Radioresistant Innate Immunity in SAVI Tissue-Specific Autoinflammation

SAVI 组织特异性自身炎症中的抗辐射先天免疫

• 批准号: 10752556
• 负责人: Katherine A. Fitzgerald
• 金额: $ 25.13万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-04 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10752556
• 关键词: Address; Adhesions; Alleles; Area; Autoimmune; Autoimmune Diseases; Automobile Driving; Bone Marrow Stem Cell; Bronchus-Associated Lymphoid Tissue; Cell Death; Cell Line; Cell Separation; Cells; Chimera organism; Cytosol; DNA; Data; Development; Disease; Embryo; Endothelial Cells; Endothelium; Exclusion; Exhibits; Fibrosis; Follow-Up Studies; Gene Expression; Genes; Genetic Transcription; Goals; Hematopoietic; Heterozygote; Host Defense; IRF3 gene; Immune; In Vitro; Individual; Infection; Inflammation; Inflammatory; Innate Immune Response; Interferons; Interstitial Lung Diseases; Knock-in; Lesion; Link; Lung; Lung diseases; Lymphatic Endothelial Cells; Lymphocyte; Lymphopenia; Molecular; Monitor; Mus; Mutation; Myelogenous; Myeloid Cells; Natural Immunity; Pathogenicity; Pathology; Pathway interactions; Patients; Peripheral; Phenotype; Play; Population; Pulmonary Fibrosis; Pulmonary Inflammation; Radiation Chimera; Rag1 Mouse; Reporter; Research; Research Personnel; Resistance; Respiratory Failure; Rheumatoid Arthritis; Role; STING agonists; Series; Stimulator of Interferon Genes; Stromal Cells; Systemic Scleroderma; T-Lymphocyte; TBK1 gene; Tamoxifen; Tissues; Vascular Diseases; Vascular Endothelial Cell; adaptive immunity; autoinflammation; autoinflammatory; autoinflammatory diseases; cell type; cellular transduction; cytokine; design; effective therapy; fibrotic lung disease; functional outcomes; gain of function mutation; gene function; human disease; in vivo; infancy; insight; interferon alpha receptor; lymph nodes; microbial; mouse model; mutant; neutrophil; novel; promoter; pulmonary function; radioresistant; reconstitution; recruit; severe COVID-19; skin lesion; stem cells; therapeutic target; tool; transcriptome; treatment strategy; Address; Adhesions; Alleles; Area; Autoimmune; Autoimmune Diseases; Automobile Driving; Bone Marrow Stem Cell; Bronchus-Associated Lymphoid Tissue; Cell Death; Cell Line; Cell Separation; Cells; Chimera organism; Cytosol; DNA; Data; Development; Disease; Embryo; Endothelial Cells; Endothelium; Exclusion; Exhibits; Fibrosis; Follow-Up Studies; Gene Expression; Genes; Genetic Transcription; Goals; Hematopoietic; Heterozygote; Host Defense; IRF3 gene; Immune; In Vitro; Individual; Infection; Inflammation; Inflammatory; Innate Immune Response; Interferons; Interstitial Lung Diseases; Knock-in; Lesion; Link; Lung; Lung diseases; Lymphatic Endothelial Cells; Lymphocyte; Lymphopenia; Molecular; Monitor; Mus; Mutation; Myelogenous; Myeloid Cells; Natural Immunity; Pathogenicity; Pathology; Pathway interactions; Patients; Peripheral; Phenotype; Play; Population; Pulmonary Fibrosis; Pulmonary Inflammation; Radiation Chimera; Rag1 Mouse; Reporter; Research; Research Personnel; Resistance; Respiratory Failure; Rheumatoid Arthritis; Role; STING agonists; Series; Stimulator of Interferon Genes; Stromal Cells; Systemic Scleroderma; T-Lymphocyte; TBK1 gene; Tamoxifen; Tissues; Vascular Diseases; Vascular Endothelial Cell; adaptive immunity; autoinflammation; autoinflammatory; autoinflammatory diseases; cell type; cellular transduction; cytokine; design; effective therapy; fibrotic lung disease; functional outcomes; gain of function mutation; gene function; human disease; in vivo; infancy; insight; interferon alpha receptor; lymph nodes; microbial; mouse model; mutant; neutrophil; novel; promoter; pulmonary function; radioresistant; reconstitution; recruit; severe COVID-19; skin lesion; stem cells; therapeutic target; tool; transcriptome; treatment strategy

The cGas/STING cytosolic DNA sensing pathway detects microbial DNA and plays a critical role in host defense. Growing evidence indicates that self-DNA that accumulates in the cytosol also engages cGAS to incite inflammation. In addition, a series of gain-of-function mutations that result in constitutive activation of STING have now been associated with a debilitating auto-inflammatory disease known as SAVI (STING-Associated- Vasculopathy with onset in Infancy). These patients suffer from severe vasculitic lesions and progressive interstitial lung disease (ILD), and frequently succumb to treatment-resistant ILD-associated with fibrosis. Similar to the lung disorders associated with other inflammatory or fibrotic lung diseases, very little is known about the pathogenic mechanisms in these patients. Thus, a better understanding of the mechanisms responsible for these conditions is needed in order to develop more effective therapies. We have recently developed a murine model for the most common SAVI mutation, STINGV154M (VM), and shown that these mice recapitulate the human disease by a variety of criteria, including the development of inflammatory/fibrotic lung disease. These VM mice provide a unique experimental tool for exploring the cell types and molecular mechanisms responsible for the initiation and progression of fibrotic lung disease. Rag1-deficiency completely rescues VM mice from both lung inflammation and lung fibrosis pointing to a critical role of the adaptive immunity in VM ILD. The role of STING has been focused on myeloid cells and VM mice have an expanded and activated myeloid/neutrophil compartment. However, radiation chimera studies have identified a key role for radioresistant cells in lung fibrosis as lethally irradiated VM mice reconstituted with WT bone marrow stem cells develop extremely severe lung disease even though these chimeras completely lack any VM-derived hematopoietic cells. Preliminary data implicate lymphatic endothelial cells (LECs) as initiators of BALT formation in VM mice. The goal of this application is to further explore the radioresistant innate immune cells that are directly or indirectly activated by the VM mutation and define the mechanisms by which these cells promote lung inflammation. Our approach will involve: (1) the in vivo analysis of a novel VM conditional KI line crossed to endothelial-restricted Cre deleter strains and other relevant controls, (2) scRNAseq comparison of stromal populations including lymphatic and vascular endothelial cells isolated from WT and VM mice; and (3) analysis of endothelial cells from VM and WT mice following in vitro activation by the VM mutation or more conventional STING agonists. There is an urgent need to identify better therapies for patients afflicted with autoimmune and autoinflammatory lung disorders and the studies proposed in this application should provide critical insights that will enable us to design the most relevant therapeutic targets. Further, these studies provide an opportunity to study the impact of STING activation on stromal cell types more generally, an emerging area of research as STING activity in endothelial cells has recently been linked to the development of severe COVID19.

CGA/STING胞质DNA感应途径检测微生物DNA并在宿主防御中起关键作用。 越来越多的证据表明，积聚在细胞质中的自动-DNA也使CGA煽动 炎。此外，一系列功能收益突变导致刺激的本构激活 现在已经与一种衰弱的自发性疾病有关 血管病，婴儿期发作）。这些患者患有严重的血管病变和进行性疾病 间质性肺疾病（ILD），并且经常屈服于与纤维化相关的耐药性ILD相关。相似的 对于与其他炎症或纤维化肺部疾病相关的肺部疾病，关于 这些患者的致病机制。因此，更好地理解负责这些机制的机制 为了开发更有效的疗法，需要条件。我们最近开发了一个鼠模型 对于最常见的SAVI突变，StingV154M（VM），并表明这些小鼠概括了人类 通过多种标准疾病，包括炎症/纤维化肺部疾病的发展。这些VM小鼠 提供独特的实验工具，用于探索负责的细胞类型和分子机制 纤维化肺部疾病的起始和进展。 RAG1缺乏完全从两种肺部救出VM小鼠 炎症和肺纤维化表明自适应免疫在VM ILD中的关键作用。刺的作用 已专注于髓样细胞和VM小鼠具有扩​​展和激活的髓样/中性粒细胞 车厢。然而，辐射嵌合体研究已经确定了辐射耐药细胞在肺纤维化中的关键作用 随着致命的辐照的VM小鼠与WT骨髓干细胞重构发生极度严重的肺 尽管这些嵌合体完全缺乏任何VM衍生的造血细胞。初步数据 暗示淋巴内皮细胞（LEC）作为VM小鼠的Balt形成的启动者。目标的目标 应用是进一步探索直接或间接激活的放射性先天免疫细胞 VM突变并定义了这些细胞促进肺部炎症的机制。我们的做法意愿 涉及：（1）新型VM条件Ki线的体内分析交叉至内皮限制的CRE DELERE 菌株和其他相关对照，（2）基质种群的SCRNASEQ比较，包括淋巴和 从WT和VM小鼠分离的血管内皮细胞； （3）分析来自VM和WT的内皮细胞 VM突变或更常规的刺痛激动剂在体外激活后的小鼠。有紧急 需要为患有自身免疫和自身炎症肺部疾病的患者确定更好的疗法 本应用程序中提出的研究应提供关键见解，使我们能够设计最多 相关的治疗靶标。此外，这些研究提供了研究刺伤影响的机会 更一般地对基质细胞类型激活，这是内皮中的刺激活性的新兴领域 细胞最近与严重的Covid19的发展有关。