APPROACHES TO CORRECT DISEASES WITH GENETICALLY MODIFIED CELLS OF HUMAN SKIN

利用转基因人类皮肤细胞治疗疾病的方法

• 批准号: 6272209
• 负责人: GERALD Gene KRUEGER
• 金额: $ 3.95万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-12-20 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6272209
• 关键词: artificial skin; athymic mouse; biopsy; congenital ichthyosis; disease /disorder model; enzyme deficiency; fibroblasts; gene expression; gene therapy; genetic disorder; genetic markers; genetic transduction; histology; human tissue; immunocytochemistry; keratinocyte; laboratory rat; method development; natural gene amplification; nonhuman therapy evaluation; radioimmunoassay; skin; skin disorder; skin transplantation; sulfatases; tissue /cell culture; transfection /expression vector; ultrasound blood flow measurement

For many genetic disorders, the abnormal, as well as the normal gene have been defined, cloned, and expressed in vitro. These advances in molecular genetics have made it possible to correct genetic disease by bringing the non-defective gene into the afflicted host via somatic cells (somatic cell gene therapy). This approach appears to be particularly suitable to heritable skin diseases of acquired malformation and abnormal morphogenesis, e.g., Darier's disease, xeroderma pigmentosa, epidermolysis bullosa, and those disorders which are characterized by a deficiency of a plasma protein, e.g., hemophilia, diabetes mellitus, etc. A central issue, now at the forefront of the biomedical community, focuses on how best to introduce these non-defective genes into afflicted individuals. It is our thesis that correction by genetically modifying cells of the skin has relatively low risk, has broad application and thus has considerable clinical utility. We are persuaded that the feasibility, limitations and methods for applicability can be answered preclinically by transplanting genetically modified human skin equivalents or components thereof to athymic rodents and by experimentally analyzing selected parameters of such skin as a function of time. Specific aims: 1. Development of the preclinical technology for transplanting genetically modified skin to humans. a. Replacing the existing epidermis of human skin grafted onto athymic rodents with an epidermis generated from genetically modified keratinocytes. b. Generating a genetically modified skin-equivalent carrying normal or genetically modified fibroblasts or keratinocytes which can be transplanted onto athymic rodents. c. Generating a dermal matrix impregnated with genetically modified fibroblasts that can be placed subcutaneously in rodents, and possibly intradermally in humans. 2. To determine the long-term functionality of skin containing these genetically modified components; specifically the capacity for a dose response, the production by and stability of the transfected gene, and the effect of transfection on structure and function inherent to skin. 3. To determine whether recessive X-linked ichthyosis, can be corrected with keratinocytes or fibroblasts transfected with the gene for steroid sulfatase, and transplanted to athymic rodents. These experiments are designed to define the potential and the limitations of this technology.

对于许多遗传疾病，异常和正常基因具有 被定义，克隆并在体外表达。 这些分子的进步 遗传学使通过带来纠正遗传疾病成为可能 通过体细胞（体细胞）进入受伤的宿主的非缺陷基因 基因治疗）。 这种方法似乎特别适合 遗传性畸形和异常的皮肤疾病 形态发生，例如，达里尔氏病，心虫色素，表皮分解 Bullosa以及那些以不足的特征的疾病 血浆蛋白，例如血友病，糖尿病等。一个中心问题， 现在处于生物医学界的最前沿，着重于如何最好 将这些非缺陷基因引入受苦的个体。 这是我们的 通过基因修饰皮肤细胞进行纠正的论文具有 相对较低的风险，具有广泛的应用，因此具有相当大的 临床实用程序。 我们被说服了可行性，局限性和 可以通过移植来临时回答适用性的方法 转基因的人皮肤等效因素或其成分 通过实验分析此类选定参数 皮肤随时间的作用。 具体目的： 1。开发用于基因移植的临床前技术 对人类的改性皮肤。 一个。替换嫁接到无关的人皮的现有表皮 啮齿动物具有由基因修饰产生的表皮 角质形成细胞。 b。产生一种遗传改性的皮肤等效性，携带正常或 可以移植的转基因成纤维细胞或角质形成细胞 进入胸膜啮齿动物。 c。生成用遗传改性的皮肤基质 可以皮下置于啮齿动物的成纤维细胞，甚至可能 在人类中受皮内。 2。确定包含这些皮肤的长期功能 基因修饰的成分；特别是剂量的能力 反应，转染基因的稳定性和稳定性以及 转染对皮肤固有的结构和功能的影响。 3。为了确定是否可以纠正隐性X连接的鱼质病 用角质形成细胞或成纤维细胞用基因转染类固醇 硫酸酶，并移植到无胸腺啮齿动物。 这些实验是 旨在定义该技术的潜力和局限性。