Neuroprotective Engineering Based on Innate Responses to Stroke

基于对中风的先天反应的神经保护工程

• 批准号: 1403036
• 负责人: Shu Liu
• 金额: $ 49.99万
• 依托单位: Northwestern University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1403036&HistoricalAwards=false
• 关键词: Neuroprotective; Engineering; Based; Innate; Responses

PI: Liu, Shu Q. Proposal Number: 1403036 Institution: Northwestern UniversityTitle: Neuroprotective Engineering Based on Innate Responses to StrokeStroke is a prevalent disorder commonly caused by arterial plaques that block blood flow to the brain, resulting in brain injury, depression, mental retardation, and/or paralysis. The injured brain is often associated with bone-like structure formation, known as brain calcification, a process disrupting the brain structure and intensifying brain injury. To date, it remains poorly understood how stroke causes brain calcification and there are few approaches effective for prevention of brain calcification. In this application, the investigators intend to elucidate the role of a cell membrane-associated family of calcium-carrying molecules known as annexins in the induction of brain calcification in a mouse model of stroke. These molecules may move from the cell membrane to the intracellular contractile filaments when brain cells are injured to cause calcium deposition or calcification, as these molecules carry calcium ions. The investigators have discovered a liver-produced molecule known as trefoil factor 3 that potentially protects the injured brain from calcification by blocking annexin deposition. The significance of this discovery is that trefoil factor 3 may be potentially used as a drug to prevent brain calcification and injury in patients with stroke. In this project, the investigators will develop an engineering strategy for boosting trefoil factor 3 production in a mouse model of stroke by delivery of the trefoil factor 3 gene or protein and test the efficacy of the engineering approach for brain protection against annexin-dependent calcification. If successful, trefoil factor 3 can be produced by a biotechnology approach and applied to human patients with stroke to prevent brain calcification, thereby reducing brain injury and functional deficits. This effort may potentially lead to a reduction in stroke-induced human morbidity and mortality. In addition to these scientific aspects, the investigators will devote efforts to establish a new undergraduate education model integrating independent research into lecture topics. This form of education will allow students to understand scientific concepts from hands-on experience and to be more engaged in cutting-edge research, enhancing students creativity and capability of solving real-world problems. Cerebral ischemia or ischemic stroke is a prevalent disorder commonly caused by cerebral artery thrombosis and/or atherosclerosis, resulting in cerebral injury and neurological deficits including depression, mental retardation, and/or paralysis. Ischemic stroke is often associated with cerebral calcification or hydroxyapatite deposition, a process disrupting neuronal structure and intensifying cerebral injury. To date, the mechanisms of cerebral calcification remain elusive and few approaches have been established for protecting the cerebrum from calcification. The investigators have found that a cell membrane-associated family of calcium-carrying molecules known as annexins may translocate from the cell membrane to the cytoskeletal microfilaments in ischemic neurons to facilitate hydroxyapatite formation. Furthermore, a liver-produced endocrine molecule known as trefoil factor 3 (TFF3) is upregulated in response to stroke, potentially protecting the ischemic cerebrum from calcification by blocking annexin deposition. In the proposed research, the investigators intend to achieve three aims: (1) evaluate the role of annexins A2, A3, and A5 in ischemic cerebral calcification and injury; (2) assess the role of TFF3 in protection of the ischemic cerebrum from annexin-dependent calcification and injury; and (3) establish neuroprotective engineering strategies based on the mechanisms of TFF3 action for maximizing protection against cerebral calcification in stroke. In a mouse model of ischemic stroke, the role of annexins will be evaluated by using siRNA-mediated loss-of-annexin and recombinant annexin-based gain-of-annexin approaches; the anti-calcification role of TFF3 will be tested by using a TFF3-/- mouse model with or without recombinant TFF3 administration; and the role of TFF3 in interference with annexin binding to neuronal micro-filaments will be assessed by molecular binding assays in the presence or absence of TFF3. Protective engineering approaches will be established based on TFF3 gene transfection and controlled TFF3 protein delivery technologies to boost TFF3 expression following ischemic stroke. These investigations will provide a foundation for understanding the mechanisms of ischemic cerebral calcification and establishing engineering technologies for protection against ischemic cerebral calcification and injury, thus potentially reducing stroke-induced human morbidity and mortality.

PI：Liu，Shu Q. 提案编号：1403036机构：西北大学标题：基于对中风的先天反应的神经保护工程中风是一种常见的疾病，通常由动脉斑块阻塞血液流向大脑引起，导致脑损伤，抑郁症，智力低下和/或瘫痪。受伤的大脑通常与骨样结构的形成有关，称为大脑钙化，这是一个破坏大脑结构并加剧脑损伤的过程。到目前为止，人们对中风如何导致脑钙化仍然知之甚少，并且几乎没有有效预防脑钙化的方法。在本申请中，研究人员打算阐明细胞膜相关的钙携带分子家族（称为膜联蛋白）在中风小鼠模型中诱导脑钙化中的作用。当脑细胞受损时，这些分子可以从细胞膜移动到细胞内收缩丝，引起钙沉积或钙化，因为这些分子携带钙离子。研究人员发现了一种肝脏产生的分子，称为三叶因子3，它可能通过阻止膜联蛋白沉积来保护受伤的大脑免受钙化。这一发现的意义在于三叶因子3可能被用作预防中风患者脑钙化和损伤的药物。在这个项目中，研究人员将开发一种工程策略，通过传递三叶因子3基因或蛋白质来促进中风小鼠模型中三叶因子3的产生，并测试工程方法对大脑保护膜联蛋白依赖性钙化的功效。如果成功，三叶因子3可以通过生物技术方法生产，并应用于中风患者，以防止脑钙化，从而减少脑损伤和功能缺陷。这一努力可能会导致减少中风引起的人类发病率和死亡率。除了这些科学方面，研究人员将致力于建立一个新的本科教育模式，将独立研究融入讲座主题。这种教育形式将使学生从实践经验中理解科学概念，并更多地参与前沿研究，提高学生的创造力和解决现实问题的能力。脑缺血或缺血性卒中是一种常见的疾病，通常由脑动脉血栓形成和/或动脉粥样硬化引起，导致脑损伤和神经功能缺损，包括抑郁症、智力迟钝和/或瘫痪。缺血性中风通常与脑钙化或羟基磷灰石沉积有关，这是一个破坏神经元结构并加剧脑损伤的过程。迄今为止，脑钙化的机制仍然是难以捉摸的，很少有方法已经建立了保护大脑钙化。研究人员发现，一个与细胞膜相关的钙携带分子家族，称为膜联蛋白，可以从细胞膜转移到缺血神经元中的细胞骨架微丝，以促进羟基磷灰石的形成。此外，肝脏产生的内分泌分子三叶因子3（TFF 3）在中风时上调，可能通过阻断膜联蛋白沉积来保护缺血性大脑免受钙化。在这项研究中，研究人员打算实现三个目标：（1）评估膜联蛋白A2、A3和A5在缺血性脑钙化和损伤中的作用;（2）评估TFF 3在保护缺血性脑免受膜联蛋白依赖性钙化和损伤中的作用;（3）建立基于TFF 3作用机制的神经保护工程策略，以最大限度地保护脑卒中中的脑钙化。在缺血性中风的小鼠模型中，膜联蛋白的作用将通过使用siRNA介导的膜联蛋白缺失和基于重组膜联蛋白的膜联蛋白获得方法来评估; TFF 3的抗钙化作用将通过使用TFF 3-/-小鼠模型在施用或不施用重组TFF 3的情况下进行测试; TFF 3在干扰膜联蛋白与神经元微丝结合中的作用将在存在或不存在TFF 3的情况下通过分子结合测定来评估。基于TFF 3基因转染和受控TFF 3蛋白递送技术将建立保护性工程方法，以促进缺血性卒中后TFF 3的表达。这些研究将为了解缺血性脑钙化的机制和建立预防缺血性脑钙化和损伤的工程技术提供基础，从而潜在地降低中风引起的人类发病率和死亡率。