Determining the conserved molecular mechanisms contributing to inflammation during Sepsis

确定脓毒症期间导致炎症的保守分子机制

• 批准号: 10400670
• 负责人: Susan Carpenter
• 金额: $ 37.42万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA CRUZ
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-14 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10400670
• 关键词: ATAC-seq; Antisense RNA; Bacteremia; Bacteria; Binding; Biological Models; Bone Marrow; CRISPR/Cas technology; Cause of Death; Cells; Centers for Disease Control and Prevention (U.S.); Chemicals; Chimera organism; Chromatin; Clinical; Clinical Treatment; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Cytosol; Data; Endotoxic Shock; Escherichia coli; Future; Gastric Adenocarcinoma; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genome; Hospitals; Human; Human Genome; Immune; Immune response; Infection; Infiltration; Inflammation; Knock-out; Knockout Mice; Lead; Length; Mediating; Molecular; Mus; Natural Immunity; Nucleotides; Pathway interactions; Patients; Persons; Phenotype; Play; Proteins; RNA; RNA Binding; Regulation; Resistance; Role; Sepsis; Septic Shock; Shock; Small Interfering RNA; Testing; Therapeutic; Therapeutic Intervention; Transcript; Transgenes; Transgenic Animals; Transgenic Organisms; Untranslated RNA; design; drug development; in vivo; insight; knock-down; knockout animal; macrophage; malignant stomach neoplasm; monocyte; mouse model; neutrophil; new therapeutic target; overexpression; progenitor; response; transcriptome sequencing

Project Summary Worldwide it is estimated that over 6 million people die each year as a result of sepsis. There are few clinical treatment options for patients, therefore, it is critical to determine the molecular mechanisms that occur during sepsis in order to identity new targets for therapeutic intervention. Here we identify the long noncoding RNA, GAPLINC, as a conserved gene between human and mice that is highly expressed in macrophages. We show that GAPLINC knockouts are resistant to LPS induced septic shock. The overall aim of this proposal is to determine how GAPLINC contributes to the immune response that leads to septic shock. In Aim 1 we will utilize our genetic mouse models to expand on our initial findings and determine what impact knockout or overexpression of GAPLINC has in response to gram negative induced sepsis in vivo. In Aim 2 we will determine the molecular mechanisms utilized by GAPLINC to influence immune genes. We will identify the minimal region within GAPLINC required to impact gene expression. We will determine the complexes GAPLINC makes either with RNA or proteins to function and finally we will identify any structural features within GAPLINC that mediates these interactions. In Aim 3 we will determine if GAPLINC is functionally conserved in humans by studying its role in controlling immune genes in primary human monocyte derived macrophages. This project will enable us to better understand the complex mechanisms that are at play during bacterial induced sepsis. By focusing on GAPLINC we will identify a new layer of regulation during sepsis providing us with new avenues for future drug development.

项目摘要 据估计，全世界每年有600多万人死于败血症。那里 对于患者来说，临床治疗选择很少，因此，确定分子水平至关重要 在脓毒症期间发生的机制，以确定治疗干预的新靶点。 在这里，我们发现长的非编码RNA，GAPLINC，是人类和人类之间的一个保守基因 在巨噬细胞中高表达的小鼠。我们证明了GAPLINC基因敲除是具有抗性的 对内毒素诱导的感染性休克有明显的保护作用。这项提案的总体目标是确定GAPLINC如何 参与导致感染性休克的免疫反应。在目标1中，我们将利用我们的 在我们初步发现的基础上扩展小鼠遗传模型，并确定基因敲除或 在体内，GAPLINC的过表达对革兰氏阴性脓毒症有反应。在AIM 2 我们将确定GAPLINC影响免疫基因的分子机制。 我们将确定GAPLINC中影响基因表达所需的最小区域。我们会 确定GAPLINC与RNA或蛋白质形成的复合体以发挥作用，最后我们 将确定GAPLINC内调解这些互动的任何结构特征。在AIM 3中 我们将通过研究GAPLINC在人类中的作用来确定它在人类中是否在功能上是保守的 控制原代人单核细胞来源的巨噬细胞的免疫基因。这个项目将 使我们能够更好地了解在细菌感染过程中发挥作用的复杂机制 诱发性败血症。通过关注GAPLINC，我们将确定脓毒症过程中的一层新的调节 为我们未来的药物开发提供了新的途径。