MRC Laboratory for Molecular Cell Biology, University College London, London, U.K.
UCL:Eisai Therapeutic Innovation Group, Translational Research Office, University College London, London, U.K.
Biochem J. 2021 Dec 10;478(23):4099-4118. doi: 10.1042/BCJ20210508.
Mitochondrial dysfunction is implicated in Parkinson disease (PD). Mutations in Parkin, an E3 ubiquitin ligase, can cause juvenile-onset Parkinsonism, probably through impairment of mitophagy. Inhibition of the de-ubiquitinating enzyme USP30 may counter this effect to enhance mitophagy. Using different tools and cellular approaches, we wanted to independently confirm this claimed role for USP30. Pharmacological characterisation of additional tool compounds that selectively inhibit USP30 are reported. The consequence of USP30 inhibition by these compounds, siRNA knockdown and overexpression of dominant-negative USP30 on the mitophagy pathway in different disease-relevant cellular models was explored. Knockdown and inhibition of USP30 showed increased p-Ser65-ubiquitin levels and mitophagy in neuronal cell models. Furthermore, patient-derived fibroblasts carrying pathogenic mutations in Parkin showed reduced p-Ser65-ubiquitin levels compared with wild-type cells, levels that could be restored using either USP30 inhibitor or dominant-negative USP30 expression. Our data provide additional support for USP30 inhibition as a regulator of the mitophagy pathway.
线粒体功能障碍与帕金森病（PD）有关。E3 泛素连接酶 Parkin 的突变可导致青少年发病的帕金森病，可能是通过损害线粒体自噬。抑制去泛素化酶 USP30 可能会抵消这种作用，从而增强线粒体自噬。本研究使用不同的工具和细胞方法，旨在独立证实 USP30 的这一作用。本文报道了选择性抑制 USP30 的其他工具化合物的药理学特征。研究了这些化合物、siRNA 敲低和显性负 USP30 过表达对不同与疾病相关的细胞模型中线粒体自噬途径的影响。USP30 的敲低和抑制导致神经元细胞模型中 p-Ser65-泛素化水平和线粒体自噬增加。此外，携带 Parkin 致病性突变的患者来源成纤维细胞与野生型细胞相比，p-Ser65-泛素化水平降低，而使用 USP30 抑制剂或显性负 USP30 表达可恢复该水平。我们的数据为 USP30 抑制作为线粒体自噬途径的调节剂提供了额外的支持。
