Technische Universität Braunschweig, Institut für Biochemie, Biotechnologie und Bioinformatik, Department Medizinische Biotechnologie, Braunschweig 38106, Germany.
Faculty of Medicine, Carl-Ludwig-Institute of Physiology, Leipzig University, Leipzig 04103, Germany.
Brain. 2023 May 2;146(5):1812-1820. doi: 10.1093/brain/awac497.
N-methyl-D-aspartate receptor (NMDAR) encephalitis is the most common subtype of autoimmune encephalitis characterized by a complex neuropsychiatric syndrome usually including memory impairment. Patients develop an intrathecal immune response against NMDARs with antibodies that presumably bind to the amino-terminal domain of the GluN1 subunit. The therapeutic response to immunotherapy is often delayed. Therefore, new therapeutic approaches for fast neutralization of NMDAR antibodies are needed. Here, we developed fusion constructs consisting of the Fc part of immunoglobulin G and the amino-terminal domains of either GluN1 or combinations of GluN1 with GluN2A or GluN2B. Surprisingly, both GluN1 and GluN2 subunits were required to generate high-affinity epitopes. The construct with both subunits efficiently prevented NMDAR binding of patient-derived monoclonal antibodies and of patient CSF containing high-titre NMDAR antibodies. Furthermore, it inhibited the internalization of NMDARs in rodent dissociated neurons and human induced pluripotent stem cell-derived neurons. Finally, the construct stabilized NMDAR currents recorded in rodent neurons and rescued memory defects in passive-transfer mouse models using intrahippocampal injections. Our results demonstrate that both GluN1 and GluN2B subunits contribute to the main immunogenic region of the NMDAR and provide a promising strategy for fast and specific treatment of NMDAR encephalitis, which could complement immunotherapy.
N-甲基-D-天冬氨酸受体(NMDAR)脑炎是最常见的自身免疫性脑炎亚型,其特征是复杂的神经精神综合征,通常包括记忆障碍。患者会对 NMDAR 产生鞘内免疫反应,产生的抗体可能与 GluN1 亚基的氨基末端结构域结合。免疫治疗的治疗反应通常是延迟的。因此,需要新的治疗方法来快速中和 NMDAR 抗体。在这里,我们开发了由免疫球蛋白 G 的 Fc 部分和 GluN1 的氨基末端结构域或 GluN1 与 GluN2A 或 GluN2B 的组合组成的融合构建体。令人惊讶的是,产生高亲和力表位都需要 GluN1 和 GluN2 亚基。该构建体与两个亚基都有效地阻止了患者来源的单克隆抗体和含有高滴度 NMDAR 抗体的患者 CSF 与 NMDAR 的结合。此外,它还抑制了啮齿动物分离神经元和人诱导多能干细胞衍生神经元中 NMDAR 的内化。最后,该构建体稳定了在啮齿动物神经元中记录的 NMDAR 电流,并通过海马内注射挽救了被动转移小鼠模型中的记忆缺陷。我们的结果表明,GluN1 和 GluN2B 亚基都有助于 NMDAR 的主要免疫原性区域,并为快速和特异性治疗 NMDAR 脑炎提供了有希望的策略,该策略可以补充免疫治疗。