ENaC-α mediates lung fluid clearance and capillary barrier function in pneumonia

ENaC-α 介导肺炎中的肺液清除和毛细血管屏障功能

• 批准号: 10205151
• 负责人: Rudolf Lucas
• 金额: $ 51.57万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10205151
• 关键词: AGTR2 gene; ASIC channel; Actin-Binding Protein; Actins; Acute Lung Injury; Alveolar; Bacteria; Bacterial Pneumonia; Binding; Blood Vessels; Blood capillaries; Capillary Endothelial Cell; Cations; Cell Line; Cells; Data; Development; Edema; Endothelial Cells; Endothelium; Epithelial; Epithelial Cells; Evaluation; Female; Fluid Balance; Fostering; Functional disorder; Genetic; Goals; Human; Hybrids; Impairment; In Vitro; Individual; Infection; Ion Channel; Knockout Mice; Life; Liquid substance; Lung; Mediating; Molecular; Mus; Outcome; Pathway interactions; Peptides; Permeability; Pharmacological Treatment; Pharmacology; Phosphorylation; Pneumococcal Pneumonia; Pneumonia; Preparation; Probability; Proteins; Publishing; Recycling; Regulation; Resolution; Role; Signal Transduction; Signaling Molecule; Slice; Sodium; Sodium Channel; Streptococcus pneumoniae; Streptococcus pneumoniae plY protein; Stress Fibers; Surface; TNF gene; Testing; Toxin; Vascular Permeabilities; alveolar epithelium; epithelial Na+ channel; filamin; improved; in vivo; lung injury; male; mutant; novel; novel therapeutic intervention; overexpression; prevent; therapeutic target; uptake; vector

PROJECT SUMMARY. Pulmonary permeability edema (PPE) associated with pneumococcal pneumonia is a life-threatening condition, resulting from capillary barrier dysfunction in conjunction with impaired alveolar liquid clearance (ALC), with no proven treatment. Vectorial Na+ uptake mediates ALC. The identification of novel therapeutic approaches to simultaneously restore both ALC and barrier function represents a critical unmet need for pneumonia- associated PPE. Our main hypothesis is that the a subunit of the epithelial sodium channel (ENaC-α) represents a promising therapeutic target in PPE since it is a component of the highly-selective cation channel (HSC), which consists of ENaC-α, β and γ subunits, and of the non-selective cation channel (NSC), which contains ASIC1a and ENaC-α. Both HSC and NSC channels mediate Na+ uptake. We also propose ENaC-α as a signaling molecule that strengthens barrier function in lung capillaries in the presence of pneumococci or their pore-forming toxin pneumolysin (PLY). Our main hypothesis is that ENaC-α exerts these actions mainly by blunting phosphorylation of the actin-binding protein filamin-A. In its non-phosphorylated form, filamin-A on the one hand promotes Na+ uptake capacity in sodium channels in alveolar epithelial cells and on the other hand it prevents stress fiber formation in capillary endothelial cells. Our novel hypothesis centers on the concept that ENaC-α functions as both an ion channel component and a signaling molecule, whose specific pharmacological activation restores ALC and barrier function during pneumococcal pneumonia. We will study the effect of specific activators of NSC (MitTx), of HSC (S3969 which binds to ENaC-β, not present in NSC) or of both (TIP peptide, which binds to ENaC-α), as well as genetic depletion or overexpression of ENaC-α on Na+ uptake capacity in pneumococci- or PLY-treated alveolar epithelial cells in vitro. We will moreover investigate whether ENaC-α activation or overexpression corrects S. pneumoniae- or PLY-induced barrier dysfunction in MVEC in vitro, through inhibition of Ca2+-dependent pathways that mediate filamin-A phosphorylation. Finally, we will test our hypothesis that direct ENaC-α activation is sufficient to mediate ALC and capillary barrier function during pneumococcal pneumonia in mice and in isolated perfused human lungs. Our expected outcomes include a better characterization of the unique role of ENaC-α in ALC during pneumococcal pneumonia, demonstration of a hitherto unknown role for ENaC-α in capillary barrier regulation and evaluation of the relative role of NSC versus HSC in protection from bacterial pneumonia-induced acute lung injury. Unraveling the unique mechanisms by which ENaC-α mediates ALC and barrier function during bacterial pneumonia can foster development of a novel breakthrough treatment for pulmonary permeability edema.

项目摘要。 与肺炎球菌肺炎相关的肺渗透性水肿（PPE）是一种危及生命的疾病， 由于毛细血管屏障功能障碍与肺泡液体清除率（ALC）受损， 经过验证的治疗。Na+摄取介导ALC。确定新的治疗方法， 同时恢复ALC和屏障功能代表了肺炎尚未满足的关键需求- 相关PPE我们的主要假设是上皮钠通道α亚单位（ENaC-α） 代表PPE中有前途的治疗靶点，因为它是高选择性阳离子通道的组分 (HSC)由ENaC-α、β和γ亚基组成，以及非选择性阳离子通道（NSC）， 含有ASIC 1a和ENaC-α。HSC和NSC通道均介导Na+摄取。我们还提出了ENaC-α 作为一种信号分子，在肺炎球菌存在时加强肺毛细血管的屏障功能， 它们的成孔毒素肺炎球菌溶血素（pneumolysin，Pneumolysin）。我们的主要假设是ENaC-α主要发挥这些作用 通过钝化肌动蛋白结合蛋白细丝蛋白-A的磷酸化。在其非磷酸化形式中，细丝蛋白-A在 一方面促进肺泡上皮细胞钠通道的Na+摄取能力， 另一方面，它阻止毛细血管内皮细胞中应力纤维的形成。我们的新假设集中在 ENaC-α同时作为离子通道成分和信号分子，其特异性 药理学活化恢复肺炎球菌肺炎期间的ALC和屏障功能。我们将研究 NSC（MitTx）、HSC（S3969，结合ENaC-β，不存在于NSC中）或 两者（TIP肽，与ENaC-α结合），以及ENaC-α基因缺失或过度表达， 肺炎球菌或PLY处理的肺泡上皮细胞的Na+摄取能力。此外，我们将 研究ENaC-α激活或过表达是否纠正S.肺炎或PLY诱导的屏障 通过抑制介导细丝蛋白A的Ca 2+依赖性途径，体外MVEC功能障碍 磷酸化最后，我们将验证我们的假设，即直接ENaC-α激活足以介导ALC 以及小鼠和离体灌注的人肺中肺炎球菌肺炎期间的毛细血管屏障功能。 我们的预期结果包括更好地描述ENaC-α在ALC中的独特作用， 肺炎球菌性肺炎，ENaC-α在毛细血管屏障调节中迄今未知作用的证明 并评估NSC与HSC在预防细菌性肺炎诱导的急性炎症中的相对作用 肺损伤揭示ENaC-α介导ALC和屏障功能的独特机制， 细菌性肺炎可促进肺渗透性新突破性治疗的发展 水肿

项目成果

Impact of Endogenous Pneumococcal Hydrogen Peroxide on the Activity and Release of Pneumolysin.

• DOI: 10.3390/toxins15100593
• 发表时间: 2023-09-30
• 期刊: Toxins
• 影响因子: 4.2
• 作者: Bazant J;Ott B;Hudel M;Hain T;Lucas R;Mraheil MA
• 通讯作者: Mraheil MA

Editorial: Cytokine-Ion Channel Interactions in Pulmonary Inflammation.

社论：肺部炎症中的细胞因子-离子通道相互作用。

• DOI: 10.3389/fimmu.2018.02598
• 发表时间: 2018
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Vadász,István;Lucas,Rudolf
• 通讯作者: Lucas,Rudolf

The arginase 1/ornithine decarboxylase pathway suppresses HDAC3 to ameliorate the myeloid cell inflammatory response: implications for retinal ischemic injury.

• DOI: 10.1038/s41419-023-06147-7
• 发表时间: 2023-09-21
• 期刊: CELL DEATH & DISEASE
• 影响因子: 9
• 作者: Shosha, Esraa;Shahror, Rami A.;Morris, Carol A.;Xu, Zhimin;Lucas, Rudolf;McGee-Lawrence, Meghan E.;Rusch, Nancy J.;Caldwell, Ruth B.;Fouda, Abdelrahman Y.
• 通讯作者: Fouda, Abdelrahman Y.

Deficiency of Myeloid Pfkfb3 Protects Mice From Lung Edema and Cardiac Dysfunction in LPS-Induced Endotoxemia.

骨髓 Pfkfb3 缺乏可保护小鼠免受 LPS 诱导的内毒素血症中的肺水肿和心脏功能障碍

• DOI: 10.3389/fcvm.2021.745810
• 发表时间: 2021
• 期刊: Frontiers in cardiovascular medicine
• 影响因子: 3.6
• 作者: Xu J;Wang L;Yang Q;Ma Q;Zhou Y;Cai Y;Mao X;Da Q;Lu T;Su Y;Bagi Z;Lucas R;Liu Z;Hong M;Ouyang K;Huo Y
• 通讯作者: Huo Y

Potent anti-inflammatory activity of the lectin-like domain of TNF in joints.

• DOI: 10.3389/fimmu.2022.1049368
• 发表时间: 2022
• 期刊: FRONTIERS IN IMMUNOLOGY
• 影响因子: 7.3
• 作者: Pinto, Ana Carolina Matias Dinelly;Nunes, Rodolfo de Melo;Nogueira, Igor Albuquerque;Fischer, Bernhard;Lucas, Rudolf;Girao-Carmona, Virginia Claudia Carneiro;de Oliveira, Vivian Louise Soares;Amaral, Flavio Almeida;Schett, Georg;Rocha, Francisco Airton Castro
• 通讯作者: Rocha, Francisco Airton Castro