A high-throughput screen for inhibitors of Plk1-interacting checkpoint helicase (PICH)

Plk1 相互作用检查点解旋酶 (PICH) 抑制剂的高通量筛选

基本信息

批准号：
10557106
负责人：
Yoshiaki Azuma
金额：
$ 21.02万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10557106
关键词：
ATP phosphohydrolase Alleles Anaphase Animal Cancer Model Antineoplastic Agents Area Auxins Biochemical Biological Assay Breast Cancer Cell Cell Death Cell Differentiation process Cell Line Cells Cessation of life Characteristics Chromatin Chromatin Remodeling Factor Chromosomal Instability Chromosome Segregation Collection DNA DNA-dependent ATPase Development Dose Drug Targeting Engineering Evaluation Family Family member Frequencies Genome Goals Human Cell Line Human Engineering In Vitro Intercalating Agents Lead Malignant Neoplasms Microtubules Mitosis Mitotic Normal Cell Normal tissue morphology Outcome Patient-Focused Outcomes Phenotype Positioning Attribute Predisposition Process Proteins Research Research Subjects Role Secondary to Testing Therapeutic Tissues Transcriptional Regulation Ultrafine Validation cancer cell cancer therapy cancer type cell type cheminformatics chromosome loss cytotoxicity detection assay dosage ds-DNA helicase high throughput screening improved inhibitor new therapeutic target novel overexpression response screening small molecule small molecule inhibitor small molecule libraries therapeutic target tool translocase transmission process triple-negative invasive breast carcinoma

项目摘要

Project Summary/Abstract Recent studies have shown that PIk1-interacting checkpoint helicase (PICH) is a potential cancer therapeutic target. PICH has critical functions in mitosis for the fidelity of chromosome segregation and without PICH there is an increased frequency of Ultra-Fine DNA Bridges (UFBs) in anaphase, which results in multi-nucleation and chromosome loss. Other proteins with key functions in mitosis, and that are required for accurate chromosome segregation, have proven to be effective targets of cancer drugs. For example, drugs that target microtubule dynamics. Importantly, PICH was found to be over-expressed in breast cancer cells and depletion of PICH was found to be extremely toxic to triple negative breast cancer cells, while not affecting matched normal tissue. Because PICH is over-expressed in a range of cancers, it is likely that many cancer types require PICH function for survival. Therefore, small molecule PICH inhibitors have potential for improved patient outcomes in cancer therapy. However, no inhibitors of PICH have been identified. Here we propose to identify selective PICH inhibitors and determine their cytotoxicity towards a range of cancer types in which PICH is over- expressed. Since PICH is a dsDNA-dependent ATPase, it has an activity suitable for high-throughput screening (HTS) for inhibitors using diverse small molecule libraries. We have established biochemical purification of active PICH and we have devised strategies and generated the tools that will allow primary hits from the HTS to be efficiently subjected to secondary screening, to validate bona fide PICH inhibitors and to eliminate non-specific inhibitors. The cellular consequences of PICH inhibition will be determined to fully characterize the effects on normal and cancer cells. Cytotoxicity studies will determine which cancer types can be selectively killed by the inhibitors and we will define the dosages required. The outcomes will be selective small molecule PICH inhibitors that will be valuable research tools and will be lead compounds for development of optimally potent molecules ahead of studies in animal cancer models.

项目摘要/摘要最近的研究表明，PIK1相互作用检查点解旋酶（PICH）是一种潜在的癌症治疗目标。 PICH在有丝分裂的有丝分裂中具有关键功能是后期中超细DNA桥（UFB）的频率增加，这导致了多核和染色体损失。其他具有有丝分裂关键功能的蛋白质，这是准确染色体所必需的隔离已被证明是癌症药物的有效靶标。例如，靶向微管的药物动力学。重要的是，发现PICH在乳腺癌细胞中过表达，而Pich的耗尽为发现对三重阴性乳腺癌细胞具有极高的毒性，而不影响匹配的正常组织。因为Pich在各种癌症中都过表达，所以许多癌症类型可能需要PICH 生存的功能。因此，小分子菌抑制剂具有改善患者预后的潜力癌症治疗。但是，尚未确定PICH的抑制剂。在这里，我们建议确定选择性 PICH抑制剂并确定其细胞毒性对PICH过度的一系列癌症类型表达。由于PICH是DSDNA依赖性ATPase，因此具有适用于高通量的活性使用各种小分子文库的抑制剂筛选（HTS）。我们已经建立了生化纯化活跃的Pich，我们制定了策略，并生成了将允许主要命中的工具从HTS有效进行次要筛查，以验证真正的PICH抑制剂和消除非特异性抑制剂。 PICH抑制的细胞后果将被确定为完全表征对正常细胞和癌细胞的影响。细胞毒性研究将确定哪些癌症类型可以被抑制剂有选择地杀死，我们将定义所需的剂量。结果将是有选择性的小分子皮基抑制剂将是有价值的研究工具，将成为铅化合物在动物癌模型研究之前，最佳有效分子的发展。