USP31 promotes the inflammatory response of human lung organoids to lipopolysaccharide treatment.

IF 2.7 3区医学 Q2 CRITICAL CARE MEDICINE

SHOCK Pub Date : 2024-08-28 DOI:10.1097/SHK.0000000000002464

Wenbiao Xiao, Bingwen Lin, Ying Wang, Peng Huang, Jiandong Lin, Xiongjian Xiao

{"title":"USP31 promotes the inflammatory response of human lung organoids to lipopolysaccharide treatment.","authors":"Wenbiao Xiao, Bingwen Lin, Ying Wang, Peng Huang, Jiandong Lin, Xiongjian Xiao","doi":"10.1097/SHK.0000000000002464","DOIUrl":null,"url":null,"abstract":"Background: Acute lung injury (ALI) is a severe condition characterized by a high mortality rate, driven by an uncontrolled inflammatory response. Emerging evidence has underscored the crucial role of the ubiquitin system in ALI. However, due to their vast number, the specific functions of individual ubiquitination regulators remain unclear.Materials and methods: In this study, we established human lung organoids (HLOs) derived from human embryonic stem cells and subjected them to lipopolysaccharide (LPS) treatment to induce an inflammatory response, mimicking ALI. Subsequently, we detected the expression of inflammatory cytokines, including tumour necrosis factor alpha (TNF-α), interleukin 6, interleukin 18 (IL-18), and interleukin-1β (IL-1β), by qPCR experiments. We also detected changes in the mRNA expression of several USPs before and after HLOs treatment and thus screened for USPs that had significant changes in HLOs after LPS stimulation. After screening for USP, we silenced the USP in HLOs and then subjected them to LPS treatment, and TNF-α, IL-6, IL-18, and IL-1β expressions were detected using qPCR assays. Meanwhile, Western blot was used to detect changes in NOD-, LRR- and pyrin domain-containing 3 (NLRP3) and apoptosis-associated Speck-like protein containing a CARD (ASC) protein level in HLOs.Results: Through screening the expression of 40 ubiquitin-specific proteases (USPs), which are responsible for removing ubiquitination, we identified several USPs that exhibited differential expression in LPS-treated HLOs compared to untreated HLOs. Notably, USP31 emerged as the most significantly upregulated USP, and the knockdown of USP31 markedly attenuated the inflammatory response of HLOs to LPS treatment.Conclusions: USP31 may play a facilitating role in the inflammatory response during ALI.","PeriodicalId":21667,"journal":{"name":"SHOCK","volume":" ","pages":""},"PeriodicalIF":2.7000,"publicationDate":"2024-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"SHOCK","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1097/SHK.0000000000002464","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CRITICAL CARE MEDICINE","Score":null,"Total":0}

引用次数: 0

Abstract

Background: Acute lung injury (ALI) is a severe condition characterized by a high mortality rate, driven by an uncontrolled inflammatory response. Emerging evidence has underscored the crucial role of the ubiquitin system in ALI. However, due to their vast number, the specific functions of individual ubiquitination regulators remain unclear.

Materials and methods: In this study, we established human lung organoids (HLOs) derived from human embryonic stem cells and subjected them to lipopolysaccharide (LPS) treatment to induce an inflammatory response, mimicking ALI. Subsequently, we detected the expression of inflammatory cytokines, including tumour necrosis factor alpha (TNF-α), interleukin 6, interleukin 18 (IL-18), and interleukin-1β (IL-1β), by qPCR experiments. We also detected changes in the mRNA expression of several USPs before and after HLOs treatment and thus screened for USPs that had significant changes in HLOs after LPS stimulation. After screening for USP, we silenced the USP in HLOs and then subjected them to LPS treatment, and TNF-α, IL-6, IL-18, and IL-1β expressions were detected using qPCR assays. Meanwhile, Western blot was used to detect changes in NOD-, LRR- and pyrin domain-containing 3 (NLRP3) and apoptosis-associated Speck-like protein containing a CARD (ASC) protein level in HLOs.

Results: Through screening the expression of 40 ubiquitin-specific proteases (USPs), which are responsible for removing ubiquitination, we identified several USPs that exhibited differential expression in LPS-treated HLOs compared to untreated HLOs. Notably, USP31 emerged as the most significantly upregulated USP, and the knockdown of USP31 markedly attenuated the inflammatory response of HLOs to LPS treatment.

Conclusions: USP31 may play a facilitating role in the inflammatory response during ALI.

查看原文本刊更多论文

USP31 促进人肺器官组织对脂多糖处理的炎症反应。

背景：急性肺损伤（ALI）是一种严重的疾病，其特点是炎症反应失控，死亡率很高。新的证据强调了泛素系统在急性肺损伤中的关键作用。然而，由于泛素数量庞大，个别泛素化调节因子的具体功能仍不清楚：在本研究中，我们建立了由人类胚胎干细胞衍生的人肺器官组织（HLOs），并将其置于脂多糖（LPS）处理中诱导炎症反应，模拟 ALI。随后，我们通过 qPCR 实验检测了炎症细胞因子的表达，包括肿瘤坏死因子α（TNF-α）、白细胞介素 6、白细胞介素 18（IL-18）和白细胞介素-1β（IL-1β）。我们还检测了几种 USP 在 HLOs 处理前后 mRNA 表达的变化，从而筛选出在 LPS 刺激后 HLOs 中有显著变化的 USP。在筛选出 USP 后，我们在 HLOs 中沉默了 USP，然后对其进行 LPS 处理，并使用 qPCR 检测 TNF-α、IL-6、IL-18 和 IL-1β 的表达。同时，采用Western印迹法检测HLOs中NOD-、LRR-和含吡咯啉结构域的3（NLRP3）和含CARD的凋亡相关斯贝克样蛋白（ASC）蛋白水平的变化：结果：通过筛查负责消除泛素化的40种泛素特异性蛋白酶（USPs）的表达，我们发现了几种USPs，与未处理的HLOs相比，它们在LPS处理的HLOs中表现出不同的表达。值得注意的是，USP31 是上调最明显的 USP，敲除 USP31 能明显减轻 HLOs 对 LPS 处理的炎症反应：结论：USP31 可能在 ALI 期间的炎症反应中起到促进作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

SHOCK 医学-外科

CiteScore

6.20

自引率

3.20%

发文量

199

审稿时长

1 months

期刊介绍： SHOCK®: Injury, Inflammation, and Sepsis: Laboratory and Clinical Approaches includes studies of novel therapeutic approaches, such as immunomodulation, gene therapy, nutrition, and others. The mission of the Journal is to foster and promote multidisciplinary studies, both experimental and clinical in nature, that critically examine the etiology, mechanisms and novel therapeutics of shock-related pathophysiological conditions. Its purpose is to excel as a vehicle for timely publication in the areas of basic and clinical studies of shock, trauma, sepsis, inflammation, ischemia, and related pathobiological states, with particular emphasis on the biologic mechanisms that determine the response to such injury. Making such information available will ultimately facilitate improved care of the traumatized or septic individual.