Production of TRPV2-targeting functional antibody ameliorating dilated cardiomyopathy and muscular dystrophy in animal models
Production of TRPV2-targeting functional antibody ameliorating dilated cardiomyopathy and muscular dystrophy in animal models
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DOI：
10.1038/s41374-019-0363-1
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发表时间：
2020-01-02
影响因子：
5
通讯作者：
Kitakaze, Masafumi
中科院分区：
文献类型：
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作者：
Iwata, Yuko;Wakabayashi, Shigeo;Kitakaze, Masafumi
作者产生了一种功能性单克隆抗体，其识别并促进TRPV 2的内化，以减少TRPV 2驱动的Ca 2+内流。使用心肌病/肌营养不良动物模型，他们证实了这种抗体的有益作用，包括改善心肌细胞和骨骼肌特性中的Ca 2+处理。异常Ca 2+处理在退行性肌肉疾病（如扩张型心肌病（DCM）和肌营养不良（MD））的病理生理学中至关重要。瞬时受体电位阳离子通道亚家族V成员2（TRPV 2）是钙离子内流的候选通道，也是退行性肌肉疾病的潜在治疗靶点，目前还没有特异性抑制剂。在这项研究中，我们产生了一种单克隆抗体（命名为mAb 88 -2）和两种多克隆抗体（pAb 591和pAb 592），它们选择性地识别来自细胞外部的TRPV 2，并与成孔外门的转塔区域相互作用。这些抗体通过TRPV 2抑制培养细胞中的Ca 2+内流，并通过细胞内化显著降低质膜中的TRPV 2。我们在DCM和MD的δ-肌聚糖缺陷仓鼠（J2 N-k）模型和鼠心力衰竭的4C 30 DCM模型中评价了功能性抗体的治疗功效。通过超声心动图和组织学染色评价，腹膜内给予功能性抗体（0.5mg/kg）2周（每周一次）防止了心功能障碍的进展，并改善了从J2 N-k仓鼠分离的心肌细胞中的异常Ca 2+处理（高舒张期Ca 2+水平和小Ca 2+瞬时峰），并防止了骨骼肌损伤。此外，该抗体有效地预防了患有终末期DCM的4C 30小鼠模型中的心力衰竭。有趣的是，在心脏和骨骼肌肌膜中积累的内源性TRPV 2在抗体施用后消失。因此，新产生的抗体能够通过促进TRPV 2的细胞内化来改善DCM和MD;对人TRPV 2特异性的抗体可以显著改善患有退行性肌肉疾病的患者的治疗。
The authors generated a functional monoclonal antibody that recognizes and promotes the internalization of TRPV2 to reduce TRPV2-driven Ca2+ influx. Using cardiomyopathic/muscular dystrophic animal models, they confirmed the beneficial effects of this antibody, which includes improvements in both Ca2+ handling in cardiomyocytes and in skeletal muscle properties.Abnormal Ca2+ handling is essential in the pathophysiology of degenerative muscle disorders, such as dilated cardiomyopathy (DCM) and muscular dystrophy (MD). Transient receptor potential cation channel, subfamily V, member 2 (TRPV2) is a candidate for Ca2+ entry and a potential therapeutic target for degenerative muscle disorders, there are few specific inhibitors for TRPV2. In this study, we produced a monoclonal antibody (designated mAb88-2) and two polyclonal antibodies (pAb591 and pAb592) that selectively recognize TRPV2 from the outside of cells and interact with the turret region of the pore-forming outer gate. These antibodies inhibited Ca2+ influx via TRPV2 in cultured cells and substantially reduced TRPV2 in the plasma membrane via cellular internalization. We evaluated the therapeutic efficacy of the functional antibody in delta-sarcoglycan-deficient hamster (J2N-k) models of DCM and MD and in the 4C30 DCM model of murine heart failure. The intraperitoneal administration of the functional antibody (0.5 mg/kg) for 2 weeks (once a week) prevented the progression of cardiac dysfunction, as evaluated by echocardiography and histological staining, and improved the abnormal Ca2+ handling (high diastolic Ca2+ level and small Ca2+ transient peak) in cardiomyocytes isolated from J2N-k hamsters and prevented skeletal muscle damage. Further, the antibody effectively prevented heart failure in the 4C30 mouse model with end-stage DCM. Interestingly, endogenous TRPV2 that accumulated in the cardiac and skeletal muscle sarcolemma disappeared upon antibody administration. Thus, the newly produced antibodies are capable of ameliorating DCM and MD by promoting the cellular internalization of TRPV2; antibodies specific to human TRPV2 may substantially improve the treatment of patients with degenerative muscle diseases.