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），全球食管癌的主要亚型是 最著名的致命癌症之一。转移性ESCC的预后很沮丧，相对生存为5年 比率仅为5％。 TP53中的突变（小鼠同源性TRP53），在大约70％的ESCC中检测到 患者有助于肿瘤转移和预后不良。了解TRP53R172H-的机制 介导的转移，这是ESCC中的“热点”突变之一，我们使用了鼠标模型并进行了 从该模型产生的转移性ESCC细胞上的RNA-seq，我从中确定了刺激因子的菌落 1（CSF-1）要大大更新。虽然它与肿瘤相关的极化均参与 通过与其受体CSF-1R结合的巨噬细胞，我的数据证明了自分泌信号的存在 这可能是由含有溴d的表观遗传学读取器4（BRD4）共同调节的。这 这项研究的总体假设是CSF-1/CSF-1R自分泌信号传导是主要机制之一 TP53突变的错义可以促进ESCC的侵袭和肺转移。我将检验这个假设 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 ESCC中促进侵袭和肺转移的信号传导途径，（2）研究BRD4作为候选人共同 TRP53R172H的调节剂有助于CSF-1上调和评估的抗肿瘤效率 抑制BRD4，（3）描绘肿瘤症，以及由 ESCC患者复发的错义P53突变，并对CSF-1/CSF-1R信号的影响。完成 AIM 1，我将评估CSF-1/CSF-1R信号传导在增殖，迁移，原发性肿瘤和3D类动物的作用 形成，侵袭和肺转移，及其下游效应，通过使用体外和 互补的体内方法。我将通过研究P53-R172H的直接相互作用和 BRD4，以及他们通过剪切和跑步序列的共享DNA绑定基序。此外，我将一般删除 并用药物抑制BRD4评估其对肿瘤发生和肺转移的影响。到 完成AIM 3，我将通过基础编辑引入P53突变，并评估其在促进中的独特作用 致癌活动。我还将研究这些突变如何影响因子的转录表达 CSF-1/CSF-1R信号传导途径转移期间，并且在小鼠初级上进行单细胞ATAC-SEQ 和转移性肿瘤以评估其染色质的可及性。这项研究将阐明肿瘤细胞的内在 ESCC转移的基础机制，该机制最终将开放开发治疗的新途径 可以扩展到其他SCC的转移性ESCC患者的策略。我将发展分析技能 思考，技术方法，科学交流，授予和手稿写作以及与 通过我的赞助商，咨询委员会和研究生计划获得的支持和资源，这将是 完成我长期以来的职业目标至关重要的是成为进行癌症研究的终身教师。