Examining the role of the Lissencephaly protein, Lis1, in dynein-based transport

检查无脑畸形蛋白 Lis1 在基于动力蛋白的运输中的作用

基本信息

批准号：
8608604
负责人：
DEANNA S SMITH
金额：
$ 30.29万
依托单位：
UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-01-01 至 2017-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8608604
关键词：
APC gene Adult Adverse effects Afferent Neurons Agonist Amyotrophic Lateral Sclerosis Animal Model Animals Axon Axonal Transport Behavioral Binding Biochemical Brain Cell Line Cerebral cortex Clinical Colon Carcinoma Complex Cyclin-Dependent Kinase 5 Defect Development Disease Dissection Dynein ATPase Embryo Event Gene Mutation Genes Genetic Glycogen Synthase Kinases Human Huntington Disease In Vitro Insulin Intervention Knockout Mice LIS1 protein Late-Onset Disorder Life Link Lithium Lithium Chloride Lysosomes Mammals Microtubule-Associated Proteins Microtubules Mind Mitochondria Molecular Monitor Mood Disorders Moods Motor Mus Mutation Nerve Neurologic Neurologic Dysfunctions Neuronal Dysfunction Neurons Non-Insulin-Dependent Diabetes Mellitus Nuclear Hormone Receptors Organelles Pathway interactions Peroxisome Proliferator-Activated Receptors Pharmaceutical Preparations Phosphorylation Phosphotransferases Plus End of the Microtubule Positioning Attribute Process Proteins Rattus Regulation Role Schizophrenia Speed Symptoms Syndrome Takeda brand of pioglitazone hydrochloride Testing Therapeutic Work base brain malformation cellular imaging design genetic manipulation inhibitor/antagonist insulin sensitizing drugs interest late disease onset lissencephaly mutant nervous system disorder novel overexpression prevent research study response retrograde transport rosiglitazone synaptic function

项目摘要

DESCRIPTION (provided by applicant): Lissencephaly, an autosomal dominant brain malformation caused by mutations in the LIS1 gene, was the first neurological disorder linked directly to cytoplasmic dynein function. This discovery led the way for molecular dissection of events regulating development of the mammalian cerebral cortex. Lis1 and a binding partner, Nudel, bind directly to dynein and regulate its activity. This interaction has been studied almost exclusively in the context of brain development. More recently, gene products associated with later-onset disorders such as schizophrenia, amyotrophic lateral sclerosis (ALS), Perry Syndrome, and Huntington's disease (HD) have been shown to interact directly with dynein pathways. The discovery of an important role for Lis1 and Nudel in regulating dynein-dependent axonal transport in cultured adult rat sensory neurons has led to the hypothesis that perturbing this regulatory network will cause neuronal dysfunction in mice. To test this, the pathological consequences of disrupting Lis1 in post-developmentally will be determined using floxed-Lis1 and Cre strains. Histological, biochemical, and behavioral studies will be carried out to determine if depletion of Lis1 post- developmentally causes neurological dysfunction. Because this could exacerbate disease symptoms in lissencephaly and other neurological disorders, these studies may provide a feasible target for clinical intervention by drugs. Possible candidates are the PPAR3 agonists Avandia and Actos, insulin sensitizers normally used to treat type 2 diabetes. Recent studies indicate that these drugs stimulate dynein in several cell lines. This is blocked by mutations in APC, a microtubule plus end associated protein typically linked to colon cancer. APC has also been found to have critical roles in neurons, influencing both synaptic function and axonal transport. Notably, the dynein response to PPAR3 agonists requires PI3K activity and is mimicked by lithium, a potent Gsk32 inhibitor used to treat mood disorders. Moreover, dynein is a target of Gsk32 in vitro and its phosphorylation results in a 5-fold increase in coimmunoprecipitation of an APC fragment suggesting that inhibition of the kinase could impact dynein interactions. Dynein distribution in sensory neurons is altered in response to PPAR3 agonists, leading to the hypothesis that PPAR3 pathways contribute to regulation of dynein-dependent axonal transport in adult neurons. This will be tested by determining the effect of pharmacological and genetic manipulation of PPAR3/Gsk32/APC pathways on organelle transport in adult rat DRG neurons and determining if crosstalk occurs between Lis1 and PPAR3 pathways.

描述（由申请人提供）：无脑畸形是一种由 LIS1 基因突变引起的常染色体显性脑畸形，是第一种与细胞质动力蛋白功能直接相关的神经系统疾病。这一发现为对哺乳动物大脑皮层发育调节事件的分子解析开辟了道路。 Lis1 和结合伴侣 Nudel 直接与动力蛋白结合并调节其活性。这种相互作用几乎完全是在大脑发育的背景下进行的研究。最近，与精神分裂症、肌萎缩侧索硬化症 (ALS)、佩里综合症和亨廷顿病 (HD) 等迟发性疾病相关的基因产物已被证明与动力蛋白通路直接相互作用。 Lis1 和 Nudel 在调节培养的成年大鼠感觉神经元中动力蛋白依赖性轴突运输中的重要作用的发现导致了这样的假设：干扰该调节网络将导致小鼠神经元功能障碍。为了测试这一点，将使用 floxed-Lis1 和 Cre 菌株确定发育后破坏 Lis1 的病理后果。将进行组织学、生化和行为研究，以确定发育后 Lis1 的缺失是否会导致神经功能障碍。由于这可能会加剧无脑畸形和其他神经系统疾病的疾病症状，因此这些研究可能为临床药物干预提供可行的目标。可能的候选药物是 PPAR3 激动剂 Avandia 和 Actos，这两种胰岛素增敏剂通常用于治疗 2 型糖尿病。最近的研究表明，这些药物可刺激多种细胞系中的动力蛋白。这一过程被 APC 突变所阻断，APC 是一种微管加末端相关蛋白，通常与结肠癌相关。 APC 还被发现在神经元中发挥着关键作用，影响突触功能和轴突运输。值得注意的是，动力蛋白对 PPAR3 激动剂的反应需要 PI3K 活性，并且可以通过锂来模拟，锂是一种用于治疗情绪障碍的有效 Gsk32 抑制剂。此外，动力蛋白是 Gsk32 的体外靶标，其磷酸化导致 APC 片段的免疫共沉淀增加 5 倍，表明抑制激酶可能影响动力蛋白相互作用。感觉神经元中的动力蛋白分布会因 PPAR3 激动剂而发生改变，从而得出这样的假设：PPAR3 通路有助于调节成体神经元中动力蛋白依赖性轴突运输。这将通过确定 PPAR3/Gsk32/APC 途径的药理学和遗传操作对成年大鼠 DRG 神经元细胞器运输的影响以及确定 Lis1 和 PPAR3 途径之间是否发生串扰来进行测试。