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.

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.

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 5th Workshop IRE on Translational Oncology was held in Rome (Italy) on 27–28 March at the IRCCS Regina Elena National Cancer Institute. This meeting entitled “The New World of RNA diagnostics and therapeutics” highlightes the significant progress in the RNA field made over the last years. Research moved from pure discovery towards the development of diagnostic biomarkers or RNA-base targeted therapies seeking validation in several clinical trials. Non-coding RNAs in particular have been the focus of this workshop due to their unique properties that make them attractive tools for the diagnosis and therapy of cancer. This report collected the presentations of many scientists from different institutions that discussed recent oncology research providing an excellent overview and representative examples for each possible application of RNA as biomarker, for therapy or to increase the number of patients that can benefit from precision oncology treatment. In particular, the meeting specifically emphasized two key features of RNA applications: RNA diagnostic (Blandino, Palcau, Sestito, Díaz Méndez, Cappelletto, Pulito, Monteonofrio, Calin, Sozzi, Cheong) and RNA therapeutics (Dinami, Marcia, Anastasiadou, Ryan, Fattore, Regazzo, Loria, Aharonov).

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.

Supplementary Materials and Methods

The global rise of single-use throw-away plastic products has elicited a massive increase in the nano/microplastics (N/MPLs) exposure burden in humans. Recently, it has been demonstrated that disposable period products may release N/MPLs with usage, which represents a potential threat to women's health which has not been scientifically addressed yet. By using polyethyl ene (PE) particles (200 nm to 9 μm), we showed that acute exposure to a high concentration of N/MPLs induced cell toxicity in vaginal keratinocytes after effective cellular uptake, as viability and apoptosis data suggest, along with transmission electron microscopy (TEM) observations. The internalised N/MPLs altered the expression of junctional and adherence proteins and the organisation of the actin cortex, influencing the level of genes involved in oxidative stress signalling pathways and that of miRNAs related to epithelial barrier function. When the exposure to PE N/MPLs was discontinued or became chronic, cells were able to recover from the negative effects on viability and differentiation/proliferation gene expression in a few days. However, in all cases, PE N/MPL exposure prompted a sustained alteration of DNA methyltransferase and DNA demethylase expression, which might impact epigenetic regulation processes, leading to accelerated cell ageing and inflammation, or the occurrence of malignant transformation.

Introduction: Extensive research underlines the critical functions of androgens in females. Nevertheless, the precise mechanisms of their action are poorly understood. Here, we review the existing literature regarding the physiological role of androgens in women throughout life. Areas covered: Several studies show that androgen receptors (ARs) are broadly expressed in numerous female tissues. They are essential for many physiological processes, including reproductive, sexual, cardiovascular, bone, muscle, and brain health. They are also involved in adipose tissue and liver function. Androgen levels change with the menstrual cycle and decrease in the first decades of life, independently of menopause. Expert opinion: To date, studies are limited by including small numbers of women, the difficulty of dosing androgens, and their cyclical variations. In particular, whether androgens play any significant role in regulating the establishment of pregnancy is poorly understood. The neural functions of ARs have also been investigated less thoroughly, although it is expressed at high levels in brain structures. Moreover, the mechanism underlying the decline of dehydroepiandrosterone (DHEA) and dehydroepiandrosterone sulfate (DHEAS) with age is unclear. Other factors, including estrogen's effect on adrenal androgen production, reciprocal regulation of ARs, and non-classical effects of androgens, remain to be determined.

Coronavirus disease 2019 (COVID-19), the pandemic infection caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), presents with an extremely heterogeneous spectrum of symptoms and signs. The clinical manifestations seem to be correlated with disease severity. COVID-19 susceptibility and mortality show a significant sex imbalance, with men being more prone to infection and showing a higher rate of hospitalization and mortality compared to women. Such variability can be ascribed to both sex-related biological factors and gender-related behavioral cues. This review will discuss the potential mechanisms accounting for sex/gender influence in vulnerability to COVID-19. Cardiovascular diseases play a central role in determining COVID-19 outcome, whether they are pre-existent or arose upon infection. We will pay particular attention to the impact of sex and gender on cardiovascular manifestations related to COVID-19. Finally, we will discuss the sex-dependent variability in some biomarkers for the evaluation of COVID-19 infection and prognosis. The aim of this work is to highlight the significance of gendered medicine in setting up personalized programs for COVID-19 prevention, clinical evaluation and treatment.

Adipose-derived mesenchymal stem cells (ASCs) represent a valid therapeutic option for clinical application in several diseases, due to their ability to repair damaged tissues and to mitigate the inflammatory/immune response. A better understanding of the underlying mechanisms regulating ASC biology might represent the chance to modulate their in vitro characteristics and differentiation potential for regenerative medicine purposes. Herein, we investigated the effects of the demethylating agent 5-azacytidine (5-aza) on proliferation, clonogenicity, migration, adipogenic differentiation and senescence of ASCs, to identify the molecular pathways involved. Through functional assays, we observed a detrimental effect of 5-aza on ASC self-renewal capacity and migration, accompanied by actin cytoskeleton reorganization, with decreased stress fibers. Conversely, 5-aza treatment enhanced ASC adipogenic differentiation, as assessed by lipid accumulation and expression of lineage-specific markers. We analyzed the involvement of the Akt/mTOR, MAPK and Wnt/β-catenin pathways in these processes. Our results indicated impairment of Akt and ERK phosphorylation, potentially explaining the reduced cell proliferation and migration. We observed a 5-aza-mediated inhibition of the Wnt signaling pathway, this potentially explaining the pro-adipogenic effect of the drug. Finally, 5-aza treatment significantly induced ASC senescence, through upregulation of the p53/p21 axis. Our data may have important translational implications, by helping in clarifying the potential risks and advantages of using epigenetic treatment to improve ASC characteristics for cell-based clinical approaches.

MicroRNAs (miRNAs) are short ( 22 nts) RNAs that regulate gene expression via binding to mRNA. MiRNAs promoting cancer are known as oncomiRs. Targeting oncomiRs is an emerging area of cancer therapy. OncomiR-21 and oncomiR-155 are highly upregulated in lymphoma cells, which are dependent on these oncomiRs for survival. Targeting specific miRNAs and determining their effect on cancer cell progression and metastasis have been the focus of various studies. Inhibiting a single miRNA can have a limited effect, as there may be other overexpressed miRNAs present that may promote tumor proliferation. Herein, we target miR-21 and miR-155 simultaneously using nanoparticles delivered two different classes of antimiRs: phosphorothioates (PS) and peptide nucleic acids (PNAs) and compared their efficacy in lymphoma cell lines. Poly-Lactic-co-Glycolic acid (PLGA) nanoparticles (NPs) containing PS and PNA-based antimiR-21 and -155 were formulated, and comprehensive NP characterizations: morphology (scanning electron microscopy), size (differential light scattering), and surface charge (zeta potential) were performed. Cellular uptake analysis was performed using a confocal microscope and flow cytometry analysis. The oncomiR knockdown and the effect on downstream targets were confirmed by gene expression (real time-polymerase chain reaction) assay. We demonstrated that simultaneous targeting with NP delivered PS and PNA-based antimiRs resulted in significant knockdown of miR-21 and miR-155, as well as their downstream target genes followed by reduced cell viability ex vivo. This project demonstrated that targeting miRNA-155 and miR-21 simultaneously using nanotechnology and a diverse class of antisense oligomers can be used as an effective approach for lymphoma therapy.