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 294,000 人患有脊髓损伤 (SCI)，其中超过 40,000 人患有脊髓损伤 退伍军人。尽管一些治疗方向在实验范式中显示出希望，但并没有 临床上存在一种可以显着逆转 SCI 相关神经功能缺损的恢复性治疗 以改善功能。处于正在转化为临床的实验性再生疗法的最前沿 人类 SCI 试验是细胞移植，从神经干细胞和间充质干细胞到施万细胞 和嗅觉鞘细胞。尽管观察到 SCI 后细胞植入的益处，但仍存在严峻的挑战 与它们的使用仍然相关，包括受损脊髓内的生存能力差，需要 非自体免疫抑制剂以及不需要的细胞分化、增殖的可能性， 植入细胞的迁移或迁移导致各种不良影响。而组合方法 已被证明可以克服其中一些缺陷，这是外源细胞的替代策略 疗法是通过外泌体囊泡（EV）刺激宿主修复。 EV 是纳米尺寸的内吞囊泡 细胞释放到周围环境中，从而允许生物分子在它们之间转移。电动汽车包含一个 各种货物，从 microRNA 到蛋白质和信号中介，可以促进细胞存活， 分化、轴突生长和髓鞘形成或抑制炎症和疤痕形成。有一种不断增长的 一致认为 EV 在调节成体神经干生态位中发挥着至关重要的作用。这些电动汽车还提供 能够被设计为表达荧光标记、靶向选择性细胞类型或加载 用于组织或靶细胞特异性递送的特定货物（例如小分子、肽和 miRNA）。 我们对细胞衍生的 EV 的理解以及实现其治疗潜力的最新进展 中风和心血管疾病等疾病扩大了EV领域。然而，它们的用途是 SCI 后的治疗方式受到限制，很大程度上仍处于起步阶段。在拟议的研究中，我们 将重点关注神经保护、神经源性和再生潜力的比较评估 EV 来自不同的亲代细胞群并在不同的细胞培养条件下。小胶质细胞 (MG) 免疫引发或生长刺激的雪旺细胞 (SC) 将评估其能力 促进亚急性 SCI 小鼠模型的修复和恢复，以回答可行性的基本问题， 交付和功效。拟议研究的目标将通过两个具体目标来实现。瞄准 1、将根据其促进神经细胞存活的能力来鉴定最有效的细胞源性EV类型 和轴突体外生长。此外，该目标还将优化其在实验性 SCI 小鼠模型中的体内递送 并评估它们对改善炎症、星形胶质细胞增生和再生相关基因的比较效果 (RAG) 表达抑制，同时促进神经发生、轴突生长和功能恢复。囊泡 SCI后最有效的EV的含量将根据其核酸含量进行表征 识别与其修复潜力相关的特定 microRNA 序列。 Aim 2 电动汽车将 专为修复性和神经源性 microRNA 的细胞特异性递送而设计。可行性和功能性 小胶质细胞和雪旺细胞衍生的 EV 工程，用于以特定 microRNA：miRNA-9 为目标的 NSC 已被证明可以改变神经干细胞的命运程序、神经发生和胶质细胞生成的限制， 分别同时促进血管生成。工程化的 EV 将使用体外测定和体内测定进行测试 实验对功能功效的影响。拟议研究的总体目标是改善 我们对细胞衍生的 EV 如何参与神经修复以及它们是否可以参与的理解 旨在进一步增强其对宿主细胞的有益作用和随后的修复作用 继SCI。