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

• 批准号: 10541284
• 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
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10541284
• 关键词: Affect; Affinity; Amino Acyl-tRNA Synthetases; Animals; Antibodies; Antibody Therapy; Antigens; Autoantibodies; Awareness; Axon; 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; base; biophysical properties; design; dimer; disease phenotype; disorder subtype; efficacy study; efficacy testing; gene therapy; human disease; in vivo; insight; motor neuron degeneration; mouse model; mutant; neurofilament; novel; novel strategies; receptor; receptor binding; remediation; response; targeted treatment; treatment duration; treatment strategy

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.

摘要 2D型Charcot-Marie-Tooth(CMT)病是由编码甘氨酰-tRNA的GARS的显性突变引起的 合成酶(GlyRS)。CMT是临床上和遗传上不同的疾病集合，在这些疾病中，外周 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 以进行性运动神经元退化、肌肉萎缩和感觉丧失为特征。没有CMT 疾病亚型有针对性的治疗，因此CMT仍然是一个未得到满足的医疗需求。基因 异质性使得单一疗法不太可能对所有形式的CMT有效。虽然基因 针对CMT、大量不同突变的单基因疾病已经出现了强有力的治疗方法 而且受每种突变影响的患者数量很少，这使得传统的基因治疗对 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蛋白接触 其本身是CMT2D补救的关键。我们之前的工作已经确定，这种化合物的毒性的一个主要来源 引起CMT的突变体GlyRS起源于细胞外空间，突变蛋白在那里异常地相互作用 与神经粘蛋白1(Nrp1)受体结合，拮抗对运动神经元维持至关重要的信号通路。 我们基于对GlyRS突变体的结构洞察设计了一种新的策略。我们发现这一点不同 CMT2D突变导致GlyRS的共同构象变化，使新的蛋白质表面暴露在 二聚体与溶液的接口。通过使用来自GlyRS二聚体界面的两种不同的多肽作为抗原 免疫后，我们成功地获得了两种单抗(每种多肽一种)。 小鼠杂交瘤可以阻断病理性的Nrp1相互作用，并显示出有希望的泛突变选择性。 这两个候选单抗将根据其生物物理性质进行表征，并用于传导 在人类疾病的小鼠模型中进行药效学、药动学和体内疗效研究 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。