Lysosomal impairment is strongly implicated in Parkinson’s disease (PD). Among the several PD-linked genes, the ATP13A2 gene, associated with the PARK9 locus, encodes a transmembrane lysosomal P5-type ATPase. Mutations in the ATP13A2 gene were primarily identified as the cause of Kufor-Rakeb syndrome (KRS), a juvenile-onset form of PD. Subsequently, an increasing list of several mutations has been described. These mutations result in truncation of the ATP13A2 protein, leading to a loss of function but surprisingly causing heterogeneity and variability in the clinical symptoms associated with different brain pathologies. In vitro studies show that its loss compromises lysosomal function, contributing to cell death. To understand the role of ATP13A2 dysfunction in disease, we disrupted its expression through a viral vector-based approach in nonhuman primates. Here, in this pilot study, we injected bilaterally into the substantia nigra of macaques, a lentiviral vector expressing an ATP13A2 small hairpin RNA. Animals were terminated five months later, and brains were harvested and compared with historical non-injected control brains to evaluate cerebral pathological markers known to be affected in KRS and PD. We characterised the pattern of dopaminergic loss in the striatum and the substantia nigra, the regional distribution of α-synuclein immunoreactivity in several brain structures, and its pathological status (i.e., S129 phosphorylation), the accumulation of heavy metals in nigral sections and occurrence of lysosomal dysfunction. This proof-of-concept experiment highlights the potential value of lentivirus-mediated ATP13A2 silencing to induce significant and ongoing degeneration in the nigrostriatal pathway, α-synuclein pathology, and iron accumulation in nonhuman primates.

溶酶体损伤与帕金森病 (PD) 密切相关。在几个 PD 连锁基因中， ATP13A2基因与 PARK9 基因座相关，编码跨膜溶酶体 P5 型 ATP 酶。 ATP13A2基因突变主要被确定为 Kufor-Rakeb 综合征 (KRS) 的病因，KRS 是一种青少年发病的帕金森病。随后，越来越多的几种突变被描述。这些突变导致 ATP13A2 蛋白截短，导致功能丧失，但令人惊讶的是，导致与不同脑部病理相关的临床症状的异质性和变异性。体外研究表明，它的缺失会损害溶酶体功能，导致细胞死亡。为了了解 ATP13A2 功能障碍在疾病中的作用，我们通过基于病毒载体的方法在非人类灵长类动物中破坏了其表达。在这项初步研究中，我们将表达 ATP13A2 小发夹 RNA 的慢病毒载体双侧注射到猕猴的黑质中。五个月后处死动物，收获大脑并与历史上未注射的对照大脑进行比较，以评估已知在 KRS 和 PD 中受影响的脑病理标志物。我们表征了纹状体和黑质中多巴胺能丧失的模式、几个脑结构中 α-突触核蛋白免疫反应性的区域分布及其病理状态（即 S129 磷酸化）、黑质切片中重金属的积累和发生溶酶体功能障碍。 这一概念验证实验强调了慢病毒介导的 ATP13A2 沉默在诱导非人灵长类动物黑质纹状体通路、α-突触核蛋白病理学和铁积累中显着且持续的退化的潜在价值。