Develop pan-specific antibody against mutant glycyl-tRNA synthetase for treating CMT2D

开发抗突变甘氨酰-tRNA合成酶的泛特异性抗体来治疗 CMT2D

基本信息

批准号：
10544795
负责人：
Xiang-Lei Yang
金额：
$ 44.38万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-01-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10544795
关键词：
Affect Affinity Amino Acyl-tRNA Synthetases Animals Antibodies Antibody Therapy Antigens Autoantibodies Awareness Binding Biological Biological Assay Biological Markers Blood Body Weight Charcot-Marie-Tooth Disease Clinical Clinical Chemistry Collection Complete Blood Count Creatine Kinase Data Development Disease Disease model Distal Drug Kinetics Electrophysiology (science)Engineering Exhibits Extracellular Space GARS gene Genetic Heterogeneity Goals Hand Strength Histology Histopathology Hybridomas Immunization Immunoglobulin G Ligase Light Maintenance Measures Medical Membrane Proteins Mendelian disorder Molecular Conformation Monitor Monoclonal Antibodies Morphology Motor Motor Neurons Mus Muscle Muscular Atrophy Mutation Myositis Nerve Neuromuscular Junction Neuropilin-1 Organ Weight Pathologic Pathology Patients Peptides Performance Peripheral Peripheral Nervous System Pharmacodynamics Phase Physiology Plasma Proteins Scheme Sensory Signal Pathway Site Source Specificity Syndrome Therapeutic Therapeutic Monoclonal Antibodies Toxic effect Work axonal degeneration biophysical properties design dimer disease phenotype disorder subtype dominant genetic mutation efficacy study efficacy testing gene therapy human model in vivo insight motor neuron degeneration mouse model mutant neurofilament novel novel strategies receptor receptor binding remediation response targeted treatment treatment duration treatment strategy

项目摘要

Abstract Charcot-Marie-Tooth (CMT) disease type 2D is caused by dominant mutations in GARS, encoding glycyl-tRNA synthetase (GlyRS). CMT is a clinically and genetically heterogeneous collection of disorders in which peripheral motor and sensory axons degenerate. The diseases specifically affect the peripheral nervous system and are characterized by progressive motor neuron degeneration, muscle atrophy, and sensory loss. None of the CMT disease subtypes have a targeted treatment, thus CMT remains an unmet medical need. The genetic heterogeneity makes it unlikely that a single therapy will be effective for all forms of CMT. Although gene therapies have emerged strongly for monogenic diseases such as CMT, the large number of different mutations involved, and the small number of patients affected by each mutation render classic gene therapy onerous for CMT2D. Identifying a causal treatment strategy applicable to different mutations in GARS would therefore be the most attractive therapeutic approach for CMT2D. Also, being able to directly engage with the GlyRS protein itself is key to CMT2D remediation. Our previous work has established that a major source of the toxicity of the CMT-causing mutant GlyRS is originated from the extracellular space, where mutant proteins aberrantly interact with Neuropilin 1 (Nrp1) receptor and antagonize a signaling pathway important for motor neuron maintenance. We designed a novel strategy based on our structural insight of the GlyRS mutants. We found that different CMT2D mutations caused a shared conformational change in GlyRS that exposes new protein surfaces at the dimer interface to solution. By using two different peptides from the dimer interface of GlyRS as antigens for immunization, we have successfully obtained two monoclonal antibodies (mAbs) (one for each peptide) from mouse hybridomas that can block the pathological Nrp1 interaction and exhibit promising pan-mutant selectivity. These two mAb candidates will be characterized for their biophysical properties, and used for conducting pharmacodynamic, pharmacokinetic, and in vivo efficacy studies in a mouse model of the human disease to evaluate whether they have sufficient biological activity to warrant further development to treat CMT2D.

抽象的 2D 型腓骨肌萎缩症 (CMT) 是由编码甘氨酰 tRNA 的 GARS 显性突变引起的合成酶（GlyRS）。 CMT 是一组具有临床和遗传异质性的疾病，其中外周血运动和感觉轴突退化。这些疾病特别影响周围神经系统，并且是其特征是进行性运动神经元变性、肌肉萎缩和感觉丧失。没有 CMT 疾病亚型有针对性的治疗，因此 CMT 仍然是一个未满足的医疗需求。遗传性异质性使得单一疗法不太可能对所有形式的 CMT 都有效。虽然基因针对 CMT 等单基因疾病的治疗方法已经出现，大量不同的突变涉及，并且受每个突变影响的患者数量很少，这使得传统的基因治疗变得繁重 CMT2D。因此，确定适用于 GARS 不同突变的因果治疗策略将是 CMT2D 最具吸引力的治疗方法。此外，能够直接与 GlyRS 蛋白结合其本身是 CMT2D 修复的关键。我们之前的工作已经确定，该物质的毒性的一个主要来源是引起 CMT 的突变 GlyRS 源自细胞外空间，突变蛋白在细胞外空间异常相互作用与 Neuropilin 1 (Nrp1) 受体结合并拮抗对运动神经元维持重要的信号通路。我们根据对 GlyRS 突变体的结构了解设计了一种新颖的策略。我们发现不同 CMT2D 突变引起 GlyRS 的共同构象变化，从而在二聚体与溶液的界面。通过使用来自 GlyRS 二聚体界面的两种不同的肽作为抗原免疫后，我们成功获得了两种单克隆抗体（mAb）（每种肽各一种）小鼠杂交瘤可以阻断病理性 Nrp1 相互作用并表现出有希望的泛突变选择性。这两种候选单克隆抗体将对其生物物理特性进行表征，并用于进行在人类疾病小鼠模型中进行药效学、药代动力学和体内功效研究评估它们是否具有足够的生物活性以保证进一步开发治疗 CMT2D。