Stapled Peptides for Protein Interaction Research and Therapeutic Targeting in Human Cancer

用于人类癌症蛋白质相互作用研究和治疗靶向的钉合肽

• 批准号: 10323091
• 负责人: Gregory Howard Bird
• 金额: $ 33.21万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-15 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10323091
• 关键词: Adult; Affinity; Apoptosis; Apoptotic; BCL2 gene; Binding; Binding Sites; Biochemistry; Biology; Cancer Biology; Cancer Patient; Cancer Relapse; Cancer Research Project; Cellular biology; Cessation of life; Chemicals; Chemistry; Chemoresistance; Child; Clinical Trials; Consultations; Cysteine; Development; Equilibrium; Family; Funding; Genetic Transcription; Goals; Human; Hydrocarbons; Investigational Therapies; Life; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Metabolism; Mining; Mitochondria; Modality; Oncogenic; Pathologic; Pediatric Oncologist; Peptides; Pharmaceutical Preparations; Preclinical Testing; Production; Protein Family; Proteins; Proteomics; Radiolabeled; Reagent; Research; Running; Shapes; Signal Transduction; Specialist; Spin Labels; Structure; Surface; Therapeutic; Translations; Variant; alpha helix; analog; anticancer research; cancer cell; cancer therapy; cellular imaging; design; in vivo imaging; innovation; insight; member; next generation; novel therapeutics; overexpression; programs; prototype; structural biology; therapeutic development; therapeutic target; tool

PROJECT SUMMARY / ABSTRACT Deregulated protein interactions contribute to the development, maintenance, and chemoresistance of human cancer. The critical contact points of cancer-causing proteins are often mediated by so-called “helix-in-groove” interactions, whereby an alpha-helical subcomponent of one protein inserts into the surface groove of another to drive oncogenic signaling. For example, BCL-2 family proteins regulate the critical balance between cellular life and death and cancer cells overexpress the anti-apoptotic members, which contain a surface groove that effectively traps the “killer” helix of pro-apoptotic members, to enforce cellular immortality. We have inserted all- hydrocarbon struts into natural alpha-helices to restore their shape, stability, and bioactivity so that they can be used as both powerful chemical tools to dissect oncogenic protein interactions and as prototype therapeutics to drug them. Over the last 15 years, I have developed special expertise in the design of stapled peptides for diverse applications in cancer research and treatment. I have generated spin-labeled analogs for NMR structural analyses, photoreactive constructs for rapid binding-site analysis by mass spectrometry, cysteine-reactive variants for covalent targeting of oncogenic proteins, fluorescent derivatives for binding affinity quantitation and cellular imaging, radiolabeled constructs for in vivo imaging, and iteratively-optimized analogs for preclinical testing and translation. The remarkable impact of stapled peptides is best reflected by their capacity to identify new cancer targets, mechanisms, and druggable binding sites and their advancement to clinical trials in adults and children with relapsed cancers. The goal of this competitive renewal application is to continue to expand the breadth and depth of stapled peptide innovation in support of critical cancer research programs that harness these reagents in proteomic discovery, structural determination, mechanism-of-action studies, and therapeutic development. Specifically, I aim to robustly support the NCI-funded R35 research program of my Unit Director, Dr. Loren Walensky, who as a chemical biologist and pediatric oncologist, focuses on characterizing the BCL-2 family interaction mechanisms that drive human cancer by neutralizing the mitochondrial apoptosis pathway. Reactivating apoptosis in cancer is essential to overcoming chemoresistance and the pathologic alpha-helical interactions of the BCL-2 family are ideally suited for interrogation by stapled peptides. In running the Stapled Peptide Design Group of the Walensky lab and Dana-Farber’s Program in Cancer Chemical Biology, I am personally responsible for developing and optimizing the chemistry that drives stapled peptide innovations, and creating a high-throughput consultation, production, purification, quantitation, and characterization workflow that is also relied upon by dozens of our internal and external collaborators. As a Research Specialist operating at the interface of chemistry, cancer biology, and experimental therapeutics, I am committed to mining the potential of next-generation stapled peptides to advance our understanding of fundamental oncogenic mechanisms and to create novel therapeutics for cancer treatment.

项目总结/摘要 失调的蛋白质相互作用有助于人类肿瘤的发展、维持和耐药性 癌致癌蛋白质的关键接触点通常由所谓的“螺旋槽”介导 相互作用，其中一种蛋白质的α-螺旋亚组分插入另一种蛋白质的表面凹槽 来驱动致癌信号。例如，BCL-2家族蛋白调节细胞间的关键平衡， 生命和死亡以及癌细胞过度表达抗凋亡成员，其包含表面凹槽， 有效地捕获促凋亡成员的"杀伤"螺旋，以增强细胞的永生性。我们把所有的- 将烃支柱转化为天然α-螺旋以恢复它们的形状、稳定性和生物活性，使得它们可以 作为强大的化学工具来剖析致癌蛋白质的相互作用，并作为原型疗法， 给他们下药在过去的15年里，我在设计钉合肽方面积累了特殊的专业知识， 在癌症研究和治疗中的多种应用。我已经生成了核磁共振结构的自旋标记类似物， 质谱法快速结合位点分析的光反应结构，半胱氨酸反应 用于致癌蛋白共价靶向的变体，用于结合亲和力定量的荧光衍生物， 细胞成像，用于体内成像的放射性标记的构建体，以及用于临床前成像的迭代优化的类似物。 测试和翻译。钉合肽的显著影响最好地反映在它们识别 新的癌症靶点、机制和药物结合位点及其在成人临床试验中的进展 和癌症复发的儿童。这种竞争性续约申请的目标是继续扩大 钉合肽创新的广度和深度，以支持关键的癌症研究计划， 这些试剂用于蛋白质组学发现、结构测定、作用机制研究和治疗 发展具体来说，我的目标是大力支持我的单位主任的NCI资助的R35研究计划， 博士化学生物学家和儿科肿瘤学家Loren Walensky专注于描述BCL-2的特征， 通过中和线粒体凋亡途径驱动人类癌症的家族相互作用机制。 在癌症中重新激活细胞凋亡对于克服化疗耐药性和病理性α-螺旋肿瘤是必不可少的。 BCL-2家族的相互作用理想地适合于通过钉合肽进行询问。在运行装订 我是瓦伦斯基实验室的肽设计小组和丹娜-法伯癌症化学生物学项目的负责人， 亲自负责开发和优化推动钉合肽创新的化学物质， 创建高通量咨询、生产、纯化、定量和表征工作流程， 也是我们数十位内部和外部合作者的依赖。作为研究专家， 化学，癌症生物学和实验治疗学的接口，我致力于挖掘潜力， 下一代钉合肽，以促进我们对基本致癌机制的理解， 来创造新的癌症治疗方法。