Filamin A in TSC

TSC 中的细丝蛋白 A

• 批准号: 9204864
• 负责人: Angelique Bordey
• 金额: $ 36.64万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-01 至 2021-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9204864
• 关键词: Actins; Address; Affect; Alpha Cell; Binding; Binding Proteins; Blood - brain barrier anatomy; Brain; Cells; Cellular biology; Charge; Child; Cognitive; Combined Modality Therapy; Complex; Cortical Malformation; Data; Defect; Dendrites; Dendritic Spines; Development; Disease; Electroporation; Epilepsy; Excision; F-Actin; FRAP1 gene; Frequencies; Goals; Grant; Hereditary Disease; Hyperactive behavior; Individual; Lesion; Life; Light; MAPK3 gene; Medial; Membrane; Mendelian disorder; Modeling; Motor Seizures; Mutation; Neurons; Operative Surgical Procedures; Pain; Pathogenicity; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacology; Plasmids; Prefrontal Cortex; Proto-Oncogene Proteins c-akt; Publishing; Refractory; Reporting; Seizures; Signal Transduction; Sirolimus; Site; Synapses; Synaptic Transmission; TSC1 gene; TSC1/2 gene; TSC2 gene; Testing; Therapeutic; Time; Tuberous sclerosis protein complex; Vertebral column; base; biophysical properties; brain malformation; crosslink; experimental study; filamin; hippocampal pyramidal neuron; in utero; in vivo; inhibitor/antagonist; knock-down; malformation; migration; mouse model; neonate; neuron development; new therapeutic target; novel; novel therapeutics; olfactory bulb; overexpression; prevent; public health relevance; receptor; small hairpin RNA; tool; transcription factor

 DESCRIPTION (provided by applicant): This proposal will test the hypothesis that decreasing Filamin A (FLNA) levels or function in tuberous sclerosis complex (TSC) prevents cortical malformations and associated seizure activity. TSC is caused by mutations in TSC1 or TSC2 leading to mTOR complex 1 (mTORC1) hyperactivity and cortical malformations associated with seizures and worsening of cognitive and psychiatric deficits. The mTORC1 inhibitor rapamycin is the only therapeutic option, does not rescue all defects, and has mild to life-threatening complications emphasizing the need to find novel drug targets. We recently reported that the level of the actin cross-linking molecule FLNA is increased in Tsc1null neurons and that this increase is responsible for dendrite abnormalities. In addition, FLNA regulates the migration of cortical neurons during development. These findings are attractive with respect to TSC for the following reasons: (1) Neuronal dysmorphogenesis (including dendritic abnormality) and stalled migration are hallmarks of TSC-related cortical malformations. (2) FLNA increase in Tsc1null neurons as well as in cells expressing a constitutively active Rheb (the mTORC1 canonical activator) was not mTORC1-dependent but rather ERK1/2-dependent, opening a novel pharmacological option for a possible combination therapy, and (3) Our preliminary data show that normalizing FLNA levels using shRNA or administrating the new FLNA modulator, PTI-125, during development prevents neuronal misplacement and dysmorphogenesis in our new model of TSC-related cortical malformations. To address our hypothesis we have the following three aims. In Aim 1, we will determine whether FLNA controls development of cortical pyramidal neurons in vivo and whether decreasing FLNA levels during development prevents cortical malformations. In Aim 2, we will examine whether there is a time-window in neonates for treatments aimed at preventing cortical malformations and reducing or eliminating associated seizure activity. Our new murine model of focal cortical malformations is associated with a high rate of daily convulsive seizures. Finally in Aim 3, we will investigate how increased FLNA levels leads to dysmorphogenesis and stalled migration, which may identify novel FLNA binding partners involved in cortical defects in TSC. Most experiments will use in utero electroporation to selectively manipulate the development of layer 2/3 cortical pyramidal neurons. This is a 2 PIs grant. The Bordey lab will be in charge of Aim 1 and 2, and the Calderwood lab will be in charge of Aim 3. Both labs will heavily interact on a weekly basis due to the need for tool (plasmid) development and the need for cell biology and in vivo experiments in Aim 2 and 3, respectively. Dr. Bordey is an expert on neuronal development and TSC, and Dr. Calderwood is a cell biologist with extensive expertise on FLNA.

 描述（由适用提供）：该提案将检验以下假设：结节性硬化症复合物（TSC）中降低Filemin a（FLNA）水平或功能可防止皮质畸形和相关的癫痫发作活性。 TSC是由TSC1或TSC2中的突变引起的，导致MTOR复合物1（MTORC1）多动和皮质畸形与癫痫发作有关，并担心认知和精神病缺陷。 MTORC1抑制剂雷帕霉素是唯一的治疗选择，并未挽救所有缺陷，并且危及生命的并发症是轻度的，强调需要寻找新的药物靶标。我们最近报道说，TSC1NULL神经元中肌动蛋白交联的分子FLN​​A的水平升高，并且这种增加导致树突异常。此外，FLNA调节发育过程中皮质神经元的迁移。由于以下原因，这些发现在TSC方面具有吸引力：（1）神经元畸形发生（包括树突异常），而停滞的迁移是与TSC相关的皮质畸形的标志。 （2）TSC1NULL神经元以及表达组成型活性Rheb（MTORC1规范激活剂）的细胞的FLNA并不依赖MTORC1依赖性，而是ERK1/2依赖性，为可能的组合疗法提供了新的药物，并且（3）我们的shrna级别（3） PTI-125在发育期间，在我们的新的TSC相关皮质畸形模型中阻止了神经元的肌电症和畸形发生。为了解决我们的假设，我们有以下三个目标。在AIM 1中，我们将确定FLNA在体内控制皮质锥体神经元的发展以及在发育过程中降低FLNA水平是否阻止皮质畸形。在AIM 2中，我们将检查新生儿中是否有一个时间窗口治疗旨在防止皮质畸形，减少或消除相关的癫痫发作活性。我们新的局灶性皮质畸形模型与每日抽搐癫痫发作的高率有关。最终，在AIM 3中，我们将研究增加的FLNA水平如何导致畸形发生和停滞的迁移，这可以鉴定出与TSC中皮质缺陷有关的新型FLNA结合伴侣。大多数实验将用于子宫电穿孔中，以选择性地操纵2/3层皮质金字塔神经元的发展。这是2个PIS赠款。 Bordey Lab将负责AIM 1和2，Calderwood Lab将负责AIM 3。由于需要工具（质粒）开发以及对AIM 2和3的细胞生物学和体内实验的需求，这两个实验室都将每周进行一次大量的相互作用。 Bordey博士是神经元发展和TSC的专家，Calderwood博士是一名在FLNA方面拥有广泛专业知识的细胞生物学家。