Regulation of Neural Stem Cells and Neurogenesis by Autophagy Genes

自噬基因对神经干细胞和神经发生的调节

• 批准号: 10434019
• 负责人: JUN-LIN GUAN
• 金额: $ 37.92万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10434019
• 关键词: Ablation; Adult; Affect; Aging; Alzheimer' s Disease; Animals; Autophagocytosis; Biochemical; Biological; Brain; C-terminal; Cell Maintenance; Cell model; Cell physiology; Cells; Complex; Data; Defect; Development; Disease; Disease Progression; Dominant-Negative Mutation; Family; Functional disorder; Funding; Future; Generations; Genes; Genetic; Goals; Growth; Homeostasis; Huntington Disease; Hydrogen Peroxide; Hypoxia; In Vitro; Injury; Knock-in; Knock-in Mouse; Knockout Mice; Laboratories; Lead; Maintenance; Metabolic; Modeling; Molecular; Mus; Mutagenesis; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Organ; Oxidative Stress; PTK2 gene; Parkinson Disease; Phosphorylation; Play; Process; Proteins; Regulation; Risk Factors; Role; TBK1 gene; age related; aged; cell age; conditional knockout; design; effective therapy; gain of function; in vivo; insight; juvenile animal; mouse model; nerve stem cell; neurogenesis; neuromechanism; neuroregulation; prevent; receptor; response; stem cells; synergism

In adult brains, neural progenitor/stem cells (NSCs) are responsible for the generation of new neurons for the maintenance of the existing circuitry and after injuries. Deficiency in NSC maintenance and/or neurogenesis contributes to both developmental defects and neurodegenerative diseases for which aging is a major risk factor. The long-term goal of the proposed studies is to determine the molecular and cellular mechanisms of NSC regulation in both young and aged animals, which can lead to the development of effective therapies for neurodegenerative diseases. Autophagy is a highly conserved cellular process for degradation of bulk cytoplasmic materials for maintenance of cellular homeostasis, and dysfunctions in autophagy have been implicated in various diseases, including neurodegeneration and other age-related disorders. FIP200 (FAK-family Interacting Protein of 200 kDa) was initially identified in our laboratory and subsequently shown as one component of the ULK1/Atg13/FIP200 complex essential for the induction of autophagy. In the previous funding period, we found that, deletion of Fip200, but not other autophagy genes Atg5, Atg7 and Atg16L1, led to defective NSC maintenance and neurogenesis, suggesting a potential role for the non-canonical function of FIP200 in NSC regulation through controlling p62 aggregate formation. In additional prelim studies, we obtained rigorous genetic evidence that non-canonical function of FIP200 is required for NSC maintenance and neurogenesis by generation and analyses of another unique mouse model with FIP200-4A mutation in NSCs that blocks autophagy function of FIP200 specifically. Additionally, we found that the FIP200 C-terminal region (FIP200-CT) could interact with p62, consistent with recent studies showing its importance in degrading p62 aggregates. Moreover, we found that FIP200 can regulate TBK1 activation, which can phosphorylate p62 to regulate its degradation, and that FIP200 also interacts with the TBK1 adaptor, AZI2. In a second set of prelim studies, we analyzed the role of another autophagy gene Beclin1 in NSCs employing new Becn1 KI mice and found that increased autophagy protected NSC pool and their neurogenesis in aged Becn1 KI mice without affecting NSC in young mice. Lastly, we obtained additional prelim data suggesting that oxidative stress may synergize with autophagy-deficiency to promote p62 aggregate formation. Building upon these preliminary and prior studies, we propose to 1). investigate the mechanisms of non-canonical functions of FIP200 and potential synergy with its autophagy function in the regulation of NSCs, 2). analyze the role and mechanisms of enhanced autophagy to prevent NSC pool decline and promote neurogenesis in Becn1 KI mice, and 3). explore the role and mechanisms of NSC maintenance and neurogenesis in autophagy-deficiency mice after oxidative insult and during aging. Together, these studies will significantly advance our understanding of the regulation of NSC and neurogenesis by autophagy genes that may contribute to future design of effective therapies for neurodegenerative and other related diseases.

在成年大脑中，神经祖细胞/干细胞（NSC）负责产生新的神经元 维护现有电路和受伤后。 NSC维护和/或缺乏 神经发生有助于发育缺陷和神经退行性疾病 对于哪个衰老是主要的危险因素。 拟议研究的长期目标是确定分子和细胞 年轻动物和老年动物的NSC调节机制，这可能导致发展 神经退行性疾病的有效疗法。自噬是一种高度保守的细胞过程 散装细胞质材料的降解，用于维持细胞体内稳态和功能障碍 自噬已与各种疾病有关，包括神经变性和其他年龄有关 疾病。最初在我们的实验室中鉴定 随后显示为ULK1/ATG13/FIP200复合物的一个组成部分。 自噬。在上一个资金期间，我们发现删除FIP200，但没有其他自噬基因 ATG5，ATG7和ATG16L1导致NSC维持和神经发生有缺陷，这表明了潜在的作用 FIP200在NSC调节中通过控制p62骨料形成的非典型功能。在 其他预定研究，我们获得了严格的遗传证据，表明FIP200的非规范功能为 NSC维持和神经发生的必需品以及另一种独特的小鼠模型的分析 NSC中的FIP200-4A突变，可阻止FIP200的自噬功能。另外，我们发现 FIP200 C末端区域（FIP200-CT）可以与p62相互作用，这与最近的研究一致 它在降解p62骨料中的重要性。此外，我们发现FIP200可以调节TBK1激活， 可以磷酸化p62调节其降解，并且FIP200也与TBK1适配器相互作用， Azi2。在第二组初步研究中，我们分析了另一个自噬基因beclin1在NSC中的作用 采用新的BECN1 KI小鼠，发现自噬受保护的NSC池及其神经发生增加 在年龄的BECN1 KI小鼠中，没有影响年轻小鼠的NSC。最后，我们获得了其他预定数据 表明氧化应激可能会随着自噬缺陷促进p62骨料形成而协同作用。 在这些初步和先前的研究的基础上，我们建议1）。研究FIP200非典型函数的机理以及其在NSCS的调节中的自噬功能的潜在协同作用，2）。 分析增强自噬的作用和机制，以防止NSC池的下降和促进 BECN1 Ki小鼠的神经发生，3）。探索NSC维护的作用和机制 氧化侮辱和衰老期间自噬缺陷小鼠的神经发生。这些研究将在一起 显着提高我们对自噬基因对NSC和神经发生的调节的理解 可能有助于对神经退行性和其他相关疾病的有效疗法的未来设计。