MONOCLONAL ANTIBODIES TO HUMAN EPIDERMIS

人类表皮单克隆抗体

• 批准号: 3155939
• 负责人: LOWELL Alan GOLDSMITH
• 金额: $ 39.57万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3155939
• 关键词: acne; cell differentiation; cell growth regulation; cell membrane; cellular immunity; clinical trials; complementary DNA; congenital ichthyosis; cytogenetics; drug resistance; epidermal growth factor; gel mobility shift assay; gene expression; genetic library; genetic mapping; genetic promoter element; genome; human genetic material tag; human subject; human therapy evaluation; human tissue; immunogenetics; in situ hybridization; karyotype; keratin; keratinocyte; keratoacanthoma; keratosis follicularis; messenger RNA; molecular biology; monoclonal antibody; pathologic keratinization; pharmacogenetics; protein glutamine gamma glutamyltransferase; psoriasis; retinoids; skin; skin disorder chemotherapy; skin neoplasms; surface antigens; transcription factor; vitamin therapy

The general hypothesis is that there are discrete defects in the normal pathway of epidermal differentiation related to the phenotypic changes associated with genetic and acquired diseases of the epidermis, such as psoriasis, the ichthyoses and Darier's disease. The long-term focus of this research is defining those defects ultimately leading to safe and effective therapies for those disorders. Retinoids have profound effects on the normal and diseased epidermis and detailed studies of their action on three different representative steps in epidermal maturation is planned; differentiation limiting or modifying events involving a new human homeobox-containing gene Hox 1.1; modulation of the epidermal growth factor receptor (EGFR); and control of late events in terminal differentiation catalyzed by intracellular transglutaminases. The detailed specific aims are: 1) Determine the molecular mechanism regulating the response of the EGFR in cervical carcinoma cells ME-180 treated with retinoic acid (RA). The half-life of mRNA will be determined in control cells and after treatment with RA. Regulatory regions of the EGFR and other retinoid inducible genes will be transfected into ME-180 cells which are then treated with RA. A promoter from the EGFR gene of ME-180 cells will be ligated to CAT reporter gene constructs and transfected into normal keratinocytes and other epithelial cells and the response to RA quantitated. The mRNAs for retinoid nuclear receptors will be measured. Transcription factors controlling the RA response will be determined by gel retardation and footprinting assays. 2) Determining the role of human Hox 1.1 in normal and altered epidermal differentiation. The expression of Hox 1.1 in normal and altered epidermal differentiation. The expression of Hox 1.1 in normal and altered epidermal differentiation. The expression Hox 1.1 will be determined, by using in situ hybridization, in human fetal skin samples of different gestational ages and in human skin biopsies of normals, and patients with acne, psoriasis and genetics disorders of keratinization, and in patients treated with systemic and topical RA. Genomic clones for Hox 1.1 will be isolated and the 5' untranslated region of the Hox 1.1 gene will be characterized and sequenced. The detailed chromosomal mapping of the gene will continue. 3) Determine the genetic, biochemical and physiological relationships between the epidermal transglutaminases. Using the current transglutaminase cDNA clones from a human epidermis cDNA library full length cDNA will be produced and genomic transglutaminase characterized, especially its 5' nontranslated region. The promoter and enhancer sequences will be characterized looking for sequences known to be found in other epidermal genes responsive to retinoids and other pharmacological agents.

一般的假设是在正常情况下有离散的缺陷 与表型变化相关的表皮分化途径 与遗传性和获得性表皮疾病有关的，例如 牛皮癣、鱼鳞病和达里尔病。长期关注的焦点 这项研究正在定义那些最终导致安全和 这些疾病的有效治疗方法。维甲酸有深远的影响 关于正常和病变的表皮及其作用的详细研究 对表皮成熟的三个不同代表步骤进行了规划； 区分限制或修改涉及新人类的事件 同源异型盒基因HOX 1.1；表皮生长因子的调节 受体(EGFR)；终末分化中晚期事件的控制 由细胞内转谷氨酰胺酶催化。详细的具体目标 主要内容有：1)确定调控细胞反应的分子机制。 维甲酸(RA)对宫颈癌细胞ME-180中EGFR的影响 MRNA的半衰期将在对照细胞中和之后确定。 类风湿关节炎的治疗。EGFR和其他类维甲酸的调节区 可诱导基因将被导入ME-180细胞，然后 用类风湿因子治疗。ME-180细胞的EGFR基因启动子将是 连接到猫报告基因构建体并转染正常 角质形成细胞和其他上皮细胞与类风湿关节炎的反应 量化的。将测量维甲酸核受体的mRNAs。 控制RA反应的转录因子将通过凝胶确定 发育迟缓和足迹分析。2)确定人类HOX的作用 1.1正常和异常的表皮分化。HOX基因的表达 1.1正常和异常的表皮分化。HOX基因的表达 1.1正常和异常的表皮分化。表达方式Hox 1.1将通过使用原位杂交技术在人胎儿皮肤中检测到 不同胎龄的样本和人类皮肤活检组织中的 正常人，以及痤疮、牛皮癣和遗传性疾病患者 角质化，以及全身和局部RA治疗的患者。 将分离HOX 1.1的基因组克隆和5‘非翻译区 将对HOX 1.1基因进行鉴定和测序。详细的 该基因的染色体图谱将继续。3)确定基因， 表皮之间的生化和生理关系 转谷氨酰胺酶。利用现有的转谷氨酰胺酶基因克隆 人表皮全长c DNA文库的构建及基因组 转谷氨酰胺酶的特性，特别是它的5‘非翻译区。 启动子和增强子序列将被鉴定以寻找 已知在其他表皮基因中发现的序列对 维甲酸和其他药理药剂。