EPHB2-Dependent Signaling: A New Molecular Paradigm in Barrett's Neoplasia

EPHB2 依赖性信号传导：巴雷特瘤形成的新分子范式

• 批准号: 10713941
• 负责人: Andrew Edward Blum
• 金额: $ 40.81万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-20 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10713941
• 关键词: 3-Dimensional; Acceleration; Animal Model; Barrett Esophagus; Barrett' s carcinogenesis; Barrett' s neoplasia; Bile Acids; Binding; Binding Proteins; Biological; Biological Markers; Biopsy; Cell Line; Cell Lineage; Cells; Chemopreventive Agent; Complex; Coupled; Development; Disease; Disease Progression; Distal; Dysplasia; EPHB2 gene; Ephrin-B2; Epithelium; Esophageal Adenocarcinoma; Esophageal Squamous Cell; Esophageal injury; Esophageal mucous membrane; Esophagus; Etiology; Exposure to; Family suidae; Gastroenterology; Genetic Transcription; Gland; Goals; Human; In Situ; In Vitro; Inflammation; Inflammatory; Inherited; Injury; Interleukin-1 beta; Intestines; Lesion; Link; Malignant - descriptor; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Metaplasia; Modeling; Molecular; Mucous Membrane; Mus; NF-kappa B; Neoplasms; Organoids; Pathogenesis; Pathologic; Pathway Analysis; Pathway interactions; Patients; Phenotype; Phosphorylation; Phosphotransferases; Predisposition; Receptor Protein-Tyrosine Kinases; Reflux; Regulation; Risk; Risk Factors; Rodent Model; Role; Signal Transduction; Source; Submucosa; Testing; Therapeutic; Time; Tissues; Transcriptional Regulation; Transitional Cell; United States; c-myc Genes; cancer prevention; carcinogenesis; clinical implementation; design; evidence base; genome-wide; in vivo; injury and repair; molecular shape; mouse model; novel; phosphoproteomics; preclinical study; precursor cell; progenitor; programs; promoter; single-cell RNA sequencing; stem cells; synergism; transcription factor; transcriptome sequencing; transcriptomics; tumor growth; ubiquitin-protein ligase; wound healing

PROJECT SUMMARY/ABSTRACT The etiology of Barrett's esophagus (BE), a molecularly complex disorder of the distal esophagus, remains elusive. Patients with BE are at an increased risk of developing esophageal adenocarcinoma (EAC), a lethal, increasingly prevalent, and the most common esophageal malignancy in the U.S. Our long-term objective is to identify the causative mechanisms underlying the onset and malignant progression of BE, and to develop evidence-based biomarkers and chemopreventive/therapeutic strategies for subsequent clinical implementation. Project 3 of this program is based on our recent discovery of EphB2 Receptor Tyrosine Kinase as a novel promoter of Barrett's neoplasia. RNA sequencing coupled with pathway modeling in treatment-naïve patient biopsies showed significant hyperactivation of EphB2 signaling in the vast majority (>90%) of BE and EAC lesions. Phenotypic assessments in vitro and in vivo demonstrated EphB2 signaling to be essential for the viability of EAC and dysplastic/BE cells. Subsequent mechanistic studies identified c-MYC as a novel target of EphB2 signaling, while single-cell RNA sequencing of esophageal progenitor cells further confirmed a strong association of EphB2 activation with columnar/BE-like cell lineage. Collectively, our study implicates EphB2 signaling for the first time in BE-EAC pathogenesis, and further reveals a novel link between EphB2 and MYC in any disease context. Expanding on these provocative findings, we now propose to fully characterize the biologic role of EphB2 in Barrett's neoplasia as follows: In Aim 1, we will determine whether EphB2 activation is a consequence of, or a pre-requisite for, columnar/BE metaplasia development. We will test whether activation of EphB2 signaling in putative BE-progenitor cells promote columnar/BE-like transformative state. In Aim 2, we will determine the pathologic consequences of EphB2 activation in vivo in the esophagus, using a constitutively- active EphB2 kinase (Ephb2F613D) mouse model that we recently developed. We will specifically test if EphB2 activation induces BE-like metaplasia and dysplasia/cancer in the esophagus; either spontaneously, or upon exposure to reflux injury, and/or in the setting of esophageal inflammation. We will further characterize the key molecular programs disrupted by EphB2 activation in the esophagus. In Aim 3, we will determine the downstream and upstream regulatory cascades of EphB2 signaling. Collectively, our proposed studies will delineate the role of EphB2 signaling for the first time, uncovering novel driver mechanisms of BE-EAC pathogenesis.

项目概要/摘要 巴雷特食管 (BE) 是一种分子复杂的远端食管疾病，其病因学仍不清楚 难以捉摸。 BE 患者患食管腺癌 (EAC) 的风险增加，这是一种致命的、 日益流行，并且是美国最常见的食管恶性肿瘤。我们的长期目标是 确定 BE 发病和恶性进展的致病机制，并制定 基于证据的生物标志物和后续临床实施的化学预防/治疗策略。 该项目的项目3是基于我们最近发现的EphB2受体酪氨酸激酶作为一种新颖的 巴雷特瘤形成的促进者。 RNA 测序与未接受治疗患者的通路建模相结合 活检显示绝大多数 (>90%) BE 和 EAC 中 EphB2 信号传导显着过度激活 病变。体外和体内表型评估表明 EphB2 信号对于 EAC 和发育不良/BE 细胞的活力。随后的机制研究将 c-MYC 确定为新的靶点 EphB2信号传导，而食管祖细胞的单细胞RNA测序进一步证实了强烈的 EphB2 激活与柱状/BE 样细胞谱系的关联。总的来说，我们的研究表明 EphB2 首次在 BE-EAC 发病机制中发现 EphB2 和 MYC 之间的新联系 任何疾病背景。扩展这些令人兴奋的发现，我们现在建议全面描述生物特征 EphB2 在 Barrett 瘤形成中的作用如下：在目标 1 中，我们将确定 EphB2 激活是否是一个 柱状/BE化生发育的结果或先决条件。我们将测试是否激活 假定的 BE 祖细胞中的 EphB2 信号传导促进柱状/BE 样转化状态。在目标 2 中，我们将 使用组成型方法确定食管体内 EphB2 激活的病理后果 我们最近开发的活性 EphB2 激酶 (Ephb2F613D) 小鼠模型。我们将专门测试EphB2是否 激活会诱导食管中BE样化生和发育不良/癌症；要么自发地，要么根据 暴露于反流损伤和/或食管炎症的情况下。我们将进一步描述关键 食道中 EphB2 激活破坏了分子程序。在目标 3 中，我们将确定下游 和 EphB2 信号传导的上游调控级联。总的来说，我们提出的研究将描述该角色 首次对 EphB2 信号传导进行研究，揭示了 BE-EAC 发病机制的新驱动机制。