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

• 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
• 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

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/摘要 该应用程序的总体目标是确定影响影响的共享机制 癌性病变，例如化生。我们的初步和发布的数据，表明伪造或SO- 在stallch中称为SPEM型Metaplasia是由构成的再生计划的表现。 大规模伤害。化生细胞本身的特征是“区分”表型 它们表达类似胚胎的或早期发育标记，这些标志物已扩散以修复组织损伤。我们 假设化生/再生过程是由于胚胎样群体的扩展所推动的 祖细胞（EPC）。 EPC在很大程度上来自成熟的秘书，它像祖细胞一样。 我们称为“ paligenenise”的逐步配置的蜂窝程序。 在这里，我们将显示通过NF2（MERLIN）和下游转录因子的河马信号传导的初步数据 YAP1/TAZ可能是EPC扩展的关键调制器。在AIM 1中，我们将测试必要的/充分性 在小鼠模型以及人类和小鼠类器官中的胃代PLASIA中的NF2和YAP1/TAZ的of。我们将表演 基于发现的RNA-seq实验，以发现新的河马靶向调节EPC和Metaplasia。目标 2我们将研究这些河马途径成分如何与paligeneniso的阶段相互作用 表征，它们是否可以克服第1至2阶段以及2和2之间的分子检查点 3。我们还将确定河马信号如何与配置的，paligenisois的基因IFRD1相互作用， 当细胞上调MTORC1以重新进入第3阶段的细胞周期时，我们将表明需要抑制P53 paligenisis。在AIM 3中，我们将测试是否通过河马途径增加EPC的形成和Metaplasia 通过将河马突变体与：1）诱变型MNU相结合来增加肿瘤发生；或2）额外的胃癌 - 相关的突变等位基因P53和CDH1；或3）通过增加人类胃癌的慢性感染 - 幽门螺杆菌诱发的细菌。 实验设计为与我们的生物统计学家Yan Yan博士合作提供适当的动力。 最先进的成像（例如，飞行器的活电池，在类符号上共享共焦； fib-sem 3-d超微结构 纳米学）将与詹姆斯·菲茨帕特里克（James Fitzpatrick）博士一起在我们的机构成像核心中进行；器官支撑， 基因编辑将与布莱尔·麦迪逊（Blair Madison）博士和我们共享的Organoid Core合作；生物信息学 包括与数据存储库的协同作用在内的分析将是通过我们与Bo Zhang博士的合作来指导的。 再生医学中心的机构生物信息学核心。