Targeting Blood-CNS-Barrier in ALS via Apolipoprotein A1

项目摘要摘要 肌萎缩侧索硬化症(ALS)是一种脑部广泛运动神经元变性的致死性疾病。 脊髓。进行性血液-中枢神经系统屏障损害(B-CNS-B)是另一种疾病 机制。中枢神经系统毛细血管内皮细胞(EC)损伤已在ALS啮齿动物模型和 肌萎缩侧索硬化患者。我们展示了静脉(Iv)移植人骨髓来源的好处。 内皮祖细胞(HBM-EPC)对SOD1小鼠功能性疾病结局和运动神经元的影响 通过减轻受损屏障的损害来建立ALS模型。移植的HBM-EPC也可能 通过释放细胞外小泡(EV)和促进退变的内皮细胞通过 运送货物蛋白质。载脂蛋白A1(ApoA1)被确定为最丰富的高表达 EVS中的蛋白质，可能是EC靶向再生的治疗活性成分。我们展示了 在类似ALS的病理条件下，ApoA1在体外提高了EC的存活率。这个项目的目的是 确定载脂蛋白A1是否促进ALS的内皮动态平衡，从而导致B-CNS-B修复。这个 该项目的重大科学进步证明了ApoA1政府阐明的 B-CNS-B修复过程中的修复过程并促进G93A SOD1突变小鼠的运动神经元存活。 此外，通过评估载脂蛋白A1的影响来确定B-CNS-B修复的修复机制 是这个项目的新奇之处。拟议研究的一个重要方面是在有症状的情况下B-CNS-B恢复 存在屏障损伤的ALS小鼠模型。目标1将确定单次静脉注射的治疗效果 ApoA1对有症状的ALS小鼠B-CNS-B修复作用的行为学检测 疾病结局(目标1A)、功能性屏障修复(目标1B)、神经胶质细胞状态(目标1C)和运动神经元 生存(目标1D)。目的2探讨载脂蛋白A1介导的症状性血管修复的机制(S) 通过检测该蛋白对ALS小鼠血管内皮细胞完整性的影响。这一目标将解决 ApoA1蛋白通过胞质PI3K/Akt途径阻断下游信号传导。的影响 抑制细胞内信号传递的研究结果将与目标1分目标中描述的结果相同。我们的 确定ApoA1给药有效性的实验设计是一种高度翻译和创新的设计 基于机制的B-CNS-B损伤修复方法积极的项目结果将证明 ApoA1蛋白在修复肌萎缩侧索硬化症患者B-CNS-B功能中的作用连 如果目标1的结果是否定的，通过抑制范式探测目标2中的载脂蛋白AI将揭示新的基于载脂蛋白AI的 优化蛋白质治疗的方法。这项研究代表了一种风险相对较低、但回报较高的 肌萎缩侧索硬化症血管修复的创新蛋白质介导疗法，从而促进转化为临床 适用于ALS患者。

Project Summary Abstract Amyotrophic lateral sclerosis (ALS) is a fatal disease of widespread motor neuron degeneration in the brain and spinal cord. Progressive impairment of the blood-CNS--barrier (B-CNS-B) represents an additional disease mechanism. Capillary endothelial cell (EC) damage in the CNS has been shown in ALS rodent models and in ALS patients. We demonstrated benefits of intravenously (iv) transplanted human bone marrow-derived endothelial progenitor cells (hBM-EPCs) on functional disease outcomes and motor neurons in an SOD1 mouse model of ALS by attenuating damage to the compromised barrier. Transplanted hBM-EPCs may also exert positive effects by release of extracellular vesicles (EVs) and facilitate restoration of degenerated ECs through delivery of cargo proteins. Apolipoprotein A1 (ApoA1) was determined as the most abundant high-expression protein in EVs and may represent a therapeutically active component for EC-targeted regeneration. We showed that ApoA1 enhanced EC survival in an ALS-like pathologic condition in vitro. The purpose of this project is to determine whether ApoA1 facilitates endothelium homeostasis leading to B-CNS-B repair in ALS. The significant scientific advance of this project is the demonstration that ApoA1 administration elucidates reparative processes in B-CNS-B restoration and promotes motor neuron survival in G93A SOD1 mutant mice. Also, the determination of reparative mechanisms underlying B-CNS-B restoration by assessing ApoA1 effects is the novelty of this project. An important aspect of the proposed study is B-CNS-B restoration in a symptomatic mouse model of ALS with existing barrier damage. Aim 1 will establish therapeutic efficacy of a single iv administration of ApoA1 into symptomatic ALS mice of both genders on B-CNS-B repair by examining behavioral disease outcomes (Aim 1A), functional (Aim 1B) barrier repair, glial cells status (Aim 1C), and motor neuron survival (Aim 1D). Aim 2 will determine the mechanism(s) of ApoA1-mediated vascular repair in symptomatic ALS mice by examining the pathway of this protein on endothelium integrity. This aim will address activity of the ApoA1 protein by impeding the downstream signaling through the cytosolic PI3K/Akt pathway. The effects of inhibiting intracellular signaling will be examined with the same outcomes as described in Aim 1 Sub-aims. Our experimental design to determine the efficacy of ApoA1 administration is a highly translational and innovative mechanism-based approach for repairing the damaged B-CNS-B. Positive project outcomes will evidence the mechanistic role of ApoA1 protein in restoring EC function towards repair of the altered B-CNS-B in ALS. Even if Aim 1 results are negative, probing ApoAI in Aim 2 via inhibitory paradigm will reveal novel ApoAI-based approaches to optimize protein treatment. This study represents a relatively low-risk, but high-reward and innovative protein-mediated therapy for vascular repair in ALS, thereby facilitating translation into a clinical application for ALS patients.