Zhao Liyuan, Meng Fanling, Li Yingjie, Liu Sujuan, Xu Mengmeng, Chu Fan, Li Chuanzhou, Yang Xiangliang, Luo Liang
National Engineering Research Center for Nanomedicine, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, P. R. China.
Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, P. R. China.
Adv Mater. 2023 Apr;35(17):e2210879. doi: 10.1002/adma.202210879. Epub 2023 Mar 18.
Efficient therapeutic strategies that concurrently target both Aβ aggregation and oxidative stress in the Alzheimer's disease (AD) microenvironment emerge as a cutting-edge tool to combat the intricate pathogenesis of AD. Here, a multivalent nanobody conjugate with rigid, reactive oxygen species (ROS) scavenging scaffold is developed to achieve simultaneous Aβ amyloidogenesis mitigation, ROS elimination, and Aβ plaque clearance. Grafting Aβ segment (33-GLMVGGVVIA-42) into the third complementary-determining region of a parent nanobody generates an engineered nanobody NB that can recognize Aβ and inhibit its aggregation through homotypic interactions. NB is further genetically modified with a fragment of human interleukin-1β (163-VQGEESNDK-171), so that the obtained fusion nanobody NBIL can also facilitate the Aβ clearance by microglia. Linking NBIL covalently onto a rigid, ROS scavenging scaffold poly(deca-4,6-diynedioic acid) (PDDA) creates the multivalent nanobody conjugate PNBIL, which not only boosts the binding affinity between NBIL and Aβ aggregates for nearly 100 times but also possesses a long-term capability of oxidative stress alleviation, inflammation reduction, and neuron protection. PNBIL has significantly attenuated symptoms on two AD mouse models through amyloidogenesis inhibition and AD microenvironment modulation, validating that the multivalent nanobody conjugate design based on combinatory nanobody and molecular engineering is a promising approach of multi-target therapeutic strategies.
在阿尔茨海默病（AD）微环境中同时针对Aβ聚集和氧化应激的高效治疗策略，成为对抗AD复杂发病机制的前沿工具。在此，开发了一种具有刚性活性氧（ROS）清除支架的多价纳米抗体缀合物，以实现同时减轻Aβ淀粉样蛋白生成、消除ROS和清除Aβ斑块。将Aβ片段（33 - GLMVGGVVIA - 42）嫁接到亲本纳米抗体的第三个互补决定区，产生一种工程化纳米抗体NB，它可以识别Aβ并通过同型相互作用抑制其聚集。NB进一步用人类白细胞介素 - 1β（163 - VQGEESNDK - 171）的片段进行基因修饰，从而使获得的融合纳米抗体NBIL也能促进小胶质细胞对Aβ的清除。将NBIL共价连接到刚性的ROS清除支架聚（癸 - 4,6 - 二炔二酸）（PDDA）上，产生多价纳米抗体缀合物PNBIL，它不仅使NBIL与Aβ聚集体之间的结合亲和力提高近100倍，而且具有长期减轻氧化应激、减轻炎症和保护神经元的能力。PNBIL通过抑制淀粉样蛋白生成和调节AD微环境，在两种AD小鼠模型上显著减轻了症状，验证了基于组合纳米抗体和分子工程的多价纳米抗体缀合物设计是多靶点治疗策略的一种有前景的方法。
