Preventive and Therapeutic Effects of Bone Marrow Stem Cell-Derived Exosomes on Spine Dural Fibrosis

骨髓干细胞源性外泌体对脊柱硬膜纤维化的预防和治疗作用

• 批准号: 10350148
• 负责人: Dong Rim Seol
• 金额: $ 16.67万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-10 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10350148
• 关键词: ANXA5 gene; Adhesions; Aftercare; Apoptosis; Applications Grants; Attention; Back; Back Pain; Biological Assay; Bone Marrow Stem Cell; Cattle; Cell Culture Techniques; Cell Lineage; Cells; Chronic; Cicatrix; Clinical; Collagen; Collagen Type I; Contracts; Data; Deposition; Dura Mater; Encapsulated; Endoscopy; Extracellular Matrix; Female; Fibroblasts; Fibrosis; Future; Gel; Gene Expression; Goals; Histologic; Human; Hyaluronic Acid; Hydrogels; Immune response; Immunofluorescence Immunologic; In Vitro; Infiltration; Inflammatory Response; Injectable; Laminectomy; Liver; Lower Extremity; Lung; Mesenchymal; Mesenchymal Stem Cells; MicroRNAs; Modeling; Morphology; Myofibroblast; Nerve; Nerve compression syndrome; Operative Surgical Procedures; Outcome; Paracrine Communication; Patients; Pharmacology; Plant Roots; Prevention; Preventive; Production; Rattus; Research; Site; Smooth Muscle Actin Staining Method; Source; Spinal; Spinal Cord; Spinal nerve root structure; Spine surgery; Sprague-Dawley Rats; Stains; Stem cell transplant; Syndrome; TGFB1 gene; Testing; Therapeutic; Therapeutic Effect; Tissues; Trauma; Vertebral column; Werdnig-Hoffmann Disease; base; cell type; engineered exosomes; exosome; experience; in vivo; inhibitor; intercellular communication; male; miRNA expression profiling; migration; minimally invasive; next generation sequencing; operation; paracrine; prevent; protein biomarkers; radicular pain; recruit; reduce symptoms; spinal cord compression; stem cell exosomes; stem cells; success; symptom cluster; tissue repair; tumorigenesis; uptake; zeta potential

PROJECT SUMMARY/ABSTRACT Failed back surgery syndrome (FBSS) refers to a cluster of symptoms in spinal cord and nerve roots after spinal surgery, especially laminectomy and discectomy, and up to 40% of patients have experienced chronic nerve radicular pain and lower extremity weakness. The main cause of FBSS is the compression of dura and nerve roots due to spinal dural fibrosis, including epidural fibrosis and peridural adhesion. Multiple factors leading to dural fibrosis are the inflammatory response post-surgical trauma and excessive matrix deposition by highly activated myofibroblasts. Spinal dural fibrosis, if not properly treated by conservative management, can generally worsen, which may require surgical treatments. However, success of these surgical interventions is only 30-35%, and up to 20% of patients have more serious condition. Thus, there is a critical and urgent need to prevent and treat spinal dural fibrosis. Exosomes have attracted attention due to their great potential as an inhibitor of dural fibrosis. Damaged tissues are repaired by the paracrine signaling of exosomes rather than direct proliferation and differentiation and this paracrine effect implies that exosome therapy has a clinical advantage over stem cell transplantation in terms of immune response and tumorigenesis. Mesenchymal stromal/stem cells (MSCs) are promising sources of exosomes, and MSC-derived exosomes showed an evident reduction of fibrosis in liver and lung. The long-term goal is to develop a minimally invasive anti-fibrosis therapy for spine dural fibrosis. To achieve that goal, we propose to evaluate the effects of exosomes on reduction of alpha smooth muscle actin (α-SMA), type I collagen, and contractile activities in human myofibroblasts and the in vivo preventive effects in a rat laminectomy model. Our central hypotheses are that: exosomes have the potential to alleviate myofibroblast activities, and hydrogel will allow sustained release of exosomes and restrict fibroblast migration as a physical barrier between the dura mater and posterior tissue. The hypotheses have been devised on the basis of our preliminary data, which revealed that MSC-derived exosomes showed both preventive and therapeutic potential in exogenously induced bovine myofibroblasts by transforming growth factor beta 1 (TGF-β1). The reduction of α-SMA expression as a myofibroblast marker was notable in pre- and post-TGF-β1 treatment with exosomes. At the completion of the proposed R21 project, our expected outcomes are to define the therapeutic potential of exosomes for prevention of peridural fibrosis and to identify microRNAs (miRNA) that may regulate anti-fibrotic effects. These results will have a very important positive impact by providing preliminary data for our future R01 grant application, which will be to evaluate in vivo therapeutic effects of MSC-derived exosomes treated after induction of dural fibrosis and to synthesize target miRNAs to avoid stem cell culture as a means of exosome production. Synthetic miRNAs will be loaded in engineered exosomes for in vivo delivery, thereby replacing MSC culture as a potential means of exosome production.

项目概要/摘要 背部手术失败综合征（FBSS）是指背部手术失败后出现的一系列脊髓和神经根症状。 脊柱手术，尤其是椎板切除术和椎间盘切除术，高达 40% 的患者经历过慢性 神经根性疼痛和下肢无力 FBSS的主要原因是硬脑膜和下肢的压迫。 因脊椎神经根纤维化，包括硬膜外纤维化和硬膜外粘连等多种因素导致。 硬脑膜纤维化的主要原因是手术创伤后的炎症反应和高度的基质沉积。 激活的肌成纤维细胞，如果不采取保守治疗，通常会导致硬脊膜纤维化。 病情恶化，可能需要手术治疗，但这些手术干预的成功率仅为 30-35%， 高达20%的患者病情更为严重，因此，预防和治疗刻不容缓。 外泌体由于其作为硬脑膜抑制剂的巨大潜力而​​引起人们的关注。 纤维化是通过外泌体的旁分泌信号修复的，而不是直接增殖。 和分化以及这种旁分泌效应意味着外泌体疗法比干细胞疗法具有临床优势 间充质基质/干细胞（MSC）的免疫反应和肿瘤发生方面的移植。 有希望的外泌体来源，并且 MSC 衍生的外泌体显示出明显减少肝脏纤维化 和肺。 长期目标是开发针对脊柱硬膜纤维化的微创抗纤维化疗法来实现。 为了实现这一目标，我们建议评估外泌体对减少 α 平滑肌肌动蛋白 (α-SMA) 的影响， I 型胶原蛋白、人肌成纤维细胞的收缩活性以及大鼠体内的预防作用 我们的中心假设是：外泌体具有减轻肌成纤维细胞的潜力。 活性，水凝胶将允许外泌体持续释放并限制成纤维细胞迁移作为物理 硬脑膜和后部组织之间的屏障是根据我们的假设提出的。 初步数据显示 MSC 衍生的外泌体显示出预防和治疗潜力 在外源诱导的牛肌成纤维细胞中，转化生长因子β1（TGF-β1）的减少。 α-SMA 作为肌成纤维细胞标志物的表达在外泌体 TGF-β1 治疗前后显着。 在拟议的 R21 项目完成后，我们的预期结果是确定治疗潜力 外泌体用于预防硬膜外纤维化并鉴定可能调节抗纤维化的 microRNA (miRNA) 这些结果将为我们未来的R01提供初步数据，产生非常重要的积极影响。 拨款申请，该申请将评估 MSC 衍生的外泌体治疗后的体内治疗效果 诱导硬脑膜纤维化并合成目标 miRNA，以避免干细胞培养作为外泌体的手段 合成的 miRNA 将被加载到工程外泌体中用于体内递送，从而取代 MSC。 培养作为外泌体生产的潜在手段。