MDSC Polarization and Helicobacter-Induced Gastric Metaplasia

MDSC 极化和螺杆菌诱导的胃化生

基本信息

批准号：
10164764
负责人：
JUANITA L. MERCHANT
金额：
$ 34.09万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-15 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10164764
关键词：
Acidity Acids Acute Atrophic Atrophic Gastritis Biological Assay Biological Markers Blood Circulation Bone Marrow C57BL/6 Mouse Cell physiology Cells Chronic Environment Epithelial Cells Erinaceidae Exhibits Flow Cytometry Frequencies Future GLI gene Gastric Metaplasia Gastric Parietal Cells Gastric Tissue Gastric mucosa Gastritis Gene Expression Gene Targeting Genes Genetic Transcription Glioma Helicobacter Helicobacter Infections Homeostasis Human Hypochlorhydria ITGAM gene Immune Immunosuppressive Agents Inflammation Interferon-alpha Interferons Intestinal Metaplasia Knockout Mice LacZ Genes Lesion Ligands Malignant Neoplasms Mass Spectrum Analysis Mediating Metaplasia Metaplastic Cell MicroRNAs Molecular Molecular Profiling Mus Myelogenous Myeloid Cells Myeloid-derived suppressor cells Neoplastic Cell Transformation Pathway interactions Patients Pattern Phase Phenotype Population Population Heterogeneity Preneoplastic Conditions Preventive therapy Production Proteins Reporter Reporting Role SHH gene Signal Transduction Smooth Pursuit Stomach Stromal Cells Suppressor-Effector T-Lymphocytes Surface T-Lymphocyte Testing Transcript Transfection Transplantation Tumor Necrosis Factor Superfamily Ligands Wild Type Mouse cytokine defined contribution experimental study granulocyte human subject injured malignant stomach neoplasm monocyte morphogens nano-string neoplastic paracrine polarized cell prevent recruit response small molecule inhibitor smoothened signaling pathway spasmolytic polypeptide therapeutic target transcription factor transcriptome sequencing

项目摘要

Abstract Chronic Helicobacter infection and the resulting inflammation induces gastric metaplasia in the corpus of WT mice but not in mice null for Gli1, demonstrating that this pre-neoplastic lesion requires the induction of canonical Hedgehog signaling. Moreover, we reported that a subset of myeloid cells expressing surface markers and T cell suppressor function indicative of myeloid-derived suppressor cells (MDSCs) express Schlafen4 (Slfn4), a direct target of the Gli1 transcription factor and a known myeloid differentiation factor. Since MDSCs are immature cells, collectively our studies demonstrate that maturation of this myeloid cell subpopulation requires Gli1 and produces proinflammatory cytokines creating a gastric microenvironment favorable for metaplasia and neoplastic transformation. More recently, we have analyzed the human homologs of Slfn4, which include SLFN5 and SLFN12L. We reported that peak expression of SLFN5 in gastric tissue occurred in human subjects with intestinal metaplasia who about a decade later developed gastric cancer. We therefore considered that the polarization of myeloid cells to MDSCs prior to neoplastic transformation might predict who is more likely to develop gastric cancer and as such could provide a therapeutic target to prevent future transformation. We used RNA-Seq and Nanostring microarrays to identify transcripts and microRNAs expressed in Slfn4+-MDSCs from the stomachs of a Helicobacter-infected mouse and found that miR130b co-localized with Slfn4+ cells in the metaplastic mouse stomach. A similar result was observed for SLFN12L in the metaplastic human stomach suggesting that miR130b might identify patients with gastric metaplasia. Our preliminary results demonstrated that Slfn4 and miR130b are required to exert T-cell suppression. In addition to type 1 interferon-regulated genes identified by RNA-Seq, these Slfn4+-MDSCs also expressed several tumor necrosis factor superfamily ligands (TNFsf) and the alarmin IL-1a. Therefore we will test the hypothesis that debris from damaged gastric epithelial cells activates Damage-activated molecular pattern (DAMP) signaling, production of IFNa and polarization to MDSCs, which contribute to a metaplastic phenotype. Aim 1, we will define how DAMP signals induce polarization of Slfn4+-MDSC. In Aim 2, we will define how Slfn4 contributes to MDSC function. In Aim 3, we will define the contribution of Slfn4+-MDSCs to Helicobacter-induced metaplasia. In Aim 1, we will use a combination of flow cytometry, T cell suppression assays and transfection studies to identify the cell populations producing IFNa and the gene targets responding to this DAMP-activated cytokine. In Aim 2, we will used mass spectrometry and pull down assays to identify Slfn4 and SLFN12L interacting proteins. In Aim 3, we will conditionally delete Slfn4 from Gli1- expressing cells to define their contribution to the metaplastic changes observed after Helicobacter infection. Completion of these aims will result in a better understanding of this MDSC subpopulation that can potentially be used as a biomarker and target of small molecule inhibitors.

抽象的慢性螺旋杆菌感染和产生的炎症会诱导胃中胃las虫小鼠，但在gli1的小鼠无效中，这表明这种肿瘤前病变需要诱导规范刺猬信号。此外，我们报道了表达表面的髓样细胞的子集标记和T细胞抑制器功能指示髓样衍生的抑制细胞（MDSC）表达 Schlafen4（SLFN4），Gli1转录因子的直接靶标和已知的髓样分化因子。由于MDSC是不成熟的细胞，我们的研究表明，该髓样细胞的成熟亚种群需要GLI1并产生促炎性细胞因子创建胃微环境有利于化生和肿瘤转化。最近，我们分析了人类 SLFN4的同源物，其中包括SLFN5和SLFN12L。我们报道了胃中SLFN5的峰值表达组织发生在人类受试者中，大约十年后发展癌症。因此，我们认为髓样细胞在肿瘤之前对MDSC的极化转型可能会预测谁更有可能发展胃癌，因此可以提供治疗靶标，以防止将来转变。我们使用RNA-Seq和Nanostring微阵列来识别来自旋律感染小鼠的胃中的SLFN4+-MDSC中表达的成绩单和microRNA 并发现MiR130b与SLFN4+细胞在化生小鼠胃中共定位。类似的结果是观察到slfn12l在化生型人类胃中胃化生。我们的初步结果表明SLFN4和MIR130B需要发挥T细胞抑制。除了通过RNA-Seq鉴定的1型干扰素调节的基因外，这些SLFN4+-MDSC也表达了几个肿瘤坏死因子超家族配体（TNFSF）和警报IL-1A。因此，我们会的检验以下假设：受损胃上皮细胞的碎屑激活损伤激活的分子模式（潮湿）信号传导，IFNA的产生和对MDSC的极化，这有助于化生表型。 AIM 1，我们将定义潮湿信号如何诱导SLFN4+-MDSC的极化。在AIM 2中，我们将定义SLFN4如何贡献MDSC功能。在AIM 3中，我们将定义SLFN4+-MDSC对旋律诱导的化生。在AIM 1中，我们将使用流式细胞仪，T细胞抑制的组合测定和转染研究以鉴定产生IFNA的细胞群和基因靶标响应这种潮湿激活的细胞因子。在AIM 2中，我们将使用质谱和下拉测定法识别SLFN4和SLFN12L相互作用的蛋白质。在AIM 3中，我们将有条件地从Gli1-删除SLFN4 表达细胞以定义其对旋旋杆菌感染后观察到的化生变化的贡献。这些目标的完成将使人们更好地理解这种MDSC亚群用作小分子抑制剂的生物标志物和靶标。