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）：烧伤创面感染是导致患者发病和死亡的主要并发症。局部抗菌剂对角质形成细胞具有细胞毒性，并会影响培养皮肤替代物的植入。该提案的目标是生产一种具有抗菌活性的治疗性人类皮肤替代品，用于治疗严重烧伤患者。在过去的十年中，人们对人源性内源性抗菌肽产生了新的兴趣。人β防御素是肽家族之一，对革兰氏阳性和革兰氏阴性细菌具有广泛的抗菌特性。 NIKS 人类角质形成细胞系是遗传均一、非致瘤、无病原体的人类角质形成细胞的一致来源，这些细胞将经过基因工程改造，以表达增强水平的广谱防御素家族成员 hBD-3。我们的具体目标是 1) 开发稳定的转基因 NIKS 细胞克隆并评估单层培养物中 hBD-3 的表达水平和抗菌活性。 2) 使用稳定的 hBD-3 克隆产生器官型培养物，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进行表征。 3）为了安全和有效而进行动物移植。