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：
7471803
负责人：
FEDERICO BERNAL
金额：
$ 16.24万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7471803
关键词：
Acidic Amino Acids Acylation Affinity Affinity Labels Alkenes Amino Acids Apoptosis Apoptotic Architecture Automation BAX gene BH3 peptide Bax protein 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 Endopeptidases Environment Faculty Family Fatty Acids Fluorescence Fluorescence Polarization Generations Goals Growth Health Helix (Snails)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 ear helix 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）：凋亡和细胞周期的调节错误是癌症发病机理的关键决定因素。这些基本的细胞过程在健康和疾病中被多个蛋白质网络调节。其中，BCl-2促凋亡蛋白的BCl-2家族以及p53肿瘤抑制蛋白的蛋白质，由于它们在响应各种刺激的响应中触发凋亡和细胞周期停滞而脱颖而出。新鉴定的p53与精选的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，这些相互作用由位于其蛋白质结构中的特定肽结构域的三维分子结构决定。这些相互作用的缺陷导致凋亡和生长停滞信号通路的病理失调。这项研究的目的是使用多学科的方法来靶向综合BCL2-家庭p53凋亡信号传导途径的控制点，以研究和重新激活癌细胞中的凋亡。为此，我们建议：1。综合出价BH3，PUMA BH3和p53的多样化的碳氢化合物抑制剂库，并评估其生化和生物物理特性； 2。通过化学衍生化优化钉钉肽的细胞内递送和多功能性； 3。单独和组合评估钉书钉BID BH3，PUMA BH3和p53肽的能力，诱导细胞周期停滞和/或激活Bax诱导的体外和鼠类异种移植模型中的Bax诱导的线粒体细胞凋亡。接合蛋白质结合口袋的肽螺旋是研究和治疗癌症的一组新型化学工具。我们的初始BID BH3和p53钉蛋白肽的初始面板表现出惊人的生化和药理学特性，包括增强（ - 螺旋性，抗蛋白酶抗性和细胞渗透性与相应的未修饰的肽相比。 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研究和操纵与癌症的发病机理和维持相关的蛋白质相互作用的化合物。跨越有机化学，结构生物学，癌症生物学和临床肿瘤学专业知识的教师，合作者和学术顾问的多学科小组将为Bernal博士作为癌症化学生物学领域的独立研究人员的发展提供理想的环境。