Asthma Sample Collection Protocol: Defining the Role of Apolipoprotein Pathways in Asthma
哮喘样本采集方案:定义载脂蛋白通路在哮喘中的作用
基本信息
- 批准号:10929167
- 负责人:
- 金额:$ 187.8万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:
- 资助国家:美国
- 起止时间:至
- 项目状态:未结题
- 来源:
- 关键词:ADP Ribose TransferasesAirway DiseaseAlveolar MacrophagesApicalApolipoproteinsAsthmaAttenuatedBlood specimenBronchoalveolar LavageBronchoalveolar Lavage FluidBronchoscopyBrush CellCD14 geneCell membraneCellsChemotaxisCiliated Bronchial Epithelial CellClinical ImmunologyCollectionCytokine ReceptorsDendritic CellsEpithelial CellsEpitheliumExtrinsic asthmaFastingGenesHLA-DR AntigensHigh Density LipoproteinsHumanHypersensitivityITGAX geneImmuneInflammasomeInflammationInflammatoryInflammatory ResponseInterleukin-1LengthLigandsLinkLipidsLiquid substanceLungMAP3K7 geneMAP3K8 geneMAPK8 geneMacrophageMediatingMembraneMicrofluidicsMinorModificationMyelogenousNeutrophil ActivationNutrientPathway interactionsPeripheral Blood Mononuclear CellPhenotypePopulationProductionProtocols documentationPyroglyphidaeRANTESRegulationResearchRoleSamplingSerumSeverity of illnessSignal PathwaySignal TransductionSputumSteroidsTLR4 geneTh2 CellsTimeTissue ProcurementsTranscriptTriglyceridesTumor Necrosis Factor ReceptorVLDL receptorViraladaptive immune responseairway epitheliumairway inflammationasthmaticasthmatic patientbronchial epitheliumchemokinecytokineeosinophileosinophilic asthmaexosomehuman RNA sequencinghuman neutrophil peptide 1mRNA Expressionmouse modelnanoscaleneutrophilparticlepatient subsetsperipheral bloodpoint of carereceptorsample collectionsoluble TNF receptor type Ivesicular releasevolunteer
项目摘要
The progress achieved under this protocol is summarized as follows:
1. The structural cell populations obtained by this protocol have been utilized to characterize the expression of ARTS-1 in lung cell populations. Bronchial epithelial cells that have been obtained by bronchial brushings have been utilized to demonstrate co- localization of membrane-associated ARTS-1 and TNFR1 in the apical cell membrane of ciliated bronchial epithelial cells.
2. Bronchoalveolar lavage fluid obtained by this protocol has been utilized to identify exosome-like vesicle release as a new mechanism by which soluble cytokine receptors can be generated, independent of ectodomain cleavage by receptor sheddases. The majority of soluble type I TNF receptor (TNFR1) in bronchoalveolar lavage fluid was found to be full-length, 55-kDa, exosome-associated TNFR1, whereas the 28-kDa cleaved TNFR1 ectodomain represented only a minor fraction. Thus, release of exosome-associated TNFR1 (i.e., eTNFR1) represents an important alternative mechanism for the release of soluble TNF receptors.
3. BAL fluid from asthmatic patients has been utilized in studies investigating the ADP-ribosyltransferase-specific modification of human neutrophil peptide-1.
4. Expression of the VLDL receptor was demonstrated for the first time on circulating CD11c+/CD14-/HLA-DR+ dendritic cells. This confirmed our result in a murine model that the VLDL receptor is expressed by dendritic cells and attenuates house dust mite-induced airway inflammation by suppressing dendritic cell-mediated adaptive immune responses.
5. BAL fluid cells have been used to show that expression of CD163 is reduced on alveolar macrophages from asthmatics as compared to normal subjects.
6. Induced sputum cells have been used to develop components of a nanoscale microfluidic flow cytometer that will liquefy sputum samples for point-of-care inflammatory phenotyping of asthmatic patients.
7. Myeloid dendritic cell subsets from patients with eosinophil-high asthma were shown to have lower levels of LRP-1 expression than those from healthy nonasthmatic subjects and that a negative correlation exists between LRP-1 expression by myeloid dendritic cell subsets and peripheral blood eosinophil counts in asthmatic patients.
8. HDL particles were shown to be negatively correlated, whereas serum triglycerides were shown to be positively correlated, with blood eosinophils in atopic asthmatics. This supports the concept that serum levels of HDL and triglycerides may be linked to systemic type 2 inflammation in atopic asthma. (J Lipid Research 2017 Aug; 58(8): 1713-1721. PMID: 28655726.)
9. Human peripheral blood myeloid DC subsets from patients with eosinophilic asthma have lower LRP-1 expression than cells from healthy nonasthmatic subjects. (JACI 2018 Oct; 142(4):1066-1079.e6. DOI: 10.1016/j.jaci.2017.10.044. PMID: 29274414.)
10. Peripheral blood mononuclear cells from asthmatics were used to show that prolonged fasting blunts the NLRP3 inflammasome and Th2 cell activation in steroid-naive asthmatics as well as diminishes airway epithelial cell cytokine production. This identifies a potential role for nutrient level-dependent regulation of inflammation in asthma. (J Immunol 2018 Sep 1; 201(5):1382-1388. PMID 30021766).
11. Bronchoalveolar lavage macrophages from asthmatics were used to show that APOE can function as an endogenous, concentration-dependent pulmonary danger signal that primes and activates the NLPR3 inflammasome to secrete IL-1. This might represent a mechanism through which APOE amplifies pulmonary inflammatory responses when concentrations in the lung are increased to greater than normal levels, which can occur during viral exacerbations of HDM-induced asthma characterized by neutrophilic airway inflammation. (J Allergy Clinical Immunology 2019; 144(2):426-441.e3. DOI: https://doi.org/10.1016/jaci.2019.02.027).
12. RNA sequencing of human asthmatic bronchial brushing cells stimulated with APOE identified increased expression of mRNA transcripts encoding multiple proinflammatory genes, including CXCL5 (C-X-C motif chemokine ligand 5), an epithelial-derived chemokine that promotes neutrophil activation and chemotaxis. We subsequently characterized the APOE signaling pathway that induces CXCL5 secretion by human asthmatic small airway epithelial cells (SAECs). We showed that elevated levels of APOE in the airway may activate a TLR4/TAK1/IK/NF-B/TPL2/JNK signaling pathway that induces CXCL5 secretion by human asthmatic SAECs. These findings identify new roles for TLR4 and TPL2 in APOE-mediated proinflammatory responses in asthma. (Am J Resp Cell Mol Biol 2020; 63 (2): 185 - 197.)
在该议定书下取得的进展概述如下:
1.利用该方法获得的结构细胞群已被用来研究ARTS-1在肺细胞群中的表达。通过支气管刷检获得的支气管上皮细胞已被用来证明膜相关的ARTS-1和TNFR1在纤毛支气管上皮细胞的顶端细胞膜上的共定位。
2.通过该方法获得的支气管肺泡灌洗液已被用于鉴定外体样囊泡的释放,这是一种不依赖于受体脱落酶对胞外结构域的切割而产生可溶性细胞因子受体的新机制。支气管肺泡灌洗液中的可溶I型肿瘤坏死因子受体(TNFR1)大部分是55 kDa的外切体相关的全长TNFR1,而28 kDa裂解的TNFR1胞外区只占一小部分。因此,释放外切体相关的TNFR1(即eTNFR1)是释放可溶性肿瘤坏死因子受体的重要替代机制。
3.利用哮喘患者的肺泡灌洗液研究了ADP-核糖基转移酶对人中性粒细胞多肽-1的特异性修饰。
4.首次在循环CD11c+/CD14-/HLA-DR+树突状细胞上发现VLDL受体的表达。这证实了我们在小鼠模型中的结果,即极低密度脂蛋白受体由树突状细胞表达,并通过抑制树突状细胞介导的适应性免疫反应来减轻屋尘螨诱导的呼吸道炎症。
5.BAL液细胞实验表明,哮喘患者肺泡巨噬细胞表面CD163的表达较正常人明显降低。
6.诱导痰细胞已被用于开发纳米级微流控流式细胞仪的部件,该仪器将液化痰样本,用于哮喘患者的护理点炎性表型。
7.哮喘患者外周血嗜酸粒细胞计数与髓系树突状细胞亚群表达LRP-1呈负相关。
8.特应性哮喘患者血中高密度脂蛋白颗粒与嗜酸性粒细胞呈负相关,血清甘油三酯与嗜酸性粒细胞呈正相关。这支持了血清高密度脂蛋白和甘油三酯水平可能与特应性哮喘的系统性2型炎症有关的概念。(J Lipid Research 2017年8月;58(8):1713-1721。PMID:28655726。)
9.嗜酸粒细胞哮喘患者外周血髓系DC亚群的LRP-1表达低于健康非哮喘患者。(JACI 2018年10月;142(4):1066-1079.e6.DOI:10.1016/j.Jaci.2017.10.044.PMID:29274414。)
10.哮喘患者外周血单个核细胞的检测结果表明,长期禁食可抑制激素治疗后哮喘患者NLRP3炎性小体和Th2细胞的活化,减少呼吸道上皮细胞细胞因子的产生。这确定了哮喘中依赖营养水平的炎症调节的潜在作用。(《免疫杂志》2018年9月1日;201(5):1382-1388。PMID 30021766)。
11.哮喘患者肺泡灌洗巨噬细胞实验表明,APOE可作为内源性、浓度依赖的肺危险信号,启动和激活NLPR3炎性小体分泌IL-1。这可能代表了一种机制,通过这种机制,当肺中的浓度增加到高于正常水平时,APOE会放大肺部炎症反应,这可能发生在以中性粒细胞炎症为特征的HDM诱导的哮喘的病毒加重期间。(《过敏临床免疫学》2019;144(2):426-441.e3.DOI:https://doi.org/10.1016/jaci.2019.02.027).
12.经APOE刺激的人哮喘支气管刷毛细胞的RNA测序表明,编码多种致炎基因的mRNA转录增加,包括CXCL5(C-X-C基序趋化因子配体5),这是一种促进中性粒细胞激活和趋化的上皮源性趋化因子。随后,我们鉴定了诱导人哮喘小气道上皮细胞(SAECs)分泌CXCL5的APOE信号通路。我们发现,高水平的APOE可能激活TLR4/TAK1/IK/NF-B/Tpl2/JNK信号通路,诱导人哮喘SAEC分泌CXCL5。这些发现确定了TLR4和Tpl2在载脂蛋白E介导的哮喘促炎反应中的新作用。(Am J RESP Cell Mol Biol 2020;63(2):185-197。)
项目成果
期刊论文数量(4)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Apolipoprotein E is a concentration-dependent pulmonary danger signal that activates the NLRP3 inflammasome and IL-1β secretion by bronchoalveolar fluid macrophages from asthmatic subjects.
- DOI:10.1016/j.jaci.2019.02.027
- 发表时间:2019-08
- 期刊:
- 影响因子:14.2
- 作者:Gordon, Elizabeth M.;Yao, Xianglan;Xu, Haitao;Karkowsky, William;Kaler, Maryann;Kalchiem-Dekel, Or;Barochia, Amisha V.;Gao, Meixia;Keeran, Karen J.;Jeffries, Kenneth R.;Levine, Stewart J.
- 通讯作者:Levine, Stewart J.
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Stewart Levine其他文献
Stewart Levine的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Stewart Levine', 18)}}的其他基金
Characterization of the Role of NUCB2 in Asthma Pathogenesis
NUCB2 在哮喘发病机制中作用的表征
- 批准号:
8558006 - 财政年份:
- 资助金额:
$ 187.8万 - 项目类别:
Identification and Characterization of microRNA Genes in Asthma
哮喘中 microRNA 基因的鉴定和表征
- 批准号:
8939833 - 财政年份:
- 资助金额:
$ 187.8万 - 项目类别:
Study of Pioglitazone Hydrochloride in Severe, Refractory Asthma
盐酸吡格列酮治疗严重难治性哮喘的研究
- 批准号:
8344773 - 财政年份:
- 资助金额:
$ 187.8万 - 项目类别:
ID of Biomarkers in Exhaled Breath Condensates from Asthmatic Patients
哮喘患者呼出气体冷凝物中生物标志物的识别
- 批准号:
7734981 - 财政年份:
- 资助金额:
$ 187.8万 - 项目类别:
Development of Apolipoprotein-based Therapeutics for Asthma
基于载脂蛋白的哮喘疗法的开发
- 批准号:
8149566 - 财政年份:
- 资助金额:
$ 187.8万 - 项目类别:
ID of Biomarkers in Exhaled Breath Condensates from Asthmatic Patients
哮喘患者呼出气体冷凝物中生物标志物的识别
- 批准号:
7969044 - 财政年份:
- 资助金额:
$ 187.8万 - 项目类别:
Characterization of the Role of NUCB2 in Asthma Pathogenesis
NUCB2 在哮喘发病机制中作用的表征
- 批准号:
8344859 - 财政年份:
- 资助金额:
$ 187.8万 - 项目类别:
Identifying and Characterizing "Corticosteroid-unresponsive" Genes in Asthma
哮喘中“皮质类固醇无反应”基因的识别和特征分析
- 批准号:
8557924 - 财政年份:
- 资助金额:
$ 187.8万 - 项目类别:
Characterization of the Role of NUCB2 in Asthma Pathogenesis
NUCB2 在哮喘发病机制中作用的表征
- 批准号:
8746634 - 财政年份:
- 资助金额:
$ 187.8万 - 项目类别:
相似海外基金
EXposomic Profiling in Airway disease to uNravel Determinants of disease in Asthma (EXPAND-Asthma) Center
气道疾病暴露组分析以解开哮喘疾病的决定因素 (EXPAND-Asthma) 中心
- 批准号:
10744673 - 财政年份:2023
- 资助金额:
$ 187.8万 - 项目类别:
Bacteriophage as a predictive biomarker in chronic Pseudomonas airway disease
噬菌体作为慢性假单胞菌气道疾病的预测生物标志物
- 批准号:
10723956 - 财政年份:2023
- 资助金额:
$ 187.8万 - 项目类别:
Defining a gene expression signature of airway disease, COPD exacerbations, and response to treatment
定义气道疾病、COPD 恶化和治疗反应的基因表达特征
- 批准号:
10733573 - 财政年份:2023
- 资助金额:
$ 187.8万 - 项目类别:
Impact of Maternal Arsenic Exposure on Offspring's Epigenetic Reprogramming of Allergic Airway Disease
母亲砷暴露对后代过敏性气道疾病表观遗传重编程的影响
- 批准号:
10733607 - 财政年份:2023
- 资助金额:
$ 187.8万 - 项目类别:
The role of anaerobic microbiota in cystic fibrosis airway disease trajectory
厌氧微生物群在囊性纤维化气道疾病轨迹中的作用
- 批准号:
10716654 - 财政年份:2023
- 资助金额:
$ 187.8万 - 项目类别:
Immune Mediators of IL-22 Signaling Alter Allergic Airway Disease
IL-22 信号的免疫介质改变过敏性气道疾病
- 批准号:
10853347 - 财政年份:2023
- 资助金额:
$ 187.8万 - 项目类别:
Microbial adaptation of Pseudomonas lipid A structure in CF airway disease progress
假单胞菌脂质 A 结构在 CF 气道疾病进展中的微生物适应
- 批准号:
10722599 - 财政年份:2023
- 资助金额:
$ 187.8万 - 项目类别:
The role of anaerobic microbiota in cystic fibrosis airway disease trajectory
厌氧微生物群在囊性纤维化气道疾病轨迹中的作用
- 批准号:
10985906 - 财政年份:2023
- 资助金额:
$ 187.8万 - 项目类别:
Mentoring patient-oriented researchers in inflammatory airway disease
指导炎症性气道疾病领域以患者为中心的研究人员
- 批准号:
10657746 - 财政年份:2022
- 资助金额:
$ 187.8万 - 项目类别: