Wei Shunhui, Poore Charlene Priscilla, Verma Ravi Kumar, Low See Wee, Chen Bo, Fan Hao, Liao Ping
Calcium Signalling Laboratory, Department of Research, National Neuroscience Institute, Singapore, 308433, Singapore.
Bioinformatics Institute (BII), Agency for Science Technology and Research (A*STAR), Singapore, 138671, Singapore.
Sci Rep. 2025 Jun 5;15(1):19769. doi: 10.1038/s41598-025-05256-x.
Transient receptor potential melastatin 4 (TRPM4) channel, a monovalent cation channel, plays a crucial role in various neurological disorders. We previously showed that TRPM4 activity can be blocked by our antibodies: M4P, which targets rat TRPM4 and M4M, against a similar antigenic epitope of human TRPM4. M4P and M4M demonstrated efficacy in mitigating stroke reperfusion injury. To facilitate human application, M4M was humanized through CDR grafting, resulting in human IgG1 antibodies (Ab1-6). These antibodies were evaluated for their binding affinity, surface staining, stability, and functional inhibition of the human TRPM4 channel. Ab6 (renamed as M4H) was selected and inhibited TRPM4 currents in human brain microvascular endothelial cells under ATP depletion conditions. Importantly, Ab6 (M4H) suppressed ATP depletion-induced cell swelling, indicating its potential for managing vascular injury in ischemic brain diseases. Future studies on animal models could advance the development of novel therapies of neurological disorders with vascular injury.
瞬时受体电位褪黑素4（TRPM4）通道是一种单价阳离子通道，在各种神经系统疾病中起关键作用。我们之前表明，TRPM4的活性可被我们的抗体阻断：针对大鼠TRPM4的M4P和针对人TRPM4相似抗原表位的M4M。M4P和M4M在减轻中风再灌注损伤方面显示出疗效。为便于人类应用，通过互补决定区（CDR）移植对M4M进行人源化，得到人IgG1抗体（Ab1 - 6）。对这些抗体进行了结合亲和力、表面染色、稳定性以及对人TRPM4通道功能抑制的评估。选择了Ab6（重命名为M4H），其在ATP耗竭条件下抑制人脑微血管内皮细胞中的TRPM4电流。重要的是，Ab6（M4H）抑制了ATP耗竭诱导的细胞肿胀，表明其在管理缺血性脑疾病中的血管损伤方面具有潜力。未来对动物模型的研究可能会推动针对伴有血管损伤的神经系统疾病的新型疗法的开发。
