Dopaminergic neurodegeneration induced by Parkinson's disease-linked G2019S LRRK2 is dependent on kinase and GTPase activity
Dopaminergic neurodegeneration induced by Parkinson's disease-linked G2019S LRRK2 is dependent on kinase and GTPase activity
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帕金森病相关的 G2019S LRRK2 诱导的多巴胺能神经变性依赖于激酶和 GTP 酶活性
DOI：
10.1101/2019.12.17.879759
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发表时间：
2019
影响因子：
11.1
通讯作者：
J. Schwarz
中科院分区：
文献类型：
--
作者：
J. Milošević;S. Schwarz;V. Ogunlade;Anne K. Meyer;A. Storch;J. Schwarz
意义 LRRK2 中与帕金森病 (PD) 相关的家族突变通常会直接在激酶结构域内增加激酶活性，或通过损害 GTP 水解而间接通过 GTPase 结构域间接增加激酶活性，从而影响其酶活性。家族性 LRRK2 突变通常还会促进培养细胞中的神经元毒性，对于常见的 G2019S 突变，这些影响是激酶依赖性的。由于普遍缺乏稳健的表型，动物模型中家族性 LRRK2 突变的机制尚不确定。我们的研究证明了激酶和 GTP 酶活性在介导 G2019S LRRK2 在啮齿动物中的神经退行性作用中的重要作用，强调两者都是有希望的 PD 治疗靶点。富含亮氨酸重复激酶 2 (LRRK2) 的突变是迟发性常染色体显性家族性帕金森病 (PD) 的最常见原因。 LRRK2 既充当激酶又充当 GTP 酶，并且已知 PD 相关突变会影响这两种酶活性。虽然 PD 相关 LRRK2 突变通常可在培养模型中诱导神经元损伤，但这些致病作用的机制仍不确定。含有家族性 LRRK2 突变的啮齿动物模型通常缺乏强大的 PD 样神经退行性表型。在这里，我们在成年大鼠中开发了一种稳健的 PD 临床前模型，通过向大脑递送重组腺病毒载体来诱导，该载体具有神经元特异性表达含有最常见 G2019S 突变的人 LRRK2。在此模型中，G2019S LRRK2 诱导黑质多巴胺能神经元的强烈变性，这是 PD 的病理标志。引入稳定的激酶失活突变或施用选择性激酶抑制剂 PF-360 可减轻 G2019S LRRK2 诱导的神经变性。药理学激酶抑制提供的神经保护是由一种不寻常的机制介导的，涉及大脑中相对于内源性 LRRK2 的人 LRRK2 蛋白的强烈不稳定。我们的研究进一步表明，G2019S LRRK2 诱导的多巴胺能神经变性至关重要需要正常的 GTP 酶活性，因为增加 GTP 水解或损害 GTP 结合活性的假设检验突变虽然通过不同的机制提供了神经保护。综上所述，我们的数据表明 G2019S LRRK2 通过依赖于激酶和 GTP 酶活性的机制诱导体内神经变性。我们的研究提供了 LRRK2 相关 PD 的稳健啮齿动物临床前模型，并提名激酶抑制和 GTP 酶活性调节作为有希望的疾病缓解治疗靶点。
Significance Parkinson’s disease (PD)-linked familial mutations in LRRK2 impact its enzymatic activity by commonly increasing kinase activity, either directly within the kinase domain or indirectly via the GTPase domain by impairing GTP hydrolysis. Familial LRRK2 mutations also commonly promote neuronal toxicity in cultured cells, and for the common G2019S mutation, these effects are kinase dependent. The mechanisms underlying familial LRRK2 mutations in animal models are uncertain, due to the general lack of robust phenotypes. Our study demonstrates important roles for kinase and GTPase activities in mediating the neurodegenerative effects of G2019S LRRK2 in rodents, highlighting both as promising therapeutic targets for PD. Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common cause of late-onset, autosomal-dominant familial Parkinson’s disease (PD). LRRK2 functions as both a kinase and GTPase, and PD-linked mutations are known to influence both enzymatic activities. While PD-linked LRRK2 mutations can commonly induce neuronal damage in culture models, the mechanisms underlying these pathogenic effects remain uncertain. Rodent models containing familial LRRK2 mutations often lack robust PD-like neurodegenerative phenotypes. Here, we develop a robust preclinical model of PD in adult rats induced by the brain delivery of recombinant adenoviral vectors with neuronal-specific expression of human LRRK2 harboring the most common G2019S mutation. In this model, G2019S LRRK2 induces the robust degeneration of substantia nigra dopaminergic neurons, a pathological hallmark of PD. Introduction of a stable kinase-inactive mutation or administration of the selective kinase inhibitor, PF-360, attenuates neurodegeneration induced by G2019S LRRK2. Neuroprotection provided by pharmacological kinase inhibition is mediated by an unusual mechanism involving the robust destabilization of human LRRK2 protein in the brain relative to endogenous LRRK2. Our study further demonstrates that G2019S LRRK2-induced dopaminergic neurodegeneration critically requires normal GTPase activity, as hypothesis-testing mutations that increase GTP hydrolysis or impair GTP-binding activity provide neuroprotection although via distinct mechanisms. Taken together, our data demonstrate that G2019S LRRK2 induces neurodegeneration in vivo via a mechanism that is dependent on kinase and GTPase activity. Our study provides a robust rodent preclinical model of LRRK2-linked PD and nominates kinase inhibition and modulation of GTPase activity as promising disease-modifying therapeutic targets.
DOI：
10.1073/pnas.0507360102
发表时间：
2005-11-15
影响因子：
11.1
作者：
West, AB;Moore, DJ;Dawson, TM
通讯作者：
Dawson, TM