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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

基本信息

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
项目类别：
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&apos 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

项目摘要

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器官。持续的肌成纤维细胞激活和相关的过量细胞外基质蛋白（ECM）沉积是该病的标志。造成这种过度纤维化反应的机制仍然难以捉摸。尽管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 SSc 的并发症。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缩短关键转录本。最近的证据表明 APA 的一个关键调节因子是裂解因子 Im 25（CFIm25）。 CFIm25 缺失导致 3' UTR 缩短。截断的 3' UTR 尾部通常会缺失 microRNA 结合位点并逃避 microRNA 介导的基因抑制。我们的初步数据表明 CFlm25 在 SSc 患者的纤维化皮肤和小鼠真皮纤维化模型中下调。击倒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，并有助于发病机制科学科学士。基于这些初步数据，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和 ECM 通过 3' UTR 缩短。该假设将在以下具体目标中得到检验：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。这一目标将阐明 CFIm25 耗竭对关键纤维化途径的下游影响。目标二：确定 CFIm25 下调机制并评估其治疗潜力真皮纤维化的目标。 TGFβ介导的miR-203下调及其后续机制将阐明 miR-203 对 CFIm25 的抑制。该目标将描述导致的上游事件 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。连续真皮成纤维细胞和皮肤来自早期 SSc 患者和匹配对照的样本将使用新型 RNA 测序进行检查技术。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。这可以导致发现放大 SSc 纤维化反应的关键机制，并最终确定一个全新的目标来治疗患有这种潜在破坏性疾病的人。