Targeting Blood-CNS-Barrier in ALS via Apolipoprotein A1

项目摘要 肌萎缩侧索硬化症（ALS）是一种致命的疾病，广泛的运动神经元变性的大脑， 脊髓血液-CNS-屏障（B-CNS-B）的进行性损害代表另一种疾病 机制CNS中的毛细血管内皮细胞（EC）损伤已经在ALS啮齿动物模型和中枢神经系统中显示。 ALS患者我们证明了静脉（iv）移植人骨髓源性 内皮祖细胞（hBM-EPCs）对SOD 1小鼠功能性疾病结局和运动神经元的影响 ALS模型通过减弱对受损屏障的损害。移植的hBM-EPCs也可能发挥 通过释放细胞外囊泡（EV）产生积极作用，并通过以下方式促进退化的EC的恢复： 货物蛋白的递送。载脂蛋白A1（ApoA 1）被确定为最丰富的高表达 蛋白质，并且可以代表用于EC靶向再生的治疗活性组分。我们展示 ApoA 1在体外ALS样病理条件下增强EC存活。该项目的目的是 确定ApoA 1是否促进ALS中导致B-CNS-B修复的内皮稳态。的 该项目的一个重大科学进展是ApoA 1给药阐明了 B-CNS-B恢复中的修复过程并促进G93 A SOD 1突变小鼠运动神经元的存活。 此外，通过评估ApoA 1效应确定B-CNS-B修复的修复机制 是这个项目的新奇。这项研究的一个重要方面是在有症状的患者中进行B-CNS-B恢复。 存在屏障损伤的ALS小鼠模型。目的1将确定单次静脉注射的治疗效果 将ApoA 1给药到两种性别的有症状的ALS小鼠中对B-CNS-B修复的影响， 疾病结局（Aim 1A）、功能性（Aim 1B）屏障修复、胶质细胞状态（Aim 1C）和运动神经元 存活率（Aim 1D）。目的2将确定ApoA 1介导的血管修复的机制， ALS小鼠通过检测这种蛋白对内皮完整性的途径。这一目标将涉及 ApoA 1蛋白通过阻碍下游信号通过胞浆PI 3 K/Akt途径。的影响 抑制细胞内信号传导将被检查，结果与目标1子目标中所述相同。我们 确定ApoA 1给药疗效的实验设计是一种高度转化和创新的方法， 修复受损的B-CNS-B的基于机制的方法。积极的项目成果将证明 ApoA 1蛋白在ALS中恢复EC功能以修复改变的B-CNS-B中的机制作用。甚至 如果Aim 1结果为阴性，则通过抑制范例探测Aim 2中的ApoAI将揭示新的基于ApoAI的 优化蛋白质处理的方法。这项研究代表了一个相对低风险，但高回报， 创新的蛋白质介导的ALS血管修复治疗，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.