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（由申请方提供）：前脑室管膜下区（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在中枢神经系统中通常被细胞接触抑制，在中风诱导的神经元损失后大大增加。这帮助我们发现了一个新的粘着斑激酶（FAK）-JNK信号通路，该通路通过配体与神经胶质细胞中的DvE5整联蛋白结合而激活，并有效地抑制其CNTF表达。重要的是，用药理学FAK抑制剂的全身治疗刺激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介导的中风诱导的CNTF，这可能是抵消中风诱导的LIF和炎症在第一周。在体外，玻连蛋白抑制CNTF和刺激LIF，可能代表一个血管源性调节神经发生。在这里，我们将定义这些途径在体内，使用现有的转基因小鼠和药理学药物在成年小鼠的组合。具体来说，目标1a将确定是否玻连蛋白抑制CNTF表达通过DvE5整合素FAK途径在SVZ星形胶质细胞在正常条件下。这将通过在玻连蛋白-/-小鼠中的脑内拯救实验来完成，注射针对Dv和E5整联蛋白亚基的整联蛋白阻断抗体，使用E5整联蛋白-/-和诱导型星形胶质细胞FAK-/-小鼠，并确定对祖细胞与神经干细胞（NSC）的影响。目的1b将测试这是否是由JNK和/或其他途径介导的，使用脑内注射药理学抑制剂和JNK-/-小鼠，同时测量对CNTF表达和神经发生的影响。目的2a将确定卒中诱导的血源性玻连蛋白的渗漏是否导致LIF表达增加以刺激NSC自我更新。这将在大脑中动脉闭塞（MCAO）模型中通过静脉注射玻连蛋白-/- 小鼠目的2b将确定是否相同的整合素-FAK通路在MCAO后发挥作用，以及CNTF和LIF是否可以通过FAK抑制剂调节。目的2c将确定LIF是否参与抑制神经发生或为后来CNTF诱导的神经发生对SVZ进行预处理。目标3a。我们将测试是否全身性FAK抑制剂治疗更有效地诱导SVZ CNTF和神经发生时，开始后，炎症已经解决后MCAO，或目标3b时，间歇给予。FAK通过与D2多巴胺受体不同的机制起作用，我们发现D2多巴胺受体通过增加星形胶质细胞CNTF来增加SVZ神经发生。因此，Aim 3c将结合联合收割机FAK抑制剂和D2激动剂，以最大化CNTF表达和MCAO后的神经发生，并确定这是否允许更多的新神经元填充损伤组织。将通过行为测试评估这些治疗对感觉运动功能的潜在影响。