Inflammation mediated evolving changes in synapse structural integrity and dynamics in vivo in a mouse model of multiple sclerosis

多发性硬化症小鼠模型体内炎症介导的突触结构完整性和动态变化

• 批准号: 10411921
• 负责人: Rebecca Lynn Gillani
• 金额: $ 17.34万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-15 至 2023-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10411921
• 关键词: Address; Affect; Animals; Architecture; Area; Atrophic; Autopsy; Award; Axon; Biometry; Brain; Chimeric Proteins; Chronic; Clinical; Communication; Communities; Complement; Demyelinations; Dendritic Spines; Development; Disease; Disease model; Excitatory Synapse; Experimental Autoimmune Encephalomyelitis; Experimental Models; Functional disorder; General Hospitals; Goals; Image; Imaging technology; Immune; Immunology; Impaired cognition; Individual; Inflammation; Inflammatory; Inhibitory Synapse; K-Series Research Career Programs; Label; Lead; Leadership; Lesion; Literature; Location; Massachusetts; Mediating; Mentors; Methodology; Methods; Modeling; Monitor; Multiple Sclerosis; Multiple Sclerosis Lesions; Mus; Myelin; Nerve Degeneration; Nerve Fibers; Neurodegenerative Disorders; Neurodevelopmental Disorder; Neurologist; Neuronal Plasticity; Neurons; Outcome; Pathology; Pathway interactions; Persons; Physiological; Play; Population Heterogeneity; Postdoctoral Fellow; Pre-Clinical Model; Process; Proteins; Pyramidal Cells; Relapse; Research; Resolution; Rodent; Rodent Model; Role; Scaffolding Protein; Series; Shapes; Stroke; Structure; Symptoms; Synapses; Synaptic Receptors; Techniques; Technology; Testing; Time; Training; Trees; Visual Cortex; career development; cell motility; cellular imaging; density; disability; efficacy evaluation; experience; experimental study; fluorophore; gray matter; hippocampal pyramidal neuron; in vivo; in vivo imaging; in vivo two-photon imaging; innovation; interest; mouse model; multidisciplinary; multiple sclerosis patient; nervous system disorder; neuroimmunology; neuroinflammation; skills; therapeutic target; two-photon; white matter; young adult

This is an application for a K08 award for Dr. Rebecca Gillani, a neurologist at the Massachusetts General Hospital. She has rigorous prior training in neuroplasticity in rodent models of a neurodevelopmental disorder and stroke. In her initial post-doctoral experience, she has gained expertise in in vivo two-photon imaging technology working on a project to define basic principles of circuit wiring in the visual cortex. She now aims to take these state-of-the-art techniques for in vivo two-photon imaging of neurons and synapses and apply them to the study of neurodegeneration in rodent preclinical models of multiple sclerosis, her area of clinical interest. The goal of this career development award is to provide her the skills to make the transition from the study of physiologic circuit wiring and plasticity, to the independent study of disease models of neuroinflammation with training as follows: 1) Multi-disciplinary mentoring from experts in neurodegeneration, neuroimmunology, and in vivo two-photon imaging; 2) methodology for experimental autoimmune encephalomyelitis mouse model of multiple sclerosis; 3) didactic training in immunology and biostatistical methodology; and 4) communication and leadership career development. Her mentor Brian Bacskai is expert in in vivo two-photon imaging in neurodegenerative diseases, and her co-mentor Elly Nedivi is expert in in vivo two-photon imaging of neuron and synapse structure. Her advisors include experts in neurodegeneration, Rudolph Tanzi, and experts in neuroimmunology, Francisco Quintana and Staci Bilbo. While there is an immense literature and ongoing effort to define the immune mechanisms underlying multiple sclerosis pathology, a critical area of unmet need is defining the specific mechanisms of neuronal and synaptic dysfunction and degeneration during the course of multiple sclerosis. These neurodegenerative changes are a key determinant of disability in multiple sclerosis. It is likely that inflammation in early multiple sclerosis does not immediately lead to synapse loss. Instead there is more likely a series of evolving changes in synapse structural integrity and dynamics with the eventual outcome of synapse loss. In Aim 1, Dr. Gillani will leverage state-of-the- art in vivo two-photon imaging technologies for longitudinal monitoring of all the excitatory and inhibitory connections onto individual cortical pyramidal neurons in a living mouse in a model of multiple sclerosis. In Aim 2, Dr. Gillani will use new methods for reversible expansion and super-resolution imaging of cellular architecture to analyze for the presence of important synaptic proteins and inflammatory factors implicated in synaptic pathology in multiple sclerosis, by observing the same neurons and synapses imaged in live animals in Aim 1. The studies she proposes will provide a comprehensive picture of how diverse populations of synapses across the entire dendritic tree of cortical pyramidal cells undergo dynamic changes over 8 weeks in a mouse model of multiple sclerosis. This research will be the basis of a R01 application in year 4 of this proposed K08 award.

这是马萨诸塞州总医院神经科医生丽贝卡·吉拉尼 (Rebecca Gillani) 博士的 K08 奖项申请。她之前接受过神经发育障碍和中风啮齿动物模型的神经可塑性方面的严格培训。在最初的博士后经历中，她获得了体内双光子成像技术的专业知识，致力于定义视觉皮层电路布线的基本原理的项目。她现在的目标是利用这些最先进的神经元和突触体内双光子成像技术，并将其应用于多发性硬化症啮齿动物临床前模型的神经变性研究，这是她临床感兴趣的领域。该职业发展奖的目标是为她提供从生理电路布线和可塑性研究过渡到神经炎症疾病模型独立研究的技能，并接受以下培训：1）神经退行性疾病、神经免疫学和体内双光子成像专家的多学科指导； 2) 多发性硬化症实验性自身免疫性脑脊髓炎小鼠模型的方法学； 3）免疫学和生物统计方法学的教学培训； 4) 沟通和领导力职业发展。她的导师 Brian Bacskai 是神经退行性疾病体内双光子成像专家，她的合作导师 Elly Nedivi 是神经元和突触结构体内双光子成像专家。她的顾问包括神经退行性疾病专家鲁道夫·坦齐 (Rudolph Tanzi) 以及神经免疫学专家弗朗西斯科·金塔纳 (Francisco Quintana) 和斯塔西·比尔博 (Staci Bilbo)。虽然有大量的文献和持续的努力来定义多发性硬化症病理学背后的免疫机制，但未满足需求的一个关键领域是定义多发性硬化症过程中神经元和突触功能障碍和变性的具体机制。这些神经退行性变化是多发性硬化症残疾的关键决定因素。早期多发性硬化症的炎症很可能不会立即导致突触丧失。相反，突触结构完整性和动力学更有可能发生一系列不断演变的变化，最终导致突触损失。在目标 1 中，吉拉尼博士将利用最先进的体内双光子成像技术，对多发性硬化症模型中活体小鼠的单个皮质锥体神经元的所有兴奋性和抑制性连接进行纵向监测。在目标 2 中，Gillani 博士将使用细胞结构可逆扩张和超分辨率成像的新方法，通过观察目标 1 中在活体动物中成像的相同神经元和突触，分析与多发性硬化症突触病理学有关的重要突触蛋白和炎症因子的存在。她提出的研究将全面了解整个树突树上不同突触群体的情况。 在多发性硬化症小鼠模型中，皮质锥体细胞在 8 周内经历动态变化。这项研究将成为拟议的 K08 奖项第 4 年 R01 申请的基础。