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.

项目总结/摘要 越来越多的证据表明，与其他神经系统疾病一样， 发育障碍，有功能失调的突触连接。这些神经元连接缺陷 可能导致TSC的症状，如认知缺陷，自闭症和癫痫。然而，缺陷 人类神经元的突触发育和功能仅从动物模型研究中提出 以及最近一些使用人类神经元的研究，这些研究导致了相互矛盾的报告。直接解决这些问题 基本问题，并帮助解决相互矛盾的结果，我们将比较的发展和功能， TSC患者神经元与其校正的对照对应物的人皮层神经元突触。皮质 神经元从TSC患者产生的人诱导多能干细胞（hiPSC）分化 细胞及其工程化的同基因对应物。利用一系列形态测量和分子信号 分析，我们将研究结构和功能的突触前和突触后隔室的控制， 混合基因型培养物中的TSC 2突变突触。我们最近令人惊讶的发现表明，TSC 2的功能 不依赖于生长锥中的mTORC来直接调节细胞骨架和控制轴突导向， 促使我们考虑类似的机制可能在树突棘发育中起作用。我们还将测试 突触传递和网络活动如何在TSC 2患者神经元中改变。从长远来看， 我们相信我们的研究可能有助于确定TSC患者的关键药物靶点。