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: 10689716
• 负责人: Gizem Efe
• 金额: $ 4.68万
• 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/10689716
• 关键词: 3-Dimensional; ATAC-seq; Advisory Committees; Affect; Autocrine Communication; Award; Binding; Bromodomain; CSF1R gene; Cancer Etiology; Cell Line; Cells; Chromatin; Clinical; 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; Institution; Invaded; M cell; Macrophage; Macrophage Colony-Stimulating Factor; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of esophagus; Manuscripts; Mediating; Mediator; Medical center; Mentorship; Metastatic Neoplasm to the Lung; Missense Mutation; Modeling; Molecular Conformation; Mus; Mutate; Mutation; Neoplasm Metastasis; Oncogenic; Organoids; Outcome; Pathway interactions; Patients; Positioning Attribute; Postdoctoral Fellow; Pre-Clinical Model; Primary Neoplasm; Prognosis; Proliferating; Property; Reader; Recurrence; 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 editing; cancer type; career; career development; collaborative environment; effective therapy; gain of function; improved; in vivo; in vivo Model; metastatic esophageal; migration; mortality; mouse model; mutant; neoplastic cell; new therapeutic target; novel therapeutic intervention; pharmacologic; programs; receptor; 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.

项目摘要 食管鳞状细胞癌（ESCC）是世界范围内食管癌的主要亚型， 最著名的致命癌症之一转移性食管鳞癌的预后很差，相对生存期只有5年 率仅为5%。TP 53（小鼠同源物Trp 53）突变，在约70%的ESCC中检测到 患者，有助于肿瘤转移和不良预后。为了了解Trp 53 R172 H-的作用机制， 介导的转移，这是ESCC中的“热点”突变之一，我们利用小鼠模型， 从该模型产生的转移性ESCC细胞上的RNA-Seq，从中我鉴定出集落刺激因子- 1（Csf-1）显著上调。虽然它通常参与肿瘤相关的极化， 巨噬细胞通过结合其受体CSF-1 R，我的数据表明存在自分泌信号 其潜在地由表观遗传阅读器含溴结构域4（BRD 4）共调节。的 本研究的总体假设是CSF-1/CSF-1 R自分泌信号传导是主要机制之一， TP 53错义突变可促进食管鳞癌的侵袭和肺转移。我将检验这个假设 （1）阐明Trp 53 R172 H介导的CSF-1/CSF-1 R在细胞内的作用 （2）研究BRD 4在ESCC侵袭和肺转移中的作用， Trp 53 R172 H的调节子，其有助于CSF-1上调并评估Trp 53 R172 H的抗肿瘤功效。 抑制BRD 4，和（3）描述肿瘤发生，以及表观遗传/转录组学状态介导的 错义p53突变在ESCC患者中复发并影响CSF-1/CSF-1 R信号传导。完成 目的1：探讨CSF-1/CSF-1 R信号通路在肿瘤细胞增殖、迁移、原发性肿瘤和三维类器官中的作用 形成、侵袭和肺转移，以及其下游效应物， 补充体内方法。我将通过研究p53-R172 H和p53-R172 H之间的直接相互作用来实现目标2。 BRD 4，以及它们通过CUT&RUN-Seq共享的DNA结合基序。另外，我会从基因上删除 和BRD 4抑制剂来评价它们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，并评估它们在促凋亡中的不同作用。 致癌活性。我还将研究这些突变如何影响转录因子的表达， 转移过程中CSF-1/CSF-1 R信号通路，并在小鼠原发性肝癌细胞上进行单细胞ATAC-Seq。 和转移性肿瘤来评估它们的染色质可及性。这项研究将阐明肿瘤细胞的内在 ESCC转移的潜在机制，这将最终开辟新的途径，在发展治疗 转移性ESCC患者的策略，可以扩展到其他SCC。我将培养分析能力 思维，技术方法，科学交流，赠款和手稿写作，以及与 通过我的赞助商、咨询委员会和研究生项目获得的支持和资源， 这对实现我长期以来的职业目标，成为一名从事癌症研究的终身教职人员至关重要。