Maternal programming of the stem cell-basophil axis for asthma

哮喘干细胞-嗜碱性粒细胞轴的母体编程

• 批准号: 10441348
• 负责人: Magdalena Maria Gorska
• 金额: $ 68.96万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-20 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10441348
• 关键词: ATAC-seq; Address; Allergens; Allergic; Asthma; Basophils; Biological Markers; Blood specimen; Bone Marrow; Cell Lineage; Cells; Child; Childhood Asthma; Chronic; Data; Development; Diesel Exhaust; Embryo; Embryo Transfer; Enhancers; Environment; Environmental Exposure; Exposure to; Female; Future; Generations; Hematopoiesis; Hematopoietic Stem Cell subsets; Hematopoietic stem cells; Histamine Release; Human; IL3 Gene; IL3RA gene; IL4 gene; IgE; Immune response; Immune system; In Vitro; Injections; Interleukin-3 Receptor; Lead; Link; Longevity; Lymphoid Cell; Maternal Exposure; Measures; Mediating; Memory; Modeling; Molecular; Mothers; Mus; Myeloid Cells; Null Lymphocytes; PTPRC gene; Pathway interactions; Phenotype; Placenta; Population; Predisposition; Prevention strategy; Process; Production; Radioactivity; Radiolabeled; Regulator Genes; Regulatory Element; Reporter; Reporting; Risk; Role; Seeds; Serum; Site; Source; Stem cell transplant; System; Testing; Tissues; Untranslated RNA; Up-Regulation; asthma model; base; chromatin remodeling; cigarette smoke; congenic; cytokine; diagnostic strategy; early embryonic stage; epidemiology study; experimental study; high risk; imprint; indexing; maternal serum; meetings; mouse model; novel diagnostics; offspring; overexpression; particle; particle exposure; pregnant; prevent; programs; promoter; pup; self-renewal; stem cells; synergism; transcriptome; transcriptome sequencing; transmission process; treatment strategy

Project Summary/Abstract A growing number of epidemiological studies link childhood asthma with maternal environmental exposures with the strongest evidence provided for diesel exhaust and cigarette smoke. How maternal exposures lead to asthma in offspring is unknown. To address this, we developed a mouse model of asthma susceptibility in pups by exposing their mothers to diesel exhaust particles (DEP). Our data suggests that in this DEP-driven model, asthma-predisposing information is transmitted in part by a subset of offspring long-term hematopoietic stem cells (LT-HSCs), in the form of a stem cell memory. This subset distinguishes itself by expression of the IL3 receptor and is programmed for enhanced generation of basophils by the synergistic action of DEP-induced IL1b and IL3. One key mechanism of the synergism between IL1b and IL3 is the ability of IL1b to increase stem cell sensitivity to IL3 which is in part due to IL1b-mediated upregulation of IL3R b chain. In addition to expanding the basophil lineage, IL3 and IL1b are likely to enhance its capacity to promote asthma. Previous studies show that IL1b potentiates basophil histamine release and IL3 potently stimulates their IL4 production, sowing the seed for type-2 immune response. Consistent with this, in DEP offspring, basophils overexpress IL4. The IL3/IL1b-primed, expanded, IL4-overexpressing basophil population becomes a key driver of the type- 2 immune response and asthma upon allergen challenge. Basophil depletion in DEP pups prevents generation of allergen-specific IgE and development of asthma. This is in contrast with data obtained in plain allergen- based models in which basophils are redundant for production of IgE and asthma. Our overarching hypothesis is that maternal exposure to DEP induces asthma susceptibility in offspring through IL1b and IL3 driven programming of the hematopoietic stem cell-basophil axis. In Aim 1, by performing stem cell transplantation experiments, we will establish links between DEP IL3R+ LT-HSCs, increased basopoiesis and predisposition to asthma. We will define molecular basis of DEP IL3R+ LT-HSC programming by analyzing their transcriptomes (RNA-seq) and chromatin remodeling at gene regulatory sites (ATAC-seq). In Aim 2 we will study importance of IL1b and IL3 in programming of the HSC-basophil axis for asthma induction. In our model, IL1b and IL3 are increased in offspring and maternal tissues, including maternal serum. We propose that maternal IL1b and IL3 are important at early stages of embryonic hematopoiesis, when HSCs undergo their initial divisions and embryonic sources of IL1b and IL3 are scarce. Offspring-derived IL1b and IL3 complete the HSC programming process, sustain basopoiesis at a high level, and contribute to basophil activation after maternal cytokines fade away. To define how maternal IL1b and IL3 are transferred to embryos, study their roles and roles of offspring- encoded IL1b and IL3, we will perform serum transfers, injections with radiolabeled cytokines, and combine IL1b or IL3 KO strategies with embryo transfers. In Aim 3 we will use human blood samples to study IL1b and IL3 guided basophil development in childhood asthma.

项目摘要/摘要 越来越多的流行病学研究将儿童哮喘与母性环境暴露联系起来 提供了最有力的证据，以证明柴油排气和香烟烟雾。产妇暴露如何导致 后代哮喘是未知的。为了解决这个问题，我们开发了幼犬哮喘敏感性的鼠标模型 通过将母亲暴露于柴油排气颗粒（DEP）。我们的数据表明，在这个Dep驱动模型中， 哮喘特征的信息部分是由后代长期造血茎的子集传播的 细胞（LT-HSC），以干细胞记忆的形式。该子集通过IL3的表达来区分自己 受体并通过DEP诱导的协同作用进行编程以增强嗜碱性粒细胞的生成 IL1B和IL3。 IL1b和IL3之间协同作用的一种关键机制是IL1B增加的能力 干细胞对IL3的敏感性部分是由于IL1B介导的IL3R B链上调。此外 扩大嗜碱性血统IL3和IL1B可能会增强其促进哮喘的能力。以前的 研究表明，IL1B增强了嗜碱性组胺释放，IL3有效刺激其IL4产生，即 播种种子以进行2型免疫反应。与此相一致，在Dep后代，嗜碱性粒细胞过表达 IL4。 IL3/IL1B结构的，扩展的，过表达IL4的嗜碱性嗜碱性人群成为该类型的关键驱动力 2对过敏原挑战的免疫反应和哮喘。嗜碱性嗜碱性贫民窟的消耗可防止产生 过敏原特异性IgE和哮喘的发育。这与普通过敏原中获得的数据相反 基于嗜碱性粒细胞多余的基于基于IgE和哮喘的模型。我们的总体假设 是通过IL1B和IL3驱动的后代促使后代的哮喘易感性是否引起了母体的暴露 造血干细胞 - 寄生虫轴的编程。在AIM 1中，通过进行干细胞移植 实验，我们将建立DEP IL3R+ LT-HSC之间的联系，增加的子宫昆虫和倾向 哮喘。我们将通过分析其转录组来定义DEP IL3R+ LT-HSC编程的分子基础 （RNA-SEQ）和染色质重塑基因调节位点（ATAC-SEQ）。在AIM 2中，我们将研究重要性 HSC-Basophil轴编程中的IL1B和IL3用于哮喘诱导。在我们的模型中，IL1B和IL3是 后代和母体组织（包括母血清）的增加。我们提出了母体IL1B和IL3 在胚胎造血的早期阶段很重要，当HSC经历其最初的分裂和 IL1B和IL3的胚胎来源很少。后代衍生的IL1B和IL3完成HSC编程 过程，在孕产妇细胞因子逐渐消失后，维持高水平的嗜碱性肺泡，并导致嗜碱性嗜碱性激活 离开。为了定义母体IL1B和IL3如何转移到胚胎，研究其后代的作用和作用 编码的IL1B和IL3，我们将进行血清转移，注射放射性标记的细胞因子，并结合 具有胚胎转移的IL1B或IL3 KO策略。在AIM 3中，我们将使用人类血液样本研究IL1B和 IL3指导儿童哮喘的嗜碱性嗜碱性发展。