STTR: Herpes Based RNAi Vectors

STTR：基于疱疹的 RNAi 载体

• 批准号: 7226932
• 负责人: Stephen Chang
• 金额: $ 10.83万
• 依托单位: SIMPLX PHARMACEUTICALS
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7226932
• 关键词: Affect; Afferent Neurons; American; Animal Model; Antibodies; Back; Behavioral; Cell Nucleus; Cell membrane; Cell physiology; Cells; Chemistry; Chronic inflammatory pain; Development; Disease; Endosomes; Fluorescence; Gene Expression; Gene Proteins; Gene Silencing; Gene Transduction Agent; Genes; Genetic Transcription; Goals; Green Fluorescent Proteins; HSV vector; Herpes Labialis; Herpesviridae; Hyperalgesia; Hypersensitivity; Image Analysis; Immunohistochemistry; In Vitro; Individual; Infectious Skin Diseases; Inflammation; Inflammatory; Intention; Laboratories; Maintenance; Measures; Mechanics; Mediating; Methods; Mus; Nerve; Neurons; Nociceptors; Numbers; Pain; Patients; Peripheral; Pharmacologic Substance; Phase; Polymerase Chain Reaction; Population; Primates; Principal Investigator; Process; Protein Isoforms; Protein Overexpression; Proteins; Purpose; RNA; RNA Interference; RNAi vector; Recombinants; Residual state; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Science; Simplexvirus; Skin; Small Business Technology Transfer Research; Sodium Channel; Sodium Channel Blockers; Specificity; Spinal Ganglia; Stimulus; Structure; Technology; Testing; Time; Tissues; Toxic effect; Transgenes; Virus; Work; base; behavior measurement; chronic pain; enhanced green fluorescent protein; gene therapy; immunoreactivity; improved; in vivo; inflammatory pain; interest; knock-down; neuronal cell body; neurotropic; programs; protein expression; recombinant virus; success; targeted delivery; tool; transgene expression; vector; voltage

DESCRIPTION (provided by applicant): Chronic pain and hypersensitivity associated with inflammation affects millions of Americans. Recently, it has become apparent that a critical mechanism in developing and maintaining inflammatory pain is the overexpression of certain sodium channels in the sensory neurons that innervate the inflamed tissue. Thus, pharmaceutical companies have for some years attempted to develop selective blockers of these sodium channels, but without success. A more promising avenue for the treatment of chronic inflammatory pain might be a gene therapy approach that selectively knocks down the expression of the overexpressed sodium channels. One of the most promising approaches to selectively knocking down or silencing gene expression has been the development of small inhibitory RNAs. RNAi's, and particularly small buttonhook RNAs (shRNA), have shown great promise in producing long duration silencing of aberei1t gene expression. The primary limitation of this technology has been the selective targeting of these molecules to the tissue and cells of interest, in this case, the sensory neurons that mediate pain in inflamed structures. For a number of years, recombinant herpes simplex viruses (those that cause cold sores) have been used as vectors for selective delivery of transgenes to the nuclei of pain sensing neurons. Thus herpes viruses are naturally neurotropic, and an estimated 85% of the US population have these viruses in our sensory neurons at anyone time. The toxicity of these viruses is remarkably low naturally, and the recombinant viruses in use for gene therapy purposes are non-replicating, thus, removing residual toxicity. Thus, recombinant herpes viruses, encoding a transgene of interest, applied to peripheral tissue, e.g., skin, are taken up by the terminals of sensory neurons, transported back to the cell bodies, and exist as endosomes in the nucleus of these cells, presenting the transgene to the transcription machinery of the cell. Work in rodents and primates has demonstrated prolonged (> 20 wks) transgene expression using these means, limited to the cells that innervate the treated tissue. In this Phase I project, it is our intention to use herpes vectors to deliver shRNAs directed toward silencing the expression of particular sodium channels to the sensory neurons innervating the inflamed tissue of mice. It is our long term goal to develop herpes based gene therapy vectors for the treatment of inflammatory and other chronic pain states in patients.

描述（由申请人提供）：与炎症相关的慢性疼痛和超敏反应影响数百万美国人。最近，已经变得明显的是，发展和维持炎性疼痛的关键机制是在支配发炎组织的感觉神经元中某些钠通道的过表达。因此，制药公司多年来一直试图开发这些钠通道的选择性阻断剂，但没有成功。一个更有希望的治疗慢性炎性疼痛的途径可能是基因治疗方法，选择性地敲低过表达的钠通道的表达。选择性敲低或沉默基因表达的最有前途的方法之一是开发小抑制性RNA。RNAi，特别是小的buttonhook RNA（shRNA），已经显示出很大的希望，在产生长时间沉默的aberei1t基因表达。该技术的主要局限性是这些分子选择性靶向感兴趣的组织和细胞，在这种情况下，是在发炎结构中介导疼痛的感觉神经元。多年来，重组单纯疱疹病毒（引起唇疱疹的病毒）一直被用作载体，用于选择性地将转基因传递到痛觉神经元的核中。因此，疱疹病毒是天然嗜神经的，估计85%的美国人口在任何时候都在我们的感觉神经元中携带这些病毒。这些病毒的毒性自然是非常低的，并且用于基因治疗目的的重组病毒是非复制的，因此消除了残留毒性。因此，将编码感兴趣的转基因的重组疱疹病毒应用于外周组织，例如，皮肤中的转基因被感觉神经元的末梢吸收，转运回细胞体，并作为内体存在于这些细胞的细胞核中，将转基因呈递给细胞的转录机构。在啮齿动物和灵长类动物中的工作已经证明使用这些方法延长（> 20周）转基因表达，仅限于神经支配被处理组织的细胞。在这个I期项目中，我们的目的是使用疱疹病毒载体来递送shRNA，该shRNA定向沉默特定钠通道到支配小鼠发炎组织的感觉神经元的表达。我们的长期目标是开发基于疱疹的基因治疗载体，用于治疗患者的炎症和其他慢性疼痛状态。