Identification of protein targets of Smo signaling in cholinergic neurons that degenerate in Alzheimer’s Disease.

本申请寻求对最近资助的R21 AG065682-01(授予日期)的行政补充支持 2020年9月10日)。额外的资金将使阿尔茨海默病(AD)相关大脑的检查成为可能 在我们的其他帕金森氏病(PD)重点分析中的核，而不扩大范围、方法或 该基金资助的技术。 我们资助的研究重点是Sonic Hedgehog(Shh)，一种胆碱能神经元营养因子，在 帕金森病患者基底节的细胞和神经化学维持及其病理学。我们之前发现， 中脑多巴胺神经元(DAN)释放Shh，它们用Shh与胆碱能神经元进行交流 (Cn)纹状体神经元。与新的成像和临床前发现一致的是，CN神经元变性 除了PD中的DAN，我们还发现有条件地消融来自DAN的Shh(ShhDAN-/-)会导致成人 起病性神经退行性变。我们的结果表明，增强CN中Shh效应器的活性可以促进 CN在帕金森病中的存活和功能。这笔资金旨在确定CN中ShhDAN信号的蛋白质靶点 纹状体用于开发可以保护CN在帕金森病中不变性的药物。此工作已启用 通过对小鼠CN蛋白质组的体内代谢标记和ShhDAN的鉴定 依赖的翻译后修饰通过质谱学。 我们的发现和方法也与AD相关，主要有三个原因：(1)Dan也在 基底节至基底前脑胆碱能神经元，在AD时变性。(2)近期影像 有证据表明AD患者的DAN显著丢失。因此，降低ShhDAN水平可能会 导致AD相关的CN退行性变。(3)与此相一致，我们在初步结果中发现ShhDAN-/- 小鼠在基底前脑显示出明显的CN丢失。总而言之，这些发现表明， 基底前脑中央核的ShhDAN信号可促进其存活。 在这里，我们寻求资金，使我们能够将基底前脑中与AD相关的组织纳入我们的CN- 特定的蛋白质组学分析。额外的AD重点分析可能会确定ShhDAN的目标 基底前脑CN独有的信号以及纹状体和基底核相关的CN靶标 前脑。因此，补充支助将深化对从基金获得的成果的解释。 Grant，此外，还有可能在AD相关的脑核中发现促进CN存活的新靶点。

This application seeks administrative supplement support for recently funded R21 AG065682-01 (award date 09/10/2020). The additional funding will allow the examination of Alzheimer’s Disease (AD) relevant brain nuclei in our otherwise Parkinson’s Disease (PD) focused analysis without expanding the scope, approach, or technology of the funded grant. Our funded research focuses on the role of Sonic Hedgehog (Shh), a cholinergic neuron trophic factor, in the cellular and neurochemical maintenance of the basal ganglia and its pathology in PD. We previously found that dopamine neurons of the mesencephalon (DAN) release Shh, which they use to communicate with cholinergic (CN) neurons in the striatum. Consistent with new imaging and preclinical findings that CN neurons degenerate in addition to DAN in PD, we found that the conditional ablation of Shh from DAN (ShhDAN-/-) results in adult onset neurodegeneration of CN. Our results suggests that boosting Shh effector activity in CN could promote CN survival and function in PD. The funded grant seeks to identify protein targets of ShhDAN signaling in CN of the striatum for the development of drugs that could protect CN from degeneration in PD. This work is enabled in our lab through in vivo metabolic labeling of mouse CN proteomes and the identification of ShhDAN dependent post-translational modifications via mass spectrometry. Our findings and approaches are also relevant to AD for three main reasons: (1) DAN also project outside of the basal ganglia to cholinergic neurons of the basal forebrain which degenerate in AD. (2) Recent imaging evidence reveals a significant loss of DANs in AD patients. Reduced levels of ShhDAN might therefore contribute to AD related CN degeneration. (3) Concordant, we found in preliminary results that ShhDAN-/- mice show a significant loss of CN in the basal forebrain. Together these findings suggest that effectors of ShhDAN signaling in CN of the basal forebrain could promote their survival. Here we seek funds that would allow us to include AD relevant tissues from the basal forebrain in our CN- specific proteomic analysis. The additional AD focused analysis would potentially identify targets of ShhDAN signaling unique to CN of the basal forebrain as well as targets relevant CN of both the striatum and basal forebrain. Thus, supplemental support would both deepen the interpretation of results obtained from the funded grant and in addition, potentially identify novel targets promoting CN survival in AD relevant brain nuclei.