Targeting Blood-CNS-Barrier in ALS via Apolipoprotein A1

通过载脂蛋白 A1 靶向 ALS 中的血液中枢神经系统屏障

• 批准号: 10680237
• 负责人: CESARIO V BORLONGAN
• 金额: $ 22.49万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10680237
• 关键词: ALS patients; Age; Amyotrophic Lateral Sclerosis; Animal Model; Antiinflammatory Effect; Apolipoprotein A-I; Astrocytes; Behavioral; Blood; Blood Vessels; Bone Marrow; Brain; Capillary Endothelial Cell; Cell Survival; Cell Therapy; Cell Transplantation; Cell physiology; Cells; Clinical Trials; Competence; Data; Disease; Disease Outcome; Disease Progression; Endothelial Cells; Endothelium; Ensure; Experimental Designs; Future; Gender; Goals; Histology; Homeostasis; Human; Immunohistochemistry; Impairment; In Vitro; Intravenous; Longevity; Mediating; Microglia; Microvascular Permeability; Motor Neurons; Mus; Mutant Strains Mice; Natural regeneration; Neurodegenerative Disorders; Neuroglia; Outcome; PIK3CG gene; Pathogenesis; Pathogenicity; Pathologic; Pathology; Pathway interactions; Patients; Process; Proteins; Publications; Random Allocation; Recovery of Function; Reporting; Research Design; Risk; Rodent Model; Role; Safety; Scientific Advances and Accomplishments; Signal Transduction; Spinal Cord; Symptoms; Testing; Therapeutic; Therapeutic Effect; Tissues; Translating; Translations; Transplantation; Treatment Efficacy; Widespread Disease; amyotrophic lateral sclerosis therapy; astrogliosis; blood damage; cell injury; clinical application; efficacy evaluation; endothelial stem cell; experimental study; extracellular vesicles; high reward; in vitro Model; in vivo; innovation; insight; intravenous administration; laboratory experiment; motor neuron degeneration; mouse model; neuronal survival; neurovascular; novel; novel therapeutic intervention; primary endpoint; protein expression; regenerative; repair strategy; repaired; reparative process; replication factor C; restoration; superoxide dismutase 1; therapeutic evaluation; translational study; wortmannin

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.

项目概要 摘要 肌萎缩性脊髓侧索硬化症（ALS）是一种致命的疾病，是大脑和大脑中广泛的运动神经元变性。 脊髓。血液-CNS-屏障 (B-CNS-B) 的进行性损伤代表另一种疾病 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 ALS 患者。我们证明了静脉注射 (iv) 移植人骨髓的益处 内皮祖细胞 (hBM-EPC) 对 SOD1 小鼠功能性疾病结果和运动神经元的影响 通过减弱对受损屏障的损害来建立 ALS 模型。移植的hBM-EPCs也可能发挥作用 通过释放细胞外囊泡 (EV) 产生积极作用，并通过促进退化 EC 的恢复 货物蛋白的输送。载脂蛋白 A1 (ApoA1) 被确定为最丰富的高表达蛋白 EV 中的蛋白质，可能代表 EC 靶向再生的治疗活性成分。我们展示了 ApoA1 增强了体外 ALS 样病理条件下 EC 的存活率。该项目的目的是 确定 ApoA1 是否促进内皮稳态，从而导致 ALS 中的 B-CNS-B 修复。这 该项目的重大科学进展是 ApoA1 管理阐明的证明 B-CNS-B 恢复过程中的修复过程并促进 G93A SOD1 突变小鼠运动神经元存活。 此外，通过评估 ApoA1 效应来确定 B-CNS-B 恢复的修复机制 是这个项目的新颖之处。拟议研究的一个重要方面是有症状的 B-CNS-B 恢复 具有现有屏障损伤的 ALS 小鼠模型。目标 1 将确定单次静脉注射的治疗效果 将 ApoA1 注射到有症状的两性 ALS 小鼠中，通过检查行为来修复 B-CNS-B 疾病结果（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）。目标 2 将确定 ApoA1 介导的有症状血管修复的机制 通过检查这种蛋白质对 ALS 小鼠内皮完整性的影响途径。这一目标将涉及 ApoA1 蛋白通过胞浆 PI3K/Akt 途径阻碍下游信号传导。的影响 抑制细胞内信号传导将得到与目标 1 子目标中描述的相同结果的检查。我们的 确定 ApoA1 给药功效的实验设计是一种高度转化和创新的方法 基于机制的方法修复受损的 B-CNS-B。积极的项目成果将证明 ApoA1 蛋白在恢复 EC 功能以修复 ALS 中改变的 B-CNS-B 中的机制作用。甚至 如果 Aim 1 结果是否定的，通过抑制范式探测 Aim 2 中的 ApoAI 将揭示基于 ApoAI 的新方法 优化蛋白质处理的方法。这项研究是一项风险相对较低但回报较高的研究 用于 ALS 血管修复的创新蛋白质介导疗法，从而促进转化为临床 ALS 患者的应用。

项目成果

Attenuation of amyotrophic lateral sclerosis via stem cell and extracellular vesicle therapy: An updated review.

通过干细胞和细胞外囊泡疗法减轻肌萎缩侧索硬化症：最新综述。

• 发表时间: 2023-12
• 作者: Lockard, Gavin;Gordon, Jonah;Schimmel, Samantha;Sayed, Bassel El;Monsour, Molly;Garbuzova;Borlongan, Cesar V
• 通讯作者: Borlongan, Cesar V