Targeting Schwann cell exosomes for treating neuropathic pain

靶向雪旺细胞外泌体治疗神经性疼痛

• 批准号: 10222806
• 负责人: WENDY M. CAMPANA
• 金额: --
• 依托单位: VA SAN DIEGO HEALTHCARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-10-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10222806
• 关键词: Address; Affect; Afferent Neurons; American; Biology; Blood; Calcium Signaling; Cells; Chronic; Clinical; Cognitive; Communication; Data; Dependence; Development; Emergency Situation; Environment; Exposure to; Foundations; General Population; Goals; Growth; Growth Cones; In Vitro; Injury; Ligation; Link; Maintenance; Measures; Membrane; Methods; MicroRNAs; Nerve; Nerve Regeneration; Nervous System Physiology; Nervous System Trauma; Neurites; Neuroglia; Neurons; Neuropathy; Nucleic Acids; Outcome; PC12 Cells; Pain; Pain Disorder; Pain management; Pathogenesis; Peripheral; Peripheral Nerves; Peripheral Nervous System; Peripheral nerve injury; Pharmacology; Phenotype; Physiology; Plasma; Play; Process; Property; Proteins; Protocols documentation; RNA; Rattus; Recovery; Refractory; Refuse Disposal; Regenerative capacity; Regenerative research; Research; Research Design; Research Project Grants; Research Proposals; Risk; Rodent; Role; Schwann Cells; Sensory; Signal Pathway; Signal Transduction; Small RNA; Source; Spinal; Stream; Surface; System; Technology; Testing; Therapeutic; Thermal Hyperalgesias; Trauma; Untranslated RNA; Veterans; Work; alternative treatment; base; behavioral study; biomaterial compatibility; chronic neuropathic pain; chronic pain; chronic pain management; combat injury; cost; debilitating pain; dorsal horn; exosome; experimental study; extracellular vesicles; first responder; high risk; improved; in vivo; inhibitor; injured; innovation; limb injury; miRNA expression profiling; molecular marker; nano-string; nanovesicle; novel; opioid epidemic; opioid therapy; pain outcome; pain processing; pain signal; painful neuropathy; prevent; programs; regeneration following injury; rehabilitation research; repaired; research and development; research vision; response; response to injury; service programs; severe injury; transcriptome; wounded soldier

It is increasingly evident that Schwann cells (SCs) in the peripheral nervous system (PNS) function as a unit with neurons to regulate sensory function. When the PNS is injured in trauma, SCs become activated for repair. This involves dramatic phenotypic transformation. If this process is abnormal or inhibited, peripheral nerve injury can result in chronic debilitating pain, a problem observed in the general population, including numerous Veterans. Currently available treatment options for chronic neuropathic pain are very limited and fraught with dependency concerns. To address this unmet clinical need, we have developed a novel method for capturing plasma-derived SC exosomes (SCDEs) from rodents that regulate key cellular mechanisms in pain processing. These SCDEs recapitulate some of the bioactivity observed in exosomes collected in medium from cultured primary SCs. The major goal of this research project is to determine whether we can exploit the activity of SCDEs to improve pain outcomes following peripheral nerve injury. To accomplish our goals, three Specific Aims are proposed. In Specific Aim 1, we will determine whether a continuum of neuropathic SCDEs or naïve SCDEs regulate neuronal sprouting and calcium signaling in DRG neurons. We will apply advance technologies including single cell RNA transcriptome profiling so that we can define the response of specific neuronal subpopulations in the DRG to SCDEs. Central to this Aim is our discovery that neuropathic, but not naïve, SCDEs are capable of inducing robust neurite outgrowth that may contribute to maintenance of chronic pain states. We hypothesize that by targeting these SCDEs we will identify key mechanisms for improving pain outcomes following peripheral nerve injury therapeutically. In Specific Aim 2, our goal is to discover novel SCDE cargo, and specifically noncoding microRNAs, that are expressed in neuropathic paradigms. This discovery-based Aim establishes a microRNA (miR) signature in SCDEs that may be associated with pain producing and pain alleviating properties. Using NanoString™ technology to measure miRs in SCDEs, we identified a pain alleviating miR-142-3p in naïve SCDEs. The proposed work should identify the SC source (unmyelinated, myelinated, activated or repair phenotype) in nerves that contributes to SCDEs. In Specific Aim 3, we will assess whether SCDE phenotypes regulate neuropathic pain in vivo. This Aim is based on our exciting preliminary data demonstrating that naïve SCDEs delivered systemically inhibit thermal hyperalgesia after partial nerve ligation. SCDEs have intrinsic features, such as the stability, biocompatibility and stealth capacity when circulating in the blood stream. Studies using miR mimics and inhibitors are planned to discover mechanisms conferred by SCDEs to DRG neurons. We consider this project highly innovative because we target SCs and sensory neurons for treating chronic pain.

越来越明显的是，周围神经系统（PNS）中的雪旺细胞（SC）作为一个单位发挥作用 神经元来调节感觉功能。当 PNS 在外伤中受伤时，SC 就会被激活进行修复。这 涉及戏剧性的表型转变。如果这个过程异常或受到抑制，周围神经损伤可能会发生 导致慢性衰弱性疼痛，这是普通人群（包括许多退伍军人）中观察到的问题。 目前针对慢性神经性疼痛的可用治疗方案非常有限并且充满依赖性 的担忧。为了解决这一未满足的临床需求，我们开发了一种捕获血浆来源的新方法 来自啮齿动物的 SC 外泌体 (SCDE) 调节疼痛处理的关键细胞机制。这些 SCDE 概括了在培养的原代 SC 的培养基中收集的外泌体中观察到的一些生物活性。这 该研究项目的主要目标是确定我们是否可以利用 SCDE 的活性来改善疼痛 周围神经损伤后的结果。为了实现我们的目标，提出了三个具体目标。在 具体目标 1，我们将确定神经病理性 SCDE 或幼稚 SCDE 的连续体是否调节神经元 DRG 神经元中的萌芽和钙信号传导。我们将应用包括单细胞RNA在内的先进技术 转录组分析，以便我们可以定义 DRG 中特定神经元亚群的反应 SCDE。这一目标的核心是我们发现神经病性而非幼稚的 SCDE 能够诱导 强大的神经突生长可能有助于维持慢性疼痛状态。我们假设通过 针对这些 SCDE，我们将确定改善周围神经后疼痛结果的关键机制 治疗上的损伤。在特定目标 2 中，我们的目标是发现新颖的 SCDE 货物，特别是非编码货物 microRNA，以神经病理范式表达。这一基于发现的目标建立了一种 microRNA SCDE 中的 (miR) 特征可能与产生疼痛和减轻疼痛的特性有关。使用 NanoString™ 技术用于测量 SCDE 中的 miR，我们在实验中发现了一种缓解疼痛的 miR-142-3p SCDE。拟议的工作应确定 SC 来源（无髓鞘、有髓鞘、激活或修复） 表型）在导致 SCDE 的神经中。在具体目标 3 中，我们将评估 SCDE 表型是否 调节体内神经病理性疼痛。这个目标是基于我们令人兴奋的初步数据，这些数据表明天真 SCDE 递送可全身抑制部分神经结扎后的热痛觉过敏。 SCDE 具有内在的 特性，例如在血流中循环时的稳定性、生物相容性和隐形能力。研究 计划使用 miR 模拟物和抑制剂来发现 SCDE 赋予 DRG 神经元的机制。我们 我们认为这个项目具有高度创新性，因为我们针对 SC 和感觉神经元来治疗慢性疼痛。