The LRRK2 kinase substrates Rab8a and Rab10 contribute complementary but distinct disease-relevant phenotypes in human neurons.
The LRRK2 kinase substrates Rab8a and Rab10 contribute complementary but distinct disease-relevant phenotypes in human neurons.
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LRRK2 激酶底物 Rab8a 和 Rab10 在人类神经元中贡献互补但不同的疾病相关表型。
DOI：
10.1101/2023.04.30.538317
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
LaVoie,MatthewJ
中科院分区：
文献类型：
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作者：
Mamais,Adamantios;Sanyal,Anwesha;Fajfer,Austin;Zykoski,CatherineG;Guldin,Michael;Riley-DiPaolo,Alexis;Subrahmanian,Nitya;Gibbs,Whitney;Lin,Steven;LaVoie,MatthewJ
LRRK2 基因突变导致家族性帕金森病，表现为多形性神经病理学，可能涉及 α-突触核蛋白或 tau 积累。LRRK2 突变被认为与 LRRK2 激酶活性的致病性增加有关。小 RAB GTP 酶的一个子集已被鉴定为 LRRK2 底物，LRRK2 依赖性磷酸化导致 RAB 失活。我们使用 CRISPR-Cas9 基因组编辑从深度表型的健康对照系中生成了一系列新的等基因 iPSC 系，这些 iPSC 系缺乏两种经过充分验证的 LRRK2 底物 RAB8a 和 RAB10。 NGN2 诱导的神经元的全面表征揭示了 RAB8a 和 RAB10 缺乏对溶酶体 pH 和高尔基体组织的相反影响，RAB8a 和 RAB10 消融分别对 α-突触核蛋白和 tau 产生单独的影响。我们的数据表明，RAB8a 或 RAB10 基因失活在很大程度上具有拮抗作用，这为人类疾病中致病性 LRRK2 突变导致的生化失活的病理特征提供了离散的见解。
Mutations in theLRRK2gene cause familial Parkinson's disease presenting with pleomorphic neuropathology that can involve α-synuclein or tau accumulation.LRRK2mutations are thought to converge upon a pathogenic increase in LRRK2 kinase activity. A subset of small RAB GTPases has been identified as LRRK2 substrates, with LRRK2-dependent phosphorylation resulting in RAB inactivation. We used CRISPR-Cas9 genome editing to generate a novel series of isogenic iPSC lines deficient in the two most well-validated LRRK2 substrates, RAB8a and RAB10, from deeply phenotyped healthy control lines. Thorough characterization of NGN2-induced neurons revealed opposing effects of RAB8a and RAB10 deficiency on lysosomal pH and Golgi organization, with isolated effects of RAB8a and RAB10 ablation on α-synuclein and tau, respectively. Our data demonstrate largely antagonistic effects of genetic RAB8a or RAB10 inactivation, which provide discrete insight into the pathologic features of their biochemical inactivation by pathogenicLRRK2mutation in human disease.