Mechanisms Governing Expansion of Embryonic Progenitor Cells (EPCs) inMetaplasia

化生中胚胎祖细胞 (EPC) 扩张的控制机制

基本信息

批准号：
10438015
负责人：
Jason C Mills
金额：
$ 40.15万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-10 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10438015
关键词：
3-Dimensional Adult Alleles Apoptosis Architecture Autophagocytosis Bacteria Bioinformatics Bypass CDH1 gene Cause of Death Cell Cycle Cell Differentiation process Cell Proliferation Cells Characteristics Chief Cell Chronic Collaborations Conserved Sequence Cytoskeleton Data Development Dose Doxycycline Embryo Event FRAP1 gene Gastric Metaplasia Genes Genetic Goals Helicobacter pylori Human Hydroxychloroquine Image Inflammation Injury Internal Ribosome Entry Site LATS1 gene Lead Lesion Lysosomes Malignant Neoplasms Mediator of activation protein Metaplasia Metaplastic Cell Methylnitrosourea Mitotic Molecular Mus Mutagens Mutant Strains Mice Natural regeneration Neurofibromin 2 Nuclear Oncogenes Organ Organoids Pathway interactions Pharmaceutical Preparations Pharmacology Phenotype Phosphotransferases Physiologic pulse Population Process Proliferating Property Publishing Regenerative Medicine Resolution Risk Role Signal Transduction Sirolimus Smooth Pursuit Stomach Stomach Neoplasms Stress TP53 gene Tamoxifen Technology Testing Time base cancer risk data repository design experimental study gastric organoids gastric tumorigenesis imaging modality mTOR Inhibitor malignant stomach neoplasm mouse model mutant new therapeutic target paligenosis premalignant prevent progenitor programs regenerative scaffold stem cells stomach body synergism tissue repair transcription factor transcriptome sequencing tumor tumor progression tumorigenesis

项目摘要

ABSTRACT/SUMMARY The Overall Goal of this application is to identify shared, conserved mechanisms that induce precancerous lesions like metaplasia. Our preliminary and published data indicate that pseudopyloric or so- called SPEM-type metaplasia in stomach is the manifestation of a conserved regeneration program induced by large-scale injury. The metaplastic cells themselves are characterized by a “dedifferentiated” phenotype wherein they express embryonic-like or early developmental markers that proliferate to repair the tissue damage. We hypothesize that the metaplastic/regenerative process is fueled by expansion of a population of embryonic-like progenitor cells (EPCs). EPCs arise in large part from mature secretory that become progenitor-like by following a stepwise, conserved cellular program we call “paligenosis”. Here, we will show preliminary data that Hippo signaling via Nf2 (Merlin) and downstream transcription factors YAP1/TAZ may be a critical, conserved modulator of EPC expansion. In Aim 1, we will test necessity/sufficiency of Nf2 and Yap1/Taz in gastric metaplasia in mouse models and in human and mouse organoids. We will perform discovery based RNA-Seq experiments to uncover new Hippo targets modulating EPCs and metaplasia. In Aim 2 we will look at how these Hippo pathway components interact with the stages of paligenosis we have characterized, whether they can overcome molecular checkpoints between Stages 1 and 2 and between 2 and 3. We will also determine how Hippo signaling interacts with the conserved, paligenosis-dedicated gene Ifrd1, which we will show is required to suppress p53 as cells upregulate mTORC1 to reenter the cell cycle in Stage 3 paligenosis. In Aim 3, we will test whether increasing EPC formation and metaplasia via the Hippo pathway increases tumorigenesis by combining Hippo mutants with: 1) the mutagen MNU; or 2) additional gastric-cancer- related mutant alleles p53 and Cdh1; or 3) by increasing chronic inflammation with the human gastric-cancer- predisposing bacterium H pylori. Experiments were designed to be appropriately powered in collaboration with our biostatistician, Dr. Yan Yan. State-of-the-art imaging (eg. AiryScan live-cell, confocal on organoids; FIB-SEM 3-D ultrastructural nanotomography) will be performed with Dr. James Fitzpatrick in our institutional imaging core; organoid support, and gene editing will be in collaboration with Dr. Blair Madison and our shared organoid core; bioinformatic analysis including synergy with data repositories will be via our collaboration with Dr. Bo Zhang, who directs the institutional bioinformatics core for the Center for Regenerative Medicine.

摘要/总结该应用程序的总体目标是识别诱导的共享的、保守的机制我们的初步和发表的数据表明，假幽门或类似的癌前病变。胃中被称为 SPEM 型化生的现象是由以下因素诱导的保守再生程序的表现：大规模损伤。化生细胞本身具有“去分化”表型的特征。它们表达类似胚胎或早期发育的标记物，这些标记物可以增殖以修复组织损伤。化生/再生过程是由类胚胎群体的扩张所推动的祖细胞 (EPC) 大部分来自成熟的分泌细胞，通过以下方式变成祖细胞样。一个逐步的、保守的细胞程序，我们称之为“paligenosis”。在这里，我们将展示 Hippo 通过 Nf2 (Merlin) 和下游转录因子进行信号传导的初步数据 YAP1/TAZ 可能是 EPC 扩展的关键、保守调节剂。在目标 1 中，我们将测试必要性/充分性。我们将在小鼠模型以及人类和小鼠类器官中研究 Nf2 和 Yap1/Taz 在胃化生中的作用。基于发现的 RNA 测序实验旨在发现调节 EPC 和化生的新 Hippo 靶标。 2 我们将研究这些 Hippo 通路成分如何与我们所拥有的 paligenosis 阶段相互作用表征，它们是否可以克服第 1 阶段和第 2 阶段以及第 2 阶段和第 2 阶段之间的分子检查点 3. 我们还将确定 Hippo 信号传导如何与保守的、古变专用基因 Ifrd1 相互作用，我们将证明，当细胞上调 mTORC1 以重新进入第 3 阶段的细胞周期时，需要抑制 p53 在目标 3 中，我们将测试是否通过 Hippo 途径增加 EPC 形成和化生。通过将 Hippo 突变体与以下物质结合来增加肿瘤发生：1) 诱变剂 MNU；或 2) 额外的胃癌- 相关突变等位基因 p53 和 Cdh1；或 3) 通过增加人类胃癌的慢性炎症- 诱发幽门螺杆菌。我们与生物统计学家 Yan Yan 博士合作，设计了适当的实验。最先进的成像（例如 AiryScan 活细胞、类器官共聚焦；FIB-SEM 3-D 超微结构纳米断层扫描）将在我们的机构成像核心支持中与 James Fitzpatrick 博士一起进行；基因编辑将与 Blair Madison 博士和我们共享的生物信息学核心合作；分析（包括与数据存储库的协同作用）将通过我们与张博博士的合作进行，张博博士是该项目的负责人。再生医学中心的机构生物信息学核心。