P20, 'MOLECULAR SHORTSTOP' FOR INFLAMATORY LUNG DISEASES

P20，炎症性肺病的“分子捷径”

• 批准号: 6076035
• 负责人: KENNETH L BRIGHAM
• 金额: $ 10.69万
• 依托单位: GENERX+, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-20 至 2001-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6076035
• 关键词: antiinflammatory agents; cystic fibrosis; drug design /synthesis /production; gene therapy; inflammation; interleukin 6; interleukin 8; liposomes; lung disorder; recombinant proteins; respiratory epithelium; transcription factor; transfection; transfection /expression vector

DESCRIPTION: (Adapted from the Investigator's Abstract) In preliminary studies of the mechanism of the inflammatory response in airway epithelial cells expressing the cystic fibrosis defect, it was found that normal airway epithelial cells turn off production of IL-6 and IL-8 when stimulated with TNF by increasing production of p20, a truncated isoform of the transcription factor C/EBPbeta (NF-IL6) which inhibits transcriptional activation by the full length C/EBPbeta protein. In contrast, airway epithelial cells expressing the CF defect do not turn off production of these pro-inflammatory cytokines when stimulated with TNF and do not increase production of p20. C/EBPbeta is a ubiquitous transcription factor and is thought to play a pivotal role in regulating the inflammatory response. If this imbalance between p20 and C/EBPbeta synthesis can be generalized to other diseases with excessive inflammation, then p20 could be a potent and general anti-inflammatory agent which could be delivered either as a protein or as a gene based therapeutic agent. It is hypothesized that p20, a truncated form of the transcription factor C/EBPbeta (a.k.a. NF-IL6), is a potent general inhibitor of inflammation in a variety of lung cell types and will be an effective anti-inflammatory therapeutic in several diseases of the lungs characterized by dysregulation of the inflammatory response. During Phase I of this program it is proposed: 1) to determine whether transfection of lung cells with a plasmid expression vector containing the gene encoding p20 driven by a CMV promoter (pCMVp20) will attenuate TNF or IL-1 stimulated production of IL-6 and IL-8; 2) to develop methods for delivering the p20 protein intracellularly using a liposome delivery system and determine whether delivery of the protein will attenuate TNF or IL-1 stimulated production of IL-6 and IL-8; 3) to determine whether intracellular delivery and inhibitory activity of p20 can be achieved by creating a recombinant fusion protein consisting of p20 and a 12 amino acid "membrane translocating polypeptide" which has been shown capable of escorting other proteins into several cell types. This concept could provide a new therapeutic approach to a host of inflammatory lung diseases, including idiopathic pulmonary fibrosis, ARDS, cystic fibrosis and asthma. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE

描述：(改编自《调查者摘要》)初步研究中 呼吸道上皮细胞炎症反应机制的研究进展 表现为囊性纤维化缺损者，发现正常呼吸道 在肿瘤坏死因子刺激下，上皮细胞停止产生IL-6和IL-8 通过增加p20的产量，p20是转录的截短亚型 抑制转录激活的C/EBPβ因子(NF-IL6) 长度C/EBPβ蛋白。相比之下，表达该蛋白的呼吸道上皮细胞 Cf缺陷在以下情况下不会关闭这些促炎细胞因子的产生 用肿瘤坏死因子刺激，不增加p20的产生。C/EBP测试版是一个 普遍存在的转录因子，被认为在 调节炎症反应。如果p20和p20之间的这种不平衡 C/EBPβ合成可推广到其他疾病 炎症，那么p20可能是一种有效的通用抗炎剂。 它既可以作为蛋白质传递，也可以作为基于基因的治疗 探员。假设p20，转录的截断形式 因子C/EBPbeta(又名核因子-IL-6)，是一种有效的一般炎症抑制物 能在多种肺细胞类型中起到有效的抗炎作用 治疗几种肺部疾病，其特征是调节功能失调 炎症反应。在该计划的第一阶段，建议：1) 确定用质粒表达载体转染肺细胞 包含由CMV启动子(PCMVp20)驱动的编码p20的基因将 减弱肿瘤坏死因子或白介素1刺激产生的白介素6和白介素8；2)发展 利用脂质体将p20蛋白输送到细胞内的方法 传递系统，并确定传递的蛋白质是否会减弱 肿瘤坏死因子或白介素1刺激IL-6和IL-8的产生；3)确定 P20的细胞内传递和抑制活性可以通过以下途径实现 构建由p20和12个氨基酸组成的重组融合蛋白 已被证明具有护航能力的“膜转运多肽” 其他蛋白质转化成几种细胞类型。这一概念可以提供一种新的 治疗一系列炎症性肺部疾病的方法，包括 特发性肺纤维化、ARDS、囊性纤维化和哮喘。 建议的商业应用：不可用