Targeting CNTF to increase adult forebrain neurogenesis

靶向 CNTF 增加成人前脑神经发生

• 批准号: 8722255
• 负责人: THEO HAGG
• 金额: $ 29.93万
• 依托单位: EAST TENNESSEE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8722255
• 关键词: Acute; Adult; Agonist; Astrocytes; Blocking Antibodies; Blood; Blood Vessels; Brain; Cell Proliferation; Cells; Ciliary Neurotrophic Factor; Clinical Trials; Data; Disease; Dopamine D2 Receptor; Extravasation; Fibrinogen; Focal Adhesion Kinase 1; Funding; Genetic; Grant; In Vitro; Inflammation; Injection of therapeutic agent; Integrin Signaling Pathway; Integrins; LIF gene; Ligand Binding; Ligands; MAPK8 gene; Malignant Neoplasms; Measures; Mediating; Middle Cerebral Artery Occlusion; Modeling; Molecular Target; Mus; Nervous system structure; Neuroglia; Neurons; Pathway interactions; Pharmaceutical Preparations; Pharmacological Treatment; Play; Production; Prosencephalon; Role; Sensorimotor functions; Signal Pathway; Stem cells; Stroke; Testing; Tissues; Transgenic Mice; Treatment Protocols; Vitronectin; Work; behavior test; in vivo; inhibitor/antagonist; injured; interest; kinase inhibitor; motor function improvement; nerve stem cell; nervous system disorder; neuroblast; neurogenesis; neuron loss; preconditioning; progenitor; public health relevance; research study; self-renewal; subventricular zone; treatment effect

DESCRIPTION (provided by applicant): Ciliary neurotrophic factor (CNTF) in the subventricular zone (SVZ) of the forebrain promotes neurogenesis in adult mice. We found that CNTF is normally repressed in the CNS by cell contact and increases greatly after stroke-induced neuron loss. This helped us to discover a new focal adhesion kinase (FAK)-JNK signaling pathway that is activated by ligand binding to DvE5 integrin in glia and potently inhibit their CNTF expression. Importantly, systemic treatment with pharmacological FAK inhibitors stimulates SVZ CNTF and neurogenesis. Our data show that stroke-induced neurogenesis is mediated entirely by stroke-induced CNTF, which may be counteracted by stroke-induced LIF and inflammation over the first week. In vitro, vitronectin represses CNTF and stimulates LIF, perhaps representing a vascular-derived regulator of neurogenesis. Here we will define these pathways in vivo, using a combination of available transgenic mice and pharmacological drugs in adult mice. Specifically, Aim 1a will determine whether vitronectin represses CNTF expression through the DvE5 integrin-FAK pathway in SVZ astrocytes under normal conditions. This will be done by intracerebral rescue experiments in vitronectin-/- mice, injecting integrin blocking antibodies against Dv and E5 integrin subunits, using E5 integrin-/- and inducible astroglial FAK-/- mice and defining effects on progenitors vs. neural stem cells (NSCs). Aim 1b will test whether this is mediated by JNK and/or other pathways, using intracerebral injections of pharmacological inhibitors and JNK-/- mice, while measuring effects on CNTF expression and neurogenesis. Aim 2a will determine whether stroke-induced leakage of blood-derived vitronectin causes increased LIF expression to stimulate NSC self-renewal. This will be done in a middle cerebral artery occlusion (MCAO) model by i.v. injections of vitronectin in vitronectin-/- mice. Aim 2b will determine whether the same integrin-FAK pathways play a role after MCAO, and whether CNTF and LIF can be regulated by FAK inhibitors. Aim 2c will define whether LIF is involved in repressing neurogenesis or is preconditioning the SVZ for later CNTF-induced neurogenesis. Aim 3a. Wwill test whether systemic FAK inhibitor treatment is more effective in inducing SVZ CNTF and neurogenesis when started after inflammation has resolved following MCAO, or Aim 3b when given intermittently. FAK acts via a different mechanism than the D2 dopamine receptor, which we showed increases SVZ neurogenesis by increasing astroglial CNTF. Therefore, Aim 3c will combine FAK inhibitor and D2 agonist to maximize CNTF expression and neurogenesis after MCAO, and determine whether this allows more new neurons to populate the injured tissue. The potential effects of these treatments on sensorimotor function will be assessed by behavioral tests.

描述（由申请人提供）：前脑室内室内区（SVZ）中的睫状神经营养因子（CNTF）促进成年小鼠的神经发生。我们发现，CNTF通常通过细胞接触在中枢神经系统中被抑制，并在中风引起的神经元丧失后大大增加。这有助于我们发现新的局灶性粘附激酶（FAK）-JNK信号传导途径，该途径通过配体在神经胶质中与DVE5整合素的结合激活并有效抑制其CNTF表达。重要的是，用药理学FAK抑制剂进行全身治疗会刺激SVZ CNTF和神经发生。我们的数据表明，中风诱导的神经发生完全是由中风诱导的CNTF介导的，这可能会在第一周在中风诱导的LIF和炎症中抵消。在体外，玻璃纤维抑制CNTF并刺激LIF，也许代表了神经发生的血管衍生的调节剂。在这里，我们将在成年小鼠中使用可用的转基因小鼠和药理学药物的组合来定义这些途径。具体而言，AIM 1A将确定在正常条件下SVZ星形胶质细胞中的DVE5整合蛋白-FAK途径是否通过DVE5整联蛋白-FAK途径抑制CNTF表达。这将通过使用E5整合素 - / - / - 和诱导的星形胶质FAK - / - 小鼠和对祖细胞（NSCS）的效果（NSC）来完成，从而通过脑中救援实验在玻璃素蛋白 - / - 小鼠中进行了整联蛋白阻断抗体，对DV和E5整合蛋白亚基进行了抗体。 AIM 1b将使用药理学抑制剂和JNK - / - 小鼠的脑内注射JNK和/或其他途径介导的，同时测量对CNTF表达和神经发生的影响。 AIM 2A将确定中风诱导的血液来源玻璃纤维泄漏是否会导致LIF表达增加以刺激NSC自我更新。这将在I.V.中的大脑中部动脉闭塞（MCAO）模型中完成。注射玻霉素 - / - 老鼠。 AIM 2B将确定相同的整联蛋白FAK途径是否在MCAO之后起作用，以及CNTF和LIF是否可以受到FAK抑制剂的调节。 AIM 2C将定义LIF是否参与抑制神经发生或正在预处理SVZ以进行后来CNTF诱导的神经发生。目标3a。 WWILL测试系统性FAK抑制剂治疗是否在炎症后启动后诱导SVZ CNTF和神经发生更有效，在MCAO后炎症后解决，或者在间歇性地给予时AIG 3B。 FAK通过与D2多巴胺受体不同的机制起作用，我们显示出通过增加星形胶质细胞CNTF来增加SVZ神经发生。因此，AIM 3C将结合FAK抑制剂和D2激动剂，以最大化MCAO后CNTF表达和神经发生，并确定这是否允许更多的新神经元填充受伤的组织。这些处理对感官功能的潜在影响将通过行为测试评估。