MONOCLONAL ANTIBODIES TO HUMAN EPIDERMIS

一般的假设是，在正常情况下， 表型变化相关的表皮分化途径 与表皮的遗传性和获得性疾病相关，例如 牛皮癣、鱼鳞病和达里尔病。 长期关注 这项研究正在定义这些缺陷，最终导致安全， 有效的治疗方法。 类维生素A具有深远的影响 对正常和病变表皮的作用， 计划在表皮成熟的三个不同的代表性步骤; 涉及新人类的分化限制或修饰事件 含同源框基因Hox 1.1;表皮生长因子的调节 受体（EGFR）;和控制终末分化中的晚期事件 由细胞内的转氨酶催化。 详细的具体目标 1）确定调节免疫应答的分子机制。 维甲酸（RA）对宫颈癌细胞ME-180 EGFR表达的影响 mRNA的半衰期将在对照细胞中测定，并且在转染后测定。 治疗RA。EGFR和其他类维生素A的调节区 将诱导型基因转染到ME-180细胞中， 用RA治疗。 来自ME-180细胞的EGFR基因的启动子将被 连接到CAT报告基因构建体并转染到正常 角质形成细胞和其他上皮细胞与对RA的反应 定量。 将测量类维生素A核受体的mRNA。 控制RA反应的转录因子将通过凝胶电泳测定。 阻滞和足迹分析。 2)确定人类Hox的作用 1.1正常和改变的表皮分化。 Hox的表达 1.1正常和改变的表皮分化。 Hox的表达 1.1正常和改变的表皮分化。 Hox的表达 1.1将通过使用原位杂交在人胎儿皮肤中测定 不同胎龄的样本和人类皮肤活检中的样本 正常人和痤疮、银屑病和遗传性疾病患者， 角化，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，并将5'非翻译区 将对Hox 1.1基因进行表征和测序。 的详细 基因的染色体定位将继续进行。 3)确定基因， 表皮细胞之间的生理生化关系 转氨酶 使用目前的转氨酶cDNA克隆， 将产生人表皮cDNA文库全长cDNA并将其基因组化 以转氨酶为特征，特别是其5'非翻译区。 启动子和增强子序列将被表征，寻找 已知在其他表皮基因中发现的序列， 类维生素A和其它药理学试剂。

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.