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: 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
• 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

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）：本提案是对II阶段NIDA尖端基础研究奖（CEBRA）的竞争更新。它的目的是开发一种新的疗法，以缓解持续的疼痛。持续的疼痛（3个月以上）是美国人常见的未解决的健康问题。在我们对神经性疼痛（疼痛途径中的病理神经元信号传导）的理解中的最新考虑包括免疫细胞和神经胶质（星形胶质细胞，小胶质细胞，卫星和Schwann）在疼痛相关的隔室中的贡献，例如脊髓索脊髓背侧角，脊髓脑膜，脊髓脑膜脑膜相关的脑膜囊型基质基质和Dorsal and and and and and andans and andans and andans and andans and and andans and andal and andal andal andlia。脊髓神经胶质通过释放良好的促炎细胞因子介导病理疼痛。抗炎细胞因子白细胞介素10（IL-10）有效抑制促炎性细胞因子作用。在当前赠款期间，数据强烈支持IL-10的IL-10基因递送的脊柱亚蛛网膜下腔（IT）基因递送可防止动物模型中的病理疼痛。长时间（3个多个月）在2个顺序注射非病毒载体时可缓解疼痛，其中第二个注射必须编码IL-10（质粒DNA编码IL-10； pDNA-IL-10）。第一次注射是将脊柱亚蛛网膜下腔隔室敏感的第二个注射，从而产生IL-10依赖性的长持续疼痛缓解。神经胶质，巨噬细胞和/或树突状细胞的强大积累是敏化的组成部分。在敏化间隔期间，IL-10蛋白信号传导对于长期IL-10基因治疗是必需的。我们假设几种免疫相互关联的病因，包括趋化性，有丝分裂和吞噬作用在致敏和IL-10转基因摄取中起关键作用。因此，激活的（趋化和/或有丝分裂）免疫细胞和神经胶质可能部分负责对PDNA-IL-10摄取的敏感反应。在减少剂量配方时，通过FDA批准的合成聚合物可以改善pDNA-IL-10药物递送。但是，临床试验需要进一步改进。可以利用识别解剖区域，细胞类型和细胞活性，以进一步改善聚合物脊柱IL-10靶向基因递送。本提案的目的很简单：（1）确定敏化期间疼痛相关区域的细胞/生化反应； （2）检查在长期基因表达和缓解疼痛的敏化过程中，在敏化过程中重要的细胞/生化特征是否也需要； （3）使用plga-pDNA-IL-10制剂进一步改善IT基因递送，其中包括在敏化和长持续基因表达过程中重要的因子共同释放，从而可以从单个注射中实现持久的疼痛缓解。 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）。注射间隔的特征是脊柱亚蛛网膜下腔室对第二次注射的敏化期，该注射会产生IL-10依赖性的长持续性疼痛缓解，并且可能涉及免疫相互关联的病因，包括趋化性，有符合性，有差点和吞噬作用。尽管临床试验需要进一步改进，但初步研究表明，通过减少剂量配方，单次注射后，由FDA批准的合成聚合物改善了PDNA-IL-10药物的递送。