Reactivation of the p53-mediated pro-Apoptotic Pathway Using Stapled Peptides

使用钉合肽重新激活 p53 介导的促凋亡途径

基本信息

批准号：
7934322
负责人：
FEDERICO BERNAL
金额：
$ 9.68万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2011-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7934322
关键词：
Acidic Amino Acids Acylation Affinity Affinity Labels Alkenes Amino Acids Apoptosis Apoptotic Architecture Automation BAX gene BH3 peptide Binding Biochemical Biological Biological Assay Biology Bromodeoxyuridine Cancer Biology Cancer cell line Cell Communication Cell Cycle Cell Cycle Arrest Cell Separation Cell membrane Cell physiology Cells Cellular Assay Chemicals Clinical Oncology Co-Immunoprecipitations Collaborations Complex Defect Detection Development Disease Doctor of Philosophy Employee Strikes Environment Faculty Family Fatty Acids Fluorescence Fluorescence Polarization Generations Goals Growth Health Hydrocarbons Image In Vitro Lead Ligands Maintenance Malignant Neoplasms Mediating Mentors Microscopic Mitochondria Modification Molecular Structure Mus Organic Chemistry Pathogenesis Pathologic Pathway interactions Peptide Hydrolases Peptide Library Peptides Permeability Phase Property Protein Binding Protein p53 Proteins Reaction Reader Reading Recombinants Regulation Research Research Personnel Resistance Roentgen Rays Role Scheme Side Signal Pathway Site Specificity Stimulus Structure TP53 gene Techniques Tertiary Protein Structure Therapeutic Tumor Suppressor Proteins Universities Xenograft Model affinity labeling analog annexin A5 base cancer cell caspase-3 crosslink cytochrome c design high throughput screening in vivo interdisciplinary approach member multidisciplinary mutant next generation novel programs prototype research study response structural biology tool

项目摘要

DESCRIPTION (provided by applicant): Faulty regulation of apoptosis and the cell cycle are critical determinants of cancer pathogenesis. These essential cellular processes are modulated, in health and disease, by multiple protein networks. Among these, the BCL-2 family of pro- and anti-apoptotic proteins, as well as the p53 tumor suppressor protein, stand out due to their essential roles in triggering apoptosis and cell cycle arrest in response to diverse stimuli. Newly identified interactions between p53 and select BCL-2 members, such as BAX and BCL-XL, impact the survival of cells, and these interactions are dictated by the three-dimensional molecular structure of specific peptide domains located within their protein architecture. Defects in these interactions lead to pathologic deregulation of apoptotic and growth arrest signaling pathways. The goal of this research is to use a multidisciplinary approach to target the control points of the integrated BCL2-family-p53 apoptotic signaling pathway in an effort to investigate and reactivate apoptosis in cancer cells. To that end, we propose to: 1. Synthesize diversified hydrocarbon-stapled peptide libraries of BID BH3, PUMA BH3, and p53 and evaluate their biochemical and biophysical properties; 2. Optimize the intracellular delivery and versatility of stapled peptides through chemical derivatization; 3. Evaluate the ability of stapled BID BH3, PUMA BH3, and p53 peptides, singly and in combination, to induce cell cycle arrest and/or activate BAX-induced mitochondrial apoptosis in vitro and in murine xenograft models. Peptide helices that engage protein binding pockets hold promise as a novel set of chemical tools to study and treat cancer. Our initial panels of BID BH3 and p53 stapled peptides demonstrate striking biochemical and pharmacologic properties, including enhanced (-helicity, protease resistance, and cell permeability compared to the corresponding unmodified peptides. Importantly, the stapled peptides recapitulate the biological activity of the native protein domains in vitro and in vivo. The recently identified intersection of the BCL-2 family and p53 pathways at the BAX control point of apoptosis opens new opportunities to develop next-generation stapled peptides to induce cell cycle arrest synergistically and reactivate apoptosis in cancer cells. The research proposed here aims to apply novel synthetic approaches such as hydrocarbon-stapling to reinforce natural peptide ligands relevant to deregulated BCL-2 and p53 pathways, thereby providing alternative tool compounds to study and manipulate protein interactions relevant to the pathogenesis and maintenance of cancer. A multidisciplinary group of faculty mentors, collaborators, and academic advisors with expertise spanning organic chemistry, structural biology, cancer biology, and clinical oncology will provide an ideal environment for Dr. Bernal's development as an independent investigator in the field of cancer chemical biology.

描述（由申请人提供）：细胞凋亡和细胞周期的错误调节是癌症发病机制的关键决定因素。在健康和疾病中，这些重要的细胞过程受到多种蛋白质网络的调节。其中，促凋亡蛋白和抗凋亡蛋白的 BCL-2 家族以及 p53 肿瘤抑制蛋白因其在响应不同刺激而触发细胞凋亡和细胞周期停滞中发挥重要作用而脱颖而出。新发现的 p53 和 BCL-2 成员（例如 BAX 和 BCL-XL）之间的相互作用会影响细胞的存活，而这些相互作用是由位于其蛋白质结构内的特定肽域的三维分子结构决定的。这些相互作用的缺陷导致细胞凋亡和生长停滞信号通路的病理失调。本研究的目标是利用多学科方法来靶向整合的 BCL2-家族-p53 凋亡信号通路的控制点，以研究和重新激活癌细胞的凋亡。为此，我们建议： 1. 合成 BID BH3、PUMA BH3 和 p53 多样化的碳氢化合物肽库，并评估其生化和生物物理特性； 2.通过化学衍生化优化钉合肽的细胞内递送和多功能性； 3. 评估装订的 BID BH3、PUMA BH3 和 p53 肽（单独或组合）在体外和小鼠异种移植模型中诱导细胞周期停滞和/或激活 BAX 诱导的线粒体凋亡的能力。与蛋白质结合袋结合的肽螺旋有望成为研究和治疗癌症的一套新型化学工具。我们最初的 BID BH3 和 p53 钉合肽组表现出惊人的生化和药理学特性，包括与相应的未修饰肽相比增强的螺旋性、蛋白酶抗性和细胞通透性。重要的是，钉合肽概括了天然蛋白质结构域的生物活性最近发现的 BCL-2 家族和 p53 通路在细胞凋亡的 BAX 控制点的交叉点为开发下一代钉合肽来诱导提供了新的机会。本文提出的研究旨在应用碳氢钉合等新型合成方法来增强与失调的 BCL-2 和 p53 通路相关的天然肽配体，从而提供替代工具化合物来研究和操纵。与癌症的发病机制和维持相关的蛋白质相互作用。一个由教师导师、合作者和学术顾问组成的多学科小组，其专业知识涵盖有机化学、结构生物学、癌症生物学和临床肿瘤学，将为 Bernal 博士作为癌症化学生物学领域的独立研究者的发展提供理想的环境。