Antimicrobial Gene Therapy

抗菌基因治疗

• 批准号: 6460477
• 负责人: GEORGE T.J HUANG
• 金额: $ 13.5万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-15 至 2004-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6460477
• 关键词: SCID mouse; antimicroorganism antibody; biological models; biotechnology; cell line; defensins; enzyme linked immunosorbent assay; gene therapy; genetic models; histology; immunocytochemistry; implant; in situ hybridization; infection; microorganism; microorganism immunology; model design /development; nonhuman therapy evaluation; peptides; tissue engineering; wound healing

DESCRIPTION: Increasing resistance to conventional antibiotics has focused attention on alternative anti-infectious therapies. Antimicrobial peptides are promising new agents that have a low susceptibility to microbial resistance mechanisms. Unlike conventional antibiotics, antimicrobial peptides are encoded by single genes and can be introduced into infected tissues by gene therapy approaches. The ultimate goal of this study is to introduce antimicrobial peptide genes into explanted human cells, and to use these augmented cells to enhance host defense mechanisms against infection. The immediate goal is to develop a model system, in which the human-Beta-defensin-2 (HBD-2) gene will be expressed in HBD-2 negative cells; its antimicrobial effect will be tested in vitro and in vivo. The HBD-2 gene was chosen because, unlike most known defensins, it does not require tissue-specific processing. A retrovirus carrying the HBD-2 gene has permitted successful transduction of mouse fibroblast NIH/3T3 cells to secrete functional antimicrobial peptides in vitro. Additionally, successful transduction of various other cell types to secrete HBD-2 has been accomplished. While antibacterial activity of HBD-2 in vitro has been clearly demonstrated, limited information is available regarding HBD-2 function in vivo. Using a novel approach, there are plans to test whether secreted HBD-2 is functional in vivo. The central hypothesis is that this antimicrobial gene therapy is effective in enhancing host innate immunity against infection. One short-term objective is to test whether the transduced cells will defend against infection in vivo using mouse models. The Specific Aims include: 1) to determine the temporal expression and the antimicrobial effects of HBD-2 in transplanted cells in vivo in a mouse model. This Aim will utilize SCID mice to grow HBD-2 expressing tumors which model will allow quantitative measurement of the antimicrobial effect of HBD-2 in vivo. 2) To establish a tissue-engineered wound healing model to test antimicrobial gene therapy in mice. This Aim will establish a model that simulates a clinical situation for testing the application of this antimicrobial gene therapy approach. Successful completion of these Aims will be the first step in establishing in vivo animal study models to explore the future applicability of antimicrobial gene therapy. The proposed approach is intended to provide potential benefit in clinical applications in three ways: 1) transplanted antimicrobial secreting cells can be used in wounded, engineered or infected tissues to prevent and/or fight against infections; 2) these cells may reduce the need for conventional antibiotics; and 3) these cells may provide long-term augmentation of the innate immune system for immunocompromised patients.

描述：对传统抗生素耐药性的增加已成为焦点 关注替代抗感染疗法。抗菌肽是 对微生物耐药性具有低敏感性的有前景的新制剂 机制。与传统抗生素不同，抗菌肽是编码的 由单基因组成，可以通过基因治疗引入受感染的组织 接近。本研究的最终目标是引入抗菌剂 将肽基因植入外植的人类细胞中，并利用这些增强的细胞 增强宿主抵抗感染的防御机制。近期目标是 开发一个模型系统，其中人类β-防御素-2 (HBD-2) 基因将被 在HBD-2阴性细胞中表达；其抗菌效果将在 体外和体内。选择 HBD-2 基因是因为，与大多数已知的不同 防御素，它不需要组织特异性处理。逆转录病毒 携带HBD-2基因已成功转导小鼠 成纤维细胞 NIH/3T3 细胞在体外分泌功能性抗菌肽。 此外，成功转导各种其他细胞类型以分泌 HBD-2已经完成。 HBD-2的体外抗菌活性 已明确证明，有关 HBD-2 的可用信息有限 在体内发挥作用。计划使用一种新颖的方法来测试是否 分泌的HBD-2在体内有功能。中心假设是，这 抗菌基因治疗可有效增强宿主先天免疫 防止感染。一个短期目标是测试转导是否 使用小鼠模型，细胞将在体内抵御感染。具体 目的包括：1) 确定时间表达和抗菌药物 HBD-2 对小鼠模型体内移植细胞的影响。这个目标将 利用 SCID 小鼠培养表达 HBD-2 的肿瘤，该模型将允许 定量测量HBD-2体内抗菌作用。 2) 至 建立组织工程伤口愈合模型测试抗菌基因 对小鼠的治疗。该目标将建立一个模拟临床的模型 测试该抗菌基因疗法应用的情况 方法。 成功完成这些目标将是建立 体内动物研究模型探索抗菌药物的未来适用性 基因疗法。拟议的方法旨在提供潜在的好处 临床应用分为三种方式：1）移植性抗菌分泌 细胞可用于受伤、工程或感染的组织以预防和/或 对抗感染； 2）这些电池可能会减少对传统电池的需求 抗生素； 3）这些细胞可以长期增强 免疫功能低下患者的先天免疫系统。