High troughput screen for small molecule inhibitors of Ran regulated functions

Ran 调节功能的小分子抑制剂的高通量筛选

• 批准号: 7965774
• 负责人: Petr Kalab
• 金额: $ 0.63万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7965774
• 关键词: Aneuploidy; Antimitotic Agents; BRCA1 gene; Binding; Cancer Cell Growth; Cell Cycle; Cells; Chemicals; Chromatin; Chromosomes; Clinical Trials; Complex; Development; Dissociation; Drug Delivery Systems; Embryo; Fluorescence Resonance Energy Transfer; Funding; Genes; Genomics; Guanosine Triphosphate Phosphohydrolases; HMMR gene; Human; Importins; Interphase; Libraries; Link; Malignant Neoplasms; Mediating; Meiosis; Mitosis; Mitotic; Mitotic spindle; Monomeric GTP-Binding Proteins; Mus; Nuclear Envelope; Nuclear Export; Nuclear Import; Nuclear Pore; Pathway interactions; Phase; Protein Dephosphorylation; RNA Interference; Regulation; Research; Running; System; TACC3 gene; Testing; Therapeutic; Undifferentiated; United States National Institutes of Health; Work; alpha Karyopherins; aurora-A kinase; base; cancer cell; cancer therapy; human STK6 protein; inhibitor/antagonist; interest; karyopherin alpha 2; kinase inhibitor; nucleocytoplasmic transport; receptor; sensor; small molecule; survivin; tool; tumor

A number of the spindle assembly factors (SAFs) which are regulated by Ran GTPase during mitosis have variety of well recognized and intensively studied (but often not well understood) connections to cancer: BRCA1, HURP, TPX2, Aurora A, TACC3, survivin, RHAMM, cdk11. Also the levels of Ran are highly increased in many human tumors and unlike in non-cancer derived cells, the growth of cancer-derived cells is inhibited by Ran RNAi. Arguably the best characterized mechanism potentially linking Ran to cancer is the RanGTP, importin beta and importin alpha1 regulating the activation of Aurora A through its binding to TPX2. RanGTP is required for the release of TPX2 from its inhibitory complex with importin alpha1-importin beta. The importin-freeTPX2 binds Aurora A, protecting it from dephosphorylation and thus supporting sustained Aurora A kinase activity towards is many mitotic targets. More than 30 Aurora A kinase inhibitors are being developed as potential cancer therapeutics and several of them entered phase I and II clinical trials. However, the deletion of Aurora A gene is lethal in embryos and the heterozygous mice develop significantly higher number of tumors and aneuploidy. Complete inhibition of the Aurora A could therefore potentially induce de novo aneuploidy and cancer. We developed fluorescence resonance energy transfer (FRET)-based sensors for the RanGTP-induced importin alpha1-importin beta dissociation. These sensors were successfully tested at the NIH Chemical Genomics Center (NCGC), Rockville, MD, as applicable in quantitative highthroughput screen (qHTS) for small molecule inhibitors of the key steps involved in the Ran-regulated importin alpha1 mitotic function. We are currently applying for funding to perform the qHTS with a library containing 300 000 compounds. In a longer term, we will work towards developing the hits from the screen into antimitotic drugs suitable for cancer treatment.

由Ran GTweek调节的许多主轴装配系数（SAF） 在有丝分裂过程中有各种各样的公认和深入研究（但往往不是很好）， 与癌症的联系：BRCA 1、HURP、TPX 2、Aurora A、TACC 3、生存素、RHAMM、cdk 11。 此外，与非癌症不同，Ran的水平在许多人类肿瘤中高度增加 在癌源细胞中，癌源细胞的生长被Ran RNAi抑制。可以说是最好的 潜在地将Ran与癌症联系起来的特征性机制是RanGTP、importin β和 importin alpha 1通过与TPX 2结合调节Aurora A的活化。RanGTP是 从其与输入素α 1-输入素的抑制复合物中释放TPX 2所需的 β的不含输入蛋白的TPX 2与Aurora A结合，保护其免于去磷酸化， 支持持续的Aurora A激酶活性朝向许多有丝分裂靶点。30多 Aurora A激酶抑制剂正被开发为潜在的癌症治疗剂， 他们进入了I期和II期临床试验。但是，Aurora A基因的缺失是致命的 在胚胎和杂合子小鼠中， 非整倍性因此，Aurora A的完全抑制可能会诱导新生 非整倍性和癌症。我们开发了基于荧光共振能量转移（FRET）的 用于RanGTP诱导的输入蛋白α 1-输入蛋白β解离的传感器。这些传感器 在NIH化学基因组学中心（NCGC，Rockville，MD）成功进行了测试（如适用） 在定量高通量筛选（qHTS）小分子抑制剂的关键步骤 参与RAN调节的输入素α 1有丝分裂功能。我们目前正在申请 资助进行qHTS与库包含300 000化合物。从长远来看，我们 将致力于将筛选出的热门药物开发成适用于 癌症治疗