Human cytomegalovirus (CMV) is a globally widespread pathogen associated with significant morbidity in immunocompromised individuals. Despite its clinical importance, no licensed vaccine is currently available. This study aimed to design a rational multi-epitope vaccine candidate targeting CMV using an integrative approach combining immunoinformatics and structural biology. Viral proteins were screened to identify epitopes with high affinity for B cells, cytotoxic T cells (CTLs), and helper T cells (HTLs) using the Immune Epitope Database (IEDB). Selected epitopes were filtered according to their antigenicity and toxicity and then assembled into a chimeric construct incorporating an immunostimulatory adjuvant. The designed vaccine was evaluated for its physicochemical properties, validated by Ramchandran and ERRAT analyses. Molecular modeling demonstrated strong and stable interactions with key innate immunity receptors, including TLR7 and TLR9, interactions confirmed by molecular dynamics simulations. In silico immune simulation predicted a robust and durable immune response, characterized by high levels of IgM and IgG, as well as significant activation of CD4 + and CD8 + lymphocytes and innate immunity components. These results highlight the potential of the proposed multi-epitope construct as a promising vaccine candidate against HCMV. However, experimental validation is essential to confirm its immunogenicity, safety, and translational applicability.

Background: Colorectal cancer (CRC) remains one of the leading causes of cancer-related mortality worldwide. Increasing evidence suggests that intestinal microbial dysbiosis contributes to colorectal tumorigenesis by reshaping host molecular signaling and the tumor immune microenvironment. However, the molecular mechanisms linking microbiome-associated alterations to host regulatory networks and disease progression remain incompletely understood. Objective: This study aimed to identify the microbiome-associated molecular regulators and immune modulators involved in colorectal cancer through an integrative multi-omics systems biology approach. Methods: Publicly available transcriptomic datasets were analyzed to identify differentially expressed genes, followed by functional enrichment, protein-protein interaction network construction, hub gene prioritization, immune infiltration profiling, survival analysis, and multi-omics characterization. Results: The identified hub genes represent hypothesis-generating host candidates for further mechanistic and clinical validation. These genes occupied key positions within host regulatory networks and were significantly associated with adverse clinical outcomes and altered CD8⁺ T-cell infiltration, suggesting their involvement in immune remodeling within the tumor microenvironment. Multi-omics characterization demonstrated that PTEN and SMAD4 alterations were predominantly associated with genomic deletions, whereas SMAD2 dysregulation was associated with transcriptomic and gene-dosage variation. Although the overall mutational burden of the identified hub genes was not significantly associated with disease-free survival (p = 0.878), these molecular regulators showed associations with immune-cell infiltration and CRC-related pathways. Conclusions: Collectively, our findings provide a system-level framework describing the associations between dysbiosis-relevant host pathways, CRC-related regulatory networks, and immune responses.

The extensive evolutionary diversity and continuous mutational adaptation of betacoronaviruses represent a major challenge to the development of broadly protective countermeasures. This study applied an evolution-informed Viroinformatics framework to systematically identify conserved and immune targets with predicted lower mutational vulnerability across betacoronaviruses. The workflow integrated comparative sequence conservation and phylogenetic analysis with immunoinformatics-based epitope screening, HLA population coverage, mutational escape profiling, structural modeling, molecular docking, and molecular dynamics simulations. Conserved cytotoxic T-lymphocyte and helper T-lymphocyte epitopes were prioritized based on immunogenicity, antigenicity, safety, HLA binding potential, evolutionary conservation, and predicted resistance to mutational escape. Combined MHC class I and II analysis predicted a theoretical global population coverage of 99.92%, which requires experimental validation. The resulting 409-amino-acid multiepitope construct was predicted to be antigenic, non-allergenic, and physiochemically stable. Structural validation showed that 99.7% of the residues were located within favored or additionally allowed regions. Molecular docking demonstrated favorable interaction of the vaccine construct with TLR7, while a 100 ns molecular dynamics simulation supported persistence of the predicted vaccine-receptor association. Overall, these findings provide a computational framework for prioritizing evolutionarily conserved immune targets and developing pre-emptive candidate epitopes showing predicted cross-betacoronavirus conservation within the analyzed dataset. Experimental studies evaluating HLA binding, cellular and humoral immunogenicity, recombinant expression, cross-reactivity, and in vivo protection are required to validate the predicted immunological potential and determine whether the proposed construct can provide broad protection against diverse betacoronaviruses.