Biology subjects

Mishra, L.

Publications and source records attributed to Mishra, L..

4 recordsLinked to original sources

bioRxiv2026

Trifluoperazine exhibits broad-spectrum antiviral activity against arboviruses

The recurrent outbreaks and geographical expansion of mosquito-borne arboviruses pose a significant challenge to public health worldwide. The disease outcome for arboviral infections ranges from acute febrile illness to severe conditions such as encephalitis, hemorrhagic shock, and mortality. Current treatment options for these viruses are limited to supportive care, necessitating an urgent need for a safe and effective broad-spectrum antiviral. In this study, we have identified Trifluoperazine (TFP), an FDA-approved antipsychotic, as a potent broad-spectrum antiviral against Japanese encephalitis Virus (JEV), Dengue virus (DENV) and Chikungunya virus (CHIKV) infections. The antiviral effect of TFP was also seen in the animal models of JEV and CHIKV with significantly reduced disease severity. Mechanistically, TFP treatment increased the phosphorylation of eIF2a and induced an adaptive ER stress response in diverse cell types. Alleviation of TFP-induced ER stress by chemical chaperone 4PBA abolished the antiviral activity of the drug and rescued virus replication in cells. The robust in vitro and in vivo efficacy of the drug against arboviruses highlights the potential for repurposing TFP as a broad-spectrum antiviral candidate.

microbiology↗

bioRxiv2025

Endophilin mediated endocytosis and Epidermal growth factor receptor govern Japanese encephalitis virus entry and infection in neuronal cells

Several trafficking pathways are operational at the plasma membrane, and both clathrin-dependent, and clathrin-independent endocytosis (CIE), can serve as virus entry portals. Our research has shown that the neurotropic flavivirus: Japanese encephalitis virus (JEV), infects neuronal cells via CIE. Here we have identified and characterized two essential host-factors for JEV trafficking in neuronal cells: Endophilin & Epidermal Growth Factor Receptor (EGFR). Through quantitative estimation of viral RNA copy number, we demonstrate that JEV entry in neuronal cells, was blocked by knock-down of Endophilin A isoforms, while activation of Endophilin-mediated endocytosis using a specific inhibitor of GSK-3{beta} enhanced virus entry. Deletion mutants of Endophilin showed an essential role of SH3, BAR, H0 domains for virus entry. High resolution fluorescence imaging of virions showed overlap with Endophilin A2 puncta. Virus entry led to rearrangements of the actin cytoskeleton, and was highly sensitive to any pharmacological actin perturbation. Virus endocytosis activated EGFR, and the specific kinase inhibitors Erlotinib, and Gefitinib, reduced virus entry and replication in cultured cells and mouse primary cortical neurons. Silencing of EGFR, competitive inhibition with receptor ligand EGF, and EGFR specific antibodies significantly impaired JEV binding and entry, indicating the crucial role of its ligand binding domain for virus attachment/receptor interaction. EGFR colocalized with virions at early time-points of infection, and the ED3 domain of the JEV-envelope protein showed specific interaction with EGFR through BLI. Our study provides evidence for JEV entry in neuronal cells through an endocytic pathway involving Endophilin and EGFR.

microbiology↗

bioRxiv2023

Methotrimeprazine exerts antiviral and neuroprotective effects in Japanese encephalitis virus infection through activation of adaptive ER stress and autophagy

Japanese encephalitis virus (JEV) is the leading global cause of virus-induced encephalitis. Its pathogenesis is driven by a combination of neuronal cell death and neuroinflammation. We hypothesized that pharmacological upregulation of autophagy could exert a neuroprotective antiviral effect, and tested a panel of forty-two FDA-approved drugs that were shown to induce autophagy. Four drugs were tested in the JE mouse model based on in vitro protective effects on neuronal cell death, inhibition of viral replication, and anti-inflammatory effects in microglial cells. The antipsychotic phenothiazines Methotrimeprazine (MTP) and Trifluoperazine (TFP) showed a significant survival benefit with reduced virus titers in the brain, prevention of blood-brain barrier (BBB) breach, and inhibition of neuroinflammation. Both drugs were potent mTOR-independent autophagy flux inducers. Mechanistically MTP inhibited SERCA channel functioning, thereby resulting in rise in cytosolic calcium levels, and induction of a unique adaptive ER stress response. In virus infected drug treated cells, there was a strong transcriptional downregulation of type I interferon and interferon-stimulated genes and upregulation of cholesterol metabolic pathway genes. The drugs exerted an autophagy-dependent antiviral effect at the level of JEV protein translation/replication complex formation in diverse cell types. Inhibition of inflammatory cytokine/chemokine release from mouse microglial cells was partly autophagy-dependent. Our study suggests that MTP exerts a combined antiviral and anti-inflammatory effect in JEV infection, and has therapeutic potential to be repurposed for JE treatment.

microbiology↗

bioRxiv2018

Baseline human gut microbiota profile in healthy people and standard reporting template

A comprehensive knowledge of the types and ratios of microbes that inhabit the healthy human gut is necessary before any kind of pre-clinical or clinical study can be performed that attempts to alter the microbiome to treat a condition or improve therapy outcome. To address this need we present an innovative scalable comprehensive analysis workflow, a healthy human reference microbiome list and abundance profile (GutFeelingKB), and a novel Fecal Biome Population Report (FecalBiome) with clinical applicability. GutFeelingKB provides a list of 157 organisms (8 phyla, 18 classes, 23 orders, 38 families, 59 genera and 109 species) that forms the baseline biome and therefore can be used as healthy controls for studies related to dysbiosis. The incorporation of microbiome science into routine clinical practice necessitates a standard report for comparison of an individuals microbiome to the growing knowledgebase of \"normal\" microbiome data. The FecalBiome and the underlying technology of GutFeelingKB address this need. The knowledgebase can be useful to regulatory agencies for the assessment of fecal transplant and other microbiome products, as it contains a list of organisms from healthy individuals. In addition to the list of organisms and abundances the study also generated a list of contigs of metagenomics dark matter. In this study, metagenomic dark matter represents sequences that cannot be mapped to any known sequence but can be assembled into contigs of 10,000 nucleotides or higher. These sequences can be used to create primers to study potential novel organisms. All data is freely available from https://hive.biochemistry.gwu.edu/gfkb and NCBIs Short Read Archive.

microbiology↗