Role of Sialidase in Pulmonary Host Defenses and Acute Lung Injury

唾液酸酶在肺宿主防御和急性肺损伤中的作用

• 批准号: 7866681
• 负责人: Alan S. Cross
• 金额: $ 46.82万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7866681
• 关键词: Accounting; Acids; Acute Lung Injury; Address; Adherence; Adhesions; Adult Respiratory Distress Syndrome; Affect; Affinity; Alveolar; Animal Model; Aspirate substance; Bacteria; Binding; Blood capillaries; Breathing; Cathepsin G; Cell Adhesion; Cell Adhesion Molecules; Cell Communication; Cell Surface Receptors; Cell surface; Cells; Cessation of life; Charge; Complex; Coupled; Data; Defense Mechanisms; Diffuse; Elastases; Embryo; Endothelial Cells; Endothelium; Environment; Epithelium; Equilibrium; Event; Excision; Free Radicals; Gases; Glycoconjugates; Host Defense; Human; ICAM1 gene; ITGAM gene; ITGB2 gene; Immune response; In Vitro; Infection; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Injury; Invaded; Lead; Length of Stay; Leukocyte Elastase; Leukocyte Trafficking; Leukostasis; Ligands; Lung; Mammalian Cell; Mediating; Microbe; Modeling; Modification; Molecular; Mus; N-Acetylneuraminic Acid; Neuraminidase; Organ; PECAM1 gene; Pathway interactions; Pattern recognition receptor; Peptide Hydrolases; Peptides; Permeability; Phosphorylation; Principal Investigator; Process; Production; Protein Isoforms; Proteins; Reactive Oxygen Species; Regulation; Role; Sialic Acids; Sialyltransferases; Signal Transduction; Site; Stimulus; Surface; Syndrome; Testing; Tissues; Toll-like receptors; Tyrosine Phosphorylation; base; cadherin 5; capillary; collagenase; cytokine; in vivo; in vivo Model; inhibitor/antagonist; interstitial; kidney cell; killings; lung injury; macromolecule; microbial; migration; neutrophil; novel therapeutic intervention; overexpression; programs; response; restoration; sialylation; toll-like receptor 4; trafficking

DESCRIPTION (provided by applicant): Polymorphonuclear leukocytes (PMN) protect the lung against microbial infection; however, an excessive or prolonged influx of PMNs may cause tissue damage. Acute lung injury is characterized by widespread inflammatory changes, including diffuse PMN infiltration, with injury to both lung epithelium and endothelium. We established that the removal of sialyl residues from the surface of PMNs and endothelial cells (EC) by an endogenous PMN sialidase is critical to normal PMN trafficking. Further, we find that desialylation of the Toll-like receptor (TLR) 4 complex enhances LPS-induced cytokine production, while restoration of sialyl residues by sialyltransferases (ST) may restore the basal state. The overall hypothesis to be tested is that sialidases and STs regulate innate immune responses in the lung, and the balance achieved will dictate whether or not the innate immune response develops into acute lung injury. To address this central hypothesis, we propose the following: In Specific Aim 1 we will define which form(s) of PMN sialidase(s) (neul and neu3) regulates adherence to and migration across human lung microvascular EC by overexpression and knockdown of specific neu proteins. In Specific Aim 2 we will define the substrates for PMN sialidase in PMNs and ECs and assess how their sialylation status affects PMN adhesion in vitro and trafficking in vivo. Since PMNs induce signaling cascades in ECs, in Specific Aim 3 we will examine whether an "appropriate" level of PMN sialidase-induced tyrosine phosphorylation of EC proteins opens the EC-EC paracellular pathway sufficiently for transendothelial migration without protein leak, while at levels that exceed this threshold, if there will be loss of EC barrier function. A "two-hit" model of acute lung injury has been proposed whereby a combination of LPS and chemotactic peptide promotes lung injury. In Specific Aim 4, we will examine the role of sialidase in sensitizing the pulmonary vasculature to circulating LPS and lung injury. The molecular mechanism(s) by which desialylation promotes signaling through the TLR4/MD2/CD14 complex will be examined, as well as the ability of STs to downregulate this response. The ability of sialidase inhibitors to reverse acute lung injury will be studied in vivo in a murine model of pulmonary leukostasis. Since inhibitors of sialidase are commercially available, these studies will provide the experimental rationale for testing a new therapeutic approach to the treatment of lung injury.

描述（由申请人提供）：多形核白细胞（PMN）保护肺免受微生物感染；但是，过多或长时间的PMN流入可能会导致组织损伤。急性肺损伤的特征是广泛的炎症变化，包括弥漫性PMN浸润，并损伤肺上皮和内皮。我们确定，通过内源性PMN唾液酸酶从PMN和内皮细胞表面（EC）从PMN和内源性PMN唾液酸酶的表面清除siAllyl残基对于正常的PMN运输至关重要。此外，我们发现Toll样受体（TLR）4络合物的脱酰化增强了LPS诱导的细胞因子的产生，而siAllyltransferases（ST）恢复了siAllyl残基可能会恢复基础状态。要检验的总体假设是，唾液酸酶和STS调节肺中先天免疫反应，实现的平衡将决定先天免疫反应是否会导致急性肺损伤。为了解决这一中心假设，我们提出以下内容：在特定目的1中，我们将定义哪种形式的pmn sialidase（s）（neul和neu3）通过过表达和敲除特定的neu蛋白来调节人类肺微血管EC的粘附和迁移。在特定的目标2中，我们将定义PMN和EC中PMN唾液酸酶的底物，并评估其靠囊状态的状态如何影响PMN的体外和体内运输。由于PMN在EC中诱导信号级联反应，因此在特定的目标3中，我们将检查EC蛋白的PMN Sialidase诱导的酪氨酸磷酸化水平是否足以打开EC-EC细胞细胞paracelluarlular途径，以足以使跨内皮迁移无蛋白质泄漏，而在这种水平上，该水平超过了这种thehshold，如果超过了Ec barrybarlesarlesarlesar的水平。已经提出了一种急性肺损伤的“两次冲击”模型，通过该模型，LPS和趋化肽的结合促进了肺损伤。在特定的目标4中，我们将研究唾液酸酶在使肺脉管系统对循环LP和肺损伤中敏感中的作用。将检查通过TLR4/MD2/CD14络合物促进信号传导的分子机制，以及STS下调该响应的能力。唾液酸酶抑制剂逆转急性肺损伤的能力将在肺白细胞的鼠模型中在体内进行研究。由于唾液酸酶的抑制剂是商业上可用的，因此这些研究将为测试一种用于治疗肺损伤的新治疗方法的实验原理。

项目成果

Changes in polysialic acid expression on myeloid cells during differentiation and recruitment to sites of inflammation: role in phagocytosis.

• DOI: 10.1093/glycob/cwu050
• 发表时间: 2014-09
• 期刊: Glycobiology
• 影响因子: 4.3
• 作者: N. Stamatos;Lei Zhang;A. Jokilammi;J. Finne;Wilbur H. Chen;Abderrahman El-Maarouf;A. Cross;K. Hankey

Neuraminidase 1-mediated desialylation of the mucin 1 ectodomain releases a decoy receptor that protects against Pseudomonas aeruginosa lung infection.

神经氨酸酶 1 介导的粘蛋白 1 胞外域的去唾液酸化会释放一种诱饵受体，从而防止铜绿假单胞菌肺部感染。

• DOI: 10.1074/jbc.ra118.006022
• 发表时间: 2019
• 期刊: The Journal of biological chemistry
• 作者: Lillehoj,ErikP;Guang,Wei;Hyun,SangW;Liu,Anguo;Hegerle,Nicolas;Simon,Raphael;Cross,AlanS;Ishida,Hideharu;Luzina,IrinaG;Atamas,SergeiP;Goldblum,SimeonE
• 通讯作者: Goldblum,SimeonE

Potential role for alternatively activated macrophages in the secondary bacterial infection during recovery from influenza.

• DOI: 10.1016/j.imlet.2011.10.009
• 发表时间: 2012-01-30
• 期刊: IMMUNOLOGY LETTERS
• 影响因子: 4.4
• 作者: Chen, Wilbur H.;Toapanta, Franklin R.;Shirey, Kari Ann;Zhang, Lei;Giannelou, Angeliki;Page, Carly;Frieman, Matthew B.;Vogel, Stefanie N.;Cross, Alan S.
• 通讯作者: Cross, Alan S.

MUC1 ectodomain is a flagellin-targeting decoy receptor and biomarker operative during Pseudomonas aeruginosa lung infection.

• DOI: 10.1038/s41598-021-02242-x
• 发表时间: 2021-11-22
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Verceles AC;Bhat P;Nagaria Z;Martin D;Patel H;Ntem-Mensah A;Hyun SW;Hahn A;Jeudy J;Cross AS;Lillehoj EP;Goldblum SE
• 通讯作者: Goldblum SE