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Role of defensin receptor signaling in IL-1beta release

防御素受体信号传导在 IL-1β 释放中的作用

基本信息

批准号：
8078868
负责人：
JISHU SHI
金额：
$ 18.32万
依托单位：
KANSAS STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8078868
关键词：
Adjuvant Affinity Affinity Chromatography Animals Area Autoimmune Diseases Bacterial Infections Bacterial Toxins Binding Binding Proteins Biology Ca(2+)-Calmodulin Dependent Protein Kinase Calcium/calmodulin-dependent protein kinase Caspase-1 Cell Surface Receptors Cell membrane Cells Conflict (Psychology)Confocal Microscopy Confusion Cytosol Defensins Development Disease Enzymes Epithelial Cells Exocytosis Fibroblast Growth Factor 2 Goals Golgi Apparatus Human Infection Inflammatory Interleukin-12 Interleukin-18 Lead Leukocytes Life Link Lysosomes Mediating Medical Research Membrane Proteins Mitogen-Activated Protein Kinase Kinases Modeling Molecular Molecular Target Multivesicular Body Pathway interactions Peptides Phospholipase C Process Production Protein Kinase Protein Kinase C Proteins Proteomics Receptor Signaling Role Septic Shock Signal Pathway Signaling Protein Small Interfering RNA System Testing antimicrobial peptide cell type cytokine extracellular human neutrophil peptide 1 in vivo inhibitor/antagonist insight macrophage microbial monocyte neutrophil novel novel therapeutic intervention prevent public health relevance receptor tool

项目摘要

DESCRIPTION (provided by applicant): Overproduction of IL-12 is associated with autoimmune diseases and microbial infections. Newly synthesized leaderless proIL-12 cannot be efficiently secreted from activated monocytes. However, when LPS-primed monocytes are further stimulated with extracellular ATP, they rapidly release large amounts of mature IL-12 and proIL-12. Thus far, the mechanism by which IL-12 is released from human monocytes is not understood. The overall goal of this proposal is to determine how IL-12 is released by investigating the molecular mechanisms by which ?-defensins block the release of IL-12 from human monocytes. Human ?-defensins (HNP-1 and HD-5), a group of antimicrobial peptides produced by neutrophils and epithelial cells, are the only inhibitors that block the release but not the processing of proIL-12 by caspase-1 in human monocytes. This proposal does not investigate the in vivo role of defensins. It only uses defensins as a tool to identify novel molecular targets for IL-12 blockade. Using photo-affinity and confocal microscopy, we have found that defensins bind to cell-membrane-associated proteins in human monocytes. We will test the hypothesis that IL-12 release and the externalization of proIL-12 and secretory lysosomes from human monocytes are regulated by distinct signaling proteins which can be inhibited by ?-defensins via an enzyme- linked receptor. To accomplish that, we have identified two specific aims: Specific Aim One: Identify the receptor that is responsible for defensin-mediated inhibition of IL-12 release from human monocytes. Using photo-affinity purification and proteomic approaches, we will identify defensin-binding proteins (DBPs) in human monocytes. We will then identify the defensin receptor by determining the effect of siRNA-mediated knockdown of these DBPs on defensin blockade of IL-12 release. Specific Aim Two: Define the signaling pathway essential for defensin receptor-mediated inhibition of IL- 12 release. We will determine whether protein kinase C, phospholipase C, Ca2????dependent protein kinase, and MAP kinases are involved in defensin blockade of IL-12 release and the externalization of proIL-12 and secretory lysosomes from human monocytes. Successful completion of this project will have broad impacts on both basic and translational medical research. If our hypotheses are correct, it will not only bring much clarity to the field of IL-12 biology, but also provide novel insights to the ER/Golgi-independent secretory pathway used by many other important leaderless proteins, including IL-18, IL-33, MIF, and FGF-2. The proposed studies will also lead to the discovery of novel molecular targets for IL-12 blockade and may lead to the development of new therapeutic approaches to prevent and treat many life-threatening microbial infections and inflammatory diseases. PUBLIC HEALTH RELEVANCE: Defensins are peptides produced by white blood cells and epithelial cells. Using defensins as a tool, we will determine how proinflammatory cytokine IL-12 is released from human monocytes. The proposed studies will lead to the discovery of novel molecular targets for IL-12 blockade and may lead to the development of new therapeutic approaches to prevent and treat many life-threatening microbial infections and inflammatory diseases.

描述（由申请人提供）：IL-12的过度产生与自身免疫性疾病和微生物感染相关。新合成的无前导proIL-12不能从活化的单核细胞有效分泌。然而，当LPS引发的单核细胞进一步用细胞外ATP刺激时，它们迅速释放大量成熟IL-12和proIL-12。到目前为止，IL-12从人单核细胞释放的机制尚不清楚。这项建议的总体目标是通过研究IL-12的分子机制来确定IL-12是如何释放的。防御素阻断人单核细胞释放IL-12。人类？防御素（HNP-1和HD-5）是由嗜中性粒细胞和上皮细胞产生的一组抗菌肽，是唯一阻断人单核细胞中caspase-1对proIL-12的释放而不是加工的抑制剂。该提议没有研究防御素的体内作用。它仅使用防御素作为鉴定IL-12阻断的新分子靶点的工具。利用光亲和和共聚焦显微镜，我们已经发现，防御素结合细胞膜相关蛋白在人类单核细胞。我们将检验以下假设：IL-12的释放以及proIL-12和分泌性溶酶体从人单核细胞的外化受不同信号蛋白的调节，这些信号蛋白可以被？防御素通过酶联受体。为了实现这一目标，我们确定了两个具体目标：具体目标一：确定负责防御素介导的抑制人单核细胞释放IL-12的受体。利用光亲和纯化和蛋白质组学方法，我们将确定防御素结合蛋白（DBPs）在人类单核细胞。然后，我们将通过确定siRNA介导的这些DBP的敲低对防御素阻断IL-12释放的影响来鉴定防御素受体。具体目标二：确定防御素受体介导的IL-12释放抑制所必需的信号通路。我们将确定是否蛋白激酶C，磷脂酶C，钙？依赖性蛋白激酶和MAP激酶参与IL-12释放的防御素阻断和proIL-12和分泌性溶酶体从人单核细胞的外化。该项目的成功完成将对基础和转化医学研究产生广泛的影响。如果我们的假设是正确的，它将不仅带来了IL-12生物学领域的清晰度，而且还提供了新的见解ER/高尔基体非依赖性分泌途径使用的许多其他重要的无前导蛋白，包括IL-18，IL-33，MIF和FGF-2。拟议的研究还将导致发现IL-12阻断的新分子靶点，并可能导致开发新的治疗方法来预防和治疗许多危及生命的微生物感染和炎症性疾病。公共卫生相关性：防御素是由白色血细胞和上皮细胞产生的肽。使用防御素作为工具，我们将确定促炎细胞因子IL-12是如何从人单核细胞释放的。拟议的研究将导致发现IL-12阻断的新分子靶点，并可能导致开发新的治疗方法来预防和治疗许多危及生命的微生物感染和炎症性疾病。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Comparison of host immune responses to homologous and heterologous type II porcine reproductive and respiratory syndrome virus (PRRSV) challenge in vaccinated and unvaccinated pigs.

DOI：
10.1155/2014/416727
发表时间：
2014
期刊：
BioMed research international
影响因子：
0
作者：
Li X;Galliher-Beckley A;Pappan L;Trible B;Kerrigan M;Beck A;Hesse R;Blecha F;Nietfeld JC;Rowland RR;Shi J
通讯作者：
Shi J

IL-1β promotes stemness and invasiveness of colon cancer cells through Zeb1 activation.