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

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.

cGas/STING 胞质 DNA 传感通路可检测微生物 DNA，并在宿主防御中发挥关键作用。 越来越多的证据表明，在细胞质中积累的自身 DNA 也会参与 cGAS 的激发 炎。此外，一系列功能获得性突变导致 STING 的组成型激活 现在已与一种称为 SAVI 的衰弱性自身炎症性疾病（STING-相关- 婴儿期发病的血管病）。这些患者患有严重的血管炎病变并且进行性进展 间质性肺疾病 (ILD)，并且经常死于与纤维化相关的难治性 ILD。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，人们知之甚少 这些患者的致病机制。因此，更好地理解造成这些现象的机制 需要条件才能开发更有效的疗法。我们最近开发了一种小鼠模型 对于最常见的 SAVI 突变 STINGV154M (VM)，并表明这些小鼠重现了人类 根据多种标准对疾病进行诊断，包括炎症/纤维化肺病的发展。这些虚拟机老鼠 提供了一个独特的实验工具来探索细胞类型和分子机制 纤维化肺病的发生和进展。 Rag1缺陷完全拯救了双肺VM小鼠 炎症和肺纤维化表明适应性免疫在 VM ILD 中发挥着关键作用。 STING 的作用 一直专注于骨髓细胞，VM 小鼠具有扩​​大和激活的骨髓/中性粒细胞 隔间。然而，辐射嵌合体研究已经确定了抗辐射细胞在肺纤维化中的关键作用 因为用 WT 骨髓干细胞重组的受到致命辐射的 VM 小鼠出现了极其严重的肺病 即使这些嵌合体完全缺乏任何 VM 衍生的造血细胞，也会导致疾病。初步数据 暗示淋巴内皮细胞（LEC）是 VM 小鼠中 BALT 形成的引发剂。此举的目标 应用是进一步探索直接或间接激活的抗辐射先天免疫细胞 VM 突变并定义这些细胞促进肺部炎症的机制。我们的方法将 涉及：(1) 与内皮限制性 Cre 删除子杂交的新型 VM 条件 KI 系的体内分析 菌株和其他相关对照，（2）基质群体的 scRNAseq 比较，包括淋巴和 从WT和VM小鼠中分离的血管内皮细胞； (3) VM 和 WT 内皮细胞分析 VM 突变或更传统的 STING 激动剂体外激活后的小鼠。有紧急情况 需要为患有自身免疫性和自身炎症性肺部疾病的患者找到更好的治疗方法 本申请中提出的研究应该提供重要的见解，使我们能够设计出最 相关的治疗靶点。此外，这些研究提供了研究 STING 影响的机会 更广泛地讲，基质细胞类型的激活是内皮细胞中 STING 活性的一个新兴研究领域 最近，细胞与严重的新冠病毒（COVID19）的发展有关。