Leucine rich repeat kinase 2 and dominantly inherited Parkinson disease

我们在这个项目中的主要目的是了解LRRK 2的许多不同结构域的突变如何导致显性遗传性帕金森病。我们一直在特别寻找在分子的许多不同功能域中发现的多个突变的共同效应。在当前阶段，我们已经证实了先前的数据，即突变型LRRK 2在急性表达时对培养中的细胞具有毒性。与加州大学旧金山分校的Steve Finkbeiner合作，我们发现突变型LRRK 2对细胞具有毒性，其方式取决于α-突触核蛋白。这种毒性作用在多个突变之间共享，并且当LRRK 2的非活性（激酶死亡）版本取代活性版本时最小化，尽管一些作用与蛋白质水平而不是活性本身相关。 这些结果支持以下论点：虽然不同的突变影响LRRK 2分子的不同部分，因此具有不同的分子效应，但它们都具有一些导致有害效应的收敛特性。我们最近发现LRRK 2与一系列其他蛋白质相互作用，其中包括两种已被提名为帕金森病遗传风险候选基因的Rab 7 L1和GAK。后续的蛋白质复合物的功能，我们发现，LRRK 2与Rab 7 L1和GAK合作，以促进通过涉及自噬-溶酶体系统的过程去除来自trans-golgi网络的囊泡子集。重要的是，我们迄今为止检测的LRRK 2中的所有突变都促进了这种效应，这表明它是致病性变体的共同特性。我们目前正在系统中进行这项工作，这将使我们能够在内源性水平上测定突变。

Our main aim in this project is to understand how mutations across many different domains of LRRK2 cause dominantly inherited Parkinsons disease. We have been particularly looking for shared effects of multiple mutations that are found in many different functional domains of the molecule. In the current period, we have confirmed previous data that mutant LRRK2 can be toxic to cells in culture when expressed acutely. In collaboration with Steve Finkbeiner at UCSF, we found that mutant LRRK2 is toxic to cells in a manner that depends on a-synuclein. This toxic effect is shared between multiple mutations and is minimized when an inactive (kinase dead) version of LRRK2 is substituted for active versions, although some of the effects are related to protein level rather than activity per se. These results support the argument that although different mutations affect different parts of the LRRK2 molecule and therefore have divergent molecular effects, they all share some convergent property(s) that lead to detrimental effects. We have recently shown that LRRK2 interacts with a series of other proteins, including two that have been nominated as candidate genes for genetic risk in Parkinson's disease, Rab7L1 and GAK. Following up on the function of the complex of proteins, we found that LRRK2 cooperates with Rab7L1 and GAK to promote the removal of a subset of vesicles derived from the trans-golgi network by a process that involves the autophagy-lysosome system. Importantly, all mutations in LRRK2 that we have assayed to date promote this effect, suggesting it is a common property of pathogenic variants. We are currently following this work up in systems that should allow us to assay mutations at the endogenous level.