bims-mevinf 2025-06-15 papers

J Virol. 2025 Jun 12. e0039425

Direct pharmacological AMPK activation inhibits mucosal SARS-CoV-2 infection by reducing lipid metabolism, restoring autophagy flux and the type I IFN response.

Andrea Cottignies-Calamarte, Flora Marteau, Feifan He, Sandrine Belouzard, Jean Dubuisson, Daniela Tudor, Benoit Viollet, Morgane Bomsel.

AMP-activated protein kinase (AMPK) plays a central role in regulating cell energy balance. When activated, AMPK suppresses energy-consuming pathways, such as lipid and protein synthesis, while increasing nutrient availability through the activation of autophagy. These pathways downstream of AMPK activation contribute to SARS-CoV-2 infection, which hijacks autophagy and accumulates lipid droplets in viral factories to support viral replication. Here, we assessed the antiviral activity of the direct pan-AMPK allosteric activator MK-8722 in vitro. MK-8722 efficiently inhibited infection of Alpha and Omicron SARS-CoV-2 variants in Vero76 and human bronchial epithelial Calu-3 cells at micromolar concentration. This inhibition relied on restoring the autophagic flux, which redirected newly synthesized viral proteins for degradation, and reduced lipid metabolism, which affected viral factories. Furthermore, MK-8722 treatment increased the type I interferon (IFN-I) response. Post-infection treatment with MK-8722 was enough to inhibit efficient viral replication and restore the IFN-I response. Finally, MK-8722 treatment did not alter the SARS-CoV-2-specific CD8+ T cell response mounted upon Spike vaccination. Overall, by activating AMPK, MK-8722 acts as an effective antiviral against SARS-CoV-2 infection, even when applied post-exposure, paving the way for preclinical tests aimed at inhibiting viral replication and improving patients' symptoms.
IMPORTANCE: Coronavirus disease 2019, caused by SARS-CoV-2 infection, has led to severe acute respiratory syndrome with very high mortality. Despite available vaccines and public health measures, new SARS-CoV-2 variants emerge with increased transmissibility requiring the development of novel therapeutic strategies. Recently, the AMP-activated protein kinase (AMPK), a cellular energy sensor, has emerged as a potential broad-spectrum antiviral target, as AMPK can modulate the intracellular environment in turn impeding viral replication. This study aims to evaluate the potential of pharmacological activation of AMPK to inhibit SARS-CoV-2 infection and replication. Our findings demonstrate that AMPK activation induces significant alterations in host cellular lipid metabolism that disrupt viral factories essential for SARS-CoV-2 replication. Furthermore, by enhancing autophagy, a process crucial for the degradation and clearance of viral particles, AMPK activation facilitates the elimination of the virus. Therefore, targeting AMPK signaling pathways could offer a promising therapeutic approach for the treatment of SARS-CoV-2 infections.

Keywords: AMP-activated protein kinase; SARS-CoV-2; antiviral agents; autophagy; lipid metabolism

DOI: https://doi.org/10.1128/jvi.00394-25

bioRxiv. 2025 Jun 03. pii: 2025.06.02.657371. [Epub ahead of print]

Multi-Omic Profiling Identifies Conserved Metabolic Pathways Critical for SARS-CoV-2 Variants Infection.

Scotland E Farley, Jennifer E Kyle, Helene Jahn, Lisa Bramer, Paul Piehowski, Athena Shepmoes, Brooke Ld Kaiser, Sarai M Williams, Josie G Eder, Carsten Schultz, Fikadu Tafesse.

The rapid evolution of SARS-CoV-2 has led to the emergence of numerous variants with en-hanced transmissibility and immune evasion. Despite widespread vaccination, infections persist, and the mechanisms by which SARS-CoV-2 reprograms host metabolism remain in-completely understood. Here, we investigated whether virus-induced lipid remodeling is con-served across variants and whether changes in lipid abundance correlate with alterations in lipid biosynthetic enzymes. Using global untargeted lipidomics and quantitative proteomics, we analyzed A549-ACE2 cells infected with the Delta (B.1.617.2) or Omicron (B.1.1.529) var-iants and compared them to cells infected with the ancestral WA1 strain. In parallel, we conducted quantitative proteomics to assess virus-induced changes in the host proteome. Our results reveal that SARS-CoV-2 drives a remarkably consistent pattern of metabolic re-wiring at both the lipidomic and proteomic levels across all three variants. We mapped changes in the expression of host metabolic enzymes and compared these to corresponding shifts in lipid abundance. This integrative analysis identified key host proteins involved in virus-mediated lipid remodeling, including fatty acid synthase (FASN), lysosomal acid lipase (LIPA), and ORM1-like protein 2 (ORMDL2). Together, these findings highlight conserved metabolic dependencies of SARS-CoV-2 variants and underscore host lipid metabolism as a potential target for broad-spectrum antiviral strategies.

DOI: https://doi.org/10.1101/2025.06.02.657371

Vet Microbiol. 2025 Jun 03. pii: S0378-1135(25)00194-4. [Epub ahead of print]307 110559

PERK-mediated eIF2α phosphorylation suppresses porcine epidemic diarrhea virus replication by attenuating global protein synthesis and inducing IFN-Ⅰ production.

Gan Yang, Yu Zhao, Haoran Guo, Pei Zhou, Ling Dong, Wentao Li, Qigai He.

Porcine epidemic diarrhea virus (PEDV) is one of the most important porcine pathogens for which no preventive and antiviral treatment measures are available. A pervious study revealed that the unfolded protein response (UPR) induced by endoplasmic reticulum (ER) stress can be utilized to inhibit PEDV replication. Here, we demonstrated that the UPR suppresses the replication of multiple genotypes of PEDV in both Vero and swine testis (ST) cells, primarily through activation of the PERK-eIF2α branch among the three UPR pathways. The PERK-eIF2α pathway inducers CCT020312 and salubrinal efficiently inhibited the replication of multiple genotypes of PEDV in both Vero and ST cells, whereas the inhibitor AMG PERK 44 promoted PEDV replication. Furthermore, we found that PERK-eIF2α arm-mediated inhibition of PEDV replication is caused by phosphorylated eIF2α-induced attenuation of global protein translation. Additionally, phosphorylated eIF2α promotes NF-κB signaling activation and facilitates to the production of IFN-Ⅰ, eliciting innate immunity to suppress viral replication. These data show that PERK-eIF2α pathway dampens the replication of multiple genotypes of PEDV, suggesting that this target may be exploited to develop as a broad-spectrum anti-PEDV drugs.

Keywords: Cellular translation; Eukaryotic initiation factor 2α; Interferon; Nuclear factor kappa B; Porcine epidemic diarrhea virus; Protein kinase R like endoplasmic reticulum kinase

DOI: https://doi.org/10.1016/j.vetmic.2025.110559

Int J Mol Sci. 2025 May 29. pii: 5209. [Epub ahead of print]26(11):

A Plant-Based Dietary Supplement Exhibits Significant Effects on Markers of Oxidative Stress, Inflammation, and Immune Response in Subjects Recovering from Respiratory Viral Infection: A Randomized, Double-Blind Clinical Study Using Vitamin C as a Positive Control.

Bruno Fink, John M Hunter, Zbigniew Pietrzkowski, Richard Fink, Coy Brunssen, Henning Morawietz, Boris Nemzer.

Respiratory viruses continue to present serious health challenges to human wellness. Growing evidence suggests that the more severe and damaging effects and symptoms of influenza, rhinovirus (RV), respiratory syncytial virus (RSV), and COVID-19 may primarily result from their common ability to disorganize the body's healthy immune response. The simultaneous over-stimulation of several reactive oxygen species (ROS) pathways and concurrent suppression of bioavailable Nitic Oxide (NO) contribute to an immune disbalance that can lead to cellular oxidative distress and an excessive inflammatory response. This study evaluated the real-time, acute ability of a single, orally administered 50 mg encapsulated dose of a plant-based dietary supplement ("PB-Blend"), compared to 1000 mg of Vitamin C as a positive control, to modulate multiple ROS associated with a dampened immune response, as well as NO and other markers of inflammation, in a cohort recovering from a moderate course of COVID-19. This randomized, double-blind study was performed on 28 individuals 18-24 days after a moderate COVID-19 infection. Participants were orally supplemented with a single encapsulated dose of either 50 mg of PB-Blend or 1000 mg Vitamin C as a positive control. Changes in the levels of bioavailable NO (measured as circulating NOHb) were assessed, as well as the ex vivo cellular formation of mitochondrial, NOX2-, iNOS-, and TNFα-dependent ROS. All parameters were measured in real time before ingestion (baseline), and then at 30, 60, 120, and 180 min after administration. ROS were measured using a portable electron paramagnetic resonance (EPR) spectrometer. Inflammatory, immunity (hsCRP and TNFα plasma levels), interleukin (IL1, IL6, IL8, and IL10), cytokine (IFNγ, TNFα, and NF-κB), and immunoglobulin (IgA, IgM, IgG, and IgE) profiles were also followed. In addition to laboratory and cell function investigations, we performed clinical cardio ergometry, blood O2 saturation, and respirometry examinations. As hypothesized, the collected baseline data from this study group confirmed that mitochondrial, NOX2, and iNOS enzymatic systems were strongly involved in the generation of ROS at 18-24 days following a positive COVID-19 PCR test. Acute single-dose supplementation of 50 mg PB-Blend had a multifunctional impact on ROS and significantly inhibited the following: (a.) mitochondrial ROS levels by up to 56%; (b.) iNOS by up to 60%; and (c.) NOX2-dependent ROS generation by up to 49%. Moreover, 1000 mg Vitamin C supplementation exhibited narrower ROS-mitigating activity by solely inhibiting NOX2-dependent ROS generation by 45%. Circulating NOHb levels were significantly increased after PB-Blend administration (33%), but not after Vitamin C administration. PB-Blend and Vitamin C exhibited similar potential to reduce ex vivo high dose TNFα (200 ng/mL)-induced H2O2 formation. These results suggest that 50 mg of PB-Blend has the potential to modulate disbalanced mitochondria, iNOS, and NOX2 enzymatic systems that can be engendered during respiratory viral infection and subsequent recovery. Moreover, PB-Blend, but not Vitamin C, showed potential to upregulate bioavailable NO, which is known to decline under these conditions. Based upon these observations, PB-Blend could be considered an alternative to, or to be used in tandem with Vitamin C in applications that promote immune support and recovery during seasons of heightened respiratory viral risk (e.g., "flu season").

Keywords: COVID-19; FMD; ROS; cellular metabolic activity (CMA); circulating NOHb; dietary supplement PB-blend; endothelial dysfunction; flu season; inducible nitric oxide synthase (iNOS); influenza; phagocytic NADPH oxidase; seasonal virus; uncoupled iNOS; viral insult

DOI: https://doi.org/10.3390/ijms26115209

J Pharm Biomed Anal. 2025 Jun 03. pii: S0731-7085(25)00351-6. [Epub ahead of print]265 117010

A biomarker identified by integrated bronchoalveolar lavage fluid and lung tissue lipidomics mitigates respiratory syncytial virus infection.

Shaodong Wang, Lili Lin, Yueyue Lei, Qiuyuan Li, Linxiu Peng.

Respiratory syncytial virus (RSV) is a highly pathogenic respiratory virus that severely impacts infants under 2 years, adults over 60, and immunocompromised individuals. Currently, no safe, low-toxicity, or cost-effective therapy is available for RSV. Research indicates that RSV infection disrupts pulmonary surfactant lipid metabolism. Through lipidomic analysis of bronchoalveolar lavage (BAL) fluid and lung tissue, we observed an increase in most phosphatidylcholines (PCs) and phosphatidylethanolamines (PEs), while all phosphatidylglycerols (PGs) decreased. Additionally, all lysophospholipids in lung tissue increased. Among the altered lipids, 27 showed significant changes in both BAL fluid and lung tissue, including 15 phospholipids (7 PCs, 5 PEs, 3 PGs, and 1 phosphatidylinositol (PI)). From the decreased PGs, we identified an active lipid, PG 16:0-18:2, which correlated strongly between BAL fluid and lung tissue. Further studies confirmed that PG 16:0-18:2 significantly reduces viral load, mitigates RSV-induced lung damage, and reduces pulmonary inflammation. In vitro experiments demonstrated that PG 16:0-18:2 directly inhibits RSV replication and proliferation in epithelial cells. These findings suggest that PG 16:0-18:2 could serve as an effective and non-toxic antiviral agent, offering a promising complementary approach to RSV treatment.

Keywords: Lipidomic; PG 16:0–18:2; Pulmonary surfactant lipids; Respiratory syncytial virus

DOI: https://doi.org/10.1016/j.jpba.2025.117010

Int J Mol Sci. 2025 May 29. pii: 5244. [Epub ahead of print]26(11):

Integrated Metabolomics and Network Pharmacology to Reveal the Mechanisms of Forsythia suspensa Extract Against Respiratory Syncytial Virus.

Haitao Du, Jie Ding, Yaxuan Du, Xinyi Zhou, Lin Wang, Xiaoyan Ding, Wen Cai, Cheng Wang, Mengru Zhang, Yi Wang, Ping Wang.

To investigate the therapeutic impact of Forsythia suspensa extract (FS) on RSV-infected mice and explore its antiviral pharmacodynamic foundations. Methods: An integrated analytical approach, combining UPLC-Q-TOF/MS with network pharmacology, was employed to analyze and identify the chemical constituents in FS, particularly those exhibiting antiviral properties against RSV. The study integrated network pharmacology and metabolomics for further analysis, and molecular docking and interaction experiments were conducted to validate the pharmacodynamic mechanisms. Finally, an RSV pneumonia mouse model was employed to evaluate the therapeutic influence of FS, including pathological and immunohistochemistry assessments. Twenty-five components in FS were identified through UPLC-Q-TOF/MS analysis. Integrated network pharmacology data revealed 43 effective components and predicted 113 potential targets of FS for anti-RSV activity. Metabolomics analysis identified 14 metabolite biomarkers closely linked to RSV-induced metabolic disruptions involving pathways. Moreover, molecular docking and Biacore experiments provided additional confirmation that FS primarily exerts its effects through compounds such as rutin, quercetin, and kaempferol. Immunohistochemistry experiments demonstrated a significant reduction in the expression of relevant proteins following FS administration, affirming its capacity to ameliorate lung inflammation induced by RSV infection through the modulation of Toll-like receptor signaling pathways. The data presented in this study illustrate that FS exerts its anti-RSV effects by regulating the Toll-like receptor signaling pathway and the arachidonic acid metabolism pathway via rutin, quercetin, and kaempferol. Furthermore, the approach of combining network pharmacology with metabolomics proves to be an effective research strategy for investigating the bioactive constituents of medicinal plants and elucidating their pharmacological effects.

Keywords: Forsythia suspensa extract; Respiratory Syncytial Virus; SPR-Biacore; metabolomics; network pharmacology

DOI: https://doi.org/10.3390/ijms26115244