Molecular mechanisms of abnormal dendritic spine development and function in human neurons with TSC2 disease mutations

TSC2疾病突变人类神经元树突棘发育和功能异常的分子机制

• 批准号: 10360715
• 负责人: Timothy M Gomez
• 金额: $ 40.76万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-20 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10360715
• 关键词: Address; Affect; Animal Model; Area; Astrocytes; Axon; Biological; Biological Assay; Calcium; Cell Line; Cells; Coculture Techniques; Cognitive deficits; Collaborations; Conflict (Psychology); Cues; Cytoskeleton; Data; Defect; Dendritic Spines; Development; Disease; Disease model; Drug Screening; Elements; Engineering; Epilepsy; Fluorescence Microscopy; Genomics; Genotype; Growth Cones; Human; Image; Image Analysis; In Vitro; Label; Laboratories; Letters; Measurement; Measures; Mediating; Methods; Molecular; Morphology; Mutation; Nervous system structure; Neurites; Neurodevelopmental Disorder; Neurons; Patients; Pilot Projects; Prosencephalon; Protein Biosynthesis; Regulation; Reporting; Research; Role; Series; Signal Transduction; Site; Structure; Study models; Supporting Cell; Symptoms; Synapses; Synaptic Transmission; TSC1 gene; TSC2 gene; Testing; Therapeutic Intervention; Time; Tuberous sclerosis protein complex; Vertebral column; Work; autism spectrum disorder; axon growth; axon guidance; cortical tubers; disease-causing mutation; druggable target; experimental study; gene correction; human stem cells; induced pluripotent stem cell; loss of function; mutant; neural network; presynaptic; protein complex; quantitative imaging; synaptic function; synaptogenesis

Project Summary/Abstract Growing evidence suggests that patients with Tuberous Sclerosis Complex (TSC), like other neuro- developmental disorders, have dysfunctional synaptic connections. These defects in neuronal connectivity likely contribute to symptoms of TSC, such as cognitive deficits, autism and epilepsy. However, defective synaptic development and function by human neurons has only been suggested from animal model studies and a few recent studies using human neurons, which have led to conflicting reports. To directly address these fundamental questions and help resolve conflicting findings, we will compare the development and function of human cortical neuron synapses of TSC patient neurons with their corrected control counterparts. Cortical neurons are differentiated from human induced pluripotent stem cells (hiPSCs) generated from TSC patient cells and their engineered isogenic counterparts. Using a series of morphometric and molecular signaling assays, we will examine the structure and function of pre- and post-synaptic compartments of control and TSC2 mutant synapses in mixed genotype cultures. Our surprising recent findings suggest that TSC2 functions independent of mTORC in growth cones to directly regulate the cytoskeleton and control axon guidance, prompting us to consider similar mechanisms may function in dendritic spine development. We will also test how synaptic transmission and network activity may be altered in TSC2 patient neurons. Over the long term, we believe our research may help identify key druggable targets in patients with TSC.

项目摘要/摘要 越来越多的证据表明，与其他神经 - 发育障碍，具有功能失调的突触连接。这些缺陷在神经元连通性中 可能导致TSC的症状，例如认知缺陷，自闭症和癫痫。但是，有缺陷 人类神经元的突触发育和功能仅在动物模型研究中提出 以及最近使用人类神经元的一些研究导致报告矛盾。直接解决这些 基本问题并帮助解决相互矛盾的发现，我们将比较 TSC患者神经元的人体皮质神经元突触具有校正的对照对应物。皮质 神经元与TSC患者产生的人类诱导的多能干细胞（HIPSC）区分开 细胞及其工程的等源性对应物。使用一系列形态计量学和分子信号传导 测定，我们将检查控制前和突触后室的结构和功能， 混合基因型培养物中的TSC2突变体突触。我们令人惊讶的最近发现表明TSC2功能 在生长锥中独立于MTORC，直接调节细胞骨架和控制轴突指导， 促使我们考虑类似的机制可能在树突状棘发育中起作用。我们还将测试 在TSC2患者神经元中，突触传播和网络活性如何改变。从长远来看， 我们认为我们的研究可能有助于确定TSC患者的关键毒品目标。