A Monoclonal Human Alveolar Epithelial Cell Line (“Arlo”) with Pronounced Barrier Function for Studying Drug Permeability and Viral Infections

Advanced Science, Год журнала: 2023, Номер 10(8)

Опубликована: Фев. 7, 2023

Abstract In the development of orally inhaled drug products preclinical animal models regularly fail to predict pharmacological as well toxicological responses in humans. Models based on human cells and tissues are potential alternatives experimentation allowing for isolation essential processes biology making them accessible vitro. Here, generation a novel monoclonal cell line “Arlo,” derived from polyclonal alveolar epithelium lentivirus immortalized hAELVi via single‐cell printing, its characterization model building block future complex vitro is described. “Arlo” systematically compared primary epithelial (hAEpCs) line. show enhanced barrier properties with high transepithelial electrical resistance (TEER) ≈3000 Ω cm 2 difference (PD) ≈30 mV under air–liquid interface (ALI) conditions, that can be modulated. The grow polarized monolayer express genes relevant integrity homeostasis observed hAEpCs. Successful productive infection severe acute respiratory syndrome coronavirus (SARS‐CoV‐2) proof‐of‐principle study offers an additional, attractive application beyond biopharmaceutical experimentation.

Язык: Английский

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Silver N-heterocyclic carbene complexes are potent uncompetitive inhibitors of the papain-like protease with antiviral activity against SARS-CoV-2

RSC Medicinal Chemistry, Год журнала: 2023, Номер 14(7), С. 1260 - 1271

Опубликована: Янв. 1, 2023

The ongoing SARS-CoV-2 pandemic has caused a high demand for novel innovative antiviral drug candidates. Despite promising results, metal complexes have been relatively unexplored as agents in general and particular against SARS-CoV-2. Here we report on silver NHC with chloride or iodide counter ligands that are potent inhibitors of the papain-like protease (PLpro) but inactive 3C-like (3CLpro) another protease. Mechanistic studies selected complex confirmed zinc removal from binding domain PLpro relevant factor their activity. In addition, enzyme kinetic experiments revealed is an uncompetitive inhibitor this rare type inhibition it offers great pharmacological advantages terms selectivity. showed very low absent host cell toxicity triggered strong effects viral replication cells infected SARS-CoV-2, making them future

Язык: Английский

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Omicron‐induced interferon signaling prevents influenza A H1N1 and H5N1 virus infection

Journal of Medical Virology, Год журнала: 2023, Номер 95(3)

Опубликована: Март 1, 2023

Recent findings in permanent cell lines suggested that SARS-CoV-2 Omicron BA.1 induces a stronger interferon response than Delta. Here, we show and BA.5 but not Delta induce an antiviral state air-liquid interface cultures of primary human bronchial epithelial cells monocytes. Both subvariants caused the production biologically active types I (α/β) III (λ) interferons protected from super-infection with influenza A viruses. Notably, abortive infection monocytes was sufficient to protect virus infection. Interestingly, while influenza-like illnesses surged during wave England, their spread rapidly declined upon emergence Omicron. Mechanistically, Omicron-induced signaling mediated via double-stranded RNA recognition by MDA5, as MDA5 knockout prevented it. The JAK/STAT inhibitor baricitinib inhibited Omicron-mediated response, suggesting it is MDA5-mediated production, which activates receptors then trigger signaling. In conclusion, our study (1) demonstrates only substantial physiologically relevant models, (2) shows protects super-infection, (3) indicates comparable states.

Язык: Английский

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Phosphoglycerate mutase regulates Treg differentiation through control of serine synthesis and one-carbon metabolism

Опубликована: Янв. 30, 2025

The differentiation and suppressive functions of regulatory CD4 T cells (Tregs) are supported by a broad array metabolic changes, providing potential therapeutic targets for immune modulation. In this study, we focused on the role glycolytic enzymes in Tregs identified phosphoglycerate mutase (PGAM) as being differentially overexpressed associated with highly phenotype. Pharmacologic or genetic inhibition PGAM reduced Treg function while reciprocally inducing markers pro-inflammatory, helper 17 (Th17)-like state. was dependent contribution 3-phosphoglycerate (3PG), substrate, to de novo serine synthesis. Blocking synthesis from 3PG reversed effect polarization, exogenous directly inhibited polarization. Additionally, altering levels vivo serine/glycine-free diet increased peripheral attenuated autoimmunity murine model multiple sclerosis. Mechanistically, found that limits polarization contributing one-carbon metabolism methylation Treg-associated genes. Inhibiting both vitro autoimmune colitis. Our study identifies novel physiologic highlights interconnectivity between glycolysis, synthesis, metabolism, epigenetic regulation function.

Язык: Английский

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Phosphoglycerate mutase regulates Treg differentiation through control of serine synthesis and one-carbon metabolism

Опубликована: Янв. 30, 2025

The differentiation and suppressive functions of regulatory CD4 T cells (Tregs) are supported by a broad array metabolic changes, providing potential therapeutic targets for immune modulation. In this study, we focused on the role glycolytic enzymes in Tregs identified phosphoglycerate mutase (PGAM) as being differentially overexpressed associated with highly phenotype. Pharmacologic or genetic inhibition PGAM reduced Treg function while reciprocally inducing markers pro-inflammatory, helper 17 (Th17)-like state. was dependent contribution 3-phosphoglycerate (3PG), substrate, to de novo serine synthesis. Blocking synthesis from 3PG reversed effect polarization, exogenous directly inhibited polarization. Additionally, altering levels vivo serine/glycine-free diet increased peripheral attenuated autoimmunity murine model multiple sclerosis. Mechanistically, found that limits polarization contributing one-carbon metabolism methylation Treg-associated genes. Inhibiting both vitro autoimmune colitis. Our study identifies novel physiologic highlights interconnectivity between glycolysis, synthesis, metabolism, epigenetic regulation function.

Язык: Английский

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Discovery of anti-SARS-CoV-2 secondary metabolites from the heartwood of Pterocarpus santalinus using multi-informative molecular networking

Frontiers in Molecular Biosciences, Год журнала: 2023, Номер 10

Опубликована: Июнь 6, 2023

A pigment-depleted extract from the heartwood of Pterocarpus santalinus L. f. (PS-DE) showed promising anti-SARS-CoV-2 activity with an IC50 29.9 μg/mL in Caco-2-F03 cells. To determine potential active constituents within prior to isolation, multi-informative molecular network (MN) was applied. Therefore, separated by high-performance counter-current chromatography (HPCCC) into 11 fractions which were subsequently tested for and analysed UPLC-tandem mass spectrometry (MS2). The resulting MN combines bioactivity data MS2 data. analysis led targeted isolation seven compounds including one pterocarpan (7) reported first time as constituent P. four so far undescribed natural products (NPs) that belong compound classes arylpropanes (9), isoflavanones (10) coumestans (16) 3-arylcoumarins (17), respectively. In total, 15 synthetic isoflavonoid is structurally related metabolites activity. Thereby, two pterocarpans (-)-homopterocarpin (5) (-)-medicarpin (2), stilbene (E)-pterostilbene (1) 7-O-methylgenistein (11) a distinct antiviral values 17.2, 33.4, 34.7, 37.9 µM, respectively, no cytotoxic effects against cells (CC50 > 100 µM). addition, structure-activity relationship (SAR) proposed indicating structural requirements herein presented results support implementation networks powerful tool dereplication bioactive NPs.

Язык: Английский

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Molecular networking unveils anti-SARS-CoV-2 constituents from traditionally used remedies

Journal of Ethnopharmacology, Год журнала: 2023, Номер 319, С. 117206 - 117206

Опубликована: Сен. 30, 2023

Язык: Английский

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Validation of nuclear receptor RORγ isoform 1 as a novel host-directed antiviral target based on the modulation of cholesterol levels

Antiviral Research, Год журнала: 2023, Номер 221, С. 105769 - 105769

Опубликована: Дек. 4, 2023

Язык: Английский

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Synergism of interferon-beta with antiviral drugs against SARS-CoV-2 variants

Journal of Infection, Год журнала: 2022, Номер 85(5), С. 573 - 607

Опубликована: Июль 31, 2022

Язык: Английский

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The cell cultures in virology: from the past to the future

Journal of microbiology epidemiology immunobiology, Год журнала: 2024, Номер 101(1), С. 143 - 153

Опубликована: Март 9, 2024

The aim of the review is to give a brief characteristic cell cultures obtained from mammalian tissues and consider current possibilities prospects for their use in virology. analysis literature data presented main databases, such as Web Science, PubMed, Scopus, Elsevier, Google Scholar RSCI (as July 2023), indicates that various types are currently used virological studies. culture has number advantages over other vitro vivo methods research. provides numerous examples on development new obtaining cultivation viruses. Among them sensitive reporter systems, design which can be promising tool diagnostics existing unknown viral infections. Cell characterized potential models virology developing diagnostic test-systems antiviral drugs. An important area application substrate production culture-derived vaccines. Another aspect also highlighted, study effect viruses host immune system or mechanisms immunopathogenesis It concluded remains near future one most practical scientific

Язык: Английский

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