Functional Significance of individual p53 mutations in determining the role of estrogen receptor beta in triple negative breast cancer

个体p53突变在确定雌激素受体β在三阴性乳腺癌中的作用中的功能意义

• 批准号: 10577874
• 负责人: GOKUL M. DAS
• 金额: $ 59.16万
• 依托单位: ROSWELL PARK CANCER INSTITUTE CORP
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10577874
• 关键词: Affect; Apoptosis; Archives; Basal Cell; Binding; Breast Cancer Cell; Breast Cancer Patient; CRISPR/Cas technology; Cancer Biology; Carboplatin; Cell model; Cell physiology; Cells; Characteristics; Clinical; Clinical Data; Clustered Regularly Interspaced Short Palindromic Repeats; Codon Nucleotides; Data; Development; Diagnosis; Disease Progression; Disease stratification; Dominant-Negative Mutation; Doxorubicin; Drug Targeting; Epidermal Growth Factor Receptor; Estrogen Receptor alpha; Estrogen Receptor beta; Frequencies; Goals; Growth; Hot Spot; Human; In Vitro; Individual; Induction of Apoptosis; Invaded; Laboratories; Lead; Link; Malignant Neoplasms; Mediating; Mission; Molecular; Mutation; Neoplasm Metastasis; Oncogenic; Organoids; Pathology; Patients; Prevention; Process; Progesterone Receptors; Property; Proteins; Public Health; Receptors, Adrenergic, beta-1; Regimen; Regulation; Reporting; Research; Resistance; Role; Signal Transduction; Site; Subgroup; Survival Analysis; TP53 gene; Tamoxifen; Testing; Therapeutic; Therapeutic Agents; Tumor Suppressor Proteins; Tumor Tissue; Tumor-Derived; United States National Institutes of Health; Validation; Xenograft Model; Xenograft procedure; chemotherapy; clinical database; clinically relevant; clinically significant; gain of function; human data; human disease; in vivo; innovation; insight; knock-down; malignant breast neoplasm; mouse model; mutant; neoplastic cell; novel; novel therapeutic intervention; novel therapeutics; overexpression; patient derived xenograft model; pre-clinical; receptor expression; response; small hairpin RNA; targeted treatment; therapeutically effective; treatment response; triple-negative invasive breast carcinoma; tumor; tumor growth; tumor progression; tumorigenic

Triple negative breast cancers (TNBCs) do not express estrogen receptor-α (ERα), progesterone receptor (PR), or human epidermal growth factor receptor 2 (HER2), and therefore, none of the targeted drugs currently in use for breast cancer are effective against them. Approximately 60-80% of TNBCs express estrogen receptor-β (ERβ). However, pro- versus anti-tumorigenic capabilities of ERβ remains controversial. Another key molecular characteristic of TNBC is the high frequency (80%) of p53 mutation. In addition to losing tumor suppressor properties and exerting dominant-negative regulation over any remaining wild type p53 (WTp53), mutant p53 also acquires oncogenic gain-of-function. Increasing evidence suggests that not all mutant p53s function similarly. Although ERβ and p53 have been implicated in TNBC pathology, whether p53 has a role in the pro- versus anti-proliferative functional duality of ERβ remains an open question. The long-term goal is to understand and exploit the role of ERβ-p53 crosstalk in breast cancer for the development of better therapeutic strategies. The objective is to study how specific mutations in p53 impinges upon ERβ function in TNBC, with the prediction that specific p53 mutation will determine its role in the ERβ-mutant p53-p73 signaling axis impacting multiple aspects of tumor progression and metastasis. The hypothesis is that ERβ binds to and inhibits both WTp53 and mutant p53, leading to opposite effects on progression and therapeutic response of TNBC to agents such as Tamoxifen (Tam). The rationale for the proposed research is that understanding how ERβ elicits opposite functions in a p53 status-dependent manner will be critical to stratify TNBC patients to repurpose established therapeutic agents such as Tam to treat large percentage of TNBC patients. The specific aims are: (1 Determine the interaction of different p53 mutants with p73 and ERβ in TNBC cells; (2) Analyze the differential effects of p53 mutants on tumor progression, metastasis and therapeutic response in vivo; and (3) Evaluate the clinical significance of the ERβ-p53-p73 signaling axis. In specific aim 1, Isogenic TNBC cells expressing different combinations of ERβ and WT and p53 mutants generated using CRISPR technology and shRNA-mediated conditional knockdown will be used for analyzing the mechanisms underlying the interaction and its impact on cellular functions in vitro and tumor progression in vivo. In specific aim 2, the effect of different p53 mutations on tumor growth and metastasis will be analyzed in vivo. The clinical relevance of these studies will be evaluated using well-characterized patient derived xenografts (PDXs); patient tumor-derived organoids (PDOs); and patient tumor tissues with linked clinical database (specific aim 3). The contribution of this research is expected to be better understanding of the mechanisms by which ERβ-p53-p73 axis in the context of different p53 mutations affects the disease progression and therapeutic response. The proposal is innovative because analyzing the differential effects of different p53 mutations as part of an integrated ERβ-mutant p53-p73 signaling axis is a departure from the status quo and has the potential of developing novel therapeutic strategies against TNBC.

三阴性乳腺癌 (TNBC) 不表达雌激素受体-α (ERα)、孕激素受体 (PR)、 或人表皮生长因子受体2（HER2），因此，目前没有使用任何靶向药物 大约 60-80% 的 TNBC 表达雌激素受体-β。 (ERβ)。然而，ERβ 的促肿瘤作用与抗肿瘤作用的另一个关键分子仍然存在争议。 TNBC的特点是p53突变频率高（80%），此外还失去抑癌基因。 特性并对任何剩余的野生型 p53 (WTp53)、突变体 p53 施加显性负调控 越来越多的证据表明，并非所有突变的 p53 都具有致癌功能。 同样，虽然 ERβ 和 p53 与 TNBC 病理学有关，但 p53 是否在亲-TNBC 中发挥作用。 ERβ 与抗增殖功能的双重性仍然是一个悬而未决的问题，长期目标是了解。 并利用 ERβ-p53 串扰在乳腺癌中的作用来开发更好的治疗策略。 目的是研究 p53 的特定突变如何影响 TNBC 中的 ERβ 功能，并预测 特定的 p53 突变将决定其在 ERβ 突变体 p53-p73 信号轴中的作用，影响多种 肿瘤进展和转移方面的假设是 ERβ 结合并抑制 WTp53 和 WTp53。 p53 突变，导致 TNBC 的进展和对药物的治疗反应产生相反的影响，例如 他莫昔芬 (Tam)。拟议研究的基本原理是了解 ERβ 如何引发相反的结果。 p53 状态依赖方式的功能对于对 TNBC 患者进行分层以重新调整已建立的用途至关重要 具体目标是：（1 确定 (2)分析不同p53突变体与p73和ERβ在TNBC细胞中的相互作用； p53突变体对肿瘤进展、转移和体内治疗反应的影响；以及（3）评估临床效果； ERβ-p53-p73信号轴的重要性在具体目标1中，表达不同的同基因TNBC细胞。 使用 CRISPR 技术和 shRNA 介导生成的 ERβ 和 WT 以及 p53 突变体的组合 条件击倒将用于分析相互作用的机制及其对 具体目标2：不同p53突变对体外细胞功能和体内肿瘤进展的影响。 将在体内分析肿瘤生长和转移，评估这些研究的临床相关性。 使用充分表征的患者来源的异种移植物（PDX）和患者肿瘤来源的类器官（PDO）； 具有链接临床数据库的肿瘤组织（具体目标3）。 更好地了解不同 p53 突变背景下 ERβ-p53-p73 轴的机制 该提案具有创新性，因为它分析了疾病的进展和治疗反应。 作为整合 ERβ 突变体 p53-p73 信号轴一部分的不同 p53 突变的差异效应是 背离现状，有潜力开发针对 TNBC 的新治疗策略。