Project 2: Development of grp94-selective Inhibitors for Cancer

项目2：grp94选择性癌症抑制剂的开发

• 批准号: 8934514
• 负责人: GABRIELA CHIOSIS
• 金额: $ 30.44万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8934514
• 关键词: Address; Advanced Malignant Neoplasm; Affinity; Animals; Back; Biochemical; Biology; Blood; Breast Cancer Treatment; Cancer Biology; Cell Line; Cell membrane; Cells; Characteristics; Chemicals; Client; Clinic; Clinical; Complement; Data; Development; Disease; Disease Resistance; ERBB2 gene; Endoplasmic Reticulum; Environment; Evaluation; Family; Genes; Genetic; Glucose; Goals; Heat shock proteins; Heat-Shock Proteins 90; Homologous Gene; Housing; Integrins; Investigation; Label; Lead; Ligands; Link; Maintenance; Malignant Neoplasms; Mediating; Medical Research; Mitochondria; Molecular Chaperones; Mus; Nature; Oncogenic; Organism; Pathway interactions; Pharmaceutical Preparations; Pharmacology; Phenotype; Protein Overexpression; Protein Tyrosine Kinase; Proteins; Proteome; Reagent; Refractory; Research; Role; Signal Transduction; Solid; System; TNF receptor-associated factor 1; Therapeutic; Toll-like receptors; Translating; Translations; Yeasts; anti-cancer therapeutic; base; cancer cell; cancer therapy; cellular engineering; combinatorial; design; drug development; drug discovery; genetic approach; improved; in vivo; inhibitor/antagonist; malignant breast neoplasm; malignant phenotype; mutant; novel; overexpression; paralogous gene; pressure; programs; receptor; screening; small molecule; small molecule libraries; spatiotemporal; targeted cancer therapy; targeted treatment; therapy design; therapy resistant; tool; tumor

PROJECT SUMMARY Background and hypothesis: While grp94 expression in cancer is linked to aggressive disease and resistance to therapy, little is known on mechanisms underlying these effects. Our inability to study grp94 in genetically tractable organisms such as yeast and the unavailability of small molecule grp94 inhibitors are largely responsible for this state of affairs. Until now investigation of grp94 has mainly used mutant cell lines and gene deficient mouse studies. Although powerful, this approach is limited because it addresses phenotypic changes in the complete absence of a gene, and moreover, makes use of an engineered cellular environment. Alternative strategies that address the role and biology of grp94 in an endogenous cellular environment, where grp94 is limiting but not absent, are therefore needed. In addition to linking grp94-mediated mechanisms to the relevant disease state, i.e. tumor type, these reagents may provide evidence refractory to the currently available means. Approach: Discovery of grp94-selective ligands and chemical tools has been challenging thus far because of a high degree of analogy between the HSP90 paralogs. We here provide preliminary evidence showing that the discovery of grp94 inhibitors with log selectivity over the other paralogs is possible. Using a combination of screening an in-house generated small-molecule library and computational analyses backed by structural studies conducted with Project 3, we have identified important structural determinants that impart ligand selectivity and high affinity for grp94. We propose to use this information as a springboard for the development of grp94-directed chemical tools such as grp94 selective ligands and solid support immobilized, biotinylated and fluorescently labeled derivatives (Aim 1a) and as a starting point towards the development of drug-like grp94 inhibitors for translation to clinic as novel anti-cancer therapeutics (Aim 1b). We also propose here to use the grp94-directed toolset for the investigation of disease-specific roles of grp94 (Aim 2). Significance: In support of this investigative chemical biology approach, we provide preliminary data that confirm its power. We show that it identifies a novel role for grp94 in regulating HER2 tyrosine kinase at the plasma membrane, specifically in the case of tumors with overexpression of this protein. Also we implicate grp94 in regulating oncogenic signal transduction at the plasma membrane, indicating grp94 as a target in HER2-overexpressing breast cancer. These findings provide the blueprint for the development of grp94 inhibitors as a novel targeted therapy for the treatment of breast cancers dependent on increased signaling through plasma membrane receptors. Importantly, although HER2 was for the last two decades one of the most widely studied HSP90 onco-client protein, the mechanistic and the therapeutic understanding unveiled by our chemical biology approach was missed so far, proving further evidence to the importance of having a strong chemical toolset in addition to genetic and biochemical means for the understanding of biology.

项目摘要 背景和假设：癌症中的GRP94表达与侵略性疾病有关 对治疗的抵抗力，在这些作用的基础机制上鲜为人知。我们无法研究GRP94 诸如酵母等遗传诱导的生物和小分子GRP94抑制剂的不可用 在很大程度上负责这种情况。到目前为止，对GRP94的研究主要使用了突变细胞系 和基因缺乏小鼠研究。尽管强大，但这种方法是有限的，因为它解决了表型 完全没有基因的变化，而且还使用了工程的细胞环境。 解决GRP94在内源性细胞环境中的作用和生物学的替代策略， 因此，GRP94是有限的，但不存在。除了将GRP94介导的机制与 相关疾病状态，即肿瘤类型，这些试剂可能会为当前的证据提供难治性的证据 可用的手段。 方法：迄今为止，GRP94选择配体和化学工具的发现一直在挑战 由于HSP90旁系同源物之间的类比很高。我们在这里提供初步证据 表明可以发现对其他旁系同源物具有对数选择性的GRP94抑制剂。使用 筛选内部生成的小分子库和由支持的计算分析的组合 通过项目3进行的结构研究，我们已经确定了赋予的重要结构决定因素 配体选择性和GRP94的高亲和力。我们建议将这些信息用作跳板 开发GRP94指导的化学工具，例如GRP94选择性配体和固定支持的固体支持， 生物素化和荧光标记的衍生物（AIM 1A），作为发展的起点 类似药物的GRP94抑制剂，用于转化为诊所的新型抗癌治疗剂（AIM 1B）。我们也建议 在这里使用GRP94指导的工具集研究GRP94的疾病特异性作用（AIM 2）。 意义：为了支持这种研究性化学生物学方法，我们提供了初步数据 这证实了它的力量。我们表明，它标识了GRP94在调节HER2酪氨酸激酶中的新作用 质膜，特别是在肿瘤过表达的情况下。我们也暗示 GRP94在调节质膜上的致癌信号转导，表明GRP94作为目标 过表达HER2的乳腺癌。这些发现为开发GRP94提供了蓝图 抑制剂作为一种新型靶向疗法，用于治疗乳腺癌，取决于增加信号传导 通过质膜受体。重要的是，尽管HER2一直是过去的二十年 最广泛研究的HSP90 Onco-Client蛋白，机械和治疗理解由 到目前为止，我们的化学生物学方法尚未错过 强大的化学工具集除了遗传和生化手段以理解生物学。