Systemic Application for Injury Site Specific Delivery via Neutrophils to Treat B

通过中性粒细胞进行损伤部位特异性递送来治疗 B 的系统应用

• 批准号: 8891368
• 负责人: XUDONG J. LI
• 金额: $ 26.86万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-19 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8891368
• 关键词: Acute; Adverse effects; Air Embolism; American; Animal Experimentation; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Appointment; Area; Back; Back Pain; Binding; Biochemistry; Biocompatible; Biological; Chronic; Clinic; Clinical; Clinical Trials; Common Cold; Development; Ensure; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Epidural Injections; Equipment; Ethers; Event; Fluoroscopy; Fullerenes; Half-Life; Health; Hematoma; Hernia; Hyperalgesia; Image; In Vitro; Infection; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Injection of therapeutic agent; Injury; Intervertebral disc structure; Knock-out; Knockout Mice; Link; Low Back Pain; Measurement; Methods; Modeling; Modification; Mus; Nanoconjugate; Nanostructures; Neck; Needles; Nerve; Nurses; Operative Surgical Procedures; Outcome; Pain; Pain in lower limb; Parents; Patients; Peptides; Peripheral; Pharmaceutical Preparations; Plant Roots; Polyethylene Glycols; Property; Proteins; Proteolysis; Publishing; Puncture procedure; Radiculopathy; Research; Research Personnel; Resistance; Resolution; Risk; Role; Signal Transduction; Site; Slipped Disk; Spinal Ganglia; Steroids; Surgeon; Suspension substance; Suspensions; System; TNF gene; Technical Expertise; Testing; Therapeutic; Therapeutic Agents; Tissues; Training; Translational Research; Validation; Vasovagal Syncope; Veins; Vertebral column; Work; aqueous; base; chronic back pain; cost effective; cost effectiveness; disability; effective therapy; fMet-Leu-Phe receptor; flu; imaging agent; imaging probe; in vivo; insight; intervertebral disk degeneration; macrophage; meetings; molecular imaging; mouse model; nano; nanostructured; neutrophil; novel; novel strategies; nucleus pulposus; receptor; receptor expression; single photon emission computed tomography; small molecule; synthetic peptide

DESCRIPTION (provided by applicant): Low back pain is an endemic problem and is the second most common reason that Americans go to see their doctor, second only to the most common cold and the flu. Low back pain is also the second most common illness-related reason given for a missed workday and the most common cause of disability. In recent years, a body of research has suggested a possible role of injury-induced inflammation in low back pain caused by damaged and degenerating intervertebral discs. As such, the use of fluoroscopy-guided lumbar epidural injections of steroids to reduce the local chemo-inflammatory response has increased dramatically since their inception as a treatment in the 1950s. However, the risks of epidural injections include, but are not limited to, epidural hematoma, epidural infection, nerve root injury, dural puncture, air embolism and vasovagal syncope. Moreover, the epidural injection is technically demanding. It is usually performed under x-ray by doctors with special training. When a patient gets an appointment, the pain is likely in a more subacute or chronic state. Therefore, a cost-effective treatment for acute/chronic back pain is of great importance. Our application proposes a novel treatment by linking nano-fullerene to cFLFLF in order to specifically target activated neutrophils that migrate into the disc herniation site and treat acut back pain via peripheral vein injection, which is not technically demanding and could be performed by a nurse. Nano-fullerene, which has potent anti-oxidative and anti-inflammatory properties, without specific biological target could not only reduce hyperalgesia induced by the extrusion of nucleus pulposus tissue from herniated discs, but also eliminate the side effects associated with standard biological target-oriented drug molecules. However, an effective delivery system is needed to overcome its highly hydrophobic properties to specifically target the site of injury. The overall hypothesis of this study is that the activated neutrophils expressig formyl peptide receptor (FPR) will bind to proteolytically resistant synthetic peptide cFLFLF and that together they will infiltrate and accumulate into the site of inflammation in the herniated dic. Linking nano-fullerene to a cFLFLF (cFLFLF-PEG-fullerene) will ensure event-specific temporal and spatial delivery of the anti-inflammatory agent to the site of injury via systemic injection to treat the pain caused by excessive inflammation in and around herniated discs. In Aim 1 of the proposed study, we will characterize the infiltration of activated neutrophils into an acute disc herniation using neutrophil-specific small molecule imaging agents and immunohistology. The imaging signal will be validated using FPR knockout mice. The neutrophils and macrophages will be analyzed for expression and quantification of FPR and for the quantification of injury. In Aim 2, we will characterize the inhibitory effect of aqueous suspensions of nano-fullerene on local inflammation induced by disc herniation in vitro and in vivo. In Aim 3, we will synthesize cFLFLF-PEG-fullerene and evaluate its systemic vs. local delivery to a site of inflammation in a mouse radiculopathy model; we will also assess injury site target systemic delivery vs. non- specific systemic delivery using wild type and FPR knockout mice in a disc degeneration model induced by needle puncture. This study will provide useful insights into a novel cost-effective therapeutic strategy for acute and chronic back/neck/leg pain involving systemic injection of non-protein, nano-structured, biocompatible fullerene conjugated with cFLFLF, a neutrophil-specific binding peptide. This method is much more cost- effectiveness compared to epidural steroid injection.

描述（由申请人提供）：腰痛是一种地方病，是美国人去看医生的第二大常见原因，仅次于最常见的感冒和流感。腰痛也是导致缺勤的第二大常见疾病相关原因，也是最常见的残疾原因。近年来，大量研究表明，损伤引起的炎症可能在椎间盘受损和退化引起的腰痛中发挥作用。因此，自 20 世纪 50 年代作为一种治疗方法出现以来，使用透视引导下腰椎硬膜外注射类固醇来减少局部化学炎症反应的情况急剧增加。然而，硬膜外注射的风险包括但不限于硬膜外血肿、硬膜外感染、神经根损伤、硬膜穿刺、空气栓塞和血管迷走性晕厥。而且，硬膜外注射技术要求较高。通常由经过特殊培训的医生在 X 光检查下进行。当患者就诊时，疼痛可能处于亚急性或慢性状态。因此，针对急/慢性背痛的经济有效的治疗非常重要。我们的申请提出了一种新颖的治疗方法，将纳米富勒烯与 cFLFLF 连接起来，以特异性靶向迁移到椎间盘突出部位的活化中性粒细胞，并通过外周静脉注射治疗急性背痛，这在技术上要求不高，可以由护士进行。纳米富勒烯具有强效抗氧化和抗炎特性，无特定生物靶点，不仅可以减轻椎间盘突出髓核组织挤出引起的痛觉过敏，还可以消除标准生物靶点相关的副作用药物分子。然而，需要一种有效的递送系统来克服其高度疏水性，以特异性地靶向损伤部位。本研究的总体假设是，表达甲酰肽受体（FPR）的活化中性粒细胞将与蛋白水解抗性合成肽 cFLFLF 结合，并且它们将一起渗透并积聚到疝出的迪克的炎症部位。将纳米富勒烯连接到 cFLFLF（cFLFLF-PEG-富勒烯）将确保通过全身注射将抗炎剂在特定事件的时间和空间上递送到损伤部位 治疗椎间盘突出及其周围过度炎症引起的疼痛。在拟议研究的目标 1 中，我们将使用中性粒细胞特异性小分子成像剂和免疫组织学来表征活化的中性粒细胞浸润到急性椎间盘突出症中的情况。成像信号将使用 FPR 敲除小鼠进行验证。将分析中性粒细胞和巨噬细胞的 FPR 表达和定量以及损伤的定量。在目标 2 中，我们将在体外和体内表征纳米富勒烯水悬浮液对椎间盘突出引起的局部炎症的抑制作用。在目标 3 中，我们将合成 cFLFLF-PEG-富勒烯，并在小鼠神经根病模型中评估其对炎症部位的全身与局部递送；我们还将在针刺诱导的椎间盘退变模型中使用野生型和 FPR 敲除小鼠评估损伤部位目标全身递送与非特异性全身递送。这项研究将为急性和慢性背部/颈部/腿部疼痛的新型经济有效的治疗策略提供有用的见解，该策略涉及全身注射与 cFLFLF（一种中性粒细胞特异性结合肽）缀合的非蛋白质、纳米结构、生物相容性富勒烯。与硬膜外类固醇注射相比，该方法更具成本效益。