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.

项目摘要/摘要 Barrett食管（BE）的病因是一种远端食管的分子复杂疾病 难以捉摸。患有BE的患者患食道腺癌（EAC）的风险增加，致命， 越来越普遍，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发作和恶性进展的原因，并发展 基于证据的生物标志物和化学预防/治疗策略，用于随后的临床实施。 该程序的项目3基于我们最近发现的EPHB2受体酪氨酸激酶作为新颖的发现 巴雷特肿瘤的发起人。 RNA测序与没有治疗的患者中的途径建模 活检显示了绝大多数BE和EAC的EPHB2信号传导的显着过度激活 病变。体外和体内表型评估表明EPHB2信号传导对于 EAC和发育不良/BE细胞的生存能力。随后的机械研究将C-MYC确定为新的目标 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，我们的研究实现了EPHB2 在BE-EAC发病机理中首次发出信号，并进一步揭示了EPHB2和MYC之间的新联系 任何疾病的情况。扩大这些挑衅性发现，我们现在建议完全表征生物学 EPHB2在Barrett的肿瘤中的作用如下：在AIM 1中，我们将确定EPHB2激活是否是一个 柱状/Be Metaplasia开发的结果或先决条件的结果。我们将测试是否激活 推定的BE基因细胞中的EPHB2信号传导促进柱状/BE样变化状态。在AIM 2中，我们将 使用组成型 - 我们最近开发的活动EPHB2激酶（EPHB2F613D）小鼠模型。我们将专门测试EPHB2是否 激活诱导食管中的类似化生和异常增生/癌症。发起人，或 暴露于反映损伤和/或食管感染的情况下。我们将进一步描述钥匙 食管中EPHB2激活破坏的分子程序。在AIM 3中，我们将确定下游 EPHB2信号传导的上游调节级联。总的来说，我们提出的研究将描述角色 EPHB2信号的首次传导，揭示了BE-EAC发病机理的新型驱动器机制。