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会影响数百万美国人。最近，很明显，发育和维持炎症性疼痛的关键机制是感官神经元中某些钠通道的过表达，它神经支配了发炎的组织。因此，制药公司多年来一直试图开发这些钠渠道的选择性阻滞剂，但没有成功。治疗慢性炎症性疼痛的更有希望的途径可能是一种基因治疗方法，它有选择地击倒过表达过表达的钠通道的表达。选择性击倒或沉默基因表达的最有前途的方法之一是抑制性RNA的发展。 RNAi，尤其是小型纽扣RNA（shRNA），在产生持续时间的Aberei1t基因表达方面表现出了巨大的希望。该技术的主要局限性是将这些分子的选择性靶向到感兴趣的组织和细胞，在这种情况下，介导发炎结构疼痛的感觉神经元。多年来，重组单纯疱疹病毒（引起唇疱疹的病毒）被用作选择性递送转基因疼痛传感神经元细胞核的向量。因此，疱疹病毒自然是神经性的，估计有85％的美国人群在任何人的感觉神经元中都有这些病毒。这些病毒的毒性自然很低，用于基因治疗目的的重组病毒是不复制的，因此可以消除残留的毒性。因此，编码感兴趣的转基因的重组疱疹病毒被应用于外周组织，例如皮肤，被感觉神经元的末端吸收，转移回细胞体，并作为内体作为内体存在于这些细胞的核中，呈现这些细胞的内体，呈现转基因向细胞的转录机械。啮齿动物和灵长类动物中的工作表明使用这些手段延长（> 20 wks）的转基因表达，仅限于支配处理的组织的细胞。在此I阶段项目中，我们打算使用疱疹向量来传递用于沉默的shrnas，以使特定的钠通道表达对感官神经元的表达，从而支配小鼠发炎的组织。我们的长期目标是开发基于疱疹的基因疗法载体，用于治疗患者的炎症和其他慢性疼痛状态。