Covalent Fluorescent Probes for Cancer Cell Detection

用于癌细胞检测的共价荧光探针

• 批准号: 8294984
• 负责人: NATHANAEL Schiander GRAY
• 金额: $ 18.41万
• 依托单位: DANA-FARBER CANCER INST
• 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/8294984
• 关键词: Affinity Chromatography; BODIPY; Benzodiazepines; Biological Markers; Biotin; Boston; Cancer Biology; Cancer Center; Cancer cell line; Cell physiology; Cells; Chromosomal Rearrangement; Collaborations; Cysteine; Dana-Farber Cancer Institute; Detection; Development; Epidermal Growth Factor Receptor; Equilibrium; Event; Family; Fibroblast Growth Factor Receptors; Fluorescence; Fluorescent Probes; Gene Amplification; Gene Deletion; Goals; Gray unit of radiation dose; Imagery; Institution; Label; Libraries; Malignant Neoplasms; Mass Spectrum Analysis; Mentors; Methods; Microscopy; Persons; Phosphotransferases; Point Mutation; Population; Protein Kinase; Relative (related person); Research Personnel; Screening procedure; Site; Staining method; Stains; Training; Yang; abstracting; base; biological systems; cancer cell; career; career development; cellular engineering; covalent bond; fluorescence imaging; imaging probe; inhibitor/antagonist; kinase inhibitor; medical schools; member; novel; scaffold; sensor; skills; small molecule; tool

Abstract: Fluorescence imaging is a powerful tool that permits visualization of specific cell states within a population; however, existing methods for fluorescence labeling are not experimentally accessible for many biological systems. Furthermore, fluorescent small-molecule sensors of cell state may provide a valuable alternative with significant benefits relative to existing methods for fluorescence imaging. The overall goal of this project is to create fluorescent small molecule ATP-site directed probes that can selectively label particular kinases and serve as imaging probes of normal versus pathological cell state. Protein kinases are in many ways ideal targets for the development of selective small molecule imaging probes for use in cancer biology. This is because protein kinases are involved in most cellular processes and changes in their localization, accessibility, and abundance are associated with changes in cellular state. Protein kinases have been used as biomarkers in cancer biology because the loss of endogenous kinase regulatory mechanisms by point mutations, gene deletions, gene amplifications, and chromosomal rearrangements has been well-established as crucial events in many cancers. The specific aims of this project are to: 1) Synthesize fluorescently-tagged kinase inhibitors capable of forming covalent bonds with ATP-site cysteine residues; 2) Use microscopy-based screening ofthe compounds prepared in Aim 1 to identify compounds that are selective-probes of normal and pathological cellular states and 3) Identify the intracellular target(s) of active compounds identified in Aim 2. This proposed is the extension of an ongoing collaboration between Drs. Nathanael Gray (Dana Farber Cancer Institute), Priscilla Yang (Harvard Medical School) and Wei Zhang (UMass Boston). All three are also members of the Dana Farber/Harvard Cancer Center. One of the strong points of this proposal is that each Co-Pl is responsible for one of the three Specific Aims allowing the project to be benefited by the balanced skills and expertise that each person and their subsequent institution brings to the partnership. Because all three investigators are eariy in their careers, Drs. Gray, Yang and Zhang will benefit from career development activities and mentors who are intemal and extemal. Additionally, the Training Core will significantiy contribute to this project.

摘要：荧光成像是一种强大的工具，可允许在人群中可视化特定的细胞态。但是，对于许多生物系统，现有的荧光标记方法在实验上无法访问。此外，相对于现有的荧光成像方法，相对于现有方法，细胞态的荧光小分子传感器可以提供有价值的替代方案。总体目标 该项目是为了创建荧光小分子ATP位点的探针，该探针可以选择性地标记特定的激酶并用作正常病理细胞态的成像探针。蛋白激酶在许多方面都是用于开发用于癌症生物学的选择性小分子成像探针的理想靶标。 这是因为蛋白激酶参与了大多数细胞过程，其定位，可及性和丰度的变化与细胞状态的变化有关。蛋白激酶已被用作癌症生物学中的生物标志物，因为按点突变，基因缺失，基因扩增和染色体重排的内源激酶调节机制丧失已得到充分公认 作为许多癌症的关键事件。该项目的具体目的是：1）合成荧光标记的激酶抑制剂，能够与ATP位点半胱氨酸残基形成共价键； 2）使用基于显微镜在AIM 1中制备的化合物的基于显微镜的筛选，以鉴定正常和病理细胞状态的选择性探针的化合物，3）识别AIM 2中鉴定的活性化合物的细胞内靶标。 提出的是DRS之间正在进行的合作的扩展。 Nathanael Gray（Dana Farber癌症研究所），Priscilla Yang（哈佛医学院）和Wei Zhang（波士顿UMass）。这三个都是达纳·法伯/哈佛癌中心的成员。该提案的重点之一是，每个Co-PL都负责三个特定目标之一，允许该项目受益于每个人及其后续机构带来合作伙伴关系的平衡技能和专业知识。由于所有三位调查人员的职业生涯都含糊其范围。格雷，杨和张将受益于职业发展活动，以及牢不可知和逃离的导师。此外，训练核心将为该项目做出重要贡献。