HEAT SHOCK PROTEIN FUNCTION AS A THERAPEUTIC TARGET

描述：研究人员最近的研究表明， 格尔德霉素（GA）与一定细胞热的选择性相互作用 休克蛋白介导的抗肿瘤活性，但具体 这种相互作用在临床相关肿瘤中的分子后果 模型仍然不清楚。 在本申请中，提出了研究， 将检查GA对两个热休克的组装和功能的影响， 已知含蛋白质的多分子复合物在 许多乳腺癌的生物学;即，雌激素受体（ER）和 突变型p53。使用网织红细胞裂解物的复溶实验将 用于鉴定ER和p53复合物的特定组分 并评估药物对复杂组装动力学的影响。 在 全细胞水平，GA对稳态表达的影响，亚细胞 定位和转录反式激活活性的两个 将对目标进行审查。最后，将检查p53和ER表达 在用GA处理的SCID小鼠中生长的乳腺肿瘤异种移植物中。 这些 实验应确定药物作用的有用生物学终点， 并将允许评估分子的意义， 在动物致瘤性组织培养中观察到的影响。

DESCRIPTION: The investigator's recent studies have implicated the selective interaction of geldanamycin (GA) with certain cellular heat shock proteins in mediating anti-tumor activity, but the specific molecular consequences of this interaction in clinically relevant tumor models remain unclear. In this application, studies are proposed which will examine GA effects on the assembly and function of two heat shock protein-containing multimolecular complexes known to be important in the biology of many breast cancers; namely, the estrogen receptor (ER) and mutant p53. Reconstitution experiments using reticulocyte lysate will be used to identify the specific components of the ER and p53 complexes and assess drug effects on the dynamics of complex assembly. At the whole-cell level, GA effects on steady state-expression, subcellular localization and transcriptional transactivating activity of the two targets will be examined. Lastly, p53 and ER expression will be examined in breast tumor xenografts growing in SCID mice treated with GA. These experiments should define useful biologic endpoints for drug action in vivo and will allow assessment of the significance of the molecular effects observed in tissue culture for tumorigenicity in animals.