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: 8650963
• 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 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8650963
• 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 probe; in vivo; insight; intervertebral disk degeneration; macrophage; meetings; molecular imaging; mouse model; nano; nanostructured; neutrophil; novel; novel strategies; nucleus pulposus; public health relevance; 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）：下腰痛是一个地方性问题，是美国人去看医生的第二大最常见原因，仅次于最常见的感冒和流感。腰痛也是第二大最常见的与疾病有关的原因，这是错过的工作日和最常见的残疾原因。近年来，一项研究表明，由于受损和退化的椎间盘损伤和退化，损伤引起的炎症在腰痛中的可能作用。因此，自1950年代的治疗开始以来，使用透视镜检查的腰椎硬膜外注射以减少局部化学炎症反应，从而急剧增加。但是，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，空气栓塞和血管腔晕厥。此外，硬膜外注射在技术上是要求的。它通常由医生进行特殊培训的医生在X光检查下进行。当患者预约时，疼痛可能处于更为亚急性或慢性状态。因此，对急性/慢性背痛的经济高效治疗非常重要。我们的应用提出了一种新的治疗方法，该治疗方法是将纳米 - 富勒烯与CFLFLF联系起来，以便针对激活的中性粒细胞，这些嗜中性粒细胞迁移到椎间盘突出症状位点，并通过外周静脉注射来治疗阿科特背痛，这在技术上并不要求，并且可以由护士进行。没有特定生物学靶标的有效的抗氧化和抗炎特性的纳米 - 富勒烯不仅可以减少因髓质椎间盘挤出核组织组织从椎间盘突出而引起的痛觉过敏，还可以消除与标准生物学靶向靶向的药物分子相关的副作用。但是，需要一个有效的输送系统来克服其高度疏水性能，以专门针对损伤部位。这项研究的总体假设是激活的嗜中性粒细胞表达甲基肽受体（FPR）将与蛋白水解抗性合成肽CFLFLF结合，并且它们将共同浸润并积累到遗传性DIC中的炎症部位。将纳米 - 富勒烯与CFLFLF（cflflf-peg-fullerene）连接起 治疗由于椎间盘突出症和周围过度炎症而引起的疼痛。在拟议的研究的AIM 1中，我们将使用中性粒细胞特异性的小分子成像剂和免疫组织学来表征活化的中性粒细胞渗入急性椎间盘突出症。成像信号将使用FPR基因敲除小鼠进行验证。将分析中性粒细胞和巨噬细胞的表达和定量FPR和损伤的定量。在AIM 2中，我们将表征纳米 - 富勒烯水性悬浮液对椎间盘突出症在体外和体内引起的局部炎症的抑制作用。在AIM 3中，我们将合成CFLFLF-PEG-富勒烯，并评估其在小鼠辐射模型中的局部递送与局部递送到炎症部位；我们还将在针线穿刺引起的椎间盘变性模型中使用野生型和FPR敲除小鼠评估损伤部位的全身性传递与非特异性全身性传递。这项研究将为急性和慢性背部/颈部/腿部疼痛提供一种新颖的具有成本效益的治疗策略的见解，涉及与中性粒细胞特异性结合肽的CFLFLF结合的非蛋白质，纳米结构化的，生物相容性的富勒烯。与硬膜外类固醇注射相比，该方法具有更高的成本效益。