Genetic Analysis of Inflammatory Responses in Wild Derived Mice

野生小鼠炎症反应的遗传分析

基本信息

批准号：
10534173
负责人：
Alexander Poltorak
金额：
$ 76.97万
依托单位：
TUFTS UNIVERSITY BOSTON
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-09-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10534173
关键词：
Affect Alleles Autoimmune Biological Response Modifiers Blood Candidate Disease Gene Cells Code Compensation Complement Cytosol DNA DNA Repair DNA Viruses DNA analysis Dangerousness Data Defect Embryo Endosomes Enzymes Etiology Exhibits Failure Female Genes Genetic Genetic Recombination Goals High Prevalence Homeostasis Human IL6 gene Immune response In Vitro Inbreeding Individual Inflammation Inflammatory Inflammatory Response Inherited Interferon Type I Interferon alpha Interferons Interleukin-6 Invaded Investigation Knockout Mice Lead Link Macrophage Maps Mediating Mediator Meiotic Recombination Messenger RNA Mus Mutation Normal Statistical Distribution Nucleic Acids Outcome Pathogenicity Pathology Pathway interactions Patients Phenotype Production Progress Reports Proteins Quantitative Trait Loci Regulation Role STING1 gene Signal Transduction Stimulator of Interferon Genes Sting Injury TREX1 gene Testing Toxic effect congenic cytokine cytotoxicity differential expression gene cloning genetic analysis immunopathology in vivo insight loss of function novel pathogen positional cloning response sensor single cell analysis single-cell RNA sequencing trait

项目摘要

PROJECT SUMMARY: A central scientific question of this competing renewal is the mechanistic understanding of the inflammatory responses to cytosolic DNA. To achieve our goals, we continue using the forward genetic analysis in the wild- derived mice of the MOLF strain that diverged from classical inbred (C57BL6) mice about 1 million years ago. A phenotype that provides preliminary data for this application is the defective IFN production in MOLF macrophages in response to DNA pathogens or cytosolic DNA - a condition conferred by a hypomorphic allele of Sting. In spite of the failure to produce IFN, activated MOLF macrophages overproduce IL-6. Other wild- derived strains exhibit similar skewing of responses to DNA-viruses and DNA. Based on these data, we hypothesized that in order to avoid IFN-mediated excessive inflammation, DNA sensing pathways must have evolved to complement IFN production, perhaps with less dangerous substitutes such as IL-6. Accordingly, we will determine how wild derived mice respond to cytosolic DNA with low IFN but high IL-6--a line of investigation that will ultimately identify genes (loci) responsible for IL-6 overproduction. In the first Aim, we will test the hypothesis that retention of STING in the ER is the mechanism of switching the DNA-responses from IFN to IL-6 production. Second, we will examine the potential contribution of other DNA-sensors and pathways into STING-mediated DNA-responses in MOLF. Finally, we will use STING congenic (B6.StingMOLF/MOLF) mice, which are completely non-responsive to DNA to genetically map and identify gene(s) that confer overproduction of IL-6 in MOLF in response to DNA. In extension of genetic analysis of the trait, we will study the responses to DNA in DNAse2-/- Sting MOLF/- mice. These mice are rescued from DNAse2-/- -associated embryonic lethality with the hypomorphic allele of Sting despite high levels of inflammatory cytokines in the blood and embryonic lethality of the DNAse2-/- Sting MOLF/- females. These data suggest that some uncharacterized STING-mediated signaling exists in these mice, which will be investigated in Aim 2 of the proposal. First, we will identify the DNA-responsive cells, and use single cell RNA-sequencing analysis in these cells to identify genes with expression levels associated with the inflammatory signature and DNA- responses. Despite being confined to eQTL (expression Quantitative Trait Loci), the single cell association studies in Aim 2 will potentially reveal all associations between the phenotype and the genes, some of which will not necessarily be mapped in Aim 1 and could be completely novel. Finally, based on embryonic lethality of the DNAse2-/- Sting MOLF/- females, we will investigate potential contributions of the X-linked Tlr7 and Tlr8 into STING-mediated responses. By investigating the mechanism of DNA responses in MOLF, we hope to provide better insight into the diversity of pathologies present in human patients with interferonopathies. !

项目摘要：这种竞争更新的中心科学问题是对炎症的机械理解对胞质DNA的反应。为了实现我们的目标，我们继续在野生中使用前瞻性遗传分析大约100万年前，衍生出与古典近交（C57BL6）小鼠不同的Molf菌株的小鼠。为此应用提供初步数据的表型是MOLF中的IFN产生缺陷巨噬细胞响应DNA病原体或胞质DNA - 由型肌层赋予的疾病刺。尽管未能产生IFN，但激活的Molf巨噬细胞过量产生IL-6。其他野生衍生的菌株表现出对DNA病毒和DNA反应的相似偏斜。基于这些数据，我们假设为了避免IFN介导的过度炎症，DNA感应途径必须具有演变为补充IFN生产，也许具有危险的替代品，例如IL-6。因此，我们将确定野生派生的小鼠如何对低IFN但高IL-6的胞质DNA反应最终将确定导致IL-6过量生产的基因（基因座）的研究。在第一个目标中，我们将检验以下假设，即ER中刺的保留是从 IFN至IL-6生产。其次，我们将研究其他DNA传感器和途径的潜在贡献进入MOLF的STING介导的DNA响应。最后，我们将使用Sting Encenic（B6.StingMolf/Molf）小鼠，对DNA完全无反应，以遗传映射并识别赋予的基因响应DNA，在MOLF中生产IL-6。在特征的遗传分析扩展时，我们将研究 DNase2 - / - sting molf/ - 小鼠中对DNA的反应。这些小鼠是从DNASE2 - / - 相关的尽管在高水平 DNase2 - / - sting molf/ - 女性的血液和胚胎致死性。这些数据建议一些这些小鼠中存在未表征的刺激介导的信号传导，将在AIM 2中进行研究提议。首先，我们将确定DNA响应性细胞，并在这些细胞以鉴定具有与炎症特征和DNA-的表达水平的基因回答。尽管仅限于eqtl（表达定量性状基因座），但单细胞关联 AIM 2中的研究将有可能揭示表型与基因之间的所有关联，其中一些关联不一定在AIM 1中映射，并且可能是完全新颖的。最后，基于胚胎致死性 DNASE2 - / - sting molf/ - 女性的中，我们将研究X连锁TLR7和TLR8的潜在贡献进入刺激介导的响应。通过调查MOLF中DNA响应的机制，我们希望提供更好的洞察力，以了解干涉病例患者中存在的病理多样性。呢

项目成果

期刊论文数量（24）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Cutting Edge: Activation of STING in T Cells Induces Type I IFN Responses and Cell Death.

最前沿：T 细胞中 STING 的激活可诱导 I 型 IFN 反应和细胞死亡。

DOI：
10.4049/jimmunol.1601999
发表时间：
2017-07-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
0
作者：
Larkin B;Ilyukha V;Sorokin M;Buzdin A;Vannier E;Poltorak A
通讯作者：
Poltorak A

ZBP1 promotes inflammatory responses downstream of TLR3/TLR4 via timely delivery of RIPK1 to TRIF.

DOI：
10.1073/pnas.2113872119
发表时间：
2022-06-14
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
通讯作者：

IRAK-2 regulates IL-1-mediated pathogenic Th17 cell development in helminthic infection.

DOI：
10.1371/journal.ppat.1002272
发表时间：
2011-10
期刊：
PLoS pathogens
影响因子：
6.7
作者：
Smith PM;Jacque B;Conner JR;Poltorak A;Stadecker MJ
通讯作者：
Stadecker MJ

The PYRIN connection: novel players in innate immunity and inflammation.

DOI：
10.1084/jem.20032234
发表时间：
2004-09-06
期刊：
JOURNAL OF EXPERIMENTAL MEDICINE
影响因子：
15.3
作者：
Stehlik, C;Reed, JC
通讯作者：
Reed, JC

Constitutive Interferon Maintains GBP Expression Required for Release of Bacterial Components Upstream of Pyroptosis and Anti-DNA Responses.

DOI：
10.1016/j.celrep.2018.06.012
发表时间：
2018-07-03
期刊：
Cell reports
影响因子：
8.8
作者：
Liu BC;Sarhan J;Panda A;Muendlein HI;Ilyukha V;Coers J;Yamamoto M;Isberg RR;Poltorak A
通讯作者：
Poltorak A