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

项目摘要/摘要 去调节的蛋白质相互作用有助于人类的发展、维持和化疗耐药 癌症。致癌蛋白质的关键接触点通常由所谓的“槽中螺旋”来调节。 相互作用，由此一种蛋白质的α-螺旋亚组分插入到另一种蛋白质的表面凹槽中 来驱动致癌信号。例如，bcl2家族蛋白调节细胞间的关键平衡。 生死细胞和癌细胞过度表达抗凋亡成员，这些成员包含一个表面凹槽， 有效地捕获支持细胞凋亡的成员的“杀手”螺旋，以强制细胞永生。我们已经把所有的- 碳氢化合物伸展成天然的α-螺旋结构，以恢复其形状、稳定性和生物活性，从而使它们能够 既可用作分析致癌蛋白质相互作用的强大化学工具，也可用作治疗 给他们下药。在过去的15年里，我在设计装订多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。我已经为核磁共振结构生成了自旋标记的类似物 用质谱仪进行快速结合部位分析的光反应结构半胱氨酸反应 致癌蛋白的共价靶向突变体，结合亲和力定量的荧光衍生物和 细胞成像、体内成像的放射性标记结构和临床前的迭代优化的类似物 测试和翻译。装订的多肽的显著影响最好地反映在它们识别 新的癌症靶点、机制和可药物结合部位及其在成人临床试验中的进展 以及患有复发癌症的儿童。这项竞争性续签申请的目标是继续扩大 支持关键癌症研究项目的装订多肽创新的广度和深度 这些试剂在蛋白质组发现、结构确定、作用机制研究和治疗方面 发展。具体地说，我的目标是大力支持NCI资助的我的单位主任的R35研究计划， Loren Walensky博士是一名化学生物学家和儿科肿瘤学家，他专注于描述bcl-2的特征 通过中和线粒体凋亡途径来驱动人类癌症的家族相互作用机制。 重新激活肿瘤细胞凋亡是克服化疗耐药和病理性α-螺旋的关键 Bcl-2家族的相互作用非常适合于用装订的多肽进行询问。在运行装订的 瓦伦斯基实验室的多肽设计小组和达纳-法伯的癌症化学生物学项目，我是 亲自负责开发和优化推动装订肽创新的化学物质，以及 创建高通量的咨询、生产、纯化、量化和表征工作流程 也被我们的数十名内部和外部合作者所依赖。作为研究专家，在 化学、癌症生物学和实验治疗学的接口，我致力于挖掘潜力 以促进我们对基本致癌机制和 为癌症治疗创造新的疗法。

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.