Genetically modifed tissue engineered in vitro human models

描述（由申请人提供）：当前提案的目的是将最先进的基因修饰技术（包括慢病毒递送系统和RNA干扰技术）应用于市售组织工程化人体外模型。这些模型将包括分化的3-D表皮培养物和气管/支气管上皮培养物，以及与间充质细胞共培养的表皮和气道上皮。在I期研究中，该技术将用于生产3种类型的转基因组织工程培养产品：1）稳定添加功能基因，2）稳定沉默基因，3）稳定引入报告基因以检测基因激活。在第二阶段，该技术将应用于其他组织工程人类体外模型，并将扩大可用的基因修饰产品的库存。这些提出的市售体外模型系统将为制药工业和学术研究实验室的研究人员提供一种现成的手段，用于研究人类上皮细胞中几乎任何基因的功能基因组学，以及分化器官型状态下上皮细胞和基质细胞之间的串扰。这些模型的使用将有助于鉴定和验证用于开发用于治疗人类皮肤和气道上皮疾病的新疗法的靶标，所述人类皮肤和气道上皮疾病包括癌症、慢性伤口、起泡疾病、瘢痕形成和与哮喘和COPD相关的气道重塑。公共卫生相关性：本项目生产的基因修饰的组织工程模型将为制药行业和学术研究实验室的研究人员提供一种现成的方法，用于研究人类上皮细胞中几乎任何基因的功能基因组学，以及分化器官型状态下上皮细胞和基质细胞之间的串扰。这些模型将用于识别和验证用于治疗人类皮肤和气道上皮疾病的新型疗法的开发目标，这些疾病包括癌症，慢性伤口，水疱疾病，瘢痕形成以及与哮喘和COPD相关的气道重塑。

DESCRIPTION (provided by applicant): The goal of the current proposal is to apply state of the art gene modification technology including lentiviral delivery systems and RNA interference technology to commercially available tissue engineered human in vitro models. These models will consist of differentiated 3-D epidermal cultures and tracheal/bronchial epithelial cultures, as well as epidermal and airway epithelia co-cultured with mesenchymal cells. In Phase I research, the technology will be used to produce 3 types of genetically modified tissue engineered culture products: 1) stable addition of a functional gene, 2) stable silencing of a gene, and 3) stable introduction of a reporter for detection of gene activation. During Phase II, the techniques will be applied to additional tissue engineered human in vitro models, and the inventory of available gene modified products will be expanded. These proposed commercially available in vitro model systems will provide researchers in the pharmaceutical industry and academic research laboratories a readily available means for studying the functional genomics of nearly any gene in human epithelial cells, as well as crosstalk between epithelial and stromal cells in the differentiated organotypic state. Use of these models will aid in identification and validation of targets for development of novel therapeutics for treatment of human skin and airway epithelial diseases including cancers, chronic wounds, blistering diseases, scarring, and airway remodeling associated with asthma and COPD. Public Health Relevance: The gene modified tissue engineered models to be produced by the current project will provide researchers in the pharmaceutical industry and academic research laboratories a readily available means for studying the functional genomics of nearly any gene in human epithelial cells, as well as crosstalk between epithelial and stromal cells in the differentiated organotypic state. The models will be utilized to identify and validate targets for the development of novel therapeutics for treatment of human skin and airway epithelial diseases including cancers, chronic wounds, blistering diseases, scarring, and airway remodeling associated with asthma and COPD.