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.

项目总结 食管鳞状细胞癌(ESCC)是全世界食道癌的主要亚型 最为人所知的致命癌症之一。转移性ESCC预后不佳，相对生存期为5年。 利率只有5%。TP53(小鼠同源Trp53)突变，在大约70%的ESCC中检测到 患者易发生肿瘤转移，预后差。要了解Trp53R172H的作用机制- 介导性转移是ESCC的热点突变之一，我们利用小鼠模型并进行了 从这个模型产生的转移的ESCC细胞上的RNA-Seq，从中我发现了集落刺激因子- 1(CSF-1)显著上调。虽然它典型地参与了肿瘤相关的极化 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 这可能是由表观遗传阅读器溴结构域包含结构域4(BRD4)共同调节的。这个 本研究的主要假设是，脑脊液-1/脑脊液-1R自分泌信号是其主要机制之一。 其中，TP53错义突变可促进ESCC的侵袭和肺转移。我将检验这一假设 通过以下相互关联的特定目的：(1)阐明Trp53R172H介导的CSF-1/CSF-1R的作用 促食管鳞癌侵袭和肺转移的信号通路，(2)探讨BRD4作为候选联合基因 Trp53R172H调节因子上调CSF-1及评价其抗肿瘤作用 抑制BRD4，以及(3)描绘致瘤以及由以下因素介导的表观遗传/转录状态 食管鳞癌患者复发的p53错义突变及其对CSF-1/CSF-1R信号的影响要完成 目的1，我将评估CSF-1/CSF-1R信号在增殖、迁移、原发肿瘤和3D器官样体中的作用 通过体外和体外实验研究其形成、侵袭和肺转移及其下游效应分子。 体内互补的方法。我将通过研究P53-R172H和P53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，我会从基因上删除 并从药物上抑制BRD4，以评估其对肿瘤发生和肺转移的影响。至 完成目标3，我将通过碱基编辑引入p53突变，并评估它们在促进 致癌活性。我还将研究这些突变如何影响因子在转录水平的表达 转移过程中的CSF-1/CSF-1R信号转导途径，并在小鼠原代培养上进行单细胞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的内在 食管癌转移的潜在机制，最终将为开发治疗方法开辟新的途径 可推广到其他鳞癌的转移性鳞癌患者的治疗策略。我会培养分析能力 思考、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、顾问委员会和研究生项目提供的支持和资源，这将是 对于实现我长期的职业目标至关重要，那就是成为一名从事癌症研究的终身教职员工。