Neuron-specific nanotherapeutics for spinal cord injury repair

用于修复脊髓损伤的神经元特异性纳米疗法

• 批准号: 10352315
• 负责人: Jeoung Soo Lee
• 金额: $ 32.44万
• 依托单位: CLEMSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10352315
• 关键词: Address; Adult; Alzheimer' s Disease; Animal Model; Animals; Apoptosis; Area; Astrocytes; Autonomic Dysfunction; Axon; Behavior; Binding; Biochemical; Biochemistry; Biocompatible Materials; Biodistribution; Bladder Control; Cations; Cell Death; Central Nervous System Diseases; Chemicals; Cicatrix; Clinical; Coculture Techniques; Combined Modality Therapy; Complex; Contusions; Cyclic AMP; Cytoplasmic Granules; Data; Demyelinations; Development; Drug Delivery Systems; Electrostatics; Exhibits; Formulation; Free Radicals; Fruit; Functional disorder; Gene Delivery; Glycolic-Lactic Acid Polyester; Growth; Growth Cones; Growth Inhibitors; Healthcare; Histologic; Hydrophobicity; Hypoxia; Impairment; In Vitro; Inflammation; Injury; Intervention; Intestines; Intrathecal Injections; Ischemia; Journals; Lead; Lesion; Ligands; Locomotor Recovery; Micelles; Modality; Modeling; Motor; Myelin; Nanostructures; Nanotechnology; Natural regeneration; Nervous System Trauma; Neural Cell Adhesion Molecule L1; Neurites; Neuroglia; Neurons; Neurosurgeon; Pain; Pathway interactions; Patients; Persons; Pharmaceutical Preparations; Polymers; Process; Production; Publishing; Rattus; Recovery; Recovery of Function; Regenerative Medicine; Regenerative capacity; Research; Rolipram; Sensory; Sexual Dysfunction; Signal Pathway; Signaling Molecule; Site; Small Interfering RNA; Specificity; Spinal Cord Contusions; Spinal Injections; Spinal cord injury; Stroke; Surface; Technology; Testing; Therapeutic Effect; Trauma; Traumatic Brain Injury; Treatment Efficacy; United States; Work; age related; amphiphilicity; astrogliosis; axon injury; axon regeneration; axonal degeneration; base; central nervous system injury; chronic pain; combinatorial; cost; design; effective therapy; efficacy outcomes; excitotoxicity; extracellular; functional group; healing; hydrophilicity; inhibitor; injury and repair; innovation; knock-down; multidisciplinary; nanoparticle; nanotherapeutic; nerve supply; neuron loss; neuroprotection; new combination therapies; new growth; new technology; novel; painful neuropathy; phosphodiesterase IV; pre-clinical; respiratory; restoration; spasticity; targeted delivery; therapeutic evaluation; therapeutic target; therapeutically effective

Due to limited intrinsic healing capacity and the absence of effective therapeutic treatment, spinal cord injury (SCI) usually results in permanent loss of motor and sensory function below the level of the lesion. SCI exhibits a complex pathophysiology that presents multiple barriers to functional recovery. These include ischemia and inflammation that cause progressive neuronal and glial cell death, as well as the presence of extracellular growth inhibitors and intrinsic deficiencies in neuronal biochemistry that limit the capacity of adult axons for plasticity/regeneration. Combination therapies using treatment modalities targeting two or more of these barriers have achieved promising preclinical results, but their application is complex and often requires multiple interventions. Thus, there is a need for the development of new technologies capable of simultaneously delivering multiple bioactive molecules. The objective of this proposal is to develop neuron-specific nanotherapeutics for SCI repair. The hypothesis is that combinatorial delivery of drugs modulating the cAMP and RhoA signaling pathways will provide neuroprotection and promote functional recovery by synergistic effect in contusion SCI model. These therapeutic targets have been chosen because of their recognition as key signaling molecules implicated in multiple aspects of SCI pathophysiology including inflammation, neuronal cell death, and both extrinsic and intrinsic barriers to axonal regeneration. Our approach is based upon a novel cationic amphiphilic co-polymer, poly (lactide-co-glycolide)-graft-polyethylenimine (PgP) developed in our lab. PgP forms polymeric micelles offering three critical features for combinatorial drug delivery: 1) a hydrophobic core for loading of rolipram (Rm), a phosphodiesterase 4 inhibitor capable of restoring cAMP levels, 2) a cationic, hydrophilic shell for electrostatic binding of siRNA targeting RhoA (siRhoA), and 3) surface functional groups that will be used for conjugation of the L1 neural cell adhesion molecule to increase neuronal targeting. In preliminary studies we show that PgP/siRhoA and PgP/Rm are each capable of achieving therapeutic effects in animal injury models. In Aim 1, we will formulate neuron-specific nanotherapeutics incorporating L1 as a targeting ligand, siRhoA in novel, designed nanostructures, and Rm (L1-PgP/siRhoA/Rm) and test their efficacy in an in vitro hypoxia SCI model. In Aim 2, we will test the most promising candidate in a rat moderate contusion injury model, incorporating studies of biodistribution and repeat and delayed administration. Therapeutic efficacy outcomes will include biochemical analysis of cAMP/RhoA, histological analysis, locomotor recovery, and neuropathic pain/allydonia. The proposed work is innovative because it uses a novel material capable of simultaneously delivering two chemically different drugs in a single formulation, cAMP/RhoA as a new combination therapy, L1 targeting and novel siRNA nanostructures. The proposed work is significant, because it addresses the critical and under-studied challenge of effective therapeutic delivery and may have widespread application to other CNS injury and disorders including traumatic brain injury, Alzheimer's disease, and stroke.

由于内在愈合能力有限且缺乏有效的治疗方法，脊髓损伤 (SCI) 通常会导致病变水平以下的运动和感觉功能永久丧失。 SCI展品 复杂的病理生理学给功能恢复带来多重障碍。这些包括缺血和 导致进行性神经元和神经胶质细胞死亡以及细胞外生长的炎症 神经元生物化学的抑制剂和内在缺陷限制了成年轴突的能力 可塑性/再生性。使用针对其中两种或多种障碍的治疗方式的联合疗法 已经取得了有希望的临床前结果，但它们的应用很复杂，通常需要多个 干预措施。因此，需要开发能够同时 传递多种生物活性分子。该提案的目标是开发神经元特异性 用于 SCI 修复的纳米疗法。该假设是调节 cAMP 的药物组合递送 和RhoA信号通路将通过协同作用提供神经保护并促进功能恢复 在挫伤 SCI 模型中。选择这些治疗靶点是因为它们被认为是关键 信号分子参与 SCI 病理生理学的多个方面，包括炎症、神经元细胞 死亡，以及轴突再生的外在和内在障碍。我们的方法基于一部小说 我们实验室开发的阳离子两亲性共聚物，聚（丙交酯-乙交酯）-接枝-聚乙烯亚胺（PgP）。 PgP 形成聚合物胶束，为组合药物输送提供三个关键特征：1) 疏水性 用于装载咯利普兰 (Rm) 的核心，咯利普兰 (Rm) 是一种能够恢复 cAMP 水平的磷酸二酯酶 4 抑制剂，2) 一种阳离子， 用于静电结合靶向 RhoA (siRhoA) 的 siRNA 的亲水壳，以及 3) 表面官能团 将用于连接 L1 神经细胞粘附分子以增加神经元靶向。在 初步研究表明，PgP/siRhoA 和 PgP/Rm 均能够在以下疾病中实现治疗效果： 动物损伤模型。在目标 1 中，我们将制定以 L1 作为靶向的神经元特异性纳米疗法 配体、新颖设计的纳米结构中的 siRhoA 和 Rm (L1-PgP/siRhoA/Rm) 并测试它们在纳米结构中的功效 体外缺氧SCI模型。在目标 2 中，我们将在大鼠中度挫伤中测试最有希望的候选者 模型，结合了生物分布以及重复和延迟给药的研究。治疗效果 结果将包括 cAMP/RhoA 的生化分析、组织学分析、运动恢复和 神经性疼痛/阿利多尼亚。拟议的工作具有创新性，因为它使用了一种能够 在单一配方中同时输送两种化学性质不同的药物，cAMP/RhoA 作为一种新药物 联合疗法、L1 靶向和新型 siRNA 纳米结构。拟议的工作意义重大，因为 它解决了有效治疗传递的关键且尚未充分研究的挑战，并且可能具有广泛的应用 应用于其他中枢神经系统损伤和疾病，包括创伤性脑损伤、阿尔茨海默病和中风。

项目成果

A cationic amphiphilic co-polymer as a carrier of nucleic acid nanoparticles (Nanps) for controlled gene silencing, immunostimulation, and biodistribution.

一种阳离子两亲性共聚物，作为核酸纳米粒子 (Nanps) 的载体，用于受控基因沉默、免疫刺激和生物分布。

• 发表时间: 2020
• 作者: Halman, Justin R;Kim, Ki;Gwak, So;Pace, Richard;Johnson, M Brittany;Chandler, Morgan R;Rackley, Lauren;Viard, Mathias;Marriott, Ian;Lee, Jeoung Soo;Afonin, Kirill A
• 通讯作者: Afonin, Kirill A