Viruses exploit host genetic machinery to establish chronic infections and contribute to chronic neurodegenerative and immune disorders. By manipulating DNA methylation, histone modifications, and non-coding RNA networks, viruses such as Epstein-Barr virus (EBV), Herpes Simplex virus (HSV), Zika virus (ZIKV), and Human immunodeficiency virus (HIV) induce epigenetic changes that silence or delay the host antiviral defenses and reprogram host gene expression. For instance, EBV recruits DNA methyltransferases (DNMTs) to hypermethylate genes such as SOCS1 while ZIKV disrupts neuronal DNA methylation via heterochromatinization. Concurrently, HIV-1 encoded Tat protein disrupts H3K27 acetylation in astrocytes that potently triggers glutamate excitotoxicity. Viruses also subvert the chromatin architecture to modulate 3D interactions and reader protein recruitment, thus rewiring transcriptional programs. Such epigenetic alterations correlate with neuropathologies wherein global hypomethylation and histone acetylation contribute to Alzheimer's or Parkinson's disease. Although several mechanisms are known, the field of viral control in neuroepigenetics is underexplored. Emerging therapies that target viral-host epigenetic crosstalk through DNMT or HDAC inhibitors are promising. This review highlights how neurotropic viruses co-opt epigenetic pathways to drive neurodegeneration. We also explore biomarker-driven strategies for personalized interventions in neurodegenerative disorders thus emphasizing potential epigenetic editing tools to restore neuronal homeostasis.

Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disease driven by immune dysregulation, chronic inflammation, and loss of immune tolerance. Increasing evidence suggests that non-coding RNAs (ncRNAs), including microRNAs, long non-coding RNAs, and circular RNAs, are involved in the fine regulation of immune responses, epigenetic changes, and cytokine signaling in SLE. Their altered expression has been associated with disease activity, specific clinical phenotypes, and treatment response, supporting their potential as biomarkers. Epstein-Barr virus (EBV), a long-suspected contributor to SLE, may further shape disease through persistent immune activation, molecular mimicry, and modulation of host ncRNA pathways. In this review, we examine how EBV and ncRNA dysregulation may converge in autoreactive B cells and other immune compartments to promote the loss of tolerance and the development of clinically distinct forms of SLE. We also discuss the implications of this axis for biomarker development and emerging therapeutic strategies, including personalized immune-targeted approaches.

Background and ObjectivesEpstein-Barr virus (EBV) infection is a prerequisite for the development of multiple sclerosis (MS), yet whether EBV acts merely as a trigger at disease onset or also contributes to immune dysregulation and disease progression remains unclear. To explore potential mechanisms linking EBV to immune alterations, we performed a comprehensive analysis of EBV markers and immune-related gene expression in peripheral blood samples from therapy-na & iuml;ve persons with MS (PwMS) and healthy donors (HD) and assessed EBV transcripts in CSF cells to explore compartment-specific viral activity.MethodsPeripheral blood mononuclear cells (PBMCs) and serum from PwMS (n = 77) and HD (n = 40) were analyzed. EBV serology, DNA load, and RNA expression were assessed by ELISA, droplet digital PCR, and preamplified real-time RT-PCR, respectively. EBV RNA was also evaluated in PwMS CSF cells. Gene expression profiling of 47 immune-related genes selected for their relevance to MS was also performed in PBMCs. Data were analyzed using univariate and multivariate statistical approaches also considering demographic, clinical, and radiologic information. Exploratory factor analysis (EFA) was used to identify transcriptional signatures associated with MS.ResultsAnti-EBNA1 IgG titers were higher in PwMS. In addition, EBV RNA and DNA were more frequently detected, and viral load was increased compared with HD. Notably, EBV transcripts associated with latency II/III (LMP1, LMP2A, EBNA1, EBNA3A) and lytic reactivation (BZLF1, gp350/220) were more prevalent in PwMS. Although viral RNA was detected in only 7% of CSF samples, all positive cases showed profiles consistent with viral reactivation. Immune gene expression analysis revealed broad upregulation of cytotoxic effectors, type I interferon pathways, and chemokine signaling in PwMS. EFA identified a significantly different gene signature linking BZLF1 expression with inflammatory genes, type I interferon responses, and chemokines involved in immune cell migration, in PwMS.DiscussionOur findings support the hypothesis that EBV latency disruption and lytic reactivation contribute to immune dysregulation in MS. The association between EBV transcriptional activity and immune gene alterations may uncover potential peripheral biomarkers of EBV-driven pathology. These molecular signatures may provide insights into novel therapeutic avenues and peripheral biomarkers for MS monitoring.

MicroRNAs (miRNAs) are small, non-coding RNAs that play critical roles in post-transcriptional gene regulation and are involved in various biological processes, including cancer progression. Efficient extraction of miRNAs from cultured cells and tissue samples is crucial for downstream applications, including quantitative real-time PCR (qRT-PCR), next-generation sequencing (NGS), and microarray analysis. This chapter provides reliable protocols for extracting high-quality RNA and microRNAs from formalin-fixed paraffin-embedded (FFPE) tissues and cultured cells. It compares manual and automated methods to guide optimal kit selection. The protocols support reproducible results from challenging samples in research and clinical settings.

BACKGROUND AND OBJECTIVES:Epstein-Barr virus (EBV) infection is a prerequisite for the development of multiple sclerosis (MS), yet whether EBV acts merely as a trigger at disease onset or also contributes to immune dysregulation and disease progression remains unclear. To explore potential mechanisms linking EBV to immune alterations, we performed a comprehensive analysis of EBV markers and immune-related gene expression in peripheral blood samples from therapy-naïve persons with MS (PwMS) and healthy donors (HD) and assessed EBV transcripts in CSF cells to explore compartment-specific viral activity. METHODS:Peripheral blood mononuclear cells (PBMCs) and serum from PwMS (n = 77) and HD (n = 40) were analyzed. EBV serology, DNA load, and RNA expression were assessed by ELISA, droplet digital PCR, and preamplified real-time RT-PCR, respectively. EBV RNA was also evaluated in PwMS CSF cells. Gene expression profiling of 47 immune-related genes selected for their relevance to MS was also performed in PBMCs. Data were analyzed using univariate and multivariate statistical approaches also considering demographic, clinical, and radiologic information. Exploratory factor analysis (EFA) was used to identify transcriptional signatures associated with MS. RESULTS:Anti-EBNA1 IgG titers were higher in PwMS. In addition, EBV RNA and DNA were more frequently detected, and viral load was increased compared with HD. Notably, EBV transcripts associated with latency II/III (LMP1, LMP2A, EBNA1, EBNA3A) and lytic reactivation (BZLF1, gp350/220) were more prevalent in PwMS. Although viral RNA was detected in only 7% of CSF samples, all positive cases showed profiles consistent with viral reactivation. Immune gene expression analysis revealed broad upregulation of cytotoxic effectors, type I interferon pathways, and chemokine signaling in PwMS. EFA identified a significantly different gene signature linking BZLF1 expression with inflammatory genes, type I interferon responses, and chemokines involved in immune cell migration, in PwMS. DISCUSSION:Our findings support the hypothesis that EBV latency disruption and lytic reactivation contribute to immune dysregulation in MS. The association between EBV transcriptional activity and immune gene alterations may uncover potential peripheral biomarkers of EBV-driven pathology. These molecular signatures may provide insights into novel therapeutic avenues and peripheral biomarkers for MS monitoring.

Epstein-Barr virus (EBV) infects more than 90% of the global population and is etiologically linked to a wide spectrum of lymphoid and epithelial malignancies. Although its role as an oncogenic virus is well established, the mechanisms underlying EBV contribution to tumorigenesis remain undefined. This review revises the “hit-and-run” hypothesis in EBV-associated cancers, proposing that the virus may initiate oncogenic transformation before leaving tumour cells. The review summarizes current evidence of EBV episomal loss, integration into host chromosomes, and the challenges associated with detecting traces of viral genetic material. Recent advances in highly sensitive detection methods, such as quantitative polymerase chain reaction (PCR), RNAscope, and single-cell droplet digital PCR, have revealed viral traces in lymphomas and carcinomas previously considered as EBV-negative by conventional in situ hybridization, supporting a broader role for EBV involvement in oncogenesis. Moreover, tumours with EBV traces have similar epigenetic and mutational landscapes to EBV-positive patients, suggesting that EBV-induced alterations may continue to have an impact even after EBV loss. Despite these findings, it still remains unclear whether residual viral elements contribute to ongoing oncogenic signalling, epigenetic alterations, or immune modulation within the tumour microenvironment. Investigating these factors could improve our ability to stratify patients based on EBV status, refine diagnostic criteria, and develop more targeted treatment approaches.

Recent sero-epidemiological studies have strengthened the hypothesis that Epstein-Barr virus (EBV) may be a causal factor in multiple sclerosis (MS). Given the complexity of the EBV-host interaction, various mechanisms may be responsible for the disease pathogenesis. Furthermore, it remains unclear whether this is a disease-specific process. Here, we showed that genes encoding EBV interactors are enriched in loci associated with MS but not with other diseases and in prioritized therapeutic targets. Analyses of MS blood and brain transcriptomes confirmed a dysregulation of MS-associated EBV interactors affecting the CD40 pathway. Such interactors were strongly enriched in binding sites for the EBV nuclear antigen 2 (EBNA2) viral transcriptional regulator, often in colocalization with CCCTC binding factor (CTCF) and RNA Polymerase II Subunit A (POLR2A). EBNA2 was expressed in the MS brain. The 1.2 EBNA2 allele downregulated the expression of the CD40 MS-associated gene analogously to the CD40 MS-risk variant. Finally, we showed that the 1.2 EBNA2 allele associates with the risk of MS. This study delineates how host and viral genetic variability converge in MS-specific pathogenetic mechanisms.

Trastuzumab (TZ) resistance remains a significant challenge in the treatment of human epidermal growth factor receptor 2 (HER2)-positive epithelial ovarian cancer (EOC), necessitating novel therapeutic strategies to improve treatment efficacy. Functionalized gold nanoparticles (AuNPs) constitute a promising platform for drug delivery and the ability to enhance tumor targeting via the enhanced permeability and retention (EPR) effect. miR-200c, a well-established tumor suppressor microRNA (miRNA), plays a crucial role in inhibiting epithelial-mesenchymal transition (EMT). However, its role in modulating HER2 signaling pathways and sensitizing ovarian cancer cells to TZ remains largely unexplored. Here, we investigate for the first time the combinatorial effect of miR-200c and thiol-functionalized AuNPs (< 10 nm) loaded with TZ (AuNPs-TZ) in overcoming TZ resistance and enhancing treatment efficacy in ovarian cancer cells. Pristine AuNPs were not cytotoxic, confirming their biocompatibility as a nanocarrier for TZ delivery. AuNPs were loaded noncovalently with TZ and maintained colloidal stability to prevent aggregation while facilitating effective cellular uptake. Treatment of ovarian cancer cells overexpressing miR-200c with AuNPs-TZ significantly reduced cell viability and increased apoptosis. Immunoblot analysis showed a reduction of phosphorylated HER2 and downstream Kirsten Rat Sarcoma Virus (KRAS) signaling. Furthermore, transmission electron microscopy (TEM) demonstrated morphological changes in miR-200c-transfected ovarian cancer cells and confirmed the localization of AuNPs carrying TZs on the cell membrane and in the cytoplasm. These findings highlight the potential of AuNPs-TZ delivery combined with miR-200c as a promising therapeutic strategy to improve the response of HER2-positive EOC to TZ treatment. These results imply the need to further develop AuNP/miRNA-based combinatorial therapies as a viable nanomedicine approach for drug-resistant cancers.

The intricate interplay between the gut microbiota and the GI tract has garnered significant attention, as growing evidence has identified the inflammasome as a crucial yet underexplored master regulator in microbiota-driven diseases. Triggered by a variety of dangers, inflammasomes are supramolecular complexes that regulate immune response. A large number of bacterial-derived inducers have been characterised so far. Although structurally divergent, threats are neutralised by the inflammasome, which is then classified into three families: (1) nucleotide-binding oligomerisation domain, leucine-rich repeat-containing proteins, (2) absent in melanoma 2-like receptors and (3) pyrin. An unbalanced microbiota composition, expressed by a dysbiotic phenotype, might therefore induce undesired inflammasome activation, altering the local host homeostasis. Recent studies on the 'microbiota-inflammasome axis' have uncovered unexpected roles for inflammasome signalling in various types of GI cancer and IBD. Additionally, beyond local gut functions, microbiota influences stress responses and neurological health through aberrant secretion of inflammasome-processed cytokines, linking gut-derived signals to systemic diseases via the vagus nerve and the hypothalamic-pituitary-adrenal axis. Besides the standard experimental approaches, this complex network of interactions is now being addressed by Artificial intelligence, which emphasises the profound impact of the gut microbiota on GI health, cancer progression and brain function, opening new avenues for therapeutic intervention in GI diseases, cancer and neurological disorders. Ultimately, microbiota-inflammasome interactions manage a regulatory framework that influences inflammation, cancer progression and systemic diseases, positioning it as both a mediator and a promising therapeutic target in GI malignancies and systemic diseases of the central nervous system.

Gastrointestinal (GI) inflammation and malignancies arise from complex interactions between the host's immune responses and microbial pathogens. Epstein-Barr virus (EBV), Helicobacter pylori (H. pylori), and Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) contribute to chronic GI inflammation, immune evasion, and tumorigenesis through distinct but interconnected mechanisms. EBV, a widespread herpesvirus, establishes a latent infection in B cells and epithelial cells. It promotes gastric carcinogenesis through immune modulation, epigenetic changes, and viral microRNAs (miRNAs). H. pylori, a gastric carcinogen, induces chronic gastritis and gastric cancer (GC) through Cytotoxin-associated gene A (CagA) and Vacuolating cytotoxin gene A (VacA) virulence factors. These factors disrupt host immune responses and enhance oncogenic signaling pathways. Recent evidence also links SARS-CoV-2 to gut dysbiosis and inflammatory responses. It worsens immune dysfunction and hence potentially impacting GI pathology. EBV and H. pylori co-infections may synergistically amplify inflammatory signaling, creating a tumor-promoting microenvironment. This review emphasizes the molecular mechanisms by which these pathogens contribute to GI diseases, focusing on their immune evasion strategies and potential therapeutic targets. Understanding these interactions is essential for developing targeted interventions for infection-driven GI malignancies.

ABSTRACT:Hematological malignancies such as Burkitt lymphoma (BL), Hodgkin lymphoma (HL), and diffuse large B-cell lymphoma (DLBCL) cause significant morbidity in humans. A substantial number of these lymphomas, particularly HL and DLBCLs have poorer prognosis because of their association with Epstein-Barr virus (EBV). Our earlier studies have shown that EBV-encoded nuclear antigen (EBNA2) upregulates programmed cell death ligand 1 in DLBCL and BLs by downregulating microRNA-34a. Here, we investigated whether EBNA2 affects the inducible costimulator (ICOS) ligand (ICOSL), a molecule required for efficient recognition of tumor cells by T cells through the engagement of ICOS on the latter. In virus-infected and EBNA2-transfected B-lymphoma cells, ICOSL expression was reduced. Our investigation of the molecular mechanisms revealed that this was due to an increase in microRNA-24 (miR-24) by EBNA2. By using ICOSL 3' untranslated region-luciferase reporter system, we validated that ICOSL is an authentic miR-24 target. Transfection of anti-miR-24 molecules in EBNA2-expressing lymphoma cells reconstituted ICOSL expression and increased tumor immunogenicity in mixed lymphocyte reactions. Because miR-24 is known to target c-MYC, an oncoprotein positively regulated by EBNA2, we analyzed its expression in anti-miR-24 transfected lymphoma cells. Indeed, the reduction of miR-24 in EBNA2-expressing DLBCL further elevated c-MYC and increased apoptosis. Consistent with the in vitro data, EBNA2-positive DLBCL biopsies expressed low ICOSL and high miR-24. We suggest that EBV evades host immune responses through EBNA2 by inducing miR-24 to reduce ICOSL expression, and for simultaneous rheostatic maintenance of proproliferative c-MYC levels. Overall, these data identify miR-24 as a potential therapeutically relevant target in EBV-associated lymphomas.

The intracellular pathway of Janus kinase/signal transducer and activator of transcription (JAK/STAT) and modification of nucleosome histone marks regulate the expression of proinflammatory mediators, playing an essential role in carcinogenesis, antiviral immunity and the interaction of host proteins with Herpesviral particles. The pathway has also been suggested to play a vital role in the clinical course of the acute infection caused by severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2; known as coronavirus infection-2019), a novel human coronavirus initially identified in the central Chinese city Wuhan towards the end of 2019, which evolved into a pandemic affecting nearly two million people worldwide. The infection mainly manifests as fever, cough, myalgia and pulmonary involvement, while it also attacks multiple viscera, such as the liver. The pathogenesis is characterized by a cytokine storm, with an overproduction of proinflammatory mediators. Innate and adaptive host immunity against the viral pathogen is exerted by various effectors and is regulated by different signaling pathways notably the JAK/STAT. The elucidation of the underlying mechanism of the regulation of mediating factors expressed in the viral infection would assist diagnosis and antiviral targeting therapy, which will help overcome the infection caused by SARS-CoV-2.

High mortality has been reported in severe cases of COVID-19. Emerging reports suggested that the severity is not only due to SARS-CoV-2 infection, but also due to coinfections by other pathogens exhibiting symptoms like COVID-19. During the COVID-19 pandemic, simultaneous respiratory coinfections with various viral (Retroviridae, Flaviviridae, Orthomyxoviridae, and Picoviridae) and bacterial (Mycobacteriaceae, Mycoplasmataceae, Enterobacteriaceae and Helicobacteraceae) families have been observed. These pathogens intensify disease severity by potentially augmenting SARSCoV-2 replication, inflammation, and modulation of signaling pathways. Coinfection emerges as a critical determinant of COVID-19 severity, principally instigated by heightened pro-inflammatory cytokine levels, as cytokine storm. Thereby, in co-infection scenario, the severity is also driven by the modulation of inflammatory signaling pathways by both pathogens possibly associated with interleukin, interferon, and cell death exacerbating the severity. In the current review, we attempt to understand the role of co- infections by other pathogens and their involvement in the severity of COVID-19.

The enigmatic association of the Epstein-Barr virus (EBV) with neuropathologies is in constant heed over the last few decades. To better understand the role of membrane cholesterol in EBV infection and the pathogenesis of astroglial cells, here we have used its inhibitor, methyl-β-cyclodextrin (MβCD). The astroglial cells treated with MβCD showed a consistent decrease in ebna1 transcripts compared to untreated EBV-infected cells. Notably, lmp1 and lmp2a were significantly downregulated upon MβCD treatment. Molecules involved in EBV-mediated downstream signalling namely STAT3 and RIP, exhibited a decrease in protein levels after exposure to MβCD while NF-kB levels were found to increase. The levels of TNF-α were decreased in MβCD+EBV at 1, 2 and 4 hrs compared to EBV infection alone. Systemic changes in biomolecular fingerprints were analyzed using Raman microspectroscopy (RS). The intensity of the Raman spectra revealed an increase in triglycerides and fatty acids in the periphery of EBV-infected LN-229 cells compared to MβCD+EBV. The full width at half maximum (FWHM) which corresponds to the peak width of cholesterol, lipid and fatty acids was found to be reduced in EBV-infected samples. This indicates an increase in unique cholesterol moieties. The shift of Raman peak at 1 hr of periphery manifested a change i.e., protein to triglycerides. An opposite pattern was observed in the nucleus. The ingenuity pathway analysis revealed novel protein molecules such as UCP-2, CYP11A1, USP-22, DBH, ADM, and SMPD1 that bridge an attachment with an altered profile of cholesterol, fatty acids and triglycerides with infection-related CNS disorders. Taken together, our results underline the important role of membrane cholesterol in EBV entry/pathogenesis in astroglial cells which might further trigger/exacerbate virus-associated neuropathologies.

Renal cell carcinoma, bladder cancer, and prostate cancer are the most widespread genitourinary tumors. Their treatment and diagnosis have significantly evolved over recent years, due to an increasing understanding of oncogenic factors and the molecular mechanisms involved. Using sophisticated genome sequencing technologies, the non-coding RNAs, such as microRNAs, long non-coding RNAs, and circular RNAs, have all been implicated in the occurrence and progression of genitourinary cancers. Interestingly, DNA, protein, and RNA interactions with lncRNAs and other biological macromolecules drive some of these cancer phenotypes. Studies on the molecular mechanisms of lncRNAs have identified new functional markers that could be potentially useful as biomarkers for effective diagnosis and/or as targets for therapeutic intervention. This review focuses on the mechanisms underlying abnormal lncRNA expression in genitourinary tumors and discusses their role in diagnostics, prognosis, and treatment.

Epstein-Barr virus (EBV) is a lymphotropic herpesvirus termed human herpesvirus 4 (HHV4). It was initially identified in biopsies of Burkitt’s lymphoma, arising in the jaw and other site of the body in childhood or early adolescent individuals in sub-Saharan region. Subsequently, its tight association with other type of lymphomas has been described, and the tightest association has been seen in nasopharyngeal carcinoma (NPC), endemic with southeast Asia and southern part of China. The malignant transforming potential of EBV has been identified in immune compromised individuals; in the context all viral genomic products are expressed among which oncogenic proteins or non-coding RNAs are expressed. The interactions between cellular and viral oncoprotein as well as host gene expression regulation by the viral genetic products have been investigated in human tumors. The switch from latent form of infection to lytic phase has been studied in EBV-associated human tumors, and the modulation by intracellular signaling pathways has been known to be of importance in EBV-mediated carcinogenesis.

The British Journal of Haematology publishes original research papers in clinical, laboratory and experimental haematology. The Journal also features annotations, reviews, short reports, images in haematology and Letters to the Editor.