Putative Roles for Peptidylarginine Deiminases in COVID-19

Peptidylarginine deiminases (PADs) are a family of calcium-regulated enzymes that are phylogenetically conserved and cause post-translational deimination/citrullination, contributing to protein moonlighting in health and disease. PADs are implicated in a range of inflammatory and autoimmune conditions, in the regulation of extracellular vesicle (EV) release, and their roles in infection and immunomodulation are known to some extent, including in viral infections. In the current study we describe putative roles for PADs in COVID-19, based on in silico analysis of BioProject transcriptome data (PRJNA615032 BioProject), including lung biopsies from healthy volunteers and SARS-CoV-2-infected patients, as well as SARS-CoV-2-infected, and mock human bronchial epithelial NHBE and adenocarcinoma alveolar basal epithelial A549 cell lines. In addition, BioProject Data PRJNA631753, analysing patients tissue biopsy data (n = 5), was utilised. We report a high individual variation observed for all PADI isozymes in the patients’ tissue biopsies, including lung, in response to SARS-CoV-2 infection, while PADI2 and PADI4 mRNA showed most variability in lung tissue specifically. The other tissues assessed were heart, kidney, marrow, bowel, jejunum, skin and fat, which all varied with respect to mRNA levels for the different PADI isozymes. In vitro lung epithelial and adenocarcinoma alveolar cell models revealed that PADI1, PADI2 and PADI4 mRNA levels were elevated, but PADI3 and PADI6 mRNA levels were reduced in SARS-CoV-2-infected NHBE cells. In A549 cells, PADI2 mRNA was elevated, PADI3 and PADI6 mRNA was downregulated, and no effect was observed on the PADI4 or PADI6 mRNA levels in infected cells, compared with control mock cells. Our findings indicate a link between PADI expression changes, including modulation of PADI2 and PADI4, particularly in lung tissue, in response to SARS-CoV-2 infection. PADI isozyme 1–6 expression in other organ biopsies also reveals putative links to COVID-19 symptoms, including vascular, cardiac and cutaneous responses, kidney injury and stroke. KEGG and GO pathway analysis furthermore identified links between PADs and inflammatory pathways, in particular between PAD4 and viral infections, as well as identifying links for PADs with a range of comorbidities. The analysis presented here highlights roles for PADs in-host responses to SARS-CoV-2, and their potential as therapeutic targets in COVID-19.

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Differential organ-specific inflammatory response to progranulin in high-fat diet-fed mice

Scientific Reports ◽

10.1038/s41598-020-80940-8 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Maki Murakoshi ◽

Tomohito Gohda ◽

Eri Adachi ◽

Saki Ichikawa ◽

Shinji Hagiwara ◽

...

Keyword(s):

Adipose Tissue ◽

High Fat Diet ◽

Kidney Injury ◽

Proximal Tubules ◽

Tubular Damage ◽

Standard Diet ◽

Mrna Levels ◽

High Fat ◽

Organ Specific

AbstractProgranulin (PGRN) has been reported to bind tumor necrosis factor (TNF) receptor and to inhibit TNFα signaling. We evaluated the effect of augmentation of TNFα signaling by PGRN deficiency on the progression of kidney injury. Eight-week-old PGRN knockout (KO) and wild-type (WT) mice were fed a standard diet or high-fat diet (HFD) for 12 weeks. Albuminuria, markers of tubular damage, and renal mRNA levels of inflammatory cytokines were higher in HFD-fed KO (KO-HFD) mice than in HFD-fed WT (WT-HFD) mice. Body weight, vacuolization in proximal tubules, and systemic and adipose tissue inflammatory markers were lower in the KO-HFD mice than in the WT-HFD mice. The renal megalin expression was lower in the KO mice than in the WT mice regardless of the diet type. The megalin expression was also reduced in mouse proximal tubule epithelial cells stimulated with TNFα and in those with PGRN knockdown by small interfering RNA in vitro. PGRN deficiency was associated with both exacerbated renal inflammation and decreased systemic inflammation, including that in the adipose tissue of mice with HFD-induced obesity. Improved tubular vacuolization in the KO-HFD mice might partially be explained by the decreased expression of megalin in proximal tubules.

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Identification of novel antiviral drug combinations in vitro and tracking their development

10.1101/2020.09.17.299933 ◽

2020 ◽

Author(s):

Aleksandr Ianevski ◽

Rouan Yao ◽

Svetlana Biza ◽

Eva Zusinaite ◽

Andres Männik ◽

...

Keyword(s):

Viral Infections ◽

Human Lung ◽

Antiviral Drug ◽

A549 Cells ◽

Drug Combinations ◽

Investigational Drug ◽

Synergistic Activity ◽

Lung Epithelial ◽

Reduced Toxicity

AbstractCombination therapies have become a standard for the treatment for HIV and HCV infections. They are advantageous over monotherapies due to better efficacy and reduced toxicity, as well as the ability to prevent the development of resistant viral strains and to treat viral co-infections. Here, we identify several new synergistic combinations against emerging and re-emerging viral infections in vitro. We observed synergistic activity of nelfinavir with investigational drug EIDD-2801 and convalescent serum against SARS-CoV-2 infection in human lung epithelial Calu-3 cells. We also demonstrated synergistic activity of vemurafenib combination with emetine, homoharringtonine, gemcitabine, or obatoclax against echovirus 1 infection in human lung epithelial A549 cells. We also found that combinations of sofosbuvir with brequinar and niclosamide were synergistic against HCV infection in hepatocyte derived Huh-7.5 cells, whereas combinations of monensin with lamivudine and tenofovir were synergistic against HIV-1 infection in human cervical TZM-bl cells. Finally, we present an online resource that summarizes novel and known antiviral drug combinations and their developmental status. Overall, the development of combinational therapies could have a global impact improving the preparedness and protection of the general population from emerging and re-emerging viral threats.

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Synthetic Peptides as a Promising Alternative to Control Viral Infections in Atlantic Salmon

Pathogens ◽

10.3390/pathogens9080600 ◽

2020 ◽

Vol 9 (8) ◽

pp. 600 ◽

Cited By ~ 1

Author(s):

Constanza Cárdenas ◽

Fanny Guzmán ◽

Marisela Carmona ◽

Cristian Muñoz ◽

Luis Nilo ◽

...

Keyword(s):

Viral Load ◽

Viral Infections ◽

Antiviral Agents ◽

In Silico Analysis ◽

Infectious Pancreatic Necrosis Virus ◽

Fish Cell ◽

Promising Alternative ◽

Anemia Virus

Viral infections in salmonids represent an ongoing challenge for the aquaculture industry. Two RNA viruses, the infectious pancreatic necrosis virus (IPNV) and the infectious salmon anemia virus (ISAV), have become a latent risk without healing therapies available for either. In this context, antiviral peptides emerge as effective and relatively safe therapeutic molecules. Based on in silico analysis of VP2 protein from IPNV and the RNA-dependent RNA polymerase from ISAV, a set of peptides was designed and were chemically synthesized to block selected key events in their corresponding infectivity processes. The peptides were tested in fish cell lines in vitro, and four were selected for decreasing the viral load: peptide GIM182 for IPNV, and peptides GIM535, GIM538 and GIM539 for ISAV. In vivo tests with the IPNV GIM 182 peptide were carried out using Salmo salar fish, showing a significant decrease of viral load, and proving the safety of the peptide for fish. The results indicate that the use of peptides as antiviral agents in disease control might be a viable alternative to explore in aquaculture.

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Premobilization of CD133+ progenitors is associated with attenuated inflammation-induced pulmonary dysfunction following extracorporeal circulation in mice

Interactive Cardiovascular and Thoracic Surgery ◽

10.1093/icvts/ivaa074 ◽

2020 ◽

Vol 31 (2) ◽

pp. 210-220

Author(s):

Dan Luo ◽

Xinhao Liu ◽

Jie Zhang ◽

Lei Du ◽

Lin Bai ◽

...

Keyword(s):

Acute Kidney Injury ◽

Lung Injury ◽

Bronchoalveolar Lavage ◽

Lung Tissue ◽

Extracorporeal Circulation ◽

Kidney Injury ◽

Inflammatory Factors ◽

Pulmonary Dysfunction

Abstract OBJECTIVES Progenitor cells mobilized by granulocyte colony-stimulating factor (G-CSF) have been shown to lessen acute kidney injury induced by extracorporeal circulation (ECC). Both acute kidney injury and lung injury are characterized by endothelial dysfunction. Our goal was to examine whether and how G-CSF-mobilized progenitors with endothelial capacity may help mitigate ECC-induced pulmonary dysfunction. METHODS G-CSF (10 μg/kg/day) was administered subcutaneously to C57BL/6 mice before or at the initiation of the ECC process, after which lung injury was assessed by measuring neutrophils in the fluid from bronchoalveolar lavage and determining the pathological score in lung tissue. CD133+ progenitors were isolated and injected into C57BL/6 mice before ECC in vivo. We incubated the CD133+ cells with pulmonary monocytes or neutrophils isolated from naïve mice in vitro. RESULTS Pretreatment with G-CSF for 2 days significantly decreased the number of neutrophils in the bronchoalveolar lavage fluid, and the pathological score (P < 0.01; n = 5) improved the PaO2/FiO2 ratio [193.4 ± 12.7 (ECC without G-CSF) vs 305.6 ± 22.6 mmHg (ECC with G-CSF); P = 0.03, n = 5] and suppressed neutrophil elastase and tumour necrosis factor-α levels in the circulation; we also observed increases in both circulating and pulmonary populations of CD133+ progenitors. Similar effects were observed in animals pretreated with CD133+ progenitors instead of G-CSF before ECC. The majority of CD133+/CD45− and CD133+/CD45+ progenitors were mobilized in the lung and in the circulation, respectively. Incubating CD133+ progenitors with neutrophils or pulmonary monocytes blocked lipopolysaccharide-induced release of inflammatory factors. CONCLUSIONS Our results suggest that pretreatment of G-CSF attenuates ECC-induced pulmonary dysfunction through inhibiting the inflammatory response in lung tissue and in the circulation with associated premobilization of CD133+ progenitors.

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Inhibitor of DNA Binding 1 Is Induced during Kidney Ischemia-Reperfusion and Is Critical for the Induction of Hypoxia-Inducible Factor-1α

BioMed Research International ◽

10.1155/2016/4634386 ◽

2016 ◽

Vol 2016 ◽

pp. 1-10 ◽

Cited By ~ 8

Author(s):

Dan Wen ◽

Yan-Fang Zou ◽

Yao-Hui Gao ◽

Qian Zhao ◽

Yin-Yin Xie ◽

...

Keyword(s):

Dna Binding ◽

Ischemia Reperfusion ◽

Hypoxia Inducible Factor ◽

Kidney Injury ◽

Mrna Levels ◽

Hypoxia Inducible Factor 1 ◽

Renal Tubular ◽

Inhibitor Of Dna Binding

In this study, rat models of acute kidney injury (AKI) induced by renal ischemia-reperfusion (I/R) and HK-2 cell models of hypoxia-reoxygenation (H/R) were established to investigate the expression of inhibitor of DNA binding 1 (ID1) in AKI, and the regulation relationship between ID1 and hypoxia-inducible factor 1 alpha (HIF-1α). Through western blot, quantitative real-time PCR, immunohistochemistry, and other experiment methods, the induction of ID1 after renal I/R in vivo was observed, which was expressed mainly in renal tubular epithelial cells (TECs). ID1 expression was upregulated in in vitro H/R models at both the protein and mRNA levels. Via RNAi, it was found that ID1 induction was inhibited with silencing of HIF-1α. Moreover, the suppression of ID1 mRNA expression could lead to decreased expression and transcription of HIF-1αduring hypoxia and reoxygenation. In addition, it was demonstrated that both ID1 and HIF-1αcan regulate the transcription of twist. This study demonstrated that ID1 is induced in renal TECs during I/R and can regulate the transcription and expression of HIF-1α.

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IL-17A activated by Toll-like receptor 9 contributes to the development of septic acute kidney injury

AJP Renal Physiology ◽

10.1152/ajprenal.00313.2019 ◽

2020 ◽

Vol 318 (1) ◽

pp. F238-F247

Author(s):

Yoshitaka Naito ◽

Takayuki Tsuji ◽

Soichiro Nagata ◽

Naoko Tsuji ◽

Tomoyuki Fujikura ◽

...

Keyword(s):

Acute Kidney Injury ◽

Critical Role ◽

Cecal Ligation ◽

Kidney Injury ◽

Mrna Levels ◽

Toll Like Receptor ◽

Γδt Cells ◽

Morphological Aspects ◽

Septic Acute Kidney Injury

Toll-like receptor 9 (TLR9), which is activated by endogenously released mtDNA during sepsis, contributes to the development of polymicrobial septic acute kidney injury (AKI). However, downstream factors of TLR9 to AKI remain unknown. We hypothesized that IL-17A activated by TLR9 may play a critical role in septic AKI development. To determine the effects of TLR9 on IL-17A production in septic AKI, we used a cecal ligation and puncture (CLP) model in Tlr9 knockout ( Tlr9KO) mice and wild-type (WT) littermates. We also investigated the pathway from TLR9 activation in dendritic cells (DCs) to IL-17A production by γδT cells in vitro. To elucidate the effects of IL-17A on septic AKI, Il-17a knockout ( Il-17aKO) mice and WT littermates were subjected to CLP. We further investigated the relationship between the TLR9-IL-17A axis and septic AKI by intravenously administering recombinant IL-17A or vehicle into Tlr9KO mice and assessing kidney function. IL-17A levels in both plasma and the peritoneal cavity and mRNA levels of IL-23 in the spleen were significantly higher in WT mice after CLP than in Tlr9KO mice. Bone marrow-derived DCs activated by TLR9 induced IL-23 and consequently promoted IL-17A production in γδT cells in vitro. Knockout of Il-17a improved survival, functional and morphological aspects of AKI, and splenic apoptosis after CLP. Exogenous IL-17A administration aggravated CLP-induced AKI attenuated by knockout of Tlr9. TLR9 in DCs mediated IL-17A production in γδT cells during sepsis and contributed to the development of septic AKI.

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Effect of GDC-0449, a hedgehog (Hh) pathway inhibitor, on tumor activity and on erlotinib and cisplatin in a xenograft model of non-small cell lung cancer (NSCLC) with activated Hh pathway.

Journal of Clinical Oncology ◽

10.1200/jco.2012.30.15_suppl.e21057 ◽

2012 ◽

Vol 30 (15_suppl) ◽

pp. e21057-e21057

Author(s):

Aamir Ahmad ◽

Fazlul H. Sarkar ◽

Main Maitah ◽

Shadan Ali ◽

Seema Sethi ◽

...

Keyword(s):

Cell Motility ◽

A549 Cells ◽

Xenograft Model ◽

Nsclc Cell Line ◽

M Cells ◽

Mrna Levels ◽

Mesenchymal Transition ◽

Clonogenic Growth

e21057 Background: We have previously shown that chronic exposure of A549 NSCLC cell line to TGF-β1 can induce epithelial-mesenchymal transition (EMT) and can make these cell lines (A549-M) resistant to erlotinib, an EGFR inhibitor and cisplatin. EMT in A549-M cells was associated with significant activation of signaling through the Hh pathway. We therefore evaluated the activity of GDC-0449, an inhibitor of the Hh pathway, and assessed its activity in combination with erlotinib and cisplatin in A549-M cells both in vitro and in vivo. Methods: We assessed the effects of GDC-0449 (20 nM) on clonogenic growth (colony formation assay), cell motility (wound healing assay) and invasion (matrigel assay) in A549-M cells, and also assessed the activity of GDC-0449 in combination with erlotinib and cisplatin both in vitro (MTT assay) and in an experimental lung metastasis animal model in vivo by injecting A549-M cells through tail vein. Results: A549-M cells showed significantly higher rate of clonogenic growth, cell motility and invasion in vitro and a greater propensity to form metastases in vivo compared to parental A549 cells, and also showed higher resistance to erlotinib and cisplatin. A549-M cells showed higher mRNA levels of Shh (sonic hedgehog, a ligand of the Hh pathway) and ZEB1 and reduced levels of E-cadherin than A549 cells. GDC-0449 significantly reduced growth of A-549-M cell in vitro but had no effect on A-549 cells. GDC-0449 also reduced clonogenic growth, cell motility and invasion of A549-M cells. Pre-treatment with GDC-0449 significantly enhanced the activity of erlotinib (5 and 10 mM) and cisplatin (1mM-10mM) in A549-M cells (p < 0.05) but not in A-549 cells. GDC-0449 reduced the number of lung metastases in the xenograft model and enhanced the activity of erlotinib and cisplatin in this model. Conclusions: This data suggests that the EMT phenotype in NSCLC may be associated with the activation of the Hh signaling pathway. It also suggests that GDC-0449 has anti-tumor activity against NSCLC with activated Hh pathway, and can enhance the effects of erlotinib and cisplatin in such tumors.

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Short-term and Long-term Exposure of the MucilAir™ Model to Polycyclic Aromatic Hydrocarbons

Alternatives to Laboratory Animals ◽

10.1177/0261192919841484 ◽

2019 ◽

Vol 47 (1) ◽

pp. 9-18 ◽

Cited By ~ 3

Author(s):

Tereza Cervena ◽

Kristyna Vrbova ◽

Andrea Rossnerova ◽

Jan Topinka ◽

Pavel Rossner

Keyword(s):

Polycyclic Aromatic Hydrocarbons ◽

Lung Tissue ◽

Aromatic Hydrocarbons ◽

Messenger Rna ◽

Mrna Levels ◽

Short Term ◽

Polycyclic Aromatic ◽

Ldh Release

Cells grown in monocultures are widely used to model lung tissue. As a result of these culture conditions, these cells exhibit poor morphological similarity to those present in in vivo lung tissue. MucilAir™, a 3-D in vitro model comprising human basal, goblet and ciliated cells, represents a fully differentiated respiratory epithelium that can be used as an alternative and a more realistic system. The aim of our study was to compare the effects of short-term and long-term exposure to two polycyclic aromatic hydrocarbons (PAHs) — benzo[a]pyrene (B[a]P) and 3-nitrobenzanthrone (3-NBA) — using MucilAir as a model of human lung tissue. Two concentrations (0.1 μM and 1 μM) were tested at three time points (24 hours, 7 days and 28 days). Several aspects were assessed: cytotoxicity (lactate dehydrogenase (LDH) release), integrity of the cell layer (transepithelial electrical resistance (TEER)), induction of oxidative stress (reactive oxygen species production) and changes in the expression of selected genes involved in PAH metabolism ( CYP1A1 and AKR1C2) and the antioxidant response ( ALDH3A1, SOD1, SOD2, GPX1, CAT, HMOX1 and TXNRD1). The results showed that exposure to B[a]P caused a spike in LDH release at day 5. Exposure to 3-NBA caused a number of spikes in LDH release, starting at day 5, and a decrease in TEER after 11 days. CYP1A1 gene expression was upregulated after the 7-day and 28-day B[a]P exposures, as well as after the 24-hour and 7-day 3-NBA exposures. HMOX1 and SOD1 were downregulated after both 24-hour PAH treatments. HMOX1 was upregulated after a 1-week exposure to 3-NBA. There were no significant changes in the messenger RNA (mRNA) levels of AKR1C2, ALDH3A1, TXNRD1, SOD2, GPX1 or CAT. These results illustrate the potential use of this 3-D in vitro lung tissue model in studying the effects of chronic exposure to PAHs.

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Aquaporin-1 attenuates macrophage-mediated inflammatory responses by inhibiting p38 mitogen-activated protein kinase activation in lipopolysaccharide-induced acute kidney injury

Inflammation Research ◽

10.1007/s00011-019-01285-1 ◽

2019 ◽

Vol 68 (12) ◽

pp. 1035-1047 ◽

Cited By ~ 5

Author(s):

Bohui Li ◽

Chunmei Liu ◽

Kaihong Tang ◽

Xuening Dong ◽

Longge Xue ◽

...

Keyword(s):

P38 Mapk ◽

Inflammatory Responses ◽

Kidney Injury ◽

Protective Role ◽

Mitogen Activated Protein Kinase ◽

Regulatory Mechanisms ◽

Detection Methods ◽

Inflammatory Factors ◽

Mrna Levels

Abstract Objective This study was designed to investigate the role of AQP1 in the development of LPS-induced AKI and its potential regulatory mechanisms in the inflammatory responses of macrophages. Methods Male Wistar rats were injected intraperitoneally with LPS, and biochemical and histological renal damage was assessed. The levels of inflammatory mediators, macrophage markers and AQP1 in blood and kidney tissues were assessed by ELISA. RTPCR was used to assess changes in the relative levels of AQP1 mRNA induced by LPS. Western blot and immunofluorescence analyses were performed to assay the activation of the p38 MAPK and NF-κB pathways, respectively. The same detection methods were used in vitro to determine the regulatory mechanisms underlying AQP1 function. Results AQP1 mRNA levels were dramatically decreased in AKI rats following the increased expression of inflammatory factors. In vitro experiments demonstrated that silencing the AQP1 gene increased inflammatory mediator secretion, altered the classical activation of macrophages, greatly enhanced the phosphorylation of p38 and accelerated the translocation of NF-κB. Furthermore, these results were blocked by doramapimod, a p38 inhibitor. Therefore, these effects were mediated by the increased phosphorylation of p38 MAPK. Conclusion Our results suggest that altered AQP1 expression may be associated with the development of inflammation in AKI. AQP1 plays a protective role in modulating acute renal injury and can attenuate macrophage-mediated inflammatory responses by downregulating p38 MAPK activity in LPS-induced RAW264.7 cells. The pharmacological targeting of AQP1-mediated p38 MAPK signalling may provide a novel treatment approach for AKI.

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The fibroblast growth factor receptor mediates the increased FGF23 expression in acute and chronic uremia

AJP Renal Physiology ◽

10.1152/ajprenal.00332.2015 ◽

2016 ◽

Vol 310 (3) ◽

pp. F217-F221 ◽

Cited By ~ 19

Author(s):

Alia Hassan ◽

Karina Durlacher ◽

Justin Silver ◽

Tally Naveh-Many ◽

Ronen Levi

Keyword(s):

Acute Kidney Injury ◽

Folic Acid ◽

Growth Factor Receptor ◽

Kidney Injury ◽

Mrna Levels ◽

High Phosphorus ◽

Chronic Uremia ◽

Serum Pth

Serum FGF23 is markedly elevated in chronic kidney disease and has been associated with poor long-term outcomes. FGF23 expression is increased by activation of the FGF receptor 1 (FGFR1) in rats with normal renal function and in vitro in bone-derived osteoblast-like cells. We studied the regulation of FGF23 by FGFR1 in vivo in acute and chronic uremia in mice and rats. Folic acid-induced acute kidney injury increased calvaria FGF23 mRNA and serum FGF23 and parathyroid hormone (PTH) levels at 6 h. The FGFR1 receptor inhibitor PD173074 prevented the folic acid-induced increase in both FGF23 mRNA and serum levels but had no effect on serum PTH levels. A more prolonged uremia due to an adenine high-phosphorus diet for 14 days resulted in high levels of FGF23 mRNA and serum FGF23 and PTH. PD173074 decreased serum FGF23 and mRNA levels with no effect on PTH in the adenine high phosphorus-induced uremic rats. Therefore, a derangement in FGF23 regulation starts early in the course of acute kidney injury, is in part independent of the increase in serum PTH, and involves activation of FGFR1. It is possible that FGFR1 in the osteocyte is activated by locally produced canonical FGFs, which are increased in uremia. This is the first demonstration that activation of FGFR1 is essential for the high levels of FGF23 in acute and chronic experimental uremia.

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