GSDMD-dependent IL-1 signaling in intestinal inflammation
GSDMD-dependent IL-1 signaling in intestinal inflammation
批准号:
10223160
负责人:
Theresa Torres Pizarro
金额:
$54.17万
依托单位国家:
美国
项目类别:
财政年份:
2020
资助国家:
美国
项目状态:
未结题
起止时间:
2020-07-24 至 2025-06-30
关键词:
AblationAddressAttenuatedAutomobile DrivingBinding ProteinsBiochemicalBiochemistryBiological AssayBiopsyCASP1 geneCASP8 geneCDC37 geneCDC37 homolog proteinCell physiologyCellsCellular StructuresChronicColitisColonComplementComplexCrohn&aposs diseaseDataDiseaseElectron MicroscopyEpithelial CellsEtiologyEventExtracellular SpaceFamily memberFinancial compensationGenetic PolymorphismHelper-Inducer T-LymphocyteIL1R1 geneImmuneIn VitroInflammasomeInflammationInflammatoryInflammatory Bowel DiseasesInterleukin-1Interleukin-1 betaIntestinal SecretionsIntestinesKnowledgeLengthLipid BindingLiteratureMapsMediatingModelingMolecularMolecular ChaperonesMusMutationMyeloid CellsNonlyticPathogenesisPathogenicityPathway interactionsPatientsPeptide Signal SequencesPolyubiquitinationPredispositionPreventive measureProcessProductionProteinsProteomicsRegulationRoleSecretory VesiclesSignal TransductionSodium Dextran SulfateSourceStimulusStructureT-LymphocyteTCR ActivationTestingTissuesUbiquitinationUlcerative ColitisUnited StatesVesicleanakinrabasecell typecytokinedesigndextran sulfate sodium induced colitiseffective therapyextracellular vesiclesin vivoinflammatory disease of the intestineinsightintestinal epitheliummouse modelnovelnovel therapeutic interventionprogramspublic health relevancerapid growthreceptorrecruitresponsetargeted treatmenttherapeutic targetubiquitin-protein ligasevesicular release
中文摘要
摘要:
在美国,大约有130万人患有IBD(慢性肠炎)。病因学
IBD的发病率仍然难以捉摸,也没有预防措施或治愈方法。炎性小体及其衍生
细胞因子IL-1β作为炎症性肠病中异常调节的信号中枢正被广泛研究
疾病(IBD)。IL-1β是以无分泌信号序列的非活性前体形式合成的。最新研究
发现一种脂质结合蛋白GasderminD(GSDMD)是释放IL-1β所必需的
Caspase-1/11炎症体激活。这一突破导致了以文学为中心的文学的迅速发展
GSDMD在髓系细胞中的孔道形成和相关的嗜热活性。有趣的是,我们发现了一本小说
GSDMD在引导肠上皮细胞释放含有IL-1β的囊泡中的非解热作用
T细胞对Caspase 8(Caspase8)的反应,但对CASP1的激活无反应。通过无偏见的蛋白质组分析,我们
在肠上皮细胞中发现了一组新的GSDMD相互作用蛋白,包括NEDD4(一个E3
连接酶)和Hsp90辅助伴侣CDC37。消融GSDMD或NEDD4可消除内毒素和三磷酸腺苷诱导的
IECS生产IL-1β。值得注意的是,LPSATP刺激导致前IL-1β的多泛素化,这是
分泌并加工成成熟的IL-1β以及含有全长GSDMD、Hsp90/CDC37、
NEDD4、ATG7、Casp8,但不包括CASP1。体外泛素化实验表明,已知相互作用的NEDD4
与Lc3结合,促进货物装入分泌囊泡,催化前IL-1β的多泛素化。的确,
而GSDMD与LC3囊泡有关;GSDMD依赖于细胞外囊泡的释放(<;200
NM),其中含有GSDMD/NEDD4/IL-1β复合体。此外,
GSDMD的Asp276的失活突变,取消了其成孔和焦链活性,但没有
影响GSDMD诱导的IECs分泌IL-1β。在TH17细胞中,高度相关的辅助性T细胞亚群
肠道炎症,这种由GSDMD引导的多泛素化前IL-1β复合体(GSDMD,Hsp90,
Casp8和NEDD4)也很容易在ATP刺激和TCR激活时检测到。总而言之,
这些数据揭示了GSDMD在非髓系细胞释放IL-1β中的一种新的非嗜热性作用。致病的
在两种小鼠肠道炎症模型中,研究了GSDMD引导的IL-1β释放的作用。首先,
而多泛素化的前IL-1β、mIL-1β和GSDMD/NEDD4-分泌复合体在
葡聚糖硫酸钠(DSS)诱导的结肠炎时肠道组织中GSDMD依赖的方式,
GSDMD缺乏可减轻肠道炎症。第二,幼稚T细胞GSDMD或IL-1β缺乏
降低了它们引发肠道炎症的能力。基于这些发现,我们假设GSDMD
介导非髓系细胞和髓系细胞释放IL-1β的不同途径(引导分泌和下垂),
它们共同作用于肠道炎症的发病机制。我们将对此进行测试
通过以下目的提出假说:(1)探讨GSDMD诱导分泌的分子机制
IL-1β释放途径;(2)探讨GSDMD-IL-1β轴的细胞类型和途径特异性作用。
肠道炎症,包括GSDMD介导的嗜热性与GSDMD引导的IL-1β的分泌
肠道发炎。
英文摘要
Abstract:
About 1.3 million people suffer from IBD (chronic inflammation of the intestine) in the United States. The etiology
of IBD remains elusive and preventive measures or a cure are not available. Inflammasomes and derived
cytokines IL-1β are being intensely investigated as the signaling hub that is dys-regulated in inflammatory bowel
disease (IBD). IL-1β is synthesized as inactive pro-forms with no secretory signal sequence. Recent studies
have found that a lipid binding protein, Gasdermin D (GSDMD), is required for release of IL-1β in response to
caspase-1/11 inflammasome activation. This breakthrough led to a rapid growth of literature that focuses on the
pore forming and associated pyroptotic activity of GSDMD in myeloid cells. Interestingly, we discovered a novel
nonpyrototic role of GSDMD in guiding the release of IL-1β containing vesicles from intestinal epithelia cells and
T cells in response to Caspase 8 (Casp8) but not casp1 activation. Through unbiased proteomic analysis, we
identified a set of novel GSDMD-interacting proteins in intestinal epithelial cells (IECs), including NEDD4 (an E3
ligase) and the Hsp90 co-chaperone CDC37. Ablation of GSDMD or NEDD4 abolished LPS and ATP-induced
IL-1β production from IECs. Strikingly, LPS+ATP stimulation led to the polyubiquitination of pro-IL-1β, which was
secreted and processed into mature IL-1β along with a complex containing full-length GSDMD, Hsp90/CDC37,
NEDD4, Atg7, Casp8 but not Casp1. In vitro ubiquitination assay demonstrated that NEDD4, known to interact
with LC3 and promote cargo loading into secretory vesicles, catalyzed the polyubiquitination of pro-IL-1β. Indeed,
while GSDMD was associated with LC3+ vesicles; GSDMD-dependent release of extracellular vesicles (< 200
nm) were detected by electron microscopy, which contains the GSDMD/NEDD4/IL-1β complex. Moreover,
inactivating mutation in the Asp276 of GSDMD, which abolishes its pore-forming and pyroptotic activity, did not
impact the GSDMD-guided IL-1β secretion from IECs. In TH17 cells, a T helper cell subset highly relevant to
intestinal inflammation, this GSDMD-guided secretion of polyubiquitinated pro-IL-1β complex (GSDMD, Hsp90,
Casp8 and NEDD4) was also readily detected in response to ATP stimulation and TCR activation. Collectively,
the data revealed a novel nonpyroptotic role for GSDMD in IL-1β release from non-myeloid cells. The pathogenic
role of the GSDMD-guided IL-1β release was investigated in two mouse models of intestinal inflammation. Firstly,
while polyubiquitinated pro-IL-1β, mIL-1β and the GSDMD/NEDD4-secretary complex were induced in a
GSDMD-dependent manner in the intestinal explants in response to dextran sodium sulfate (DSS)-induce colitis,
GSDMD-deficiency attenuated the intestinal inflammation. Secondly, GSDMD or IL-1β deficiency in naïve T cells
reduced their ability to elicit intestinal inflammation. Based on these findings, we hypothesize that the GSDMD
mediates distinct pathways (guided secretion and pyroptosis) for IL-1β release in non-myeloid and myeloid cells,
which jointly contribute to the pathogenesis of intestinal inflammation in a coordinated manner. We will test this
hypothesis through the following aims: (1) Investigate the molecular mechanism for GSDMD-guided secretory
pathway for IL-1β release; (2) Investigate the cell-type and pathway-specific role of GSDMD-IL-1β axis in
intestinal inflammation, including GSDMD-mediated pyroptotic versus GSDMD-guided IL-1β secretion on the
intestinal inflammation.
期刊论文(0)
专著(0)
科研奖励(0)
会议论文
The pathogenic roles of GSDMD-dependent gut epithelium extracellular vesicles in IBD
-
批准号:10853519
-
项目类别:
-
资助金额:$4.4万
-
财政年份:2023
-
负责人:Theresa Torres Pizarro
-
依托单位:
The pathogenic roles of GSDMD-dependent gut epithelium extracellular vesicles in IBD
-
批准号:10386894
-
项目类别:
-
资助金额:$42.05万
-
财政年份:2021
-
负责人:Theresa Torres Pizarro
-
依托单位:
The pathogenic roles of GSDMD-dependent gut epithelium extracellular vesicles in IBD
-
批准号:10599251
-
项目类别:
-
资助金额:$42.05万
-
财政年份:2021
-
负责人:Theresa Torres Pizarro
-
依托单位:
The pathogenic roles of GSDMD-dependent gut epithelium extracellular vesicles in IBD
-
批准号:10211603
-
项目类别:
-
资助金额:$42.05万
-
财政年份:2021
-
负责人:Theresa Torres Pizarro
-
依托单位:
GSDMD-dependent IL-1 signaling in intestinal inflammation
-
批准号:10654589
-
项目类别:
-
资助金额:$54.17万
-
财政年份:2020
-
负责人:Theresa Torres Pizarro
-
依托单位:
GSDMD-dependent IL-1 signaling in intestinal inflammation
-
批准号:10441357
-
项目类别:
-
资助金额:$54.17万
-
财政年份:2020
-
负责人:Theresa Torres Pizarro
-
依托单位:
Histology/Imaging Core C
-
批准号:10555242
-
项目类别:
-
资助金额:$23.7万
-
财政年份:2015
-
负责人:Theresa Torres Pizarro
-
依托单位:
Enrichment Program
-
批准号:10555238
-
项目类别:
-
资助金额:$2.59万
-
财政年份:2015
-
负责人:Theresa Torres Pizarro
-
依托单位:
Histology/Imaging Core C
-
批准号:10361545
-
项目类别:
-
资助金额:$23.7万
-
财政年份:2015
-
负责人:Theresa Torres Pizarro
-
依托单位:
Enrichment Program
-
批准号:10361543
-
项目类别:
-
资助金额:$2.59万
-
财政年份:2015
-
负责人:Theresa Torres Pizarro
-
依托单位:
Mechanisms underlying sex differences in the pathogenesis of IBD
-
批准号:8517580
-
项目类别:
-
资助金额:$18.45万
-
财政年份:2012
-
负责人:Theresa Torres Pizarro
-
依托单位:
Mechanisms underlying sex differences in the pathogenesis of IBD
-
批准号:8385111
-
项目类别:
-
资助金额:$23.55万
-
财政年份:2012
-
负责人:Theresa Torres Pizarro
-
依托单位:
ANIMAL CORE
-
批准号:7491476
-
项目类别:
-
资助金额:$15.49万
-
财政年份:2007
-
负责人:Theresa Torres Pizarro
-
依托单位:
EPITHELIAL INNATE RESPONSES IN CHRONIC SAMP ILEITIS
-
批准号:7491475
-
项目类别:
-
资助金额:$20.73万
-
财政年份:2007
-
负责人:Theresa Torres Pizarro
-
依托单位:
CORE--Morphology/Imaging Core
-
批准号:7447856
-
项目类别:
-
资助金额:$25.55万
-
财政年份:2007
-
负责人:Theresa Torres Pizarro
-
依托单位:
EPITHELIAL INNATE RESPONSES IN CHRONIC SAMP ILEITIS
-
批准号:7021096
-
项目类别:
-
资助金额:$19.33万
-
财政年份:2005
-
负责人:Theresa Torres Pizarro
-
依托单位:
ANIMAL CORE
-
批准号:7021099
-
项目类别:
-
资助金额:$23.03万
-
财政年份:2005
-
负责人:Theresa Torres Pizarro
-
依托单位:
CORE--Morphology/Imaging Core
-
批准号:6797534
-
项目类别:
-
资助金额:$21.55万
-
财政年份:2004
-
负责人:Theresa Torres Pizarro
-
依托单位:
CORE--CYTOKINE/IMMUNOLOGY FACILITY
-
批准号:6652816
-
项目类别:
-
资助金额:$10.78万
-
财政年份:2002
-
负责人:Theresa Torres Pizarro
-
依托单位:
CORE--CYTOKINE/IMMUNOLOGY FACILITY
-
批准号:6651780
-
项目类别:
-
资助金额:$10.78万
-
财政年份:2002
-
负责人:Theresa Torres Pizarro
-
依托单位:
海外基金