Anti-Cancer Drugs that Target Diaphanous-related Formins

针对透明相关福明的抗癌药物

基本信息

批准号：
6863648
负责人：
ARTHUR ALBERTS
金额：
$ 16.38万
依托单位：
VAN ANDEL RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-03-01 至 2006-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6863648
关键词：
X ray crystallography antineoplastics apoptosis binding proteins breast neoplasms cell transformation cytoskeleton drug discovery /isolation guanine nucleotide binding protein guanosinetriphosphatases intermolecular interaction neoplasm /cancer pharmacology neoplastic cell paclitaxel prostate neoplasms protein purification protein structure function

项目摘要

DESCRIPTION (provided by applicant): Rationale. Taxanes, epitomized by the drug Taxol, and their derivatives represent a state-of-the-art anti-cancer therapy. They bind to tubulin and disrupt the dynamic remodeling of the microtubule cytoskeleton, leading to inhibition of cell motility and proliferation; cells respond by initiating programmed cell death. These drugs are limited clinically, however, by their side effects and tumor cell resistance. Thus, there is a need to further identify anti-cancer compounds that disrupt cytoskeletal function. Guiding Hypothesis. Rho small GTPases act as critical regulators of cellular shape during growth factor responses and cell division; their activity is also required for oncogenic transformation. Diaphanous related formins (Drf), members of the formin family of cytoskeletal regulators, are activated by Rho proteins upon binding to the Drf GTPase binding domains (GBDs). The interaction disrupts an intramolecular association between the GBD and the Dia-autoregulatory domain (DAD). Introduction of a DAD-derived peptide into cells is sufficient to deregulate Drf proteins; DAD mimics Rho binding, thereby disrupting intramolecular autoinhibition. The result is the inappropriate stabilization of the cytoskeleton. Like taxanes, DAD peptide causes cells to eventually undergo apoptosis. We hypothesize that DAD peptide represents a novel class of anti-cancer drug that targets and deregulates Drfs in cancer cells. The proposed experiments will test our guiding hypothesis. Specific Aims. (1) Identify the Diaphanous-related formin(s) that is/are targeted by DAD peptide (beginning with DAD peptide derived from the Drf3 protein) in Taxol-sensitive and -resistant breast and prostate cancer cell lines, (2) determine the structure of DAD peptide bound to its target, the GTPase binding domain, and (3) develop a screening method for identifying novel molecules that disrupt Drf autoregulation that is amenable to high-throughput screening (HTS) of drug libraries. If our hypothesis is correct, then DAD peptide will be sufficient to kill even Taxol-resistant cancer cells. DAD variants, that fail to interact with the GBD of cellular Drfs, will be inactive, and the disrupted sites will correspond to molecular determinants that define the structure of DAD bound to the GBD. Finally, DAD peptides will disrupt the GBD-DAD interaction in vitro; the assays will then be optimized, using DAD peptides and their derivatives, for screening strategies that will allow us to identify additional molecules that harbor Drf-deregulating activity.

描述（由申请人提供）：理由。紫杉醇及其衍生物代表了最先进的抗癌疗法。它们与小管蛋白结合并破坏微管细胞骨架的动态重塑，从而抑制细胞运动和增殖。细胞通过启动程序性细胞死亡做出反应。但是，这些药物在临床上受到副作用和肿瘤细胞耐药性的限制。因此，有必要进一步识别破坏细胞骨架功能的抗癌化合物。指导假设。 Rho小GTP酶在生长因子反应和细胞分裂过程中充当细胞形状的关键调节剂。它们的活性也需要进行致癌。 diaphanous formins（DRF）（DRF）是细胞骨架调节剂家族的成员，在与DRF GTPase结合结构域（GBD）结合后被Rho蛋白激活。这种相互作用破坏了GBD与Dia-AutoreRegulatoration域（DAD）之间的分子内关联。将爸爸衍生的肽引入细胞足以消除DRF蛋白。爸爸模仿Rho结合，从而破坏分子内自身抑制。结果是细胞骨架的不适当稳定。像紫杉烷一样，爸爸肽会导致细胞最终发生细胞凋亡。我们假设DAD肽代表了一类新型的抗癌药物，该药物靶向并消除癌细胞中的DRF。提出的实验将检验我们的指导假设。具体目标。（1）识别由dad肽靶向的diaphanous相关formin （从对紫杉醇敏感和耐药性的乳腺癌和前列腺癌细胞系中得出的DAD肽开始，（2）确定与其靶标结合的DAD肽的结构，GTPase结合结构域，（3）开发一种筛选方法，用于识别识别DRF自动调查的新型分子，以使库型升温（均为较高的毒品），该方法是对高剂量的识别（Hy-formentupt for to niberable for potertupt（Himenable for to tomentupt）（Hy-formentupt）。如果我们的假设是正确的，那么DAD肽就足以杀死抗紫杉醇的癌细胞。未能与细胞DRF的GBD相互作用的DAD变体将是不活跃的，而破裂的位点将对应于定义与GBD结合的dad结构的分子决定因素。最后，爸爸肽会在体外破坏GBD-DAD相互作用。然后，使用DAD肽及其衍生物进行筛选策略，将对这些测定进行优化，以使我们能够识别出携带DRF施加活性的其他分子。