Investigation of Mechanosensory Signaling in the Pathogenesis of Traumatic Brain Injury in Human iPSC-derived Cortical Brain Organoids

人 iPSC 衍生的皮质脑类器官中机械感觉信号传导在创伤性脑损伤发病机制中的研究

• 批准号: 10548870
• 负责人: Joshua Berlind
• 金额: $ 4.23万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-15 至 2023-04-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10548870
• 关键词: 3-Dimensional; 5' -AMP-activated protein kinase; Acute; Animal Model; Animals; Antisense Oligonucleotides; Anxiety; Astrocytes; Autopsy; Biochemical; Biology; Biophysical Process; Blood; Brain; Case Study; Cations; Cell Line; Cells; Chemicals; Chronic; Clinical; Complex; Data; Dementia; Diffuse Axonal Injury; Disease; Edema; Elderly; Environmental Risk Factor; Etiology; Event; Focused Ultrasound Therapy; Frontotemporal Dementia; Functional disorder; Gene Expression; General Population; Genetic; Genetic Transcription; Goals; Human; Immunoassay; Immunologic Factors; In Vitro; Incidence; Individual; Inflammation; Injury; Investigation; Measures; Mechanics; Mental Depression; Metabolic dysfunction; Military Personnel; Modeling; Molecular; Mus; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neuronal Injury; Neurons; Organoids; Outcome; Pathogenesis; Pathologic; Pathology; Patients; Phenotype; Phosphotransferases; Process; Risk; Role; Sampling; Signal Transduction; Survival Rate; Symptoms; System; TBI Patients; TDP-43 aggregation; Testing; Time; Tissue Model; Transgenic Mice; Trauma; Traumatic Brain Injury; Ultrasonics; United States; Western Blotting; Work; axon injury; blood-brain barrier disruption; cell injury; cell type; controlled cortical impact; dementia risk; disability; disease-causing mutation; hyperphosphorylated tau; improved; in vivo; in vivo Model; induced pluripotent stem cell; injured; innovation; insight; knock-down; mechanotransduction; mortality; mouse model; nerve stem cell; neuron loss; neuronal survival; neuroprotection; new therapeutic target; novel; preservation; pressure; prevent; programs; protein TDP-43; response; severe injury; tau Proteins; tau aggregation; tau-1; therapeutic target; transcriptome sequencing; transcriptomics

Project Summary/Abstract Global incidence of Traumatic Brain Injury (TBI) is on the rise, particularly among athletes, military personnel, and elderly citizens1. TBI represents a spectrum of mild to severe injuries that share many long-term pathologies and clinical symptoms such as Tau and TDP43 pathology, axonal injury, neuronal death, and increased risk of depression and neurodegenerative diseases2-6. Although end-stage pathologies are well characterized from post-mortem samples, the molecular mechanisms contributing to injury remain poorly understood in part due to the difficulty of studying live brains in human patients and the variable biophysical processes that occur between patients. As a result, available treatment options remain limited and are largely ineffective61. Previous studies of TBI conducted in animal models present with edema, inflammation, and blood-brain barrier disruption following an induced trauma6,7. Although important to the pathophysiology of TBI, this complex cascade of events complicates our ability to accurately understand cell autonomous injury mechanisms. Therefore, a reductionist system to study the response of distinct cell types to TBI would be beneficial for identifying and targeting changes that occur post-injury. To this end, we will utilize a 3-D cortical organoid culture system grown from human induced Pluripotent Stem Cells (iPSCs) to elucidate cell autonomous mechanisms of injury and degeneration caused by TBI8. We have developed a unique system of focused ultrasonic injury to mimic TBI in vitro which recapitulates key pathologic and transcriptional features of in vivo models. This allows for detailed study of both acute and chronic changes following an induced trauma, and provides a platform to integrate environmental and genetic contributions to injury while preserving human-specific biology. Using this system, we will combine bulk RNA-seq transcriptomic analysis with biochemical and immunoassays to uncover novel cell-type specific injury mechanisms. Using iPSC lines from patients with neurodegenerative diseases, we will test how TBI modulates genetically-induced disease mechanisms. Finally, we will test a mouse model of cortical controlled impact (CCI) to validate our findings in vivo. This proposal will greatly enhance our understanding of specific injury mechanisms in discrete cell types and may help to identify novel neuroprotective therapeutic targets.

项目摘要/摘要 全球创伤性脑损伤(TBI)的发病率正在上升，特别是在运动员、军事人员、 和老年公民1。脑损伤代表了一系列轻微到严重的损伤，这些损伤具有许多长期的病理特征。 和临床症状，如Tau和TDP43病理，轴突损伤，神经元死亡和增加的风险 抑郁症和神经退行性疾病2-6。尽管终末期病理的特征很好地表现为 在死后样本中，造成损伤的分子机制仍然知之甚少，部分原因是 研究人类患者活体大脑的困难以及发生在以下两种生物物理过程之间的差异 病人。因此，现有的治疗选择仍然有限，而且在很大程度上是无效的。以前的研究 在出现水肿、炎症和血脑屏障破坏的动物模型上进行脑损伤 诱发创伤6，7.尽管对脑外伤的病理生理学很重要，但这一复杂的事件级联反应 使我们准确理解细胞自主损伤机制的能力变得复杂。因此，一个简化论者 研究不同细胞类型对脑损伤的反应的系统将有助于识别和定位变化 是在受伤后发生的。为此，我们将利用从人类生长的三维皮质器官培养系统 诱导多能干细胞(IPSCs)阐明细胞损伤和退化的自主机制 由TBI8引起。我们开发了一种独特的体外模拟脑损伤的聚焦超声损伤系统，该系统 概述了体内模型的关键病理和转录特征。这样就可以对两者进行详细的研究 诱发创伤后的急性和慢性变化，并提供一个平台来整合环境和 基因对伤害的贡献，同时保存人类特有的生物学。使用这个系统，我们将合并批量 结合生化和免疫分析的RNA-seq转录分析揭示新的细胞类型特异性损伤 机制。使用神经退行性疾病患者的IPSC株，我们将测试TBI是如何调节的 基因导致的疾病机制。最后，我们将测试皮质受控撞击(CCI)的小鼠模型。 在活体内验证我们的发现。这一建议将极大地提高我们对具体伤害的理解 分离细胞类型的机制，并可能有助于确定新的神经保护性治疗靶点。