Elucidation of mutant p53-medidated mechanisms in promoting metastatic esophageal cancer

阐明突变型 p53 介导的促进转移性食管癌的机制

• 批准号: 10537915
• 负责人: Gizem Efe
• 金额: $ 4.59万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10537915
• 关键词: 3-Dimensional; ATAC-seq; Advisory Committees; Affect; Autocrine Communication; Award; Binding; Bromodomain; Cancer Etiology; Cell Line; Cells; Chromatin; Clinical; Colony-Stimulating Factor Receptors; Communication; DNA Binding; DNA Binding Domain; Data; Development Plans; Diagnosis; Disease; Epigenetic Process; Esophageal Squamous Cell Carcinoma; Esophageal carcinoma; Event; Faculty; Foundations; Future; Genetic; Genetic Transcription; Genomics; Goals; Grant; Homologous Gene; Human; In Vitro; Longitudinal Studies; M cell; Macrophage Colony-Stimulating Factor; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of esophagus; Manuscripts; Mediating; Mediator of activation protein; Medical center; Mentorship; Metastatic Neoplasm to the Lung; Metastatic to; Missense Mutation; Modeling; Molecular Conformation; Mus; Mutate; Mutation; Neoplasm Metastasis; Oncogenic; Organoids; Outcome; Pathway interactions; Patients; Pharmacology; Positioning Attribute; Pre-Clinical Model; Primary Neoplasm; Prognosis; Property; Reader; Recurrence; Regulation; Resources; Role; Scientist; Signal Pathway; Signal Transduction; Squamous cell carcinoma; Survival Rate; TP53 gene; Testing; Therapeutic; Thinking; Training; Tumor Cell Invasion; Tumor Promotion; Tumor Suppressor Proteins; Tumor-associated macrophages; Universities; Up-Regulation; Work; Writing; anticancer research; base; base editing; cancer type; career; career development; collaborative environment; effective therapy; gain of function; improved; in vivo; in vivo Model; macrophage; metastatic esophageal; migration; mortality; mouse model; mutant; neoplastic cell; new therapeutic target; novel therapeutic intervention; programs; skills; small hairpin RNA; small molecule; tenure track; transcriptome sequencing; transcriptomics; tumor; tumor growth; tumorigenesis; tumorigenic

PROJECT SUMMARY Esophageal Squamous Cell Carcinoma (ESCC), the predominant subtype of esophageal cancer worldwide is one of the most known lethal cancers. The prognosis of metastatic ESCC is dismal with a 5-year relative survival rate of only 5%. Mutations in TP53 (mouse homolog Trp53), which are detected in approximately 70% of ESCC patients, contribute to tumor metastasis and poor prognosis. To understand the mechanisms of Trp53R172H- mediated metastasis, which is one of the “hotspot” mutations in ESCC, we utilized mouse models and performed RNA-Seq on metastatic ESCC cells generated from this model, from which I identified Colony stimulating factor- 1 (Csf-1) to be significantly upregulated. While it is canonically involved in polarization of tumor-associated macrophages through binding to its receptor CSF-1R, my data demonstrate the existence of autocrine signaling that is potentially co-regulated by epigenetic reader Bromodomain-Containing Domain 4 (BRD4). The overarching hypothesis of this study is that CSF-1/CSF-1R autocrine signaling is one of the major mechanisms by which missense TP53 mutations can promote invasion and lung metastasis in ESCC. I will test this hypothesis through the following interrelated Specific Aims: (1) Elucidate the role of Trp53R172H-mediated CSF-1/CSF-1R signaling pathway in promoting invasion and lung metastasis in ESCC, (2) Investigate BRD4 as a candidate co- regulator of Trp53R172H that contributes to CSF-1 upregulation and assess the anti-tumorigenic efficacy of inhibiting BRD4, and (3) Delineate the tumorigenic, as well as epigenetic/transcriptomic states mediated by missense p53 mutations recurrent in ESCC patients and impact upon CSF-1/CSF-1R signaling. To accomplish Aim 1, I will assess the role of CSF-1/CSF-1R signaling in proliferation, migration, primary tumor and 3D organoid formation, invasion and lung metastasis, as well as its downstream effectors through utilizing in vitro and complementary in vivo approaches. I will accomplish Aim 2 by studying the direct interaction of p53-R172H and BRD4, and also their shared DNA binding motifs through CUT&RUN-Seq. Additionally, I will genetically delete and pharmacologically inhibit BRD4 to evaluate their effects on tumorigenesis and lung metastasis. To accomplish Aim 3, I will introduce p53 mutations through base editing and evaluate their distinct roles in pro- oncogenic activities. I will also investigate how these mutations affect the transcriptional expression of factors in CSF-1/CSF-1R signaling pathway during metastasis, and also conduct single-cell ATAC-Seq on mouse primary and metastatic tumors to evaluate their chromatin accessibility. This study will elucidate the tumor cell intrinsic mechanisms underlying ESCC metastasis, which will ultimately open new avenues in developing therapeutic strategies for metastatic ESCC patients that can be extended to other SCCs. I will develop skills in analytical thinking, technical approaches, scientific communication, grant and manuscript writing, and mentorship with the support and resources available through my sponsors, advisory committee and graduate program, which will be critical to accomplish my long-standing career goal to become a tenure-track faculty to conduct cancer research.

项目概要 食管鳞状细胞癌 (ESCC) 是全球食管癌的主要亚型 最知名的致命癌症之一。转移性食管鳞癌的预后很差，相对生存期为 5 年 率仅为5%。 TP53（小鼠同源物 Trp53）突变，在大约 70% 的 ESCC 中检测到 患者，导致肿瘤转移，预后不良。了解 Trp53R172H- 的机制 介导的转移，这是食管鳞癌的“热点”突变之一，我们利用小鼠模型并进行了 对从该模型生成的转移性 ESCC 细胞进行 RNA 测序，我从中识别出了集落刺激因子 - 1 (Csf-1) 显着上调。虽然它典型地参与肿瘤相关的极化 巨噬细胞通过与其受体CSF-1R结合，我的数据证明了自分泌信号的存在 它可能受到表观遗传阅读器含溴结构域 4 (BRD4) 的共同调节。这 本研究的总体假设是 CSF-1/CSF-1R 自分泌信号传导是主要机制之一 TP53错义突变可促进食管鳞癌的侵袭和肺转移。我将测试这个假设 通过以下相互关联的具体目标：（1）阐明Trp53R172H介导的CSF-1/CSF-1R的作用 信号通路促进 ESCC 侵袭和肺转移，（2）研究 BRD4 作为候选协同蛋白 Trp53R172H 的调节因子有助于 CSF-1 上调并评估其抗肿瘤功效 抑制 BRD4，以及 (3) 描述致瘤性以及表观遗传/转录组状态 ESCC 患者中反复出现的错义 p53 突变及其对 CSF-1/CSF-1R 信号传导的影响。为了完成 目标 1，我将评估 CSF-1/CSF-1R 信号传导在增殖、迁移、原发肿瘤和 3D 类器官中的作用 通过体外和实验研究其形成、侵袭和肺转移及其下游效应子 互补的体内方法。我将通过研究 p53-R172H 和 p53-R172H 的直接相互作用来实现目标 2 BRD4，以及它们通过 CUT&RUN-Seq 共享的 DNA 结合基序。另外，我会基因删除 并通过药理抑制BRD4来评估其对肿瘤发生和肺转移的影响。到 为了实现目标 3，我将通过碱基编辑引入 p53 突变，并评估它们在 pro- 致癌活动。我还将研究这些突变如何影响因子的转录表达 转移过程中的CSF-1/CSF-1R信号通路，并对小鼠原代细胞进行单细胞ATAC-Seq 和转移性肿瘤，以评估其染色质可及性。这项研究将阐明肿瘤细胞内在的 ESCC 转移的机制，最终将为开发治疗方法开辟新途径 针对转移性食管鳞癌患者的策略可以扩展到其他鳞状细胞癌。我将培养分析技能 思维、技术方法、科学交流、资助和手稿写作以及与 通过我的赞助商、咨询委员会和研究生项目提供的支持和资源，这将是 这对于实现我长期的职业目标至关重要，即成为一名进行癌症研究的终身教授。