利用人体诱导多能干细胞(iPSC)了解 2 型重症哮喘的持续气道阻塞情况

IF 0.5 4区 医学 Q4 RESPIRATORY SYSTEM Revue des maladies respiratoires Pub Date : 2024-03-01 DOI:10.1016/j.rmr.2024.01.012
E. Ahmed , F. Foisset , C. Bourdais , A. Nasri , A. Petit , S. Assou , D. Gras , P. Chanez , I. Vachier , H. Hammad , J. Vos (De) , B. Lambrecht , A. Bourdin
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引用次数: 0

摘要

导言尽管有新的生物制剂可治疗严重哮喘的炎症,但针对气道持续阻塞的治疗仍具有挑战性。Galectin-10 嗜酸性粒细胞衍生的结晶,也称为夏科-雷登结晶(CLCs),已被描述存在于重症哮喘患者气道的粘液栓中。然而,CLCs 在粘液分泌中的直接作用尚未确定。我们假设,堵塞的气道构成了一个独特的生态位,2 型免疫细胞在此与结构细胞交流,从而使疾病长期存在。我们的目标是利用诱导多能干细胞(iPSCs)建立一个新模型。方法从2型重症哮喘患者中提取了三个人类iPSCs品系(蒙彼利埃大学医院MOSAIC研究,NCT05616338),并在气液界面(i-ALI)中分化成气道上皮细胞。健康的 iPSC 系 UHOMi002-A 作为对照。在ALI培养的第21天,iPSC衍生的气道上皮在一周内每两天接受一次IL-13(10纳克/毫升)刺激,或接受重组Gal10晶体(100纳克/毫升)、IL-13和Gal10晶体或PBS(载体)的急性刺激(24小时)。结果我们成功分化了从 T2 重症哮喘患者中产生的 iPSC 株系,并在每个发育阶段都达到了较高的纯度。通过流式细胞术定量分析C-X-C Motif趋化因子受体4(CXCR4)/c-KIT双阳性细胞和叉头框A2(FOXA2)+/SRY-框转录因子17(SOX17)+免疫标记,评估每个细胞系最终内胚层的平均细胞纯度为80%。通过转录因子 NK2 Homeobox 1(NKX2.1)表达和流式细胞术检测羧基肽酶 M(CPM),腹侧前肠内胚层(vAFE)阶段的纯度被评估为 70%。慢性 IL-13 挑战和 CLC 都能诱导哮喘 iPSC 株系中 MUC5AC+ 细胞的增加和神经内分泌细胞的增加。
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Understanding persistent airway obstruction in type 2 severe asthma using human induced pluripotent stem cells (iPSCs)

Introduction

Despite the new biologics to treat inflammation in severe asthma, targeting persistent obstruction of the airways remains challenging. Galectin-10 eosinophil derived crystals, also known as Charcot-Leyden crystals (CLCs) have been described to be present in the mucus plugs in the airways of patients with severe asthma. However, a direct role for CLCs in mucus production has not been established. We hypothesize that plugged airways constitute a unique niche where type 2 immune cells communicate with structural cells to perpetuate disease. We aimed to set up a new model using induced pluripotent stem cells (iPSCs).

Methods

Three human iPSCs lines from type 2 severe asthma patients have been derived (MOSAIC study, University Hospital of Montpellier, NCT05616338) and differentiated into airway epithelium in air–liquid interface (i-ALI). The healthy iPSC line UHOMi002-A was used as a control. At day 21 of ALI culture, iPSC derived-airway epithelia were stimulated at the apical side with either IL-13 every two days (10 ng/mL) during one week, acute stimulation (24 h) with recombinant Gal10 crystals (100 ng/mL), both IL-13 and Gal10 crystals or PBS (vehicle). We aimed to evaluate the effect on i-ALI differentiation at day 30.

Results

We successfully differentiated the iPSC lines generated from the T2 severe asthma patients, and achieving a high purity rate at each developmental stages. The mean cell purity at the definitive endoderm for each cell line was > 80% assessed by flow cytometry quantification of C-X-C Motif Chemokine Receptor 4 (CXCR4)/c-KIT double positive cells and immunolabelling of Forkhead Box A2 (FOXA2)+/SRY-box transcription factor 17 (SOX17)+. Purity for ventral anterior foregut endoderm (vAFE) stage was evaluated at 70%, through Transcription Factor NK2 Homeobox 1 (NKX2.1) expression, Carboxy Peptidase M (CPM) by flow cytometry. vAFE cells from the hiPSC lines differentiated into bronchial epithelium in air–liquid interface conditions. Chronic IL-13 challenging and CLC were both able to induce an increasing of MUC5AC+ cells and also an increase of neuroendocrine cells in asthmatic iPSC lines.

Conclusion

iALI bronchial epithelium can recapitulate T2 severe asthma features in vitro, and highlighted a possible direct effect of the CLC on the airway epithelium.

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来源期刊
Revue des maladies respiratoires
Revue des maladies respiratoires 医学-呼吸系统
CiteScore
1.10
自引率
16.70%
发文量
168
审稿时长
4-8 weeks
期刊介绍: La Revue des Maladies Respiratoires est l''organe officiel d''expression scientifique de la Société de Pneumologie de Langue Française (SPLF). Il s''agit d''un média professionnel francophone, à vocation internationale et accessible ici. La Revue des Maladies Respiratoires est un outil de formation professionnelle post-universitaire pour l''ensemble de la communauté pneumologique francophone. Elle publie sur son site différentes variétés d''articles scientifiques concernant la Pneumologie : - Editoriaux, - Articles originaux, - Revues générales, - Articles de synthèses, - Recommandations d''experts et textes de consensus, - Séries thématiques, - Cas cliniques, - Articles « images et diagnostics », - Fiches techniques, - Lettres à la rédaction.
期刊最新文献
[A survey on tobacco-free areas at hospital entrances]. [Lung function during normal pregnancy]. [Smoking cessation during pregnancy]. [Deleterious effects of heated tobacco on respiratory symptoms and lung function]. [Efficacy of biologics for severe asthma on allergic comorbidities].
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