Exosomal vesicles for neuroprotection and repair after SCI

外泌体囊泡用于 SCI 后的神经保护和修复

• 批准号: 10656410
• 负责人: Mousumi Ghosh
• 金额: --
• 依托单位: MIAMI VA HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10656410
• 关键词: Address; Adult; Affect; American; Anti-Inflammatory Agents; Autonomic Dysfunction; Behavior assessment; Binding; Cardiovascular Diseases; Cell Culture Techniques; Cell Differentiation process; Cell Survival; Cell Therapy; Cell Transplantation; Cells; Chimeric Proteins; Chronic; Cicatrix; Clinic; Clinical; Clinical Treatment; Clinical Trials; Consensus; Contract Services; Creativeness; Cyclic AMP; Development; Differentiation and Growth; Disparate; Encapsulated; Endocytic Vesicle; Engineering; Environment; Exhibits; Failure; Foundations; Future; Gene Expression; Goals; Growth; Harvest; Health Care Costs; Human; Immune; Immune system; Immunologics; Immunosuppressive Agents; In Vitro; Inflammation; Innate Immune Response; Interleukin-4; Intervention; Investigation; Knowledge; Label; Locomotion; Measurement; Mesenchymal Stem Cells; Methodology; MicroRNAs; Microglia; Modality; Natural regeneration; Nature; Neuroglia; Neurologic Deficit; Neurologic Dysfunctions; Neurons; Nucleic Acids; Peptides; Persons; Pharmaceutical Preparations; Play; Population; Proliferating; Proteins; Ramp; Recovery; Recovery of Function; Repression; Role; Schwann Cells; Signal Transduction; Source; Spinal cord injury; Spinal cord injury patients; Stroke; Testing; Therapeutic; Thoracic spinal cord structure; Time; Tissues; Translating; United States; Vesicle; Veterans; angiogenesis; astrogliosis; axon growth; axon regeneration; cell type; comparative; comparative effectiveness; effective therapy; efficacy evaluation; engineered exosomes; exosome; experimental study; functional improvement; functional restoration; gait examination; gliogenesis; implantation; improved; in vitro Assay; in vivo; in vivo evaluation; infancy; loss of function; microRNA delivery; migration; motor control; mouse model; myelination; nanoscale; nanosized; nanovesicle; nerve stem cell; neural; neurogenesis; neuroinflammation; neuronal survival; neuroprotection; novel therapeutic intervention; programs; regeneration potential; regenerative therapy; repaired; reparative capacity; restoration; restorative treatment; small molecule; social; stem; stem cell fate; targeted delivery; transcriptome sequencing; vector; vesicular release

An estimated 294,000 people live with spinal cord injury (SCI) in the United States of which over 40,000 are veterans. Though several therapeutic directions have shown promise in experimental paradigms, there does not exist a restorative treatment clinically that can significantly reverse the neurological deficits associated with SCI to improve function. At the forefront of experimental regenerative therapies that are being translated to clinical trials for human SCI is the transplantation of cells, from neural and mesenchymal stem cells to Schwann cells and olfactory ensheathing cells. Though benefits are observed with cell implantation after SCI, critical challenges associated with their use remains, including poor viability within the injured spinal cord, the need for an immunosuppressant when not autologous as well as the possibility of unwanted cell differentiation, proliferation, or migration of the implantation cells leading to various undesirable effects. Whereas combinatory approaches have been demonstrated to overcome some of these deficiencies, an alternate strategy to exogenous cell therapy is to stimulate host repair through exosomal vesicles (EVs). EVs are nanosized endocytic vesicles that cells release into the immediate environment, allowing transfer of biomolecules between them. EVs contain a variety of cargoes from microRNA to proteins and signaling intermediaries that can promote cell survival, differentiation, axon growth and myelination or subdue inflammation and scar formation. There is a growing consensus that EVs play a crucial role in regulating the adult neural stem niche. These EVs also offer the capacity to be engineered to express a fluorescent label, be targeted to a selective cell type, or be loaded with specific cargoes (e.g. small molecules, peptides, and miRNAs) for tissue or targeted cell specific delivery. Recent advances in our understanding of cell derived EVs and realization of their therapeutic potential in conditions such as stroke and cardiovascular disease have expanded the EV field. However, their use as a therapeutic modality after SCI has been limited and remains largely in its infancy. In the proposed studies, we will focus on the comparative assessment of the neuroprotective, neurogenic and the regenerative potential of EVs derived from disparate parental cell populations and under different cell culture conditions. Microglia (MG) and Schwann cells (SCs), immunologically primed or growth-stimulated, will be evaluated for their capacity to promote repair and recovery in murine models of subacute SCI to answer fundamental questions of feasibility, delivery, and efficacy. The goals of the proposed study will be accomplished through two Specific Aims. In Aim 1, the most effective cell-derived EV type will be identified according to their ability to promote neural cell survival and axon growth in vitro. Further, the aim will optimize their in vivo delivery in an experimental SCI mouse model and assess their comparitive effects on ameliorating inflammation, astrogliosis and regeneration associated gene (RAG) expression repression while promoting neurogenesis, axonal growth and functional recovery. The vesicle content of the most efficacious EV after SCI will be characterized with respect to its nucleic acid content to identify specfic microRNA sequences that correlate with their reparative potential. In Aim 2 EVs will be engineered for cell-specific delivery of reparative and neurogenic microRNA. The feasibility and functionality of microglia and Schwann cell derived EV engineering for NSC targeting with a specific microRNA: miRNA-9, that has been demonstrated to alter the neural stem cell fate program, neurogenesis and the restriction of gliogenesis, respectively while promoting angiogenesis. The engineered EV will be tested using in vitro assays and, in vivo experiments for effects on functional efficacy.The overall objective of the proposed studies is to improve our understanding of how cell derived EVs may be involved in neurorepair and whether they can be engineered to further enhance their beneficial effects on host cells and subsequent reparative actions following SCI.

据估计，美国有29.4万人患有脊髓损伤(SCI)，其中超过4万人是 退伍军人。尽管几个治疗方向在实验范例中显示出了希望，但没有 临床上存在一种恢复性治疗方法，可以显著逆转与脊髓损伤相关的神经功能障碍 改善功能。处于正在转化为临床的实验性再生疗法的前沿 人类脊髓损伤的试验是将神经和间充质干细胞移植到雪旺细胞。 以及嗅鞘细胞。尽管观察到脊髓损伤后细胞移植的好处，但关键的挑战 与使用它们相关的残骸，包括损伤脊髓内的生存能力差，需要一种 免疫抑制时，非自体以及不想要的细胞分化，增殖， 或植入细胞的迁移导致各种不良影响。鉴于组合方法 已经证明可以克服其中的一些缺陷，这是一种替代外源细胞的策略 治疗方法是通过胞外小泡(EVS)刺激宿主修复。EVS是纳米大小的内吞囊泡 细胞释放到直接的环境中，允许生物分子在它们之间转移。电动汽车包含一个 从微型RNA到蛋白质，再到可以促进细胞存活的信号中介， 分化、轴突生长和髓鞘形成或抑制炎症和疤痕形成。有一个不断增长的 一致认为电动汽车在调节成人神经干细胞的利基环境中发挥着关键作用。这些电动汽车还提供 被设计成表达荧光标记的能力，针对选择性细胞类型的能力，或被加载到 特定货物(例如，小分子、多肽和miRNAs)，用于组织或靶向细胞特定递送。 细胞源性血管内皮生长因子及其治疗潜力的研究进展 中风和心血管疾病等疾病扩大了EV领域。然而，它们作为一种 脊髓损伤后的治疗方式一直有限，而且在很大程度上仍处于起步阶段。在建议的研究中，我们 将集中于神经保护、神经发生和再生潜力的比较评估 EV来自不同的亲代细胞群体，在不同的细胞培养条件下。小胶质细胞(MG) 而雪旺细胞(SCs)，无论是免疫启动的还是生长刺激的，都将被评估其能力 促进亚急性脊髓损伤小鼠模型的修复和恢复以回答可行性的基本问题， 递送，和疗效。拟议研究的目标将通过两个具体目标来实现。在AIM 1，最有效的细胞衍生EV类型将根据它们促进神经细胞存活的能力来确定 和轴突的体外生长。此外，该目标将在实验性脊髓损伤小鼠模型中优化它们的体内释放。 并评估它们在改善炎症、星形胶质细胞增生症和再生相关基因方面的比较效果 (RAG)表达抑制，同时促进神经发生、轴突生长和功能恢复。水泡 脊髓损伤后最有效的EV的含量将根据其核酸含量来表征 识别与其修复潜力相关的特定microRNA序列。在AIM中，2辆电动汽车将 专为细胞特异性递送修复和神经源性microRNA而设计。的可行性和功能性 小胶质细胞和雪旺细胞来源的EV工程用于以特定的microRNA：miRNA-9靶向NSC 已经被证明改变了神经干细胞的命运程序，神经发生和神经胶质发生的限制， 分别在促进血管生成的同时。改造后的EV将通过体外测试和体内测试进行测试 对功能效率的影响的实验。拟议研究的总体目标是改善 我们对细胞来源的EV如何参与神经修复以及它们是否可以 进一步增强其对宿主细胞的有益影响和随后的修复行动 追随SCI。