧化应激相关疾病的药物奠定基础。
