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

• 批准号: 7983425
• 负责人: ERIN Damita MILLIGAN
• 金额: $ 33.12万
• 依托单位: UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
• 依托单位国家: 美国
• 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/7983425
• 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; Data; Dendritic Cells; Devices; Dose; Drug Delivery Systems; Drug Formulations; Effectiveness; Encapsulated; Etiology; Exposure to; FDA approved; Figs - dietary; 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; viral DNA

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NIDA尖端基础研究奖（CEBRA）第二阶段的竞争性更新。其目标是专注于开发一种新的治疗持续性疼痛的方法。持续性疼痛（3个月以上）是美国人常见的未解决的健康问题。我们对神经性疼痛（疼痛通路中的病理性神经元信号传导）的理解中的最近考虑包括免疫细胞和神经胶质细胞（星形胶质细胞、小胶质细胞、卫星细胞和许旺细胞）在疼痛相关隔室（例如脊髓背角、脊膜相关蛛网膜下基质和背根神经节）中的贡献。脊髓胶质细胞通过释放特征明确的促炎细胞因子介导病理性疼痛。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。在目前的资助期间，数据强烈支持IL-10的脊髓蛛网膜下（鞘内; IT）基因递送在动物模型中预防和逆转病理性疼痛。在2次连续IT注射非病毒载体后实现长期（3+个月）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）。第一次注射用于使脊髓蛛网膜下腔对第二次注射敏感，第二次注射产生IL-10依赖性长期疼痛缓解。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。在致敏间隔期间的IL-10蛋白信号传导对于长期IL-10基因治疗是必要的。我们推测，几种免疫相关的病因，包括趋化性，有丝分裂，和吞噬作用的敏化和IL-10转基因的摄取发挥关键作用。因此，活化的（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摄取的致敏反应。FDA批准的合成聚合物在以减少的剂量制剂单次注射后改善pDNA-IL-10药物递送。然而，临床试验需要进一步改进。鉴定致敏背后的解剖学区域、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。本发明的目的是直接的：（1）鉴定致敏过程中疼痛相关区域的细胞/生化反应;（2）检查致敏过程中重要的细胞/生化特征在长时间基因表达和疼痛缓解过程中是否也是必需的;和（3）使用PLGA-pDNA-IL-10制剂进一步改善IT基因递送，所述制剂包括在致敏和长时间致敏期间重要因子的共释放，持续基因表达，使得可以从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递送有效地抑制促炎细胞因子作用，并产生非病毒载体的2次连续IT注射的长期（3+个月）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）。注射间隔的特征在于脊髓蛛网膜下腔对第二次注射的致敏期，其产生IL-10依赖性长期疼痛缓解，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。尽管临床试验需要进一步改进，但初步研究表明，FDA批准的合成聚合物在单次注射减少剂量制剂后改善了pDNA-IL-10药物递送。