Interferon Regulatory Factor 7 Links Interferon Pathway Activation to the Exaggerates Fibrotic Response in Systemic Sclerosis

干扰素调节因子 7 将干扰素通路激活与系统性硬化症中过度的纤维化反应联系起来

• 批准号: 10682192
• 负责人: Shervin Assassi
• 金额: $ 22.49万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10682192
• 关键词: Address; Attenuated; Autoimmune; Autoimmune Diseases; Bleomycin; Blood; Blood Cells; CRISPR/Cas technology; Cell Line; ChIP-seq; Data; Dermal; Development; Disease; Effector Cell; Fibroblasts; Fibrosis; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Gene Expression Regulation; Genetic Predisposition to Disease; Genetic Transcription; Goals; Human; Immune; Immune system; Immunity; In Vitro; Interferon Type I; Interferons; Knockout Mice; Link; Longitudinal cohort; MADH3 gene; Mediating; Modeling; Molecular; Molecular Profiling; Morbidity - disease rate; Mus; Organ; Pathogenesis; Pathologic; Pathway interactions; Patients; Publishing; RNA; Rheumatism; Role; Sampling; Scleroderma; Signal Transduction; Single Nucleotide Polymorphism; Skin; Skin Tissue; System; Systemic Scleroderma; Tissues; Transcript; Transforming Growth Factor beta; Up-Regulation; Work; conditional knockout; effective therapy; experimental study; in vivo; indium-bleomycin; insight; interferon regulatory factor-7; knock-down; mortality; mouse model; overexpression; peripheral blood; response; single-cell RNA sequencing; skin fibrosis; targeted treatment; transcription factor; transcriptome sequencing

Systemic sclerosis (SSc-scleroderma) is a multisystem autoimmune, fibrotic disease associated with high morbidity and mortality. Progress in the development of effective therapies for SSc has been hampered by a fragmented understanding of its pathogenesis. Although abundant evidence implicates dysregulated immunity in SSc, the mechanisms by which the immune system influences fibroblast function are not well-understood. We propose herein a multifaceted approach to elucidate the role of interferon regulatory factor 7 (IRF7) in dermal fibroblasts as a pathologic bridge between immune dysregulation and fibrosis in SSc. An interferon (IFN) gene expression signature is the most prominent peripheral blood cell transcript profile in SSc. Moreover, our large-scale gene expression studies have identified IRF7, a key transcription factor in the type I IFN pathway, as the top predicted upstream regulator of the SSc molecular profile in both skin and blood, as well as a prominent regulator of SSc fibroblasts. Our recently published work showed that IRF7 is significantly upregulated and activated in SSc skin and explanted dermal fibroblasts, and that type I IFN upregulates IRF7 expression in fibroblasts. IRF7 interacts with SMAD3 and potentiates TGFβ induced fibrosis signaling in fibroblasts. Moreover, global Irf7 knockdown attenuates dermal fibrosis in two murine dermal fibrosis models. Lastly, our recent preliminary data suggest fibroblast specific Irf7 knockdown might be sufficient for attenuating the bleomycin induced dermal fibrosis. Herein, we hypothesize that IRF7 links type I IFN pathway to the fibrotic response by potentiating the TGFβ signaling in fibroblasts which are the primary effector cells in SSc. Our primary goal is to elucidate the role of type I IFN induced IRF7 upregulation in potentiating the TGFβ canonical pathway via IRF7 interaction with SMAD3 in dermal fibroblasts. The following Specific Aims will be pursued: Aim 1: Define the fibroblast specific contribution of Irf7 depletion in murine dermal fibrosis models. The impact of fibroblast specific Irf7 depletion on dermal fibrosis in bleomycin induced and Tsk1 dermal fibrosis models will be investigated. Aim 2: Elucidate the functional effects of IRF7 on SMAD3 mediated transcriptional activity and gene expression regulation Human fibroblast cell lines with IRF7 over- expression and deletion will be generated and TGFβ-mediated SMAD3 transcriptional targets and gene expression regulation will be characterized by ChIP- and RNA-sequencing. Aim 3: Delineate the relationship between the peripheral blood cell interferon signature and IRF7 upregulation at the end-organ level in patients with SSc. The relationship between the peripheral blood IFN signature and IRF7 expression in SSc skin tissue and dermal fibroblast subpopulations will be characterized using bulk and single cell RNA sequencing. Cumulatively, this proposal can elucidate a key mechanism by which immune dysregulation leading to IRF7 activation potentiates the fibrotic response in SSc. Ultimately, this can open the door to identification of more targeted and effective treatment options for this potentially devastating disease.

全身性硬化症（SSC-Scleroderma）是一种多系统自身免疫性，纤维化疾病，与高有关 发病率和死亡率。 SSC有效疗法开发的进展已受到A的阻碍 对其发病机理的理解分散。尽管大量证据实现了免疫抑制失调的失调 在SSC中，免疫系统影响成纤维细胞功能的机制无法理解。我们 此处的提案在阐明干扰素调节因子7（IRF7）在皮肤中的作用的多方面方法 成纤维细胞作为免疫失调与SSC纤维化之间的病理桥梁。 干扰素（IFN）基因表达特征是最突出的外周血细胞转录本谱。 SSC。此外，我们的大型基因表达研究已经确定了IRF7，这是一个关键的转录因子 I型IFN途径，作为皮肤和血液中SSC分子剖面的上游调节剂的最高途径， 以及SSC成纤维细胞的突出调节剂。我们最近发表的工作表明，IRF7显着 在SSC皮肤上上调和激活 成纤维细胞中的表达。 IRF7与SMAD3相互作用，电势TGFβ诱导的纤维化信号传导 成纤维细胞。此外，在两个鼠皮肤纤维化模型中，全球IRF7敲低可减轻皮肤纤维化。 最后，我们最近的初步数据表明成纤维细胞特定的IRF7敲低可能足以减弱 博来霉素诱导皮肤纤维化。在此，我们假设IRF7链接I型IFN途径与纤维化 通过在成纤维细胞中潜在的TGFβ信号传导的反应，这是SSC中主要效应细胞的反应。我们的主要 目标是阐明I型IFN引起的IRF7上调在潜在TGFβ规范中的作用 通过IRF7与SMAD3相互作用的途径。将追求以下具体目标： 目标1：定义鼠皮肤纤维化模型中IRF7耗竭的成纤维细胞特异性贡献。这 成纤维细胞特异性IRF7部署对博来霉素诱导的皮肤纤维化和TSK1真皮纤维化的影响 模型将进行研究。目标2：阐明IRF7对SMAD3介导的功能效应 转录活性和基因表达调节具有IRF7过度的人成纤维细胞系 将产生表达和缺失，TGFβ介导的SMAD3转录靶标和基因 表达调节将以芯片和RNA测序为特征。目标3：描述关系 在最终器官水平的外周血细胞干扰特征和IRF7上调之间 患有SSC的患者。 SSC中外周血IFN签名和IRF7表达之间的关系 使用大量和单细胞RNA测序表征皮肤组织和皮肤成纤维细胞亚群。 累积地，该提案可以阐明一种关键机制，通过该机制，免疫失调导致IRF7 激活电位SSC中的纤维化反应。最终，这可以打开识别更多的大门 针对这种潜在毁灭性疾病的有针对性和有效的治疗选择。