FL2 siRNA as a Novel Therapeutic Option to Induce Spinal Cord Regeneration Following Injury

FL2 siRNA 作为诱导损伤后脊髓再生的新型治疗选择

• 批准号: 10593477
• 负责人: Lisa Ann Baker
• 金额: $ 42.57万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-24 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10593477
• 关键词: Acute; Adult; Aftercare; Bladder; Cell Culture Techniques; Chest; Cicatrix; Contusions; Data; Dorsal; Down-Regulation; Embryo; Encapsulated; Enzymes; FDA approved; Face; Foundations; Functional Regeneration; Gene Expression; Health; Hindlimb; Histologic; Histology; Inflammatory Response; Injury; Interferons; Left; Lesion; Locomotor Recovery; Microglia; Microtubules; Modeling; Molecular; Molecular Biology; Motor; Movement; Natural regeneration; Nerve Regeneration; Neurons; Pathology; Patients; Peripheral nerve injury; Persons; Pilot Projects; RNA Interference; Rattus; Recovery of Function; Regulation; Rest; Rodent; Role; Sensory; Site; Small Interfering RNA; Spinal Cord; Spinal Cord Contusions; Spinal Cord Lesions; Spinal cord injury; Testing; Therapeutic; Tissues; Transgenic Mice; Traumatic CNS injury; United States; axon growth; axon guidance; axon regeneration; cell behavior; cell motility; cell type; central nervous system injury; disabling symptom; effective therapy; efficacy evaluation; genetic regulatory protein; improved; loss of function; nanoparticle; neural circuit; novel therapeutic intervention; novel therapeutics; protein expression; regeneration following injury; response; response to injury; sensory system; spinal cord regeneration; wound

ABSTRACT Fidgetin-like 2 (FL2) is a microtubule (MT) regulatory protein that modulates MT dynamics through its putative MT-severing activity, regulating cell motility and axonal growth and guidance. Recently, we identified FL2 as a negative regulator of axonal growth, and demonstrated that targeted depletion of FL2 following peripheral nerve injury enhances functional nerve regeneration in rats. Our preliminary studies show that following injury FL2 is upregulated at the injury site in several adult tissues including the spinal cord. In pilot studies using rodent SCI models, we similarly found that local depletion of FL2 from the injury site using FL2 siRNA embedded in nanoparticles (SiFi2) improved recovery of locomotor and bladder function after thoracic contusion and compression injury. We hypothesize that FL2 negatively regulates axonal regeneration after SCI, and that downregulation of FL2 after SCI will improve functional recovery. We aim to test this hypothesis with 3 specific aims. The first aim will determine the extent and duration of FL2 silencing following SiFi2 treatment. The second aim will evaluate the functional effects of SiFi2 treatment after SCI by exploring hindlimb locomotor and sensory function after SiFi2 administration. The third aim will evaluate molecular and histological changes at the spinal cord lesion site following SiFi2 treatment by examining axonal regeneration, glial scar formation, and inflammatory response in the spinal cord tissue from Aim 2. These studies will be the first to evaluate FL2 regulation of the central nervous system (CNS) injury response, and the first to assess the therapeutic potential of using RNAi to transiently downregulate FL2 expression in order to improve functional recovery after CNS injury. While this proposal focuses specifically on SCI as a therapeutic application, we anticipate the data generated from these studies will have broader implications as they will characterize the role of a previously unstudied regulator of CNS traumatic injury response, one which can potentially be targeted to enhance regeneration following a wide range of CNS injuries.

抽象的 fidgetin样2（FL2）是一种微管（MT）调节蛋白，可通过其推定调节MT动力学 MT细分活性，调节细胞运动和轴突生长和指导。最近，我们将FL2确定为 轴突生长的负调节剂，并证明了外周神经后FL2的靶向耗竭 损伤增强了大鼠的功能神经再生。我们的初步研究表明，损伤后FL2是 在包括脊髓在内的几个成年组织中的损伤部位上调。在使用啮齿动物科学的试点研究中 模型，我们同样发现，使用嵌入在损伤部位的FL2局部耗竭 纳米颗粒（SIFI2）改善了胸骨挫伤后的运动和膀胱功能的恢复 压缩损伤。我们假设FL2在SCI后负调节轴突再生，并且 SCI后FL2的下调将改善功能恢复。我们旨在用3个特定的 目标。第一个目标将决定SIFI2处理后FL2沉默的程度和持续时间。第二个 AIM将通过探索后肢运动和感觉来评估SCI后SIFI2治疗的功能效应 SIFI2给药后的功能。第三个目标将评估脊柱的分子和组织学变化 SIFI2治疗后的脐带病变部位通过检查轴突再生，神经胶质疤痕形成和 AIM 2的脊髓组织中的炎症反应。这些研究将是第一个评估FL2的研究 中枢神经系统（CNS）损伤反应的调节，并首先评估治疗潜力 使用RNAi瞬时下调FL2表达以改善CNS后的功能恢复 受伤。虽然该提案专门针对SCI作为治疗应用，但我们预计数据 这些研究产生的将具有更广泛的含义，因为它们将表征以前的作用 中枢神经系统创伤性损伤反应的未研究调节剂，该反应有可能针对增强 在广泛的中枢神经系统损伤之后再生。