Spinal Neuroimmune Mechanisms Underlying IL-10 Gene Therapy for Pain Control

IL-10 疼痛控制基因疗法背后的脊髓神经免疫机制

基本信息

批准号：
8299615
负责人：
ERIN Damita MILLIGAN
金额：
$ 32.2万
依托单位：
UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-09-20 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8299615
关键词：
American Animal Model Anti-Inflammatory Agents Anti-inflammatory Astrocytes Award Basic Science Biochemical Biochemical Process Biological Process Cell Culture Techniques Cell Survival Cell physiology Cells Cellular Structures Chemotaxis Clinical Clinical Trials Collecting Cell DNA Data Dendritic Cells Devices Dose Drug Delivery Systems Drug Formulations Effectiveness Encapsulated Etiology Exposure to FDA approved Gene Delivery Gene Expression Gene Transfer Glycolates Goals Grant Health IL10 gene Immune In Vitro Injection of therapeutic agent Interleukin-10 Lead Mediating Microglia Mitosis Mitotic National Institute of Drug Abuse Neuroglia Neuroimmunomodulation Neurons Non-Viral Vector Pain Pain management Pathway interactions Persistent pain Phagocytosis Pharmaceutical Preparations Phenotype Plasmid Cloning Vector Play Polymers Population Procedures Process Production Proteins Rattus Reporter Reporter Genes Research Role Series Signal Transduction Signaling Protein Spinal Spinal Cord Spinal Ganglia Spinal cord posterior horn Spinal meninges Staging Syndrome Techniques Testing Therapeutic Time Transgenes Treatment Efficacy Viral Work cell type chronic neuropathic pain cytokine design dosage gene therapy improved in vivo macrophage novel strategies painful neuropathy plasmid DNA preclinical study prevent public health relevance release factor research study response therapeutic gene therapeutic transgene transgene expression uptake vector

项目摘要

DESCRIPTION (provided by applicant): The present proposal is a competing renewal of a Stage II NIDA Cutting Edge Basic Research Award (CEBRA). Its aims are focused on developing a new therapy for persistent pain relief. Persistent pain (3+ months) is a common, unresolved health problem in Americans. A recent consideration in our understanding of neuropathic pain (pathological neuronal signaling in the pain pathway) includes the contribution of immune cells & glia (astrocytes, microglia, Satellite & Schwann) in pain relevant compartments such as the spinal cord dorsal horn, spinal meninges associated subarachnoid matrix, & dorsal root ganglia. Spinal cord glia mediate pathological pain via the release of well-characterized proinflammatory cytokines. The anti-inflammatory cytokine, interleukin-10 (IL-10), potently inhibits proinflammatory cytokine actions. During the current grant period, data strongly support spinal subarachnoid (intrathecal; IT) gene delivery of IL-10 prevents & reverses pathological pain in animal models. Long-duration (3+ months) pain relief is achieved upon 2 sequential IT injections of non-viral vectors, where the 2nd injection must encode IL-10 (plasmid DNA encoding IL-10; pDNA- IL-10). The first injection serves to sensitize the spinal subarachnoid compartment to the 2nd injection that creates IL-10-dependent long-duration pain relief. A robust accumulation of glia, macrophage &/or dendritic cells are components of sensitization. IL-10 protein signaling during the sensitization interval is necessary for long-duration IL-10 gene therapy. We postulate that several immune interrelated etiologies, including chemotaxis, mitosis, & phagocytosis play critical roles for sensitization & IL-10 transgene uptake. Thus, activated (chemotactic &/or mitotic) immune cells & glia could be responsible, in part, for the sensitized response to pDNA-IL-10 uptake. An FDA-approved synthetic polymer improves pDNA-IL-10 drug delivery after a single injection at reduced dosage formulations. However, further improvement is needed for clinical trials. Identifying the anatomical region, cell type and cellular activity underlying sensitization can be exploited to further improve polymer spinal IL-10 targeted gene delivery. The aims of the present proposal are straightforward: (1) To identify the cellular/biochemical responses in pain-relevant regions during sensitization; (2) To examine whether the cellular/biochemical profiles important during sensitization are also necessary during long duration gene expression and pain relief; and (3) To further improve IT gene delivery using PLGA- pDNA-IL-10 formulations that include co-release of factors important during sensitization & long-duration gene expression such that enduring pain relief can be achieved from a single injection. PUBLIC HEALTH RELEVANCE: Spinal subarachnoid gene delivery of the anti-inflammatory cytokine, interleukin-10 (IL-10), potently inhibits proinflammatory cytokine actions and produces long-duration (3+ months) pain relief of 2 sequential IT injections of non-viral vectors, where the 2nd injection must encode IL-10 (plasmid DNA encoding IL-10; pDNA- IL-10). The inter-injection interval is characterized as a sensitization period of the spinal subarachnoid compartment to the 2nd injection that creates IL-10-dependent long-duration pain relief & may involve immune interrelated etiologies, including chemotaxis, mitosis, & phagocytosis. Although, further improvement is needed for clinical trials, preliminary studies show an FDA-approved synthetic polymer improves pDNA-IL-10 drug delivery after a single injection at reduced dosage formulations.

描述（由申请人提供）：本提案是第二阶段 NIDA 前沿基础研究奖 (CEBRA) 的竞争性更新。其目标是开发一种缓解持续疼痛的新疗法。持续疼痛（3 个月以上）是美国人常见的、尚未解决的健康问题。我们最近对神经性疼痛（疼痛通路中的病理性神经元信号传导）的理解包括免疫细胞和神经胶质细胞（星形胶质细胞、小胶质细胞、卫星细胞和雪旺细胞）在疼痛相关区室（如脊髓背角、相关脊膜）中的贡献蛛网膜下腔基质和背根神经节。脊髓神经胶质细胞通过释放明确的促炎细胞因子来介导病理性疼痛。抗炎细胞因子白细胞介素 10 (IL-10) 可有效抑制促炎细胞因子的作用。在当前资助期内，数据强烈支持脊髓蛛网膜下腔（鞘内；IT）基因递送 IL-10 可预防和逆转动物模型中的病理性疼痛。通过连续 2 次 IT 注射非病毒载体可实现长期（3 个月以上）疼痛缓解，其中第 2 次注射必须编码 IL-10（编码 IL-10 的质粒 DNA；pDNA-IL-10）。第一次注射使脊髓蛛网膜下腔对第二次注射敏感，从而产生依赖于 IL-10 的长期疼痛缓解。神经胶质细胞、巨噬细胞和/或树突细胞的大量积累是致敏的组成部分。致敏间隔期间的 IL-10 蛋白信号转导对于长期 IL-10 基因治疗是必要的。我们假设几种免疫相关的病因，包括趋化性、有丝分裂和吞噬作用，对于致敏和 IL-10 转基因摄取发挥着关键作用。因此，激活的（趋化和/或有丝分裂）免疫细胞和神经胶质细胞可能部分负责对 pDNA-IL-10 摄取的敏化反应。 FDA 批准的合成聚合物可在减少剂量的配方中单次注射后改善 pDNA-IL-10 药物输送。然而，临床试验还需要进一步改进。识别致敏背后的解剖区域、细胞类型和细胞活性可用于进一步改善聚合物脊髓 IL-10 靶向基因递送。本提案的目的很简单：（1）确定致敏过程中疼痛相关区域的细胞/生化反应； (2) 检查在致敏过程中重要的细胞/生化特征在长期基因表达和疼痛缓解过程中是否也是必要的； (3)使用PLGA-pDNA-IL-10制剂进一步改善IT基因递送，该制剂包括在致敏和长时间基因表达期间重要的因子的共同释放，从而可以通过单次注射实现持久的疼痛缓解。公共卫生相关性：抗炎细胞因子白细胞介素 10 (IL-10) 的脊髓蛛网膜下腔基因传递可有效抑制促炎细胞因子的作用，并通过 2 次连续 IT 注射非病毒药物产生长期（3 个月以上）疼痛缓解载体，其中第二次注射必须编码 IL-10（编码 IL-10 的质粒 DNA；pDNA-IL-10）。注射间隔的特征是脊髓蛛网膜下腔对第二次注射的敏化期，该敏化期产生 IL-10 依赖性长持续时间疼痛缓解，并且可能涉及免疫相关病因，包括趋化性、有丝分裂和吞噬作用。尽管临床试验还需要进一步改进，但初步研究表明 FDA 批准的合成聚合物在减少剂量配方的单次注射后可改善 pDNA-IL-10 药物输送。