使用一种可穿透血脑屏障的抗体在中枢神经系统中中和Nogo-A。

Nogo-A neutralization in the central nervous system with a blood-brain barrier-penetrating antibody.

作者信息

Joly Sandrine, Augusto Gilles, Mdzomba Baya, Meli Ivo, Vogel Monique, Chan Andrew, Pernet Vincent

机构信息

Department of Neurology, Inselspital, Bern University Hospital, University of Bern, Switzerland; Center for experimental neurology (ZEN), Bern University Hospital, University of Bern, Switzerland; Centre de recherche du CHU de Québec-Université Laval and Department of Molecular Medicine, Faculté de médecine, Université Laval, Québec, Québec, Canada; Department of Ophthalmology, Bern University Hospital, University of Bern, Switzerland; Department of Biomedical Research, University of Bern, Bern, Switzerland.

Department of Biomedical Research, University of Bern, Bern, Switzerland; Department of Immunology, Inselspital, Bern University Hospital, University of Bern, Switzerland; Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom.

出版信息

J Control Release. 2024 Feb;366:52-64. doi: 10.1016/j.jconrel.2023.12.041. Epub 2023 Dec 29.

DOI:10.1016/j.jconrel.2023.12.041

PMID:38154541

Abstract

The poor penetration of monoclonal antibodies (mAb) across the blood-brain barrier (BBB) impedes the development of regenerative therapies for neurological diseases. For example, Nogo-A is a myelin-associated protein highly expressed in the central nervous system (CNS) whose inhibitory effects on neuronal plasticity can be neutralized with direct administration of 11C7 mAb in CNS tissues/fluids, but not with peripheral administrations such as intravenous injections. Therefore, in the present study, we engineered a CNS-penetrating antibody against Nogo-A by combining 11C7 mAb and the single-chain variable fragment (scFv) of 8D3, a rat antibody binding transferrin receptor 1 (TfR) and mediating BBB transcytosis (11C7-scFv8D3). The binding of 11C7-scFv8D3 to Nogo-A and to TfR/CD71 was validated by capture ELISA and Biolayer Interferometry. After intravenous injection in mice, capture ELISA measurements revealed fast plasma clearance of 11C7-scFv8D3 concomitantly with brain and spinal cord accumulation at levels up to 19 fold as high as those of original 11C7 mAb. 11C7-scFv8D3 detection in the parenchyma indicated effective blood-to-CNS transfer. A single dose of 11C7-scFv8D3 induced stronger activation of the growth-promoting AkT/mTOR/S6 signaling pathway than 11C7 mAb or control antibody. Taken together, our results show that BBB-crossing 11C7-scFv8D3 engages Nogo-A in the mouse CNS and stimulates neuronal growth mechanisms.

摘要

单克隆抗体（mAb）难以穿透血脑屏障（BBB），这阻碍了神经疾病再生疗法的发展。例如，Nogo-A是一种在中枢神经系统（CNS）中高度表达的髓鞘相关蛋白，直接向CNS组织/液中注射11C7 mAb可中和其对神经元可塑性的抑制作用，但通过静脉注射等外周给药方式则无法实现。因此，在本研究中，我们通过将11C7 mAb与8D3的单链可变片段（scFv）相结合，构建了一种可穿透CNS的抗Nogo-A抗体，8D3是一种大鼠抗体，可结合转铁蛋白受体1（TfR）并介导BBB转胞吞作用（11C7-scFv8D3）。通过捕获ELISA和生物膜干涉技术验证了11C7-scFv8D3与Nogo-A以及TfR/CD71的结合。给小鼠静脉注射后，捕获ELISA测量结果显示，11C7-scFv8D3在血浆中快速清除，同时在脑和脊髓中的蓄积水平比原始11C7 mAb高19倍。在实质组织中检测到11C7-scFv8D3表明其实现了有效的血脑转移。单剂量的11C7-scFv8D3比11C7 mAb或对照抗体更能强烈激活促进生长的AkT/mTOR/S6信号通路。综上所述，我们的结果表明，能够穿越BBB的11C7-scFv8D3可作用于小鼠CNS中的Nogo-A并刺激神经元生长机制。