TRIM72 modulates caveolar endocytosis in repair of lung cells

Alveolar epithelial and endothelial cell injury is a major feature of the acute respiratory distress syndrome, in particular when in conjunction with ventilation therapies. Previously we showed [Kim SC, Kellett T, Wang S, Nishi M, Nagre N, Zhou B, Flodby P, Shilo K, Ghadiali SN, Takeshima H, Hubmayr RD, Zhao X. Am J Physiol Lung Cell Mol Physiol 307: L449–L459, 2014.] that tripartite motif protein 72 (TRIM72) is essential for amending alveolar epithelial cell injury. Here, we posit that TRIM72 improves cellular integrity through its interaction with caveolin 1 (Cav1). Our data show that, in primary type I alveolar epithelial cells, lack of TRIM72 led to significant reduction of Cav1 at the plasma membrane, accompanied by marked attenuation of caveolar endocytosis. Meanwhile, lentivirus-mediated overexpression of TRIM72 selectively increases caveolar endocytosis in rat lung epithelial cells, suggesting a functional association between these two. Further coimmunoprecipitation assays show that deletion of either functional domain of TRIM72, i.e., RING, B-box, coiled-coil, or PRY-SPRY, abolishes the physical interaction between TRIM72 and Cav1, suggesting that all theoretical domains of TRIM72 are required to forge a strong interaction between these two molecules. Moreover, in vivo studies showed that injurious ventilation-induced lung cell death was significantly increased in knockout (KO) TRIM72KO and Cav1KO lungs compared with wild-type controls and was particularly pronounced in double KO mutants. Apoptosis was accompanied by accentuation of gross lung injury manifestations in the TRIM72KO and Cav1KO mice. Our data show that TRIM72 directly and indirectly modulates caveolar endocytosis, an essential process involved in repair of lung epithelial cells through removal of plasma membrane wounds. Given TRIM72's role in endomembrane trafficking and cell repair, we consider this molecule an attractive therapeutic target for patients with injured lungs.

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Expression and functional implications of CCR2 expression on murine alveolar epithelial cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00079.2003 ◽

2004 ◽

Vol 286 (1) ◽

pp. L68-L72 ◽

Cited By ~ 23

Author(s):

Paul J. Christensen ◽

Ming Du ◽

Bethany Moore ◽

Susan Morris ◽

Galen B. Toews ◽

...

Keyword(s):

Epithelial Cells ◽

Lung Injury ◽

Basement Membranes ◽

Alveolar Epithelial Cells ◽

Lung Epithelial Cells ◽

Alveolar Epithelial ◽

Ccr2 Expression ◽

Lung Epithelial ◽

Functional Consequences ◽

Delayed Closure

Acute lung injury results in damage to the alveolar epithelium, leading to leak of proteins into the alveolar space and impaired gas exchange. Lung function can be restored only if the epithelial layer is restored. The process of reepithelialization requires migration of lung epithelial cells to cover denuded basement membranes. The factors that control the migration of lung epithelial cells are incompletely understood. We examined isolated murine type II alveolar epithelial cells (AECs) for expression of CC chemokine receptor 2 (CCR2) and functional consequences of the binding of the main CCR2 ligand monocyte chemoattractant protein-1 (MCP-1). We found that primary AECs bound MCP-1 and expressed CCR2 mRNA. These cells demonstrated functional consequences of CCR2 expression with migration in response to MCP-1 in chemotaxis/haptotaxis assays. Primary AECs cultured from mice lacking CCR2 did not respond to MCP-1. Monolayers of AECs lacking CCR2 demonstrated delayed closure of mechanical wounds compared with AEC monolayers expressing CCR2. Delayed closure of mechanical wounds of wild-type AECs was also demonstrated in the presence of anti-MCP-1 antibody. These data demonstrate for the first time that AECs express CCR2 and are capable of using this receptor for chemotaxis and healing of wounds. CCR2-MCP-1 interactions may be important in the process of reepithelialization after lung injury.

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Time-resolved dual RNA-Seq reveals extensive rewiring of lung epithelial and pneumococcal transcriptomes during early infection

10.1101/048959 ◽

2016 ◽

Author(s):

Rieza Aprianto ◽

Jelle Slager ◽

Siger Holsappel ◽

Jan-Willem Veening

Keyword(s):

Epithelial Cells ◽

Pneumococcal Infection ◽

Alveolar Epithelial Cells ◽

Lung Epithelial Cells ◽

Infection Model ◽

Rna Seq ◽

Time Resolved ◽

Alveolar Epithelial ◽

Lung Epithelial ◽

Host Microbe Interactions

AbstractStreptococcus pneumoniae(pneumococcus) is the main etiological agent of pneumonia. Pneumococcal pneumonia is initiated by bacterial adherence to lung epithelial cells. Infection to the epithelium is a disruptive interspecies interaction involving numerous transcription-mediated processes. Revealing transcriptional changes may provide valuable insights into pneumococcal disease. Dual RNA-Seq allows simultaneous monitoring of the transcriptomes of both host and pathogen. Here, we developed a time-resolved infection model of human lung alveolar epithelial cells byS. pneumoniaeand assessed transcriptome changes by dual RNA-Seq. Our data provide new insights into host-microbe interactions and show that the epithelial glutathione-detoxification pathway is activated by bacterial presence. We observed that adherent pneumococci, not free-floating bacteria, access host-associated carbohydrates and repress innate immune responses. In conclusion, we provide a dynamic dual-transcriptomics overview of early pneumococcal infection with easy online access (http://dualrnaseq.molgenrug.nl). Further database exploration may expand our understanding of epithelial-pneumococcal interaction, leading to novel antimicrobial strategies.Graphical Abstract

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TM4SF5-mediated CD44v8-10 splicing variant promotes survival of type II alveolar epithelial cells during idiopathic pulmonary fibrosis

Cell Death and Disease ◽

10.1038/s41419-019-1878-5 ◽

2019 ◽

Vol 10 (9) ◽

Cited By ~ 1

Author(s):

Ji Eon Kim ◽

Hye-Jin Kim ◽

Jae Woo Jung ◽

Dae-Geun Song ◽

Dasomi Park ◽

...

Keyword(s):

Epithelial Cells ◽

Idiopathic Pulmonary Fibrosis ◽

Pulmonary Fibrosis ◽

Cell Fate ◽

Alveolar Epithelial Cells ◽

Lung Epithelial Cells ◽

Type I ◽

Type Ii ◽

Alveolar Epithelial ◽

Primary Mouse

Abstract Reactive oxygen species (ROS) regulate cell fate, although signaling molecules that regulate ROS hormesis remain unclear. Here we show that transmembrane 4 L six family member 5 (TM4SF5) in lung epithelial cells induced the alternatively spliced CD44v8-10 variant via an inverse ZEB2/epithelial splicing regulatory proteins (ESRPs) linkage. TM4SF5 formed complexes with the cystine/glutamate antiporter system via TM4SF5- and CD44v8-10-dependent CD98hc plasma-membrane enrichment. Dynamic TM4SF5 binding to CD98hc required CD44v8-10 under ROS-generating inflammatory conditions. TM4SF5 and CD44v8-10 upregulated cystine/glutamate antiporter activity and intracellular glutathione levels, leading to ROS modulation for cell survival. Tm4sf5-null mice exhibited attenuated bleomycin-induced pulmonary fibrosis with lower CD44v8-10 and ESRPs levels than wild-type mice. Primary mouse alveolar epithelial cells (AECs) revealed type II AECs (AECII), but not type I, to adapt the TM4SF5-mediated characteristics, suggesting TM4SF5-mediated AECII survival following AECI injury during idiopathic pulmonary fibrosis (IPF). Thus, the TM4SF5-mediated CD44v8-10 splice variant could be targeted against IPF.

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Resveratrol inhibits Staphylococcus aureus-induced TLR2/MyD88/NF-κB-dependent VCAM-1 expression in human lung epithelial cells

Clinical Science ◽

10.1042/cs20130816 ◽

2014 ◽

Vol 127 (6) ◽

pp. 375-390 ◽

Cited By ~ 14

Author(s):

I-Ta Lee ◽

Chih-Chung Lin ◽

Chih-Kai Hsu ◽

Ming-Yen Wu ◽

Rou-Ling Cho ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Epithelial Cells ◽

Human Lung ◽

Alveolar Epithelial Cells ◽

Lung Epithelial Cells ◽

Polyphenolic Compound ◽

Alveolar Epithelial ◽

Lung Epithelial ◽

Dependent Pathway ◽

Human Lung Epithelial Cells

In the present study, we found that S. aureus induced VCAM-1 expression in human pulmonary alveolar epithelial cells via a TLR2/MyD88/NF-κB-dependent pathway, which was inhibited by treatment with the polyphenolic compound resveratrol.

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Regulation of IL-33 by Oncostatin M in Mouse Lung Epithelial Cells

Mediators of Inflammation ◽

10.1155/2016/9858374 ◽

2016 ◽

Vol 2016 ◽

pp. 1-12 ◽

Cited By ~ 8

Author(s):

Carl D. Richards ◽

Laura Izakelian ◽

Anisha Dubey ◽

Grace Zhang ◽

Steven Wong ◽

...

Keyword(s):

Epithelial Cells ◽

Alveolar Epithelial Cells ◽

Lung Epithelial Cells ◽

Oncostatin M ◽

Mouse Lung ◽

Nuclear Staining ◽

Potent Inducer ◽

Alveolar Epithelial ◽

Lung Epithelial

IL-33 modulates both innate and adaptive immune responses at tissue sites including lung and may play critical roles in inflammatory lung disease. Although IL-33 expression can be altered upon NF-Kappa B activation, here we examine regulation by Oncostatin M, a gp130 cytokine family member, in mouse lung tissue. Responses were assessed in BALB/c mouse lung at day 7 of transient overexpression using endotracheally administered adenovirus encoding OSM (AdOSM) or empty vector (AdDel70). Whole lung extracts showed induction of IL-33 mRNA (>20-fold) and protein (10-fold increase in immunoblots) by AdOSM relative to AdDel70. Immunohistochemistry for IL-33 indicated a marked induction of nuclear staining in alveolar epithelial cellsin vivo. Oncostatin M stimulated IL-33 mRNA and IL-33 full length protein in C10 mouse type 2 alveolar epithelial cells in culture in time-dependent and dose-dependent fashion, whereas IL-6, LIF, IL-31, IL-4, or IL-13 did not, and TGFβrepressed IL-33. IL-33 induction was associated with activation of STAT3, and pharmacological inhibition of STAT3 ameliorated IL-33 levels. These results indicate Oncostatin M as a potent inducer of IL-33 in mouse lung epithelial cells and suggest that an OSM/IL-33 axis may participate in innate immunity and inflammatory conditions in lung.

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Fetal lung cell-derived matrix alters distal lung epithelial ion transport

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1995.268.5.l762 ◽

1995 ◽

Vol 268 (5) ◽

pp. L762-L771 ◽

Cited By ~ 6

Author(s):

O. M. Pitkanen ◽

A. K. Tanswell ◽

H. M. O'Brodovich

Keyword(s):

Epithelial Cells ◽

Ion Transport ◽

Short Circuit ◽

Alveolar Epithelial Cells ◽

Lung Epithelial Cells ◽

Bathing Solution ◽

Type I ◽

Ussing Chambers ◽

Lung Epithelial ◽

Distal Lung

Extracellular matrix (ECM) synthesized by the fetal mesenchymal cells provides a supporting structure for the growing airways and is important for airway branching and in the differentiation of the primitive epithelium. We studied whether ECM, in addition to its structural role in lung interstitium, influences the ion transport of rat fetal distal lung epithelial cells (FDLE). FDLE monolayers were cultured on two different fetal mixed lung cell (MLC)-derived matrix preparations and studied in Ussing chambers. FDLE on MLC matrix had an increased resting equivalent short-circuit current (Ieq). Amiloride (10(-4) M apically) decreased the Ieq significantly in all the FDLE monolayers. The residual Ieq was significantly larger in FDLE grown on MLC matrix (increased by 150 and 80% under baseline and beta 2-agonist-stimulated conditions, respectively) than on control filters and filters coated with type I collagen, and type IV collagen, laminin, or fibronectin. The matrix produced by MLC isolated at an earlier gestational stage decreased the FDLE's sensitivity to amiloride. The increased amiloride-insensitive Ieq was only modestly affected by the Na+/K+/Cl- cotransport inhibitor bumetanide (10(-4) M basally) but was abolished when the [Cl-] of the bathing solution was reduced to 10 mM. These observations demonstrated that MLC elaborated ECM is able to change the nature of the ion transport of FDLE. ECM may be an important factor governing the ion transporting phenotype of fetal type II alveolar epithelial cells.

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Modulation of sodium transport in fetal alveolar epithelial cells by oxygen and corticosterone

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00218.2002 ◽

2003 ◽

Vol 284 (2) ◽

pp. L376-L385 ◽

Cited By ~ 39

Author(s):

Ulrich H. Thome ◽

Ian C. Davis ◽

Susie Vo Nguyen ◽

Brent Jay Shelton ◽

Sadis Matalon

Keyword(s):

Epithelial Cells ◽

Short Circuit ◽

Alveolar Epithelial Cells ◽

Lung Epithelial Cells ◽

Ussing Chambers ◽

Alveolar Epithelial ◽

Lung Epithelial ◽

Distal Lung ◽

Postnatal Adaptation ◽

Short Circuit Currents

Regulation of active Na+transport across fetal distal lung epithelial cells (FDLE) by corticosterone (CST), corticotropin-releasing hormone (CRH), and oxygen tension may be crucial for postnatal adaptation. FDLE isolated from 19-day rat fetuses (term: 22 days) were grown on permeable supports to confluent monolayers (duration 3 days) in 2.5, 5, 12, or 20% O2 with 5% CO2-balance N2 and mounted in Ussing chambers for measurement of short-circuit currents ( I sc). FDLE monolayers grown in 20% O2 had significantly higher levels of total I sc and of their amiloride-sensitive ( I amil) and ouabain-sensitive ( I ouab) components than hypoxic cells. Values (μA/cm2 ± SE) for 2.5–5% O2 and 20% O2 were, respectively, I sc5.3 ± 0.2 vs. 8.4 ± 0.3 ( P < 0.001), I amil 3.4 ± 0.2 vs. 4.3 ± 0.2 ( P < 0.01), and I ouab 3.4 ± 0.6 vs. 9.1 ± 0.6 ( P < 0.001). Addition of CST but not CRH to the culture medium at any O2concentration increased I amil. FDLE cells grown at 5% O2 expressed significantly lower levels of α-, β-, and γ-epithelial Na+ channel (ENaC), and of the α1-Na+-K+-ATPase, as determined by Western blotting. We conclude that higher O2concentrations increased total vectorial Na+ transport, and the function of Na+-K+-ATPase and apical amiloride-sensitive Na+ conductance, whereas CST only increased ENaC function.

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Biophysical determinants of alveolar epithelial plasma membrane wounding associated with mechanical ventilation

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00437.2012 ◽

2013 ◽

Vol 305 (7) ◽

pp. L478-L484 ◽

Cited By ~ 34

Author(s):

Omar Hussein ◽

Bruce Walters ◽

Randolph Stroetz ◽

Paul Valencia ◽

Deborah McCall ◽

...

Keyword(s):

Mechanical Ventilation ◽

Plasma Membrane ◽

Epithelial Cells ◽

Cell Injury ◽

Membrane Integrity ◽

Alveolar Epithelial Cells ◽

Interfacial Stress ◽

Interactive Effects ◽

Mechanically Ventilated ◽

Alveolar Epithelial

Mechanical ventilation may cause harm by straining lungs at a time they are particularly prone to injury from deforming stress. The objective of this study was to define the relative contributions of alveolar overdistension and cyclic recruitment and “collapse” of unstable lung units to membrane wounding of alveolar epithelial cells. We measured the interactive effects of tidal volume (VT), transpulmonary pressure (PTP), and of airspace liquid on the number of alveolar epithelial cells with plasma membrane wounds in ex vivo mechanically ventilated rat lungs. Plasma membrane integrity was assessed by propidium iodide (PI) exclusion in confocal images of subpleural alveoli. Cyclic inflations of normal lungs from zero end-expiratory pressure to 40 cmH2O produced VT values of 56.9 ± 3.1 ml/kg and were associated with 0.12 ± 0.12 PI-positive cells/alveolus. A preceding tracheal instillation of normal saline (3 ml) reduced VT to 49.1 ± 6 ml/kg but was associated with a significantly greater number of wounded alveolar epithelial cells (0.52 ± 0.16 cells/alveolus; P < 0.01). Mechanical ventilation of completely saline-filled lungs with saline (VT = 52 ml/kg) to pressures between 10 and 15 cmH2O was associated with the least number of wounded epithelial cells (0.02 ± 0.02 cells/alveolus; P < 0.01). In mechanically ventilated, partially saline-filled lungs, the number of wounded cells increased substantially with VT, but, once VT was accounted for, wounding was independent of maximal PTP. We found that interfacial stress associated with the generation and destruction of liquid bridges in airspaces is the primary biophysical cell injury mechanism in mechanically ventilated lungs.

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Single-Cell Transcriptomics Identifies Dysregulated Metabolic Programs of Aging Alveolar Progenitor Cells in Lung Fibrosis

10.1101/2020.07.30.227892 ◽

2020 ◽

Author(s):

Jiurong Liang ◽

Guanling Huang ◽

Xue Liu ◽

Forough Taghavifar ◽

Ningshan Liu ◽

...

Keyword(s):

Lipid Metabolism ◽

Epithelial Cells ◽

Lung Injury ◽

Single Cell ◽

Cell Types ◽

Alveolar Epithelial Cells ◽

Lung Epithelial Cells ◽

Alveolar Epithelial ◽

Metabolic Defects ◽

Lung Epithelial

ABSTRACTAging is a critical risk factor in progressive lung fibrotic diseases such as idiopathic pulmonary fibrosis (IPF). Loss of integrity of type 2 alveolar epithelial cells (AEC2s) is the main causal event in the pathogenesis of IPF. To systematically examine the genomic program changes of AEC2s with aging and lung injury, we performed unbiased single cell RNA-seq analyses of lung epithelial cells from either uninjured or bleomycin-injured young and old mice. Major lung epithelial cell types were readily identified with canonical cell markers in our dataset. Heterogenecity of AEC2s was apparent, and AEC2s were then classified into three subsets according to their gene signatures. Genes related to lipid metabolism and glycolysis were significantly altered within these three clusters of AEC2s, and also affected by aging and lung injury. Importantly, IPF AEC2s showed similar genomic programming and metabolic changes as that of AEC2s from bleomycin injured old mouse lungs relative to controls. Furthermore, perturbation of both lipid metabolism and glycolysis significantly changed progenitor renewal capacity in 3-Demensional organoid culture of AEC2s. Taken togather, this work identified metabolic defects of AEC2s in aging and during lung injury. Strategies to rectify these altered programs would promote AEC2 renewal which in turn improves lung repair.One sentence summaryMetabolic defects of alveolar progenitors in aging and during lung injury impair their renewal.

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Senescence associated long non-coding RNA 1 regulates cigarette smoke-induced senescence of type II alveolar epithelial cells through sirtuin-1 signaling

Journal of International Medical Research ◽

10.1177/0300060520986049 ◽

2021 ◽

Vol 49 (2) ◽

pp. 030006052098604

Author(s):

Dong Yuan ◽

Yuanshun Liu ◽

Mengyu Li ◽

Hongbin Zhou ◽

Liming Cao ◽

...

Keyword(s):

Epithelial Cells ◽

Cigarette Smoke ◽

Alveolar Epithelial Cell ◽

Alveolar Epithelial Cells ◽

Lung Epithelial Cells ◽

Sirtuin 1 ◽

Type Ii ◽

Alveolar Epithelial ◽

Non Coding Rna ◽

Long Non Coding Rna

Objective The primary aim of our study was to explore the mechanisms through which long non-coding RNA (lncRNA)-mediated sirtuin-1 (SIRT1) signaling regulates type II alveolar epithelial cell (AECII) senescence induced by a cigarette smoke-media suspension (CSM). Methods Pharmacological SIRT1 activation was induced using SRT2104 and senescence-associated lncRNA 1 (SAL-RNA1) was overexpressed. The expression of SIRT1, FOXO3a, p53, p21, MMP-9, and TIMP-1 in different groups was detected by qRT-PCR and Western blotting; the activity of SA-β gal was detected by staining; the binding of SIRT1 to FOXO3a and p53 gene transcription promoters was detected by Chip. Results We found that CSM increased AECII senescence, while SAL-RNA1 overexpression and SIRT1 activation significantly decreased levels of AECII senescence induced by CSM. Using chromatin immunoprecipitation, we found that SIRT1 bound differentially to transcriptional complexes on the FOXO3a and p53 promoters. Conclusion Our results suggested that lncRNA-SAL1-mediated SIRT1 signaling reduces senescence of AECIIs induced by CSM. These findings suggest a new therapeutic target to limit the irreversible apoptosis of lung epithelial cells in COPD patients.

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