Exosome RNA-Therapeutics to Promote CNS Myelination

外泌体 RNA 疗法促进中枢神经系统髓鞘形成

• 批准号: 9128775
• 负责人: Richard P Kraig
• 金额: $ 32.35万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9128775
• 关键词: Adult; Age; Aging; Animal Model; Autoimmune Diseases; Axon; Biological Assay; Blood; Brain; Brain Diseases; Brain Injuries; Cell Death; Cell Differentiation process; Cells; Cessation of life; Childhood; Clinical; Competence; Data; Demyelinations; Dendritic Cells; Experimental Autoimmune Encephalomyelitis; Female; Goals; Immune; Immune System Diseases; Immunosuppression; Injury; Interferon Type II; Interferons; Legal patent; Lysophosphatidylcholines; Measures; Messenger RNA; MicroRNAs; Modeling; Multiple Sclerosis; Multiple Sclerosis Lesions; Mus; Myelin; Neurons; Nose; Oligodendroglia; Outcome Measure; Peripheral; Phase; Physical activity; Process; Proteins; RNA; Rattus; Reaction; Recovery; Renal function; Research; Serum; Severity of illness; Signal Transduction; Signaling Molecule; Slice; Small RNA; Source; Therapeutic; Thick; Toxic effect; Work; aged; aging brain; base; clinical efficacy; design; environmental enrichment for laboratory animals; exosome; experience; immune activation; immunogenic; innovation; liver function; mouse model; myelination; nerve stem cell; nervous system disorder; novel therapeutics; oligodendrocyte precursor; pre-clinical; precursor cell; prevent; remyelination; repaired; research study; response; social; tumorigenesis; vesicular release; young adult

DESCRIPTION (provided by applicant): Multiple sclerosis (MS) is the most common disabling neurological disease of young adulthood. It is an incurable autoimmune disorder involving oligodendrocyte loss, demyelination, and an inability to remyelinate damaged brain. Remyelination declines with age and MS progression, where oligodendrocyte precursor cells (OPCs) are actively prevented from differentiating into myelinating cells. Current therapies are only directed at reducing demyelination via immune suppression which reduces immune competence. Importantly, no therapies exist to prevent the inevitable decline in remyelination. This project is designed to remove this void. OPC differentiation into myelinating cells is regulated by certain microRNAs (miRNAs) that are reduced in MS lesions. Notably, blood from young rats enhances remyelination of aged brain. The rationale for this work comes from pilot data showing that blood-derived exosomes, small vesicles released from cells, contain miRNAs that trigger remyelination. Non-immunogenic exosomal miRNAs from the serum of young and environmentally enriched (EE) rats enhanced remyelination. The long-term goal of our research is to develop neurotherapeutics based on understanding how EE protects brain. The overall goal of this application is to develop exosomes containing miRNAs as a therapeutic for brain remyelination. The central hypothesis is that these exosomes will enhance remyelination of brain and can be crafted into a therapy for MS. The project has four Aims: AIM 1: Establish interferon-gamma (IFN-γ) -stimulated dendritic cells (DCs) as an effective source of exosomes that mimic EE-exosomes to enhance remyelination without injury or immune reaction in slice cultures. AIM 2: Establish an efficient means of loading DC exosomes with fabricated targeted miRNA species and establish their ability to enhance remyelination without injury or immune reaction in slice cultures compared to EE-exosomes and IFN-γ-stimulated DC exosomes. AIM 3: Establish the optimal efficacy of intravenously (then nasally) delivered (Aims 1-2) exosomes to promote remyelination without injury or immune reaction in a rat lysolecithin model of MS demyelination. AIM 4: Establish the optimal efficacy of intravenously (then nasally) administered (Aims 1-2) exosomes to promote remyelination in a mouse model of MS [experimental autoimmune encephalomyelitis (EAE)] without injury or enhanced immune reaction. Outcome measures will be: 1) OPC, differentiated oligodendrocyte, and compact myelin levels, targeted miRNAs & target levels; 2) toxicity, measured by cell death, liver and renal function assays, plus oncogenesis; 3) immune activation assayed by serum and brain markers; and 4) clinical measures of disease severity. The work is innovative because it uses naturally occurring signaling molecules and vehicles for delivery of EE-like enhancement of myelination. The proposed research is significant because it will establish the preclinical feasibility and utility of EE-based miRNA-containing exosomes as a novel therapeutic to remyelinate MS- damaged brain without immune compromise.

描述（由申请人提供）：多发性硬化症（MS）是最常见的残障神经系统疾病。这是一种无法治愈的自身免疫性疾病，涉及少突胶质细胞损失，脱髓鞘和无法再生脑损伤的大脑。随着年龄和MS的进展，透明度下降，其中少突胶质细胞前体细胞（OPC）被积极防止区分髓生成细胞。当前的疗法仅针对通过免疫抑制来降低脱髓鞘，从而降低免疫能力。重要的是，没有疗法可以防止不可避免的再髓样下降。该项目旨在消除此空隙。 OPC分化为髓生成细胞受到MS病变中降低的某些microRNA（miRNA）的调节。值得注意的是，来自年轻大鼠的血液增强了老化大脑的再生。这项工作的基本原理来自试验数据，表明血液来源的外泌体，从细胞释放的小囊泡，包含触发透明性的miRNA。来自年轻和环境富集（EE）大鼠血清中的非免疫原性外泌体miRNA增强了再生。我们研究的长期目标是基于了解EE如何保护大脑的神经疗法。该应用的总体目标是开发含有miRNA作为脑透明式治疗的外泌体。中心假设是这些外泌体将增强大脑的再髓样，并可以用于MS治疗。该项目具有四个目标：目标1：建立干扰素 - 伽马（IFN-γ）刺激的树突状细胞（DC），作为模拟EE外座体的有效外泌体来源，以在切片培养物中增强无损伤或免疫反应而增强re骨。 AIM 2：与靶向的miRNA物种一起建立一种有效的手段，将DC外泌体加载，并确定与EE-诊断和IFN-γ刺激的DC外生体相比，在切片培养物中增强透明或免疫反应的能力。 AIM 3：在MS脱髓鞘的大鼠溶血素模型中，静脉内（AIMS 1-2）静脉内（AIMS 1-2）建立最佳疗效（AIMS 1-2），而无需造成损伤或免疫反应。 AIM 4：建立静脉内（然后是鼻腔）给药（AIMS 1-2）外泌体的最佳功效，以在没有受伤或增强免疫反应的MS [实验性自身免疫性脑脊髓炎（EAE）]的MS [实验性自身免疫性脑脊髓炎（EAE）]中促进透明度。结果指标将是：1）OPC，分化的少突胶质细胞和紧凑的髓磷脂水平，靶向miRNA和靶水平； 2）毒性，通过细胞死亡，肝脏和肾功能分析以及肿瘤发生测量； 3）通过血清和脑标记物测定的免疫激活； 4）疾病严重程度的临床指标。这项工作具有创新性，因为它使用自然发生的信号分子和车辆来传递类似EE的髓鞘形成。拟议的研究之所以重要，是因为它将建立含EE的miRNA外泌体的临床前的可行性和效用，作为一种新的治疗性，可在没有免疫损害的情况下延迟MS损伤的大脑。