Antunes Flavia T T, Gandini Maria A, Garcia-Caballero Agustin, Huang Sun, Ali Md Yousof, Gambeta Eder, Souza Ivana A, Harding Erika K, Ferron Laurent, Stray-Pedersen Asbjorg, Gadotti Vinicius M, Zamponi Gerald W
Department of Clinical Neurosciences, Hotchkiss Brain Institute and Alberta Children's Hospital Research Institute, Cumming School of Medicine, University of Calgary, Calgary, Canada.
Zymedyne Therapeutics , Calgary, Canada.
J Exp Med. 2025 Aug 4;222(8). doi: 10.1084/jem.20241877. Epub 2025 May 16.
Cav3.2 T-type calcium channels and their dysregulation by the deubiquitinase USP5 contribute to development of inflammatory and neuropathic pain. We report on a pediatric patient with a de novo heterozygous missense mutation R24W in USP5 who exhibits pain insensitivity. We created a CRISPR knock-in mouse harboring this mutation and performed detailed behavioral analyses in acute and chronic pain models. Heterozygous R24W mice of both sexes are resistant to acute pain and to thermal hypersensitivity in chronic inflammatory and neuropathic pain models. In contrast, only male R24W mice confer resistance to development of mechanical hypersensitivity. R24W mice lack upregulation of Cav3.2 and USP5 that is normally observed with CFA-induced inflammation. Moreover, mutant USP5 exhibits a dramatic reduction in enzymatic activity but stronger interactions with Cav3.2. Hence, R24W mutant USP5 is a critical regulator of chronic and acute pain states in humans by acting as a dominant-negative regulator of Cav3.2. Our data validate USP5 as a potential therapeutic target for chronic pain in humans.
Cav3.2 T型钙通道及其被去泛素化酶USP5的失调促成了炎症性疼痛和神经性疼痛的发展。我们报告了一名患有USP5基因新生杂合错义突变R24W的儿科患者，该患者表现出疼痛不敏感。我们创建了一只携带此突变的CRISPR基因敲入小鼠，并在急性和慢性疼痛模型中进行了详细的行为分析。在慢性炎症和神经性疼痛模型中，雌雄杂合R24W小鼠均对急性疼痛和热超敏反应具有抗性。相比之下，只有雄性R24W小鼠对机械性超敏反应的发展具有抗性。R24W小鼠缺乏通常在CFA诱导的炎症中观察到的Cav3.2和USP5的上调。此外，突变型USP5的酶活性显著降低，但与Cav3.2的相互作用更强。因此，R24W突变型USP5通过作为Cav3.2的显性负调控因子，成为人类慢性和急性疼痛状态的关键调节因子。我们的数据验证了USP5作为人类慢性疼痛潜在治疗靶点的作用。
