Sangwan Smriti, Rieder Hannah E, Moore Destaye, Khanlou Negar, Novitch Bennett, Geisberg Mark, Eisenberg David S
UCLA-DOE and Molecular Biology Institutes, Department of Biological Chemistry Los Angeles, California, USA.
Department of Pharmacology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA.
bioRxiv. 2025 May 8:2025.05.05.652290. doi: 10.1101/2025.05.05.652290.
Antibodies offer versatility as diagnostic and therapeutic tools to target specific protein epitopes. However, the transient nature of intermediate protein conformations, such as that of amyloid oligomers, poses a challenge for antibody development. We use a structure-guided approach to generate a monoclonal antibody against oligomers of Superoxide Dismutase 1 (SOD1). Mutations in SOD1 are linked to a subset of familial Amyotrophic Lateral Sclerosis (fALS), a fatal neurodegenerative disease. Based on the corkscrew-like features of non-native SOD1 oligomers previously determined, we generate an antibody specific to SOD1 oligomers. We show that the antibody, CSAb detects SOD1 oligomers, not fibrils or native SOD1, and alleviates the cytotoxic effects of SOD1 oligomers in a cell culture model of primary motor neurons. Immunohistochemical analyses of human ALS subjects show CSAb reactivity in both neuronal and non-neuronal cells. Finally, we provide evidence that CSAb reactive SOD1 oligomers are present in non-SOD1 linked fALS and sporadic ALS subjects. Together, our study provides a new probe against SOD1 oligomers and suggests that cytotoxic SOD1 oligomers are prevalent in diverse ALS genotypes.
抗体作为靶向特定蛋白质表位的诊断和治疗工具具有多功能性。然而，中间蛋白质构象的瞬态性质，如淀粉样寡聚体的构象，对抗体开发构成了挑战。我们采用结构导向方法来生成一种针对超氧化物歧化酶1（SOD1）寡聚体的单克隆抗体。SOD1中的突变与家族性肌萎缩侧索硬化症（fALS）的一个亚组相关，fALS是一种致命的神经退行性疾病。基于先前确定的非天然SOD1寡聚体的螺旋状特征，我们生成了一种对SOD1寡聚体特异的抗体。我们表明，该抗体CSAb能检测到SOD1寡聚体，而非原纤维或天然SOD1，并在原代运动神经元的细胞培养模型中减轻了SOD1寡聚体的细胞毒性作用。对人类ALS患者的免疫组织化学分析显示CSAb在神经元和非神经元细胞中均有反应性。最后，我们提供证据表明CSAb反应性SOD1寡聚体存在于非SOD1相关的fALS和散发性ALS患者中。总之，我们的研究提供了一种针对SOD1寡聚体的新探针，并表明细胞毒性SOD1寡聚体在多种ALS基因型中普遍存在。
