Laboratory for Neurobiology and Gene Therapy, Department of Neurosciences, KU Leuven, Herestraat 49 - Bus 1023, 3000, Leuven, Belgium.
Mol Neurobiol. 2019 Aug;56(8):5273-5286. doi: 10.1007/s12035-018-1449-2. Epub 2018 Dec 27.
Mutations and variations in the leucine-rich repeat kinase 2 (LRRK2) gene are strongly associated with an increased risk to develop Parkinson's disease (PD). Most pathogenic LRRK2 mutations display increased kinase activity, which is believed to underlie LRRK2-mediated toxicity. Therefore, major efforts have been invested in the development of potent and selective LRRK2 kinase inhibitors. Several of these compounds have proven beneficial in cells and in vivo, even in a LRRK2 wild-type background. Therefore, LRRK2 kinase inhibition holds great promise as disease-modifying PD therapy, and is currently tested in preclinical and early clinical studies. One of the safety concerns is the development of lung pathology in mice and non-human primates, which is most likely related to the strongly reduced LRRK2 protein levels after LRRK2 kinase inhibition. In this study, we aimed to better understand the molecular consequences of chronic LRRK2 kinase inhibition, which may be pivotal in the further development of a LRRK2 kinase inhibitor-based PD therapy. We found that LRRK2 protein levels are not restored during long-term LRRK2 kinase inhibition, but are recovered upon inhibitor withdrawal. Interestingly, LRRK2 kinase inhibitor-induced destabilization does not occur in all pathogenic LRRK2 variants and the N-terminal part of LRRK2 appears to play a crucial role in this process. In addition, we identified CK1, an upstream kinase of LRRK2, as a regulator of LRRK2 protein stability in cell culture and in vivo. We propose that pharmacological LRRK2 kinase inhibition triggers a cascade that results in reduced CK1-mediated phosphorylation of yet unidentified LRRK2 phosphorylation sites. This process involves the N-terminus of LRRK2 and ultimately leads to LRRK2 protein degradation.
LRRK2 基因中的突变和变异与帕金森病(PD)发病风险的增加密切相关。大多数致病性 LRRK2 突变显示激酶活性增加,这被认为是 LRRK2 介导的毒性的基础。因此,人们投入了大量精力开发有效的、选择性的 LRRK2 激酶抑制剂。其中一些化合物已被证明在细胞和体内都有疗效,甚至在 LRRK2 野生型背景下也是如此。因此,LRRK2 激酶抑制作为一种有希望的疾病修饰性 PD 治疗方法,目前正在进行临床前和早期临床试验。其中一个安全问题是在小鼠和非人类灵长类动物中出现肺部病变,这很可能与 LRRK2 激酶抑制后 LRRK2 蛋白水平的强烈降低有关。在这项研究中,我们旨在更好地了解慢性 LRRK2 激酶抑制的分子后果,这可能对进一步开发基于 LRRK2 激酶抑制剂的 PD 治疗方法至关重要。我们发现,在长期 LRRK2 激酶抑制期间,LRRK2 蛋白水平不会恢复,而是在抑制剂停药后恢复。有趣的是,LRRK2 激酶抑制剂诱导的不稳定性不会发生在所有致病性 LRRK2 变体中,LRRK2 的 N 端似乎在这个过程中起着关键作用。此外,我们确定 CK1,LRRK2 的上游激酶,是细胞培养和体内 LRRK2 蛋白稳定性的调节剂。我们提出,药理学 LRRK2 激酶抑制触发了一个级联反应,导致 CK1 介导的未鉴定的 LRRK2 磷酸化位点的磷酸化减少。这个过程涉及 LRRK2 的 N 端,最终导致 LRRK2 蛋白降解。