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.

项目摘要： 突变体p53-PARP-MCM途径在三阴性乳腺癌中的作用 突变体p53（MTP53）在80％的三重负乳腺癌（TNBC）中发现。有需要 增加对TNBC中MTP53信号传导的了解，以确定检测的策略 并治疗疾病。 TNBC在非洲裔美国妇女中比白人更常见 妇女以及拟议的工作将使包括AA在内的所有具有TNBC的妇女受益 人口过多。该应用建议确定 与PARP1相关的TNBC相关功能获得的致癌肿瘤MTP53 R273H （此处称为PARP）蛋白质，MCM2-7解旋酶和复制DNA。 p53 arg 273 用他的和其他热点p53错义突变蛋白代替，众所周知 在TNBC中充当致癌蛋白，但它们的功能以外的功能 转录副因素尚不清楚。 R273H MTP53以高水平存在 在许多TNBC中的染色质上 生物合成基因和染色质重塑因子，但R273H也令人难以置信 带有PARP和MCM的复制DNA。我们的研究发现了这种新颖的复制 - 相关的途径，我们称其为MTP53-PARP-MCM染色质轴。我们的初步和 已发布的数据报告，敲低MTP53 R273H减少了染色质相关的MCM2-7 和parp。在这种情况下，MTP53的敲低降低了组合的PARP抑制剂（PARPI） talazoparib加上DNA损伤剂Temozolomide介导的细胞死亡，表明 MTP53表达影响癌细胞对协同PARPI和DNA的敏感性 损坏介导的细胞杀死。我们的初步和发布的数据显示核MTP53 R273H与PARP和MCMS和MTP53 R273H共同关联，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 表达R273H MTP53的患者衍生异种移植组织中的高PARP酶活性。我们 还检测到在初步肿瘤中TNBC中MTP53和PARP的高共同点 微阵列屏幕以及通过分析癌症基因组反向相 蛋白质数据集。结果，我们考虑了解染色质相关的生物学 MTP53-PARP-MCM复合物在复制DNA上是一个询问区域，将具有 对我们对TNBC的理解，检测和治疗选择的长期影响 在各种女性人群中。 TNBC中的MTP53-PARP-MCM轴是一种创新的 途径并阐明蛋白质一起工作的机械关系 复制相关的活动将有助于我们更好地理解这种疾病。我们的假设 稳定的MTP53在使用氨基的复制应力期间与复制DNA相互作用 来自其羧基末端区域的酸。结果，MTP53-PARP-MCM轴为 参与允许增加蛋白质复合物的募集和对PARP的敏感性 抑制剂结合DNA损伤剂。我们将通过以下来检验该假设 三个目标。 目标1：阐明与TNBC相关的致癌MTP53-的作用和机制 PARP-MCM轴。 目标2：使用新颖 异种移植小鼠模型中的成像平台和多种族人类的筛选 乳腺组织微阵列使所有TNBC女性受益。 AIM 3：验证影响PARPI结果的MTP53-PARP-MCM轴的驾驶员角色 在原位小鼠模型中。 完成这项工作将提供有关MTP53如何与PARP和PARP串扰的机械见解 细胞培养和异种移植模型中染色质上的MCM蛋白，并将告知 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。