White matter damage in Alzheimer?s disease: New cellular targets and mechanisms

描述（由申请人提供）：该“探索性”应用的目标是提供对淀粉样蛋白（A <$）脆弱性的新的机制理解，以及A <$破坏维持正常髓鞘形成的关键细胞功能的途径。我们最近发现了一种新的调节途径，它提供了氧化变化和细胞信号控制之间的顺序联系。在该途径中，细胞暴露于具有促氧化活性的化学多样性物质引起的氧化状态的增加引起Fyn激酶的活化。这导致c-Cbl的激活，c-Cbl是Fyn的靶标的E3泛素连接酶。c-Cbl的激活导致其靶蛋白的泛素化，其中包括受体酪氨酸激酶（RTK）的子集。由于它们与c-Cbl的相互作用，这些RKT的降解增强，导致下游信号传导的抑制。作为这种降解的结果，下游激活的信号传导介质如Erk 1/2和Akt被抑制。正如人们可以从这种效应预测的那样，细胞分裂受到抑制，细胞存活也可能受损。我们建议现在测试的假设，Fyn/c-Cbl通路的激活在淀粉样蛋白（A？）毒性中起着重要作用。由于髓鞘损伤在AD病理学中的重要性，所提出的实验集中于A肽对少突胶质细胞及其祖细胞的影响。此外，由于Fyn/c-Cbl假说还预测暴露于亚致死浓度的促氧化剂刺激物将抑制细胞分裂，我们将进一步检验A肽对少突胶质细胞具有细胞毒性但也抑制产生它们的祖细胞分裂的假说。如果这一预测是正确的，这将表明A？既损害髓鞘形成细胞，又抑制修复所需的细胞分裂。因此，这项研究提出了一种新的分子途径，A？通过这种途径影响细胞功能。先前的一些研究表明Fyn在AD的发病机制中起重要作用。我们的研究将为Fyn激活可能破坏AD细胞功能的机制提供新的见解。目的1测试以下假设：少突胶质细胞及其祖细胞暴露于A？导致氧化还原/Fyn/c-Cbl途径的激活、作为c-Cbl靶点的RTK的降解以及来自这些RTK的下游信号事件的选择性抑制。根据A ²的类型和浓度以及检查的细胞类型，这与祖细胞分裂（亚致死剂量）的抑制以及较高浓度下祖细胞和/或少突胶质细胞死亡的诱导有关。目的2检验以下假设：Fyn/c-Cbl通路的激活在少突胶质细胞谱系中A？介导的细胞分裂抑制和/或细胞死亡诱导中具有重要功能。目的3测试的假设，即抗氧化剂和营养因子，保护免受毒性作用的A ²抑制A ²介导的氧化还原/Fyn/c-Cbl途径的激活，从而提供了一个新的潜在网站的抗氧化剂在AD中的保护作用。 公共卫生相关性：这项研究为淀粉样蛋白对阿尔茨海默病中枢神经系统造成损害的方式提供了新的见解。我们的研究确定了一种新的分子途径，淀粉样蛋白通过这种途径破坏细胞功能，对这种疾病中有髓神经束广泛损伤的发病机制有了新的认识，并且对抗氧化治疗保护免受淀粉样蛋白影响的方法有了新的认识。这项研究将有助于确定防止淀粉样蛋白毒性的新方法。

DESCRIPTION (provided by applicant): The goal of this "exploratory" application is to provide a novel mechanistic understanding of vulnerability to amyloid (A ¿) protein, and of the pathways through which A¿ disrupts function of those cells critical in maintenance of normal myelination. We have recently discovered a novel regulatory pathway that provides a sequential linkage between oxidative changes and control of cell signaling. In this pathway, increases in oxidative status caused by exposure of cells to chemically diverse substances with pro-oxidant activity cause activation of Fyn kinase. This leads to activation of c-Cbl, an E3 ubiquitin ligase that is a target of Fyn. Activation of c-Cbl leads to ubiquitylation of its target proteins, which include among them a subset of receptor tyrosine kinases (RTKs). As a result of their interaction with c-Cbl, degradation of these RKTs is enhanced, leading to a suppression of downstream signaling. As a consequence of this degradation, downstream activation of such signaling mediators as Erk1/2 and Akt are suppressed. As one would predict from such an effect, cell division is suppressed and cell survival may also be impaired. We propose to now test the hypothesis that activation of the Fyn/c-Cbl pathway plays an important role in amyloid (A¿) toxicity. The experiments proposed focus on the effects of A¿ peptides on oligodendrocytes and their progenitor cells, due to the importance of myelin damage in AD pathology. Moreover, as the Fyn/c- Cbl hypothesis also predicts that exposure to sublethal concentrations of pro-oxidant stimuli will suppress cell division, we will further test the hypothesis that A¿ peptides are cytotoxic for oligodendrocytes but also suppress division of the progenitors from which they are generated. If this prediction is correct, this would indicate that A¿ both damages myelin-forming cells and suppresses the cell division required for repair. This research thus proposes a new molecular pathway by which A¿ affects cell function. Several studies have previously suggested an important role of Fyn in the pathogenesis of AD. Our studies will provide novel insights into the mechanism by which Fyn activation may disrupt cellular function in AD. Aim 1 tests the hypothesis that exposure of oligodendrocytes and their progenitors to A¿ causes activation of the redox/Fyn/c-Cbl pathway, degradation of RTKs that are c-Cbl targets, and selective suppression of downstream signaling events from these RTKs. This is associated with, depending on the type and concentration of A¿ and the cell type examined, suppression of progenitor cell division (at sublethal doses) and induction of progenitor cell and/or oligodendrocyte death at higher concentrations. Aim 2 tests the hypothesis that activation of the Fyn/c-Cbl pathway is functionally important in A¿ -mediated suppression of cell division and/or induction of cell death in the oligodendrocyte lineage. Aim 3 tests the hypothesis that anti-oxidants and trophic factors that protect against toxic effects of A¿ suppress A¿ -mediated activation of the redox/Fyn/c-Cbl pathway, thus providing a novel potential site of action for the protective effects of anti-oxidants in AD. PUBLIC HEALTH RELEVANCE: This research provides novel insights into the means by which amyloid ¿ protein causes damage to the central nervous system in Alzheimer's disease. Our studies identify a novel molecular pathway by which amyloid ¿ protein disrupts cell function, new insights into the pathogenesis of the extensive damage to myelinated tracts in this disease, and a new understanding of means by which anti-oxidant therapy protects from the effects of amyloid ¿ protein. This research will help in identifying new means of protecting against amyloid ¿ toxicity.