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：
8618957
负责人：
DEANNA S SMITH
金额：
$ 29.53万
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/8618957
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）：Lissencephaly是由LIS1基因突变引起的常染色体显性脑畸形，是与细胞质动力蛋白功能直接相关的第一种神经系统疾病。这一发现导致了调节哺乳动物大脑皮层发育事件的分子解剖的道路。 Lis1和结合伴侣Nudel直接与动力蛋白结合并调节其活性。在大脑发育的背景下，几乎完全研究了这种相互作用。最近，与晚期发作的疾病相关的基因产物，例如精神分裂症，肌萎缩性侧面硬化症（ALS），佩里综合征和亨廷顿氏病（HD）（HD）已被证明与动力蛋白途径直接相互作用。 LIS1和NUDEL在调节培养的成年大鼠感觉神经元中调节Dynein依赖性轴突转运中的重要作用的发现导致了这样的假设，即扰动该调节网络会导致小鼠的神经元功能障碍。为了测试这一点，将使用Floxed-Lis1和Cre菌株确定破坏发育后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的耗竭是否引起神经功能障碍。因为这可能加剧脑膜炎和其他神经系统疾病的疾病症状，所以这些研究可能为药物临床干预提供了可行的靶标。可能的候选者是PPAR3激动剂Avandia和Actos，通常用于治疗2型糖尿病的胰岛素敏化剂。最近的研究表明，这些药物在几种细胞系中刺激动力蛋白。这是由APC突变阻断的，APC是一种微管加终端相关蛋白，通常与结肠癌相关。还发现APC在神经元中具有关键作用，从而影响突触功能和轴突运输。值得注意的是，对PPAR3激动剂的动力蛋白反应需要PI3K活性，并由锂（一种有效的GSK32抑制剂）模仿，用于治疗情绪障碍。此外，动力蛋白是GSK32体外的靶标，其磷酸化导致APC片段的共免疫沉淀增加了5倍，这表明激酶的抑制作用可能会影响Dynein相互作用。感觉神经元中的动力蛋白分布对PPAR3激动剂的响应改变，导致假设PPAR3途径有助于调节成年神经元中动力蛋白依赖性轴突转运。这将通过确定PPAR3/GSK32/APC途径对成年大鼠DRG神经元中细胞器转运的药理和遗传操纵的影响来测试，并确定在LIS1和PPAR3途径之间发生串扰。