Characterization of patient-derived intestinal organoids for modelling fibrosis in Inflammatory Bowel Disease.

IF 4.8 3区 医学 Q2 CELL BIOLOGY Inflammation Research Pub Date : 2024-08-01 Epub Date: 2024-06-06 DOI:10.1007/s00011-024-01901-9
Ilaria Laudadio, Claudia Carissimi, Noemi Scafa, Alex Bastianelli, Valerio Fulci, Alessandra Renzini, Giusy Russo, Salvatore Oliva, Roberta Vitali, Francesca Palone, Salvatore Cucchiara, Laura Stronati
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Abstract

Background and aims: Intestinal fibrosis is a common complication of Inflammatory Bowel Disease (IBD), namely Crohn's disease (CD) and ulcerative colitis (UC), but the precise mechanism by which it occurs is incompletely understood hampering the development of effective therapeutic strategies. Here, we aimed at inducing and characterizing an inflammation-mediated fibrosis in patient-derived organoids (PDOs) issued from crypts isolated from colonic mucosal biopsies of IBD pediatric patients and age matched-control subjects (CTRLs).

Methods: Inflammatory-driven fibrosis was induced by exposing CTRL-, CD- and UC-PDOs to the pro-inflammatory cytokine TNF-α for one day, followed by a co-treatment with TNF-α and TGF-β1 for three days. Fibrotic response was proven by analyzing inflammatory and fibrotic markers by RT-qPCR and immunofluorescence. Transcriptomic changes were assessed by RNA-sequencing.

Results: Co-treatment with TNF-α and TGF-β1 caused in CTRL- and IBD-PDOs morphological changes towards a mesenchymal-like phenotype and up-regulation of inflammatory, mesenchymal, and fibrotic markers. Transcriptomic profiling highlighted that in all intestinal PDOs, regardless of the disease, the co-exposure to TNF-α and TGF-β1 regulated EMT genes and specifically increased genes involved in positive regulation of cell migration. Finally, we demonstrated that CD-PDOs display a specific response to fibrosis compared to both CTRL- and UC-PDOs, mainly characterized by upregulation of nuclear factors controlling transcription.

Conclusions: This study demonstrates that intestinal PDOs may develop an inflammatory-derived fibrosis thus representing a promising tool to study fibrogenesis in IBD. Fibrotic PDOs show increased expression of EMT genes. In particular, fibrotic CD-PDOs display a specific gene expression signature compared to UC and CTRL-PDOs.

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用于炎症性肠病纤维化建模的患者来源肠器官组织的特征。
背景和目的:肠纤维化是炎症性肠病(IBD),即克罗恩病(CD)和溃疡性结肠炎(UC)的一种常见并发症,但人们对其发生的确切机制了解甚少,这阻碍了有效治疗策略的开发。在这里,我们的目的是诱导并描述由患者衍生的有机体(PDOs)中由炎症介导的纤维化,该有机体来自从 IBD 儿科患者和年龄匹配对照组(CTRLs)结肠粘膜活检组织中分离出的隐窝:方法:将CTRL、CD和UC-PDOs暴露于促炎细胞因子TNF-α一天,然后用TNF-α和TGF-β1联合处理三天,诱导炎症驱动的纤维化。通过 RT-qPCR 和免疫荧光分析炎症和纤维化标记物,证实了纤维化反应。RNA测序评估了转录组的变化:结果:TNF-α和TGF-β1的联合治疗导致CTRL-和IBD-PDOs形态发生变化,形成间充质样表型,炎症、间充质和纤维化标志物上调。转录组图谱分析显示,在所有肠道 PDO 中,无论疾病如何,同时暴露于 TNF-α 和 TGF-β1 会调节 EMT 基因,特别是增加参与细胞迁移正向调节的基因。最后,我们证实,与CTRL-和UC-PDOs相比,CD-PDOs对纤维化表现出特殊的反应,主要特征是控制转录的核因子上调:本研究表明,肠道 PDOs 可能会发生炎症衍生的纤维化,因此是研究 IBD 纤维形成的一种很有前途的工具。纤维化的 PDOs 显示出 EMT 基因表达的增加。与UC和CTRL-PDOs相比,纤维化的CD-PDOs尤其显示出特殊的基因表达特征。
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来源期刊
Inflammation Research
Inflammation Research 医学-免疫学
CiteScore
9.90
自引率
1.50%
发文量
134
审稿时长
3-8 weeks
期刊介绍: Inflammation Research (IR) publishes peer-reviewed papers on all aspects of inflammation and related fields including histopathology, immunological mechanisms, gene expression, mediators, experimental models, clinical investigations and the effect of drugs. Related fields are broadly defined and include for instance, allergy and asthma, shock, pain, joint damage, skin disease as well as clinical trials of relevant drugs.
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