The Role of the Mutant p53-PARP-MCM Pathway in Triple Negative Breast Cancer

突变型 p53-PARP-MCM 通路在三阴性乳腺癌中的作用

• 批准号: 10545729
• 负责人: Jill E. Bargonetti
• 金额: $ 33.85万
• 依托单位: HUNTER COLLEGE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-06 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10545729
• 关键词: ADP ribosylation; Affect; African American; African ancestry; Amino Acids; Animals; Area; Biological; Biological Markers; Biology; Breast Cancer Cell; Breast Cancer Patient; C-terminal; CRISPR/Cas technology; Caring; Cell Culture Techniques; Cell Death; Cell Line; Cells; Chromatin; Chromatin Remodeling Factor; Communities; Complex; DNA; DNA Binding; DNA Damage; DNA biosynthesis; DNA replication fork; DNA-Protein Interaction; Data; Data Reporting; Data Set; Detection; Diagnostic; Disease; Genes; Genetic Transcription; Heat shock proteins; Human; Immunohistochemistry; Implant; Knowledge; MCM Protein; MCM2 gene; Maintenance; Malignant Neoplasms; Mammary Gland Parenchyma; Mediating; Methods; Missense Mutation; Modeling; Molecular Analysis; Multiprotein Complexes; Mus; Mutation; Neoplasm Circulating Cells; Nuclear; Oncogenic; Outcome; Pathway interactions; Patients; Peptides; Phase; Poly(ADP-ribose) Polymerase Inhibitor; Poly(ADP-ribose) Polymerases; Population; Population Heterogeneity; Property; Proteins; Publishing; Race; Role; Scientist; Signal Transduction; Sterol Biosynthesis Pathway; TP53 gene; Testing; The Cancer Genome Atlas; Therapeutic; Tissue Microarray; Tissues; Tumor Promotion; Tumor Suppressor Genes; Tumor Volume; Woman; Work; Xenograft Model; Xenograft procedure; cancer cell; cancer imaging; cancer subtypes; cell killing; cofactor; gain of function; health disparity; helicase; imaging agent; imaging platform; improved; innovation; insight; knock-down; mortality; mouse model; multi-ethnic; mutant; neoplastic cell; novel; patient derived xenograft model; protein complex; recruit; replication stress; screening; synergism; temozolomide; theranostics; treatment choice; triple-negative invasive breast carcinoma; tumor

Project Summary: The Role of the Mutant p53-PARP-MCM Pathway in Triple Negative Breast Cancer Mutant p53 (mtp53) is found in 80% of triple negative breast cancers (TNBCs). There is a need to increase understanding of mtp53 signaling in TNBC in order to determine strategies to detect and treat the disease. TNBCs are more common in African American women than in white women and therefore the proposed work will benefit all women with TNBC including the AA over-represented population. This application proposes to determine the biological role of TNBC-associated gain-of-function oncogenic mtp53 R273H that is associated with PARP1 (herein referred to as PARP) protein, MCM2-7 helicases and replicating DNA. The p53 Arg 273 substituted with His, and other hotspot p53 missense mutant proteins, are well known to function as oncogenic proteins in TNBCs, but their functions outside of acting as aberrant transcription co-factors are not clearly understood. The R273H mtp53 is present in high levels on chromatin in many TNBCs and works as a co-factor to activate the transcription of sterol biosynthesis genes and chromatin remodeling factors, but incredibly R273H also co-associates with PARP and MCMs on replicating DNA. Our studies have uncovered this novel replication- associated pathway and we call it the mtp53-PARP-MCM chromatin axis. Our preliminary and published data reports that knockdown of mtp53 R273H reduces chromatin associated MCM2-7 and PARP. Knockdown of mtp53 in this setting decreases combined PARP inhibitor (PARPi) talazoparib plus DNA damaging agent temozolomide-mediated cell death, demonstrating that mtp53 expression influences the sensitivity of cancer cells to synergistic PARPi plus DNA damage mediated cell killing. Our preliminary and published data showed nuclear mtp53 R273H co-associates with PARP and MCMs and mtp53 R273H directly associates with EdU at nascent replication forks. Furthermore, we detected high mtp53 R273H, high PARP protein, and high PARP enzymatic activity in patient derived xenograft tissue expressing R273H mtp53. We also detected high co-association of mtp53 and PARP in TNBC in a preliminary tumor microarray screen as well as through analysis of The Cancer Genome Atlas reverse phase protein data sets. As a result, we consider understanding the biology of chromatin-associated mtp53-PARP-MCM complexes on replicating DNA to be an area of inquiry that will have a sustained long-term influence on our understanding, detection, and treatment choices for TNBC in diverse populations of women. The mtp53-PARP-MCM axis in TNBC is an innovative pathway and elucidating the mechanistic relationship of the proteins working together in replication-associated activities will help us to better understand the disease. Our hypothesis is that stable mtp53 interacts with replicating DNA during replication stress using amino acids from its carboxy-terminal regions. As a consequence, the mtp53-PARP-MCM axis is engaged allowing increased recruitment of protein complexes and sensitivity to PARP inhibitors combined with DNA damaging agents. We will test this hypothesis by the following three aims. Aim 1: Elucidate the role and mechanism of the TNBC-associated oncogenic mtp53- PARP-MCM axis. Aim 2: Increase our understanding of co-expression of mtp53 and PARP using novel imaging platforms in xenograft mouse models and screening of multiethnic human breast tissue microarrays to benefit all women with TNBC. Aim 3: Validate the driver role of the mtp53-PARP-MCM axis influencing PARPi outcomes in orthotopic mouse models. Completing this work will provide mechanistic insights on how mtp53 cross-talks with PARP and MCM proteins on chromatin in cell culture and xenograft models, and will inform the potential of using novel imaging agents to detect nuclear co-expression of mtp53 and PARP as informative biomarkers for TNBC.

项目概要： 突变型 p53-PARP-MCM 通路在三阴性乳腺癌中的作用 突变 p53 (mtp53) 存在于 80% 的三阴性乳腺癌 (TNBC) 中。有需要 增加对 TNBC 中 mtp53 信号传导的了解，以确定检测策略 并治疗疾病。 TNBC 在非裔美国女性中比白人更常见 女性，因此拟议的工作将使 TNBC 的所有女性受益，包括 AA 人口比例过高。本申请旨在确定以下物质的生物学作用： 与 PARP1 相关的 TNBC 相关功能获得性致癌 mtp53 R273H （本文称为 PARP）蛋白、MCM2-7 解旋酶和复制 DNA。 p53 精氨酸 273 众所周知，用 His 取代的以及其他热点 p53 错义突变蛋白 在 TNBC 中发挥致癌蛋白的作用，但它们的功能除了作为异常蛋白外 转录辅因子尚不清楚。 R273H mtp53 含量较高 位于许多 TNBC 的染色质上，并作为激活甾醇转录的辅助因子 生物合成基因和染色质重塑因子，但令人难以置信的是 R273H 也协同关联 与 PARP 和 MCM 一起复制 DNA。我们的研究发现了这种新颖的复制—— 相关途径，我们称之为 mtp53-PARP-MCM 染色质轴。我们的初步和 已发表的数据报告表明，mtp53 R273H 的敲低会减少染色质相关的 MCM2-7 和 PARP。在这种情况下 mtp53 的敲低会降低组合 PARP 抑制剂 (PARPi) 他拉佐帕尼加 DNA 损伤剂替莫唑胺介导的细胞死亡，证明 mtp53 表达影响癌细胞对协同 PARPi 加 DNA 的敏感性 损伤介导的细胞杀伤。我们的初步和公布的数据显示核mtp53 R273H 与 PARP 和 MCM 共同关联，mtp53 R273H 直接与 EdU 关联 新生的复制叉。此外，我们检测到高 mtp53 R273H、高 PARP 蛋白和 在表达 R273H mtp53 的患者来源的异种移植组织中具有高 PARP 酶活性。我们 还在初步肿瘤的 TNBC 中检测到 mtp53 和 PARP 的高度共关联 微阵列筛选以及通过癌症基因组图谱反相分析 蛋白质数据集。因此，我们考虑了解染色质相关的生物学 mtp53-PARP-MCM 复合物对 DNA 复制的影响将成为一个研究领域 对我们对 TNBC 的理解、检测和治疗选择产生持续的长期影响 在不同的女性群体中。 TNBC 中的 mtp53-PARP-MCM 轴是一款创新型 途径并阐明蛋白质协同工作的机制关系 复制相关的活动将帮助我们更好地了解这种疾病。我们的假设 是稳定的 mtp53 在复制应激期间使用氨基与复制 DNA 相互作用 来自其羧基末端区域的酸。因此，mtp53-PARP-MCM 轴是 参与允许增加蛋白质复合物的募集和对 PARP 的敏感性 抑制剂与 DNA 损伤剂相结合。我们将通过以下方式检验这个假设 三个目标。 目标 1：阐明 TNBC 相关致癌 mtp53- 的作用和机制 PARP-MCM 轴。 目标 2：利用新颖的方法加深我们对 mtp53 和 PARP 共表达的理解 异种移植小鼠模型的成像平台和多种族人类的筛选 乳腺组织微阵列使所有患有 TNBC 的女性受益。 目标 3：验证 mtp53-PARP-MCM 轴影响 PARPi 结果的驱动作用 在原位小鼠模型中。 完成这项工作将为 mtp53 如何与 PARP 串扰和 细胞培养和异种移植模型中染色质上的 MCM 蛋白，并将揭示细胞培养和异种移植模型中染色质的潜力 使用新型成像剂检测 mtp53 和 PARP 的核共表达作为信息 TNBC 的生物标志物。