Genetic Engineering of Human Skin Substitutes

人类皮肤替代品的基因工程

• 批准号: 6937010
• 负责人: ANGELA L F GIBSON
• 金额: $ 3.08万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6937010
• 关键词: artificial skin; athymic mouse; bioengineering /biomedical engineering; biotechnology; burns; cell line; defensins; keratinocyte; predoctoral investigator; tissue engineering; transfection /expression vector; wound healing; wound infection

DESCRIPTION (provided by applicant): Burn wound infection is a major complication leading to patient morbidity and mortality. Topical antimicrobials are cytotoxic to keratinocytes and compromise engraftment of cultured skin substitutes. The goal of this proposal is to generate a therapeutic human skin substitute with antimicrobial activity for use in the treatment of severely burned patients. In the past decade there has been emerging excitement regarding human derived endogenous antimicrobial peptides. One family of peptides, human beta defensins has broad antimicrobial properties against gram positive and gram negative bacteria. The NIKS human keratinocyte cell line is a consistent source of genetically uniform, non-tumorigenic, pathogen free human keratinocytes that will be genetically engineered to express enhanced levels of the broad spectrum defensin family member, hBD-3. Our specific aims are to 1) Develop stable genetically modified NIKS cell clones and evaluate expression levels of hBD-3 and antimicrobial activity in monolayer culture. 2) Produce organotypic cultures using stable hBD-3 clones and characterize based on expression levels, growth properties, ability to form stratified epidermis, and antimicrobial activity. 3) Perform animal grafting for safety and efficacy.

描述（由申请人提供）：烧伤伤口感染是导致患者发病率和死亡率的主要并发症。局部抗微生物是对角质形成细胞的细胞毒性，并妥协了培养的皮肤替代品。该提案的目的是产生具有抗菌活性的治疗性人皮替代品，用于治疗严重燃烧的患者。在过去的十年中，关于人衍生的内源性抗菌肽的兴奋。人类β防御素的一个家族具有较大的抗菌特性，抗革兰氏阳性和革兰氏阴性细菌。 NIKS人角质形成细胞系是遗传均匀，非肿瘤，无病原体的无病原体角质形成细胞的一致来源，将在基因上进行基因设计，以表达增强的广泛频谱防御素家庭成员HBD-3。我们的具体目的是1）产生稳定的转基因NIKS细胞克隆，并评估单层培养中HBD-3和抗菌活性的表达水平。 2）使用稳定的HBD-3克隆产生器官培养物，并根据表达水平，生长特性，形成分层表皮的能力和抗菌活性来表征。 3）进行动物嫁接以进行安全和功效。