Endocrine-immune-reproductive System Interactions

内分泌-免疫-生殖系统相互作用

基本信息

批准号：
8351106
负责人：
Tomoshige Kino
金额：
$ 53.38万
依托单位：
EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8351106
关键词：
AKAP13 gene Acquired Immunodeficiency Syndrome Acute-Phase Proteins Adipose tissue Agonist Amino Acids Animals Anti-Inflammatory Agents Anti-inflammatory Antigen Presentation Asthma Atherosclerosis Biological Assay Blood Pressure Burn injury C-terminal Cardiac Catecholamines Cell Nucleus Cells Characteristics Coupling Cultured Cells Cytomegalovirus Cytomegalovirus Infections Defect Dehydration Dendritic Cells Development Diabetes Mellitus Diet Disease EP300 gene Endocrine Endocrine system Estrogen Receptors Exogenous Factors Exposure to Family Fibrosis Functional disorder Genes Genetic Transcription Glucocorticoid Receptor Glucocorticoids Guanine Nucleotide Exchange Factors HIV-1 Hepatocyte Hormonal Hormones Host Defense Human Hypertension Hypothalamic structure Immune Immune system Immunity Immunologic Deficiency Syndromes Infection Inflammation Inflammatory Insulin Resistance Interferons Interleukin-10 Interleukin-11 Interleukin-12 Interleukin-6 JNK-activating protein kinase Knockout Mice Leukocytes Lipodystrophy Lymphocyte MAP Kinase Gene MAPK14 gene Manuscripts Mediating Medicine Mitogen-Activated Protein Kinase Kinases Mitogen-Activated Protein Kinases Mitogens Molecular Molecular Profiling Monomeric GTP-Binding Proteins Mus NFAT5 protein Newcastle Disease Nitric Oxide Synthase Nuclear Hormone Receptors Nuclear Receptors Organism PPAR delta PPAR gamma Pathogenesis Pathologic Plasma Play Positioning Attribute Posterior Pituitary Gland Predisposition Production Proteins Protozoa Publications Publishing Regulation Reporter Reporting Reproductive system Role STAT3 gene Serine Side Signal Transduction Signs and Symptoms Skeletal Muscle Sodium Chloride Spleen Stress Syndrome System Therapeutic Tissues Toxoplasmosis Tumor Necrosis Factor-alpha Tyrosine Phosphorylation Uremia Viral Viral Antigens Virus Virus Diseases Wild Type Mouse base cytokine extracellular histone acetyltransferase human TNF protein hypothalamic-pituitary-adrenal axis leukemia inhibitory factor mRNA Expression macrophage member monocyte oncostatin M p38 MAPK Signaling Pathway pathogen promoter receptor research study response rho scaffold small molecule transcription factor vpr Gene Products

项目摘要

We have performed several lines of experiments to examine the interactions between the endocrine and immune systems. We showed that two small molecules an IL-6 antagonist and a PPAR-delta agonist have strong anti-inflammatory activities via STAT3 inhibition. The former compound, TB-2-081, interacted with the gp130 subunit of the IL-6-type receptors and blocked the activity not only of IL-6 but also of the leukemia inhibitory factor, oncostatin M and IL-11. A major change in the cell was the inhibition of the tyrosine phosphorylation of the STAT3 transcription factor. The latter compound, GW50516, inhibited the production of several acute phase reactants by hepatic cells showing thus major anti-inflammatory activity. This effect was also mediated by inhibition of STAT3 activity at the promoter of responsive genes. Viruses, including the human immunodeficiency syndrome type-1 (HIV-1), cytomegalovirus (CMV), and Newcastle disease (NDV) viruses, are potent activators and modulators of the host immune and endocrine systems, influencing hormonal actions in host tissues, such as leukocytes, adipose tissue and skeletal muscles. These effects further contribute to viral expansion and pathogenesis. Indeed, we demonstrated that HIV-1 accessory protein Vpr suppresses PPAR-gamma activity playing a potentially important role in the development of the characteristic AIDS-associated lipodystrophy and insulin-resistance syndrome. In dendritic cells (DCs), which play a central role in the recognition and presentation of viral antigens, infection of CMV or NDV, and perhaps HIV-1, causes dramatic changes in the expression of a group of nuclear hormone receptors, including the glucocorticoid and estrogen receptors, as well as of several transcriptional co-regulators, such as p300 and p160-type histone acetyltransferase coactivators, possibly altering secretion/production of interferons and other cytokines by these cells. We have published one manuscript based on the results obtained in the mRNA expression profiling of nuclear hormone receptors and coregulators. Since NOR1, one member of NR4A group nuclear receptors, was the most highly regulated NR upon viral infection in DCs, we obtained NOR1 knockout mice from Dr. Conneely, the Baylor collage of medicine, and have examined impact of pathogen infection to the action of DCs purified from NOR1 knockout mice. DCs from these mice showed a significant defect in the response of interleukin (IL)-12 to viral infection compared to those from wild type mice, while NOR-1 potently stimulated the IL-12 p40 promoter activity in reporter assays. NOR-1 knockout mice demonstrated significant reduction of IL-12 production against infection of Toxoplasma gondii, a protozoa against which IL-12 acts as an essential component for host defense. We are currently examining details of molecular regulation of NOR-1 on IL-12 production both in the cellular and animal systems. In the same line of experiments, we found that CMV and NDV stimulated IL-10 expression in DCs, and glucocorticoids further potentiated such virus-induced expression of this cytokine. Since IL-10 inhibits synthesis of pro-inflammatory cytokines and has ability to suppress antigen presentation by DCs and monocytes, synergistic activation of IL-10 by glucocorticoids may explain why exposure to stress and subsequent activation of the HPA axis increases susceptibility to viral infection, and possibly, subsequent development of viral-associated disorders, such as asthma, atherosclerosis and caners. We found that viral infection activated the extracellular signal-regulated kinase 1 (ERK1), a member of mitogen-activated protein kinase family, which in turn phosphorylated the human GR at serine located at amino acid position 211 and modulated the transcriptional activity of GR on the IL-10 promoter. Extracellular hyperosmolarity or osmotic stress is a major threat for land organisms, and thus strongly stimulates HPA axis through secretion of ariginine vasopression from the hypothalamus/posterior lobe of the pituitary gland. In addition to this systemic response, osmotic stress also activates a cellular signaling cascade called adaptive response to extracellular hyperosmolarity: Extracellular hyperosmolarity induces and activates a Rel-homology domain-containing transcription factor, the nuclear factor of activated T-cells 5 (NFAT5), which subsequently stimulates transcription of osmotic stress-responsive genes and causes intracellular accumulation of small organic osmolytes to maintain isotonicity between the inside and outside of the cells. We previously reported an intracellular signaling cascade responsive to osmotic stress in lymphocytes: The Rho-type guanine nucleotide exchange factor (GEF) Brx or AKAP13 is essential for the osmotic stress-stimulated expression of NFAT5, and is a key component of the intracellular signaling cascade transmitting the extracellular hyperosmolarity signal to the nucleus. Osmotic stress-mediated induction of NFAT5 requires the Brx GEF domain and p38 mitogen-activated kinase (MAPK), while Brx in response to osmotic stress attracts through its C-terminal domain the cJun kinase (JNK)-interacting protein (JIP) 4, a scaffold specific to activation of the p38 MAPK cascade and NFAT5, coupling activated Rho-type small G proteins to components of the p38 MAPK signaling pathway. Importantly, this signaling system plays a critical role in the differentiation of lymphocytes in the spleen and stimulates production of TNF-alpha and other cytokines. Osmotic stress-mediated activation of this cellular signaling system may also be an important component in immune dysregulation observed in some pathologic conditions accompanied by plasma hyperosmolarity, such as diabetes mellitus, uremia, dehydration and severe burn all of which are known to develop altered cytokine production and immune-associated symptoms/signs. To further elucidate roles of Brx/NFAT5-mediated osmotic stress on the regulation of the immune system, we are now developing mice carrying specific deletion of the brx gene in DCs/monocytes/macrophages using the Cre/LoxP system. We hypothesized that Brx/NFAT5 plays a significant roles in the high-salt diet-induced decrease of nitric oxide synthase and subsequent increase of blood pressure, as recent publication indicated that NFAT5 expressed in residential macrophages participate in this BP regulatory system. Using the conditional Brx knockout mice in DCs/macrophages, we will examine contribution of Brx on high salt diet-induced hypertension, as well as on the cardiac fibrosis, which is in part mediated by inflammation.

我们进行了几组实验来检查内分泌系统和免疫系统之间的相互作用。我们发现两种小分子（IL-6 拮抗剂和 PPAR-δ 激动剂）通过 STAT3 抑制具有很强的抗炎活性。前一种化合物 TB-2-081 与 IL-6 型受体的 gp130 亚基相互作用，不仅阻断 IL-6 的活性，还阻断白血病抑制因子、制瘤素 M 和 IL-11 的活性。细胞中的一个主要变化是 STAT3 转录因子的酪氨酸磷酸化受到抑制。后一种化合物，GW50516，抑制肝细胞产生几种急性期反应物，从而显示出主要的抗炎活性。这种效应也是通过抑制响应基因启动子处的 STAT3 活性来介导的。病毒，包括人类免疫缺陷综合征 1 型 (HIV-1)、巨细胞病毒 (CMV) 和新城疫 (NDV) 病毒，是宿主免疫和内分泌系统的有效激活剂和调节剂，影响宿主组织中的激素作用，例如如白细胞、脂肪组织和骨骼肌。这些作用进一步促进病毒扩张和发病机制。事实上，我们证明了 HIV-1 辅助蛋白 Vpr 抑制 PPAR-gamma 活性，在与 AIDS 相关的特征性脂肪营养不良和胰岛素抵抗综合征的发展中发挥潜在重要作用。树突状细胞 (DC) 在病毒抗原的识别和呈递中发挥核心作用，CMV 或 NDV 以及可能还有 HIV-1 的感染会导致一组核激素受体（包括糖皮质激素）的表达发生巨大变化和雌激素受体，以及几种转录辅助调节因子，例如 p300 和 p160 型组蛋白乙酰转移酶辅助激活因子，可能会改变干扰素的分泌/产生，这些细胞产生其他细胞因子。我们根据核激素受体和共调节因子 mRNA 表达谱的结果发表了一篇手稿。由于 NOR1（NR4A 组核受体的成员之一）是 DC 中病毒感染时受最高度调节的 NR，因此我们从贝勒医学院 Conneely 博士那里获得了 NOR1 敲除小鼠，并检查了病原体感染对其作用的影响。从 NOR1 敲除小鼠中纯化的 DC。与野生型小鼠相比，这些小鼠的 DC 在白细胞介素 (IL)-12 对病毒感染的反应方面表现出显着缺陷，而 NOR-1 在报告基因检测中有效刺激了 IL-12 p40 启动子活性。 NOR-1 敲除小鼠表现出针对弓形虫感染的 IL-12 产生显着减少，弓形虫是一种原生动物，IL-12 是宿主防御的重要组成部分。我们目前正在研究细胞和动物系统中 NOR-1 对 IL-12 产生的分子调节细节。在同一系列实验中，我们发现 CMV 和 NDV 刺激 DC 中的 IL-10 表达，而糖皮质激素进一步增强了病毒诱导的这种细胞因子的表达。由于 IL-10 抑制促炎细胞因子的合成，并能够抑制 DC 和单核细胞的抗原呈递，因此糖皮质激素对 IL-10 的协同激活可以解释为什么暴露于压力和随后的 HPA 轴激活会增加对病毒感染的易感性，并可能导致随后出现与病毒相关的疾病，例如哮喘、动脉粥样硬化和癌症。我们发现病毒感染激活了细胞外信号调节激酶1（ERK1），它是丝裂原激活蛋白激酶家族的一员，反过来又磷酸化人类GR的211位氨基酸丝氨酸，并调节GR的转录活性。 IL-10启动子。细胞外高渗透压或渗透压是陆地生物的主要威胁，因此通过下丘脑/垂体后叶分泌精氨酸血管加压来强烈刺激 HPA 轴。除了这种全身反应外，渗透压还会激活细胞信号级联，称为对细胞外高渗性的适应性反应：细胞外高渗性诱导并激活含有 Rel 同源结构域的转录因子，即活化 T 细胞 5 (NFAT5) 的核因子，随后刺激渗透应激反应基因的转录，并导致细胞内小有机渗透剂的积累，以维持细胞内外的等渗性。我们之前报道了淋巴细胞中对渗透压有反应的细胞内信号级联：Rho 型鸟嘌呤核苷酸交换因子 (GEF) Brx 或 AKAP13 对于渗透压刺激的 NFAT5 表达至关重要，并且是细胞内信号级联的关键组成部分将细胞外高渗信号传递至细胞核。渗透应激介导的 NFAT5 诱导需要 Brx GEF 结构域和 p38 丝裂原激活激酶 (MAPK)，而响应渗透应激的 Brx 通过其 C 末端结构域吸引 cJun 激酶 (JNK) 相互作用蛋白 (JIP) 4，一种特异性激活 p38 MAPK 级联和 NFAT5 的支架，将激活的 Rho 型小 G 蛋白与 p38 MAPK 信号通路的成分偶联。重要的是，该信号系统在脾脏淋巴细胞的分化中发挥着关键作用，并刺激 TNF-α 和其他细胞因子的产生。渗透压介导的细胞信号系统激活也可能是免疫失调的重要组成部分，在一些伴有血浆高渗透压的病理状况下观察到，例如糖尿病、尿毒症、脱水和严重烧伤，所有这些都会导致细胞因子产生改变和免疫相关症状/体征。为了进一步阐明 Brx/NFAT5 介导的渗透压对免疫系统调节的作用，我们现在正在使用 Cre/LoxP 系统开发在 DC/单核细胞/巨噬细胞中携带 brx 基因特异性缺失的小鼠。我们假设 Brx/NFAT5 在高盐饮食诱导的一氧化氮合酶降低和随后的血压升高中发挥重要作用，因为最近的出版物表明在住宅巨噬细胞中表达的 NFAT5 参与了这一血压调节系统。使用 DC/巨噬细胞中的条件 Brx 敲除小鼠，我们将检查 Brx 对高盐饮食诱发的高血压以及部分由炎症介导的心脏纤维化的贡献。