Development of assays for HTS to identify inhibitors of a new PPI involved in cancer metastasis

开发 HTS 检测方法以鉴定参与癌症转移的新 PPI 抑制剂

• 批准号: 9311182
• 负责人: Celine DerMardirossian
• 金额: $ 44.03万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9311182
• 关键词: Actins; Adverse effects; Affinity; Area; Automobile Driving; Basement membrane; Binding; Biological; Biological Assay; Biology; Breast Cancer Cell; Breast Cancer Patient; Breast Cancer therapy; Cell surface; Cells; Cellular Structures; Cessation of life; Collection; Complex; Cytoskeleton; Data; Detection; Development; Devices; Diagnosis; Drug Targeting; Experimental Designs; Extracellular Matrix; Fluorescence; Fluorescence Resonance Energy Transfer; Focal Adhesions; Functional disorder; Future; Goals; Guanine; Guanine Nucleotide Exchange Factors; Guanosine Triphosphate Phosphohydrolases; Human; Impairment; In Vitro; Investigation; Knowledge; Laboratories; Lead; Libraries; Life; MDA MB 231; Malignant neoplasm of prostate; Metastatic breast cancer; Molecular; Neoplasm Metastasis; Nude Mice; Patients; Peptides; Pharmaceutical Preparations; Play; Process; Protein Analysis; Proteins; Research Personnel; Role; Savings; Series; Signal Pathway; Signal Transduction; Signaling Protein; Specificity; Testing; Therapeutic; Time; Toxic effect; Treatment Efficacy; Tumor Cell Migration; Vesicle; assay development; base; cancer cell; cell motility; design; experimental study; follow-up; high throughput screening; improved; in vivo; inhibitor/antagonist; innovation; leukemia; malignant breast neoplasm; migration; miniaturize; neoplastic cell; novel; outcome forecast; overexpression; prevent; protein protein interaction; protein transport; resistance mechanism; rho; screening; small molecule; small molecule inhibitor; stable cell line; time use; trafficking; tumor; tumor progression; uncontrolled cell growth

Abstract Breast cancer is a global problem, with 1.3 million new cases diagnosed and 430,000 deaths each year worldwide. Recently, researchers have begun developing exciting drugs to target the protein-protein interactions (PPIs) that regulate metastasis-driving signaling pathways. PPIs are critical for cellular signal transductions that play key roles in both normal and abnormal functions in cells. Changes in specificity and affinity of these interactions can lead to cellular malfunctions, such as uncontrolled cell growth and/or cell migration. A critical key in dissemination process is the ability of cells to coordinate signaling pathways with cytoskeleton dynamics. As critical coordinators of the cytoskeleton machinery, GTPases and their regulator GEFs are key players for cell movement. Many tumors show increased expression and/or activation of GTPases and GEFs. These proteins play fundamental roles in several aspects of cancer progression, metastasis and poor prognosis. Our long-term goals are to better understand the molecular mechanisms of GTPases in metastasis process and to device more effective, less toxic breast cancer therapies. Our strong preliminary data produced in our laboratory uncover a new protein-protein interaction critical in cell invasion and metastasis. The objective in this application is to implement a pilot high-throughput screen (HTS) to identify small molecule inhibitors of these novel binding partners that will represent potentially life-saving therapeutic leads for inhibiting cancer metastasis. Three specific aims will be developed in this proposal: 1) Development of fluorescence assay to detect this PPI; 2) Pilot screen: Complete pilot screen against a compound collection of 20,000 compounds and follow up assays; 3) Biological effects: perform initial characterization of active compounds on cell migration and invasion. Our HTS will involve an innovative approach that analyzes PPIs in cells and improve scientific knowledge about key signaling proteins in cell migration. This investigation will have substantial significance in understanding fundamental and defined features of cell motility, with implications for many areas of biology and pathophysiology.

抽象的 乳腺癌是一个全球性问题，每年新诊断出 130 万例乳腺癌，导致 43 万人死亡 全世界。最近，研究人员开始开发令人兴奋的药物来靶向蛋白质-蛋白质相互作用 （PPIs）调节转移驱动信号通路。 PPI 对于细胞信号转导至关重要 在细胞的正常和异常功能中发挥关键作用。这些的特异性和亲和力的变化 相互作用可能导致细胞功能障碍，例如不受控制的细胞生长和/或细胞迁移。一个批评的 传播过程的关键是细胞协调信号通路与细胞骨架动力学的能力。 作为细胞骨架机制的关键协调者，GTPase 及其调节器 GEF 是细胞骨架机制的关键参与者。 细胞运动。许多肿瘤显示 GTPases 和 GEF 的表达和/或激活增加。这些 蛋白质在癌症进展、转移和不良预后的多个方面发挥着重要作用。我们的 长期目标是更好地了解 GTPase 在转移过程中的分子机制并 装置更有效、毒性更小的乳腺癌疗法。我们实验室产生的强有力的初步数据 揭示一种在细胞侵袭和转移中至关重要的新蛋白质-蛋白质相互作用。本申请的目标 是实施试点高通量筛选（HTS）来识别这些新型结合的小分子抑制剂 合作伙伴将代表抑制癌症转移的潜在挽救生命的治疗线索。三 该提案将制定具体目标：1）开发荧光测定法来检测该 PPI； 2) 飞行员 筛选：针对 20,000 种化合物的化合物集合进行完整的试点筛选并进行后续分析； 3） 生物学效应：对活性化合物对细胞迁移和侵袭的影响进行初步表征。 我们的 HTS 将采用创新方法来分析细胞中的 PPI 并提高科学知识 关于细胞迁移中的关键信号蛋白。此次调查将具有重大意义 了解细胞运动的基本特征和定义特征，对生物学和生物学的许多领域都有影响 病理生理学。