A Phenotype-Optimized Zebrafish Model of Progressive Supranuclear Palsy

表型优化的进行性核上性麻痹斑马鱼模型

• 批准号: 10704465
• 负责人: Edward Alan Burton
• 金额: $ 34.16万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-15 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10704465
• 关键词: Phenotype; Optimized; Zebrafish; Model; Progressive

Abstract Progressive supranuclear palsy (PSP) is a neurodegenerative disease characterized clinically both by profound loss of balance and other neurological symptoms. Falls are central to early PSP presentation and diagnosis and lead to fractures and hospitalization. No current treatments improve balance or prevent disease progression. Strong pathological, molecular, and genetic evidence implicates deposits of the 4­repeat isoform of the micro­ tubule associated protein Tau (4R­Tau) in PSP pathophysiology. Current models struggle to link 4R­Tau pathol­ ogy to the debilitating falls that define PSP. Development of new therapies for PSP would benefit profoundly from a means to evaluate interventions in the context of the neural circuits responsible for balance. Leveraging the historical strengths of our individual laboratories, we have worked together to design a new model system to model Tau pathology in the context of balance. Our model system uses the larval zebrafish, a small vertebrate with exceptional molecular and optical accessibility and rigorously defined balance behavior. In unpublished pilot experiments have used a wide set of neurological and behavioral tests show that express­ ing 4R­Tau in zebrafish neurons gives rise to: (i) molecular and cellular pathology (ii) locomotor and oculomotor deficits (iii) systematic balance deficits that together resemble PSP. Here we propose to extend and validate this new and exciting model. In the R61 aim, to validate our model internally we will develop two complementary pipelines to assay balance behavior and establish new transgenic lines and protocols to gain temporal control of 4R­Tau expression. First, we will optimize an assay of balance with rigorously­defined quantitative behavioral endpoints suitable for effi­ cient evaluation of chemical modifiers or candidate gene targets. Next we will use a novel light­inducible trans­ gene expression system (GAVPO/UAS) for parametric control of 4R­Tau levels at particular time points. If our balance assay and transgenic lines meet defined milestones for success, we will proceed to the R33 aim. There, we will validate our model externally along three axis: First, we will assay construct validity by testing if a hu­ man pathogenic Tau mutant enhances the severity of observed phenotypes. Second we will assay face validity by evaluating the pathology, neurological and balance deficits following induction of 4R­Tau. Lastly we will as­ say predictive validity by administering chemicals known to modulate tauopathy in other model systems. Our team consists of a physician scientist specializing in movement disorders (in the clinic) and zebrafish models of neurodegeneration (in the lab) and a basic neuroscientist who has successfully used zebrafish to understand normal development and function of neural circuits responsible for balance. Together, we propose to establish and validate an innovative new “phenotype­optimized” model system to address a deep unmet clinical need: neurotherapeutic discovery for PSP in the context of its definitive balance symptoms. The work is broadly signif­ icant as it advances our ability to define general mechanisms of neurodegeneration.

摘要 进行性核上性麻痹（PSP）是一种神经退行性疾病，其临床特征为严重的 失去平衡和其他神经症状。福尔斯是PSP早期表现和诊断的核心， 导致骨折和住院。目前没有任何治疗方法可以改善平衡或预防疾病进展。 强有力的病理学、分子学和遗传学证据表明，微血管内皮细胞的4个重复亚型的沉积， 微管相关蛋白Tau（4 R-Tau）在PSP病理生理学中的作用。目前的模型难以将4 R Tau pathol 对PSP的定义是虚弱的福尔斯。PSP的新疗法的开发将大大受益 从一种手段来评估在负责平衡的神经回路的背景下的干预。 利用我们各自实验室的历史优势，我们共同设计了一个新的 模型系统来在平衡的背景下对Tau病理学进行建模。我们的模型系统使用的是斑马鱼的幼体， 一种小型脊椎动物，具有特殊的分子和光学可及性以及严格定义的平衡行为。 在未发表的试点实验中，使用了一系列广泛的神经和行为测试，结果表明， 在斑马鱼神经元中施用4 R β Tau引起：（i）分子和细胞病理学（ii）运动和眼运动 赤字（iii）系统平衡赤字，共同类似PSP。在这里，我们建议扩展和验证 这个新的令人兴奋的模型。 在R61目标中，为了在内部验证我们的模型，我们将开发两个互补的管道来分析平衡 行为并建立新的转基因系和方案以获得4 R β Tau表达的时间控制。第一、 我们将优化一种平衡测定法，该测定法具有严格定义的定量行为终点， 化学修饰剂或候选基因靶点的有效评估。接下来，我们将使用一种新型的光诱导跨膜 基因表达系统（GAVP 0/UAS）用于在特定时间点参数控制4 R β Tau水平。如果我们的 平衡测定和转基因品系满足成功的定义的里程碑，我们将继续进行R33目标。在那里， 我们将从外部沿着三个轴验证我们的模型：首先，我们将通过测试一个人是否符合条件来分析结构效度。 人致病性Tau突变体增强了观察到的表型的严重性。其次，我们将分析表面效度 通过评估4 R β Tau诱导后的病理学、神经学和平衡缺陷。最后，我们将作为 例如通过施用已知在其他模型系统中调节tau蛋白病的化学物质的预测有效性。我们 该团队由一名专门研究运动障碍（临床）和斑马鱼模型的医生科学家组成。 神经变性（在实验室）和一个基础神经科学家谁已经成功地利用斑马鱼来了解 负责平衡的神经回路的正常发育和功能。我们共同提议 并验证创新的“表型优化”模型系统，以解决深度未满足的临床需求： 在PSP明确的平衡症状的背景下，针对PSP的神经治疗发现。这项工作具有广泛的意义。 因为它提高了我们定义神经变性一般机制的能力。