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: 10210801
• 负责人: GOKUL M. DAS
• 金额: $ 56.69万
• 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/10210801
• 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; 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; Structure; 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; base; chemotherapy; clinical database; clinically relevant; clinically significant; gain of function; 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; 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.

三重负乳腺癌（TNBC）不表达雌激素受体-α（ERα），孕酮受体（PR），， 或人类表皮生长因子受体2（HER2），因此，目前正在使用的靶向药物均未 因为乳腺癌是有效的。大约60-80％的TNBC表达雌激素受体-β （ERβ）。然而，ERβ的亲抗性抗肿瘤能力仍然有争议。另一个关键分子 TNBC的特征是p53突变的高频（80％）。除了失去抑制肿瘤 在任何剩余的野生型p53（WTP53），突变体p53上的特性和主导性调节 还获得致力于功能的功能。越来越多的证据表明，并非所有突变体p53功能 尽管在TNBC病理学中已隐含ERβ和p53，但p53是否在促进 ERβ的功能双重性与抗增殖相对于增殖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在更好的治疗策略中的发展。 目的是研究p53中的特定突变如何影响TNBC中的ERβ功能，并预测 该特定的p53突变将决定其在ERβ突变剂p53-P73信号轴上影响多个的作用 肿瘤进展和转移的各个方面。假设是ERβ与WTP53和 突变p53，导致对TNBC对药物的进展和治疗反应的影响相反 他莫昔芬（TAM）。拟议研究的理由是了解ERβ如何引起相反 以p53状态依赖性方式的功能对于将TNBC患者分层为建立的复制品至关重要 TAM等治疗剂治疗大量TNBC患者。具体目的是：（1确定 TNBC细胞中不同p53突变体与p73和ERβ的相互作用； （2）分析 p53突变体的体内肿瘤进展，转移和治疗反应； （3）评估临床 ERβ-P53-P73信号轴的重要性。在特定目标1中，表达不同的同基因TNBC细胞 使用CRISPR技术和SHRNA介导的ERβ和WT和p53突变体的组合 有条件的敲低将用于分析相互作用及其对其对影响的机制 细胞在体外的体外和肿瘤进展。在特定的目标2中，不同的p53突变对 肿瘤生长和转移将在体内进行分析。这些研究的临床相关性将被评估 使用良好的患者衍生Xenographtics（PDXS）；患者肿瘤衍生的器官（PDOS）；和病人 具有连接的临床数据库的肿瘤组织（特定目标3）。这项研究的贡献预计将是 更好地理解不同p53突变中ERβ-P53-P73轴的机制 影响疾病进展和治疗反应。该提案具有创新性，因为分析了 不同p53突变作为综合ERβ突变p53-p73信号轴的一部分的差异效应是一个 偏离现状，并具有开发针对TNBC的新型治疗策略。