CFlm25 mediated alternative polyadenylation regulates fibrosis in systemic sclerosis

CFlm25介导的选择性多聚腺苷酸化调节系统性硬化症中的纤维化

• 批准号: 10616484
• 负责人: Shervin Assassi
• 金额: $ 33.98万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-05 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10616484
• 关键词: 3' Untranslated Regions; Address; Affect; Binding Sites; Biology; Bleomycin; Cells; Clinical; Collagen; Cutaneous sclerosis; Data; Deposition; Dermal; Diffuse Scleroderma; Disease; Distal; Down-Regulation; Event; Extracellular Matrix Protein Gene; Extracellular Matrix Proteins; FDA approved; Fibroblasts; Fibrosis; Genes; Goals; Human; In Vitro; Length; Link; Mediating; MicroRNAs; Modeling; Morbidity - disease rate; Mus; Myofibroblast; Names; Organ; Pathogenesis; Pathway interactions; Patients; Persons; Pharmaceutical Preparations; Poly A; Poly(A) Tail; Polyadenylation; Production; Profibrotic signal; Prognosis; RNA; RNA Processing; Regulation; Repression; Research; Rheumatism; Sampling; Scleroderma; Site; Skin; Standardization; System; Systemic Scleroderma; Tail; Technology; Therapeutic; Time; Transcript; Transforming Growth Factor beta; Translations; Up-Regulation; Work; cleavage factor; cytokine; effective therapy; experimental study; gene repression; improved; indium-bleomycin; knock-down; mRNA Translation; mortality; mouse model; new therapeutic target; novel; protein expression; response; skin fibrosis; therapeutic target; transcriptome sequencing; translational study; tsk mouse

Systemic sclerosis (SSc) is a multi-system, fibrotic disease that affects the skin and a variety of internal organs. Persistent myofibroblast activation and associated excessive extracellular matrix protein (ECMs) deposition are hallmarks of the disease. The mechanisms that account for this excessive fibrotic response remain elusive. Despite its high mortality and morbidity, there are no FDA approved medications for fibrotic complications of SSc. Our recent work indicates that a RNA-processing mechanism known as alternative polyadenylation (APA) is critical for the upregulation of profibrotic genes in dermal fibrosis through 3’ UTR tail shortening of key transcripts. Recent evidence demonstrated that a key regulator of APA is Cleavage factor Im 25 (CFIm25). The CFIm25 deletion leads to 3’ UTR shortening. A truncated 3’ UTR tail will often lack microRNA binding sites and evades the microRNA-mediated gene repression. Our preliminary data indicate CFlm25 is downregulated in fibrotic skin of SSc patients and in murine dermal fibrosis models. Knockdown of CFIm25 in normal skin fibroblasts is sufficient to promote the 3’ UTR shortening of key profibrotic genes. Moreover, the central fibrotic cytokine TGFβ suppresses CFIm25 expression through miR-203 upregulation. Overall, our data uncovered a novel mechanism by which TGFβ mediated CFIm25 downregulation leads to 3’ UTR shortening and the over-production of profibrotic factors and ECMs, and contributes to the pathogenesis of SSc. Based on this preliminary data, the main hypothesis of this project is that TGFβ and miR-203 downregulate CFIm25 in fibroblasts, resulting in dermal fibrosis by upregulation of profibrotic genes and ECMs through 3’ UTR shortening. This hypothesis will be examined in the following specific aims: Aim I: Define the fibroblast specific contribution of CFIm25 depletion in dermal fibrosis murine models. This aim will elucidate the downstream effects of CFIm25 depletion on key fibrotic pathways. Aim II: Determine the mechanisms for CFIm25 downregulation and assess their potential as therapeutic targets in dermal fibrosis. The mechanisms for TGFβ mediated miR-203 downregulation and subsequent CFIm25 repression by miR-203 will be elucidated. This aim will characterize the upstream events leading to CFIm25 depletion and will identify potential therapeutic targets. Aim III: Characterize CFIm25 in SSc human skin/fibroblasts and identify fibrotic genes dysregulated by APA. Serial dermal fibroblasts and skin samples from patients with early SSc and matched controls will be examined using a novel RNA sequencing technology. This aim will provide for the first time an unbiased, longitudinal view of CFIm25 mediated APA profile in a fibrotic disease. The proposed research links for the first time the CFIm25 mediated 3’ UTR shortening to dermal fibrosis. This can lead to discovering a key mechanism that amplifies the fibrotic response in SSc and ultimately to identifying an entirely novel target for treatment of persons with this potentially devastating disease.

系统性硬化症 (SSc) 是一种多系统纤维化疾病，影响皮肤和多种内部器官 持续的肌成纤维细胞激活和相关的细胞外基质蛋白（ECM）过多。 沉积是该疾病的标志，其机制导致了这种过度的纤维化反应。 尽管其死亡率和发病率很高，但目前还没有 FDA 批准的治疗纤维化的药物。 我们最近的工作表明，一种称为替代的 RNA 加工机制。 多腺苷酸化 (APA) 对于通过 3' UTR 尾上调真皮纤维化中的促纤维化基因至关重要 最近的证据表明 APA 的关键调节因子是切割因子 Im。 25 (CFIm25)。CFIm25 缺失会导致 3’UTR 缩短，3’UTR 尾部通常会缺失。 我们的初步数据表明，microRNA 结合位点并逃避 microRNA 介导的基因抑制。 CFlm25 在 SSc 患者的纤维化皮肤和小鼠真皮纤维化模型中下调。 正常皮肤成纤维细胞中的 CFIm25 足以促进关键促纤维化基因的 3' UTR 缩短。 此外，中枢纤维化细胞因子 TGFβ 通过上调 miR-203 抑制 CFIm25 表达。 总体而言，我们的数据揭示了一种新机制，TGFβ 介导 CFIm25 下调导致 3' UTR 缩短以及促纤维化因子和 ECM 的过度产生，并有助于发病机制 基于该初步数据，该项目的主要假设是 TGFβ 和 miR-203。 下调成纤维细胞中的 CFIm25，通过上调促纤维化基因导致真皮纤维化 以及通过 3' UTR 缩短实现的 ECM。 该假设将在以下具体目标中进行检验： 目标 I：确定真皮纤维化小鼠模型中 CFIm25 耗竭对成纤维细胞的特异性贡献。 该目标将阐明 CFIm25 耗竭对关键纤维化途径的下游影响： 确定 CFIm25 下调机制并评估其治疗潜力 TGFβ 介导的 miR-203 下调及其后续机制。 将阐明 miR-203 对 CFIm25 的抑制作用，这一目标将表征导致 CFIm25 的上游事件。 CFIm25 耗尽并将确定潜在的治疗靶点。目标 III：表征 SSc 人类中的 CFIm25。 皮肤/成纤维细胞并识别 APA 失调的纤维化基因。 来自早期 SSc 患者和匹配对照的样本将使用新型 RNA 测序进行检查 这一目标将首次提供 CFIm25 介导的 APA 的公正、纵向视图。 纤维化疾病的概况。 拟议的研究首次将 CFIm25 介导的 3' UTR 缩短与真皮纤维化联系起来。 可以导致发现放大 SSc 纤维化反应的关键机制，并最终 一个全新的目标，用于识别患有这种潜在破坏性疾病的人。

项目成果

Fibroblast Subpopulations in Systemic Sclerosis: Functional Implications of Individual Subpopulations and Correlations with Clinical Features.

系统性硬化症中的成纤维细胞亚群：个体亚群的功能意义以及与临床特征的相关性。

• 发表时间: 2023-12-24
• 作者: Zhu, Honglin;Luo, Hui;Skaug, Brian;Tabib, Tracy;Li, Yi;Tao, Yongguang;Matei, Alexandru;Lyons, Marka A;Schett, Georg;Lafyatis, Robert;Assassi, Shervin;Distler, Jörg H W
• 通讯作者: Distler, Jörg H W

Comprehensive analysis of the major histocompatibility complex in systemic sclerosis identifies differential HLA associations by clinical and serological subtypes.

对系统性硬化症中主要组织相容性复合体的综合分析通过临床和血清学亚型确定了不同的 HLA 关联。

• 发表时间: 2021-08
• 影响因子: 27.4
• 作者: Acosta;Kerick, Martin;Lopéz;Assassi, Shervin;Beretta, Lorenzo;Simeón;Ortego;International SSc Group;Proudman, Susanna M;Australian Scleroderma Interest Group (ASIG);Hunzelmann
• 通讯作者: Hunzelmann

False positive anti-Topoisomerase I (Scl-70) antibody results in clinical practice: A case series from a scleroderma referral center.

临床实践中抗拓扑异构酶 I (Scl-70) 抗体的假阳性结果：来自硬皮病转诊中心的病例系列。

• 发表时间: 2022-10
• 影响因子: 5
• 作者: Lam, Brian H;Assassi, Shervin;Charles, Julio;Taherian, Rana;Lyons, Marka A;Jandali, Bochra;Mayes, Maureen D;Skaug, Brian
• 通讯作者: Skaug, Brian

Reduced digestion of circulating genomic DNA in systemic sclerosis patients with the DNASE1L3 R206C variant.

DNASE1L3 R206C 变异的系统性硬化症患者循环基因组 DNA 的消化减少。

• 发表时间: 2023-09-01
• 作者: Skaug, Brian;Guo, Xinjian;Li, Yuanteng Jeff;Charles, Julio;Pham, Kay T;Couturier, Jacob;Lewis, Dorothy E;Bracaglia, Claudia;Caiello, Ivan;Mayes, Maureen D;Assassi, Shervin
• 通讯作者: Assassi, Shervin

Interferon regulatory factor 7 (IRF7) represents a link between inflammation and fibrosis in the pathogenesis of systemic sclerosis.

干扰素调节因子 7 (IRF7) 代表系统性硬化症发病机制中炎症和纤维化之间的联系。

• 发表时间: 2019
• 影响因子: 27.4
• 作者: Wu, Minghua;Skaug, Brian;Bi, Xiongjie;Mills, Tingting;Salazar, Gloria;Zhou, Xiaodong;Reveille, John;Agarwal, Sandeep K;Blackburn, Michael R;Mayes, Maureen D;Assassi, Shervin
• 通讯作者: Assassi, Shervin