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CHARACTERIZATION OF PSORALEN DNA PHOTOADDUCTS

补骨脂素 DNA 照片加合物的表征

基本信息

批准号：
3446428
负责人：
FRANCIS P GASPARRO
金额：
$ 5.4万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1986
资助国家：
美国
起止时间：
1986-01-01 至 1986-12-31
项目状态：
已结题

项目摘要

8-Methoxypsoralen and ultra-violet A (PUVA) therapies are used in the treatment of psoriasis and cutaneous T cell lymphoma (CTCL). Recently, a new phototherapy for CTCL, photopheresis, has been developed which may serve as a prototype of a new form of photopharmacology. The efficacy of PUVA in these diseases has been attributed to the formation of 8-MOP photoadducts in cellular DNA. The nature and quantity of these adducts in mammalian systems has not been determined, nor has their repair been fully characterized. AlphaDNA, a 92 base pair fragment widely distributed throughout the human genome, will be used to elucidate the effects of specific base sequences on photoadduct formation. Three psoralens (8-methoxypsoralen, 4'-aminomethyl-4,5',8-trimethylpsoralen, and 4',5-dimethylangelicin) will be used in comparative studies to discern the effects of base sequences on monoadduct and crosslink formation. The overall objective of the proposed studies is to determine the nature, quantity and location of the UVA-induced psoralen photoadducts in AlphaDNA. Specifically the following aspects will be examined. 1. the quantitation of psoralen photoadducts in AlphaDNA by HPLC analysis of enzymatically hydrolyzed AlphaDNA samples. AlphaDNA from PUVA treated lymphoid cells will also be analyzed in the same way. 2. the determination of base sequences in which photoadducts have formed, as well as the identity of adducts in specific DNA sites using footprinting techniques in combination with psoralen-DNA monoclonal antibodies. 3. cytofluorometric determination of the effects of PUVA on cultured lymphoid cells using both propidium iodide staining and specific psoralen-DNA monoclonal antibodies. The development of the techniques described in this proposal will eventually lead to the detailed characterization of the repair of psoralen photoadducts. Knowledge of the sites of unrepaired photoadducts will lead to a better understanding of phototherapies based on psoralen plus UVA and culminate in the rational design of new, potentially more effective, psoralens.

8-甲氧补骨脂素和紫外线A（PUVA）疗法用于治疗银屑病和皮肤T细胞淋巴瘤（CTCL）。近日一已经开发了一种新的CTCL光疗，即单采，作为一种新的药物药理学的原型。的疗效这些疾病中的PUVA归因于8-MOP的形成细胞DNA中的光加合物。这些加合物的性质和数量哺乳动物系统尚未确定，也没有完全修复表征了 AlphaDNA是一个92个碱基对的片段，广泛分布于人体内。基因组，将被用来阐明特定的碱基序列对光加合物形成。三种萜类化合物（8-甲氧基萜， 4 ′-氨甲基-4，5 ′，8-三甲基白芷烯和4 ′，5-二甲基当归素）将用于比较研究以辨别碱基序列对单加合物和交联形成。建议的总体目标研究是为了确定的性质，数量和地点的 UVA诱导的AlphaDNA中的peptide光加合物。具体公开了如下将对各个方面进行审查。 1.高效液相色谱法测定AlphaDNA中补骨脂素光加合物酶水解的AlphaDNA样本。来自PUVA处理的AlphaDNA 淋巴样细胞也将以相同的方式进行分析。 2.确定其中形成光加合物的碱基序列，以及使用足迹法鉴定特定DNA位点的加合物技术与pepsilen-DNA单克隆抗体组合。 3.细胞荧光法测定PUVA对培养的使用碘化丙啶染色和特异性 peptide-DNA单克隆抗体。本提案所述技术的发展将最终导致详细的表征修复的peptien 光加合物对未修复的光加合物位点的了解将导致更好地了解基于peptide + UVA的光疗，最终合理设计新的，可能更有效的， puppelens。