Search Results (11,588)

Background: Mycobacterium abscessus complex (MABC), comprising Mycobacterium abscessus subsp. abscessus (MABa), Mycobacterium abscessus subsp. bolletii (MABb), and Mycobacterium abscessus subsp. massiliense (MABm), is an emerging group of non-tuberculous mycobacteria with clinically significant infections and challenging treatment outcomes due to extensive antimicrobial resistance. Accurate subspecies identification and characterization of resistance- and virulence-associated determinants are essential for effective disease management. This study aimed to determine the prevalence and subspecies distribution of MABC and to characterize resistance-associated mutations and virulence factors, including biofilm formation. Methods: A total of 1110 NTM-suspected clinical samples were screened during the study period, between January 2024 and October 2025. Samples negative by GeneXpert MTB/RIF were subjected to Mycobacteria Growth Indicator Tube (MGIT) culture, followed by Ziehl–Neelsen staining and MPT64 antigen testing. Acid-fast bacilli-positive, MPT64-negative isolates were identified as NTM and analyzed using GenoType CM and NTM-DR line probe assays (LPA) for species identification and detection of resistance-associated mutations. A polymerase chain reaction (PCR) assay was optimized to differentiate MABa and MABm. All MABC clinical strains were further characterized for colony morphology (smooth and rough) and biofilm formation. Three biofilm-producing MABa strains (2 rough and 1 smooth) that were detected as macrolide-resistant by NTM-DR were subjected to whole-genome sequencing (WGS). Results: Among 1110 clinical samples, MABC was identified in 2.25% (n = 25) of cases, while other NTM species accounted for 4.41% (n = 49). Among 25 MABC clinical strains, 14 (56%) were MABm, and 11 (44%) were MABa, as confirmed by both LPA and PCR. LPA-NTM DR detected erm(41) T28 sequevar (n = 9) and C28 mutation (n = 2) among MABa strains, with one strain exhibiting aminoglycoside resistance-associated rrs mutation. Nineteen isolates displayed a smooth morphotype (MABa = 8 and MABm = 11), and six were rough (MABa = 3 and MABm = 3). Biofilm formation was observed in both smooth (n = 5) and rough (n = 4) morphotypes. WGS analysis confirmed erm(41) T28 sequevar, identified a missense mutation (A238G), and revealed genes associated with glycopeptidolipid biosynthesis. Conclusions: Our findings provide important insights into subspecies identification and genetic determinants associated with drug resistance and virulence in MABC. The biofilm-forming ability observed in both smooth and rough morphotypes emphasizes its potential role in persistence and treatment challenges, emphasizing the need for comprehensive diagnostic strategies. Full article

Background/Objective: Antimicrobial resistance poses a critical public health crisis, highlighting the urgent requirement to investigate bacterial evolutionary adaptations and pioneer alternative therapeutics. Consequently, bacteriophages and metal-based compounds are emerging as viable options to combat drug-resistant infections. Building on our finding that T4 phage resistance in E. coli B also confers adaptation to high iron(III), we used RNA-sequencing (RNA-seq) to explore bacterial gene expression in resistant and control populations. We analyzed samples from our five experimental groups—Ancestor (ANC), control (CON), phage-selected (Phage), iron(III)-selected (FE), and phage/iron(III)-selected (PF), to understand how these regimes drive transcriptional changes. Method: Total RNA was extracted using the TRIzol protocol, and sequencing libraries were prepared with the Illumina RNA Total Library Prep Kit. Sequencing was performed on the Illumina NextSeq 1000/2000 platform. Reads were aligned to the E. coli B ATCC 11303 reference genome, and pairwise comparisons between the five experimental groups were conducted to determine differential gene expression profiles. Results: Principal component analysis (PCA) showed that iron-adapted populations (FE and PF) separated distinctly from the Ancestor and control populations along PC1 (capturing 40% of the variance), while the phage-selected replicates were split, with one (Phage5) clustering with CON3 and two (Phage2, Phage4) falling closer to, but clearly separated from, the Ancestor. Differential expression analysis (P_adj < 0.05 and |log₂FC| ≥ 1) revealed extensive transcriptional rewiring, with 482 and 381 differentially expressed genes (DEGs) in the FE and PF populations, respectively, compared to the Ancestor, and 177 DEGs in the phage-selected population compared to the Ancestor. The direct pairwise comparison between the iron-selected and phage/iron-selected populations yielded zero DEGs, demonstrating that both iron-adapted populations converged on a near-identical gene expression profile regardless of their distinct genetic and evolutionary backgrounds. Conclusions: This study suggests that pmrB and arn pathway genes may serve as primary markers for resistance to iron stress. These results are significant because they demonstrate a coordinated, multi-gene defense mechanism in E. coli B against high iron(III) stress, in which the arn operon and eptA remodel lipid A and the outer membrane, while glycerol-3-phosphate metabolism and phage-shock/chaperone pathways are repressed. Full article

The escalating problem of bacterial drug resistance poses a severe threat to global public health, with Acinetobacter baumannii (A. baumannii) exhibiting particularly high resistance rates that are closely linked to its Quorum Sensing (QS) system. This narrative review synthesizes current knowledge on the A. baumannii QS system, focusing on its essential role in mediating antimicrobial resistance, as well as the latest progress in developing QS inhibitors. Key findings indicate that the A. baumannii QS system enhances bacterial survival by promoting biofilm formation, and regulating the expression of efflux pump and resistance genes. However, significant translational challenges remain, including the risk of inducing resistance, the poor bioavailability and suboptimal pharmacokinetic properties, and their reduced efficacy in complex biological systems. In conclusion, while QS inhibitors represent a promising therapeutic target, overcoming current developmental bottlenecks requires combining them with traditional antibiotics, exploring novel drug delivery strategies, and integrating cutting-edge tools to accelerate drug discovery and clinical application. Full article

Intradermal injections of anti-canine immunoglobulin E (IgE) in healthy dogs have been utilized in preclinical drug testing to evaluate the efficacy of anti-allergic drugs used to treat canine atopic dermatitis (AD). However, the molecular effects of established canine anti-allergic drugs on this acute canine IgE-mediated atopic model remain largely uninvestigated. The objective of this study was to characterize the effect of proactive oclacitinib and prednisolone treatments on the immune and skin barrier transcriptome of IgE-mediated late-phase reactions (LPRs) in an acute model of canine AD. Sixteen healthy adult research-bred beagles were randomized to receive either oclacitinib or prednisolone orally for six days, followed by an intradermal anti-canine IgE injection. Biopsies were collected 24 h post-injection for RNA isolation and sequencing; previously analyzed transcriptomes (healthy skin, saline-injected skin, IgE lesions without drug modulation) from the same colony of dogs served as controls. Administration of prednisolone and oclacitinib prior to intradermal anti-IgE injections reduced the number of differentially expressed genes (DEGs) in 24 h samples to 1251 and 1471, respectively. Both treatments resulted in a decrease in expression of several significantly upregulated T helper-(Th)1 (e.g., MX1, OAS1, STAT1), Th2 (e.g., CCL13, CCL8, IL13RA1, IL-33, IL5RA, OSM), chemokine and receptor (e.g., CCL19, CCL2, CCL3, CCR1, CCR3) genes in comparison to the untreated IgE-mediated lesions. Interestingly, only prednisolone treatment significantly reduced IL-13 upregulation, an important gene in the Th2 immune response. In conclusion, both prednisolone and oclacitinib reduced the transcriptomic changes observed in the acute lesions of the canine IgE-induced atopic dermatitis model, with prednisolone inducing a broader inhibitory immune response. Full article

This study investigated the efficacy of a modified Ovsynch (MOVS) protocol incorporating a 50% reduced dose of a gonadotropin-releasing hormone analog (GnRH_a) conjugated with chitosan nanoparticles. Thirty-two Holstein heifers under heat-stress (HS) conditions were randomly assigned to either a standard Ovsynch (SOVS) or MOVS protocol (n = 16/group). Ovarian dynamics were monitored across the protocol days (D0, D3, D7, D9, D14, and D19). Hormonal profiles, metabolic and oxidative markers, and mRNA expressions of folliculogenesis- and apoptosis-related genes were evaluated. Both protocols improved follicular recruitment (p > 0.05). However, the MOVS significantly enhanced the dominant follicle (DF) diameter on D9 (p < 0.001). On D14 and D19, CL diameters were larger in the MOVS group (p < 0.001). These findings were associated with significantly high estradiol (E₂) concentrations on D9 (p < 0.05) and progesterone (P₄) concentrations on D19 (p < 0.05). Metabolic and oxidative stress markers were increased in MOVS-treated heifers, while antioxidant markers were also improved (p < 0.05). At the molecular level, the MOVS numerically upregulated the expression of folliculogenesis-related genes (BMP15, GDF9, LHR, FSHR, and IGF-1R) and an anti-apoptotic gene (BCL2) in peripheral blood (p > 0.05). In conclusion, the MOVS protocol achieved a 50% reduction in GnRHa dose while improving ovarian, endocrine, and reproductive physiological indicators, suggesting a promising strategy for optimizing synchronization in heifers under HS conditions. Full article

Background/Objectives: Cabozantinib is a tyrosine kinase inhibitor that primarily targets MET. It has become an important drug in the treatment of renal cell carcinoma (RCC); however, many patients do not respond to cabozantinib treatment and there is no effective next-line therapy. In this study, we identified molecular-targeted drugs that exhibit synergistic effects with cabozantinib using CRISPR/Cas9 screening. Methods: A kinome-wide synthetic lethal CRISPR/Cas9 screen was used to identify target molecules using 786-o RCC cells. A library was generated, and treatment with vehicle or cabozantinib was carried out, followed by next-generation sequencing to identify candidate genes. A combination index based on the Chou–Talalay method was used to evaluate the synergistic effect of cabozantinib through cell viability assays. Xenograft assays were conducted to determine the effect in vivo. Results: CRISPR/Cas9-based screening revealed four genes (MEK1, DCLK1, DYRK3, and FGFR1) that were candidates for synthetic lethality by cabozantinib in RCC cells. We focused on MEK1 because the MEK1 inhibitor cobimetinib has been approved for melanoma treatment. In a cell proliferation assay using 786-o and A498 RCC cells, the combination of cobimetinib and cabozantinib exhibited a synergistic effect. A xenograft assay also revealed a significant synergistic effect of cobimetinib and cabozantinib. Conclusions: CRISPR/Cas9 screening identified MEK1 as a candidate for a synthetic lethal target with cabozantinib in RCC. The combined inhibition of MET/VEGFR and MEK1 suppressed compensatory MAPK reactivation and downregulated the PI3K-Akt pathway, including the survival-associated genes PPP2R3B and ATF6B, and produced significant tumor growth suppression in vivo. These findings highlight the potential of cabozantinib plus cobimetinib, an already-FDA-approved MEK inhibitor, as a readily translatable combination strategy to overcome cabozantinib resistance in RCC. Full article

Experimental evidence has shown that gallic acid (GA), a naturally occurring polyphenolic compound, and docetaxel (DTX), a taxane chemotherapeutic agent, each possess antitumor activity against multiple cancer types. Although both compounds have been investigated individually, their combined effects in triple-negative breast cancer (TNBC) have received limited attention, and the molecular basis of their interaction remains unclear. The present study examined the in vitro effects of GA and DTX in MDA-MB-231 TNBC cells while simultaneously assessing their comparative cytotoxicity in HaCaT human keratinocytes. Evaluation of treatment efficacy included measurement of cell viability by the MTT assay and assessment of drug interactions using the Chou–Talalay combination index (CI) method. Apoptosis together with cell-cycle distribution was subsequently examined using both Annexin V/PI flow cytometry and TALI^® image-based cytometry. Additional analyses included β-tubulin immunofluorescence (IF), caspase-9 immunocytochemistry, ELISA, wound-healing assays, quantitative real-time PCR, and bioinformatic analyses to investigate treatment-associated biological alterations. Combined exposure to GA and DTX produced a significant reduction in cell viability and exhibited synergistic activity in MDA-MB-231 cells. The coordinated biological response to the combined treatment was characterized by increased apoptotic cell death, arrest of the cell cycle at the G2/M phase, extensive disorganization of the β-tubulin network, and enhanced caspase-9 immunoreactivity. Beyond its effects on cell survival, the combined regimen substantially decreased the release of IL-6, IL-8, and TNF-α, limited wound-healing capacity, and reshaped the expression profile of the apoptosis- and cell cycle-related genes BCL2, BAX, CASP9, and CDKN1A. Bioinformatic analyses further revealed enrichment of apoptosis- and cell-cycle-associated pathways that were generally consistent with the experimental observations. The overall pattern of experimental responses indicates that combining GA with DTX enhances the in vitro antitumor efficacy of DTX in TNBC cells by simultaneously influencing apoptotic pathways, cell-cycle regulation, inflammatory cytokine secretion, and cellular migratory capacity. Although the bioinformatic findings provide supportive hypothesis-generating evidence, additional studies using three-dimensional models, in vivo experiments, and functional validation approaches are necessary to confirm the underlying molecular mechanisms and to further define the translational potential of this therapeutic combination. Full article

Background: Janus kinase inhibitors (JAK inhibitors) have expanded therapeutic options for rheumatoid arthritis (RA), although factors associated with treatment response in routine clinical practice remain incompletely defined. Objectives: This study aimed to evaluate clinical, treatment-related and pharmacogenetic factors associated with response to different JAK inhibitors in patients with RA. Methods: An ambispective observational real-world cohort study was conducted in patients with RA treated with tofacitinib, baricitinib, filgotinib, or upadacitinib. Disease activity was assessed at 3 and 6 months using the Disease Activity Score in 28 joints based on C-reactive protein (DAS28-CRP). Clinical response was evaluated according to European Alliance of Associations for Rheumatology (EULAR) response criteria, low disease activity (LDA), and remission thresholds. Clinical, laboratory, and treatment-related variables were collected, and selected single-nucleotide polymorphisms (SNPs) in JAK1, JAK2, and JAK3 genes were genotyped. Bivariate and multivariable analyses were performed to identify variables associated with treatment outcomes. Results: Lower baseline inflammatory burden and lower disease activity were consistently associated with higher probabilities of EULAR response, LDA, and remission across JAK inhibitors. Treatment-related factors were also associated with improved outcomes. Pharmacogenetic associations were heterogeneous and drug-specific, with the most recurrent exploratory signals involving JAK2 variants. However, these genetic findings showed variability across outcomes and time points. Conclusions: In this real-world RA cohort, clinical and treatment-related factors were the most consistent variables associated with response to JAK inhibitors. Pharmacogenetic variation within the JAK pathway, particularly involving JAK2, may contribute to drug-specific variability in response, but these findings should be considered exploratory because of the limited sample size, multiple comparisons, and sparse genotype subgroups. Larger independent studies are required before JAK genotyping can be incorporated into individualized treatment strategies. Full article

Paclitaxel (PTX), a taxane-derived chemotherapeutic agent, is frequently used in the treatment of breast cancer (BC). Its anticancer effects are primarily associated with microtubule stabilization, disruption of cell-cycle progression, and triggering of apoptotic cell death. In the present study, we investigated the effects of PTX on the expression of genes involved in cancer-related pathways, energy metabolism, and drug resistance in four molecularly distinct BC cell lines: MCF-7, BT-474, SK-BR-3, and MDA-MB-231. The half-maximal inhibitory concentrations (IC₅₀) of PTX in BC cell lines and the non-tumorigenic hTERT-HME1 breast epithelial cell line were determined by the MTT assay to assess cell cytotoxicity. BC cells were exposed to nine different concentrations of PTX for 24, 48, and 72 h to evaluate concentration- and time-dependent effects. Following treatment, total RNA was isolated and converted into cDNA, and RT-qPCR analysis was performed to investigate PTX-mediated alterations in the expression of genes associated with cancer-related pathways. The impact of PTX on the cell-cycle phase distribution and apoptotic cell death was evaluated by flow cytometry. Treatment with PTX for 48 h at concentrations of 12.60 nM in MCF-7, 5.09 nM in BT-474, 16.09 nM in SK-BR-3, and 36.66 nM in MDA-MB-231 cells reduced cell viability and increased apoptosis. PTX treatment also altered the expression of genes involved in apoptosis, cell-cycle regulation, angiogenesis, epithelial–mesenchymal transition, hypoxia-related signaling, energy metabolism, telomere maintenance, and therapy resistance. Collectively, these findings demonstrate that PTX elicits heterogeneous molecular and cellular responses across molecularly distinct BC cell lines, particularly in cell viability, apoptosis, metabolic regulation, and treatment response. These in vitro findings suggest potential molecular mechanisms that could explain why some cells are more sensitive to PTX than others, but further experimental and clinical validation is needed to confirm this. Full article

This study investigates the synergistic therapeutic potential of Rivastigmine (RVG) and Epigallocatechin (EGC) in Alzheimer’s disease (AD), a multifactorial neurodegenerative disorder characterized by neuroinflammation, oxidative stress, and dysregulated signaling pathways. Conventional therapies primarily provide symptomatic relief and target limited pathways, highlighting the need for multi-target strategies with improved efficacy and safety. An integrated in silico approach combining pharmacokinetic evaluation, network pharmacology, molecular docking, and molecular dynamics simulations is used to determine the synergistic potential of RVG and EGC. Pharmacokinetic analysis indicates favorable drug-likeness and acceptable ADME/Tox profiles for both compounds. Network pharmacology identified 146 overlapping targets associated with AD, highlighting key hub genes including NFKB1, MAPK1, STAT1, PRKACA, GRB2, LYN, and PTPN11, which are involved in neuroinflammation, synaptic signaling, and neuronal survival. Functional enrichment analysis indicated significant involvement of MAPK/ERK signaling and immune-regulatory pathways. Importantly, the PD-1/PD-L1 signaling pathway is identified as a novel mechanism connecting neuroimmune modulation with intracellular kinase-driven neurodegeneration. Molecular docking studies showed strong binding affinities of RVG and EGC toward key AD-related targets, particularly MAPK1, supported by stable hydrogen bonding and interaction profiles. Molecular dynamics simulations confirmed stable protein-ligand interactions, with EGC contributing structural stability and RVG exhibiting adaptive flexibility within the binding pocket. These results suggest that the RVG-EGC combination exhibits synergistic potential by simultaneously modulating neuroinflammatory, oxidative stress, and kinase-mediated signaling pathways. The integration of PD-1/PD-L1 and MAPK/ERK signaling provides a novel mechanistic pathway for multi-target therapeutic intervention in AD. Full article

Breast cancer is a highly heterogeneous malignancy with multiple molecular subtypes and variable treatment responses. Despite advances in endocrine therapy, HER2-targeted therapy, chemotherapy, and immunotherapy, treatment resistance and disease recurrence remain major clinical challenges. There is growing evidence that transcriptional plasticity and enhancer relinking contribute to tumor progression and treatment adaptation, highlighting the powerful role of epigenetic regulators. CREB-binding protein (CBP) and E1A-associated protein p300 (EP300) are transcriptional coactivators that regulate breast cancer enhancer activity and lineage-specific gene expression. Emerging research suggests that CBP/p300 is more of a context-dependent vulnerability point than a universal carcinogenic driver. ER-positive tumors exhibit a strong dependence on CBP/p300-mediated transcriptional programs, while the triple-negative breast cancer subgroup, including androgen receptor-positive and immunosuppressive tumors, may rely on CBP/p300-dependent signaling to maintain survival and treatment resistance. This is in contrast to their role in HER2-positive breast cancer. This review summarizes the biological functions of CBP/p300 in breast cancer and discusses subtype-specific vulnerability, biomarker-directed patient stratification, drug resistance mechanisms, rational combination strategies, and current translational challenges, emphasizing the need for precise treatment of breast cancer. Full article

Background/Objectives: Liver fibrosis, a wound-healing response to chronic liver injury characterized by excessive extracellular matrix (ECM) accumulation, represents a major global health burden with no approved anti-fibrotic therapies. Pien Tze Huang (PZH), an officially approved traditional Chinese medicine (NMPA Drug Approval No. Z35020243), has demonstrated hepatoprotective effects, yet its epigenetic mechanisms in fibrosis treatment remain unexplored. Methods: We performed the first integrated methylome–transcriptome–proteome analysis to investigate PZH’s anti-fibrotic mechanisms in a CCl₄-induced mouse model using reduced representation bisulfite sequencing (RRBS), RNA-seq, and TMT-labeled LC-MS/MS. Results: We identified 10,974 differentially methylated loci (DMLs) and 773 differentially expressed genes (DEGs) modulated by PZH treatment. Integration analysis revealed ANXA3 and CORO1A as candidate therapeutic targets exhibiting significant inverse methylation-expression correlations validated at both transcriptomic and proteomic levels. Notably, PZH treatment modulated the CRLF-CLCF1 cytokine complex and the EGR-3 transcription factor network (42/44 genes enriched), suggesting broad transcriptional reprogramming in fibrotic liver. Protein–protein interaction (PPI) analysis highlighted key gene pairs such as Dnmt1-Uhrf1, Cbfb-Runx1, and Col4a1-Col4a2, implicating PZH in epigenetic maintenance, transcription factor regulation, and ECM remodeling. Conclusions: These findings suggest mechanistic insights into PZH’s multi-target anti-fibrotic effects and offer a rationale for developing potential therapeutic targets for liver fibrosis. Full article

Background/Objectives: Antimicrobial resistance (AMR) is a major global health threat that has outpaced the development of new antibiotics. Drug repurposing has emerged as a promising strategy for identifying alternative antimicrobial therapies. Antidepressants have attracted interest because experimental studies suggest they possess off-target antimicrobial activity, although growing evidence indicates they may also promote antimicrobial resistance. This review critically examines both the therapeutic potential and risks of antidepressant–microbial interactions. Methods: A structured narrative review was conducted using literature identified from major biomedical databases. Experimental, animal, microbiome, environmental, and clinical studies were synthesized according to antidepressant class, antimicrobial mechanisms, interactions with conventional antibiotics, microbiome modulation, and antimicrobial resistance. Results: Experimental evidence demonstrates that several antidepressants inhibit bacterial and fungal pathogens through mechanisms including membrane disruption, oxidative stress induction, efflux pump inhibition, and biofilm interference. Some agents also enhance the activity of conventional antibiotics. However, current evidence is largely preclinical. Multiple studies indicate that antidepressants may also promote antimicrobial resistance through reactive oxygen species-mediated mutagenesis, efflux pump activation, horizontal gene transfer, microbiome dysbiosis, and environmental exposure. Conclusions: Antidepressants represent both a promising drug-repurposing opportunity and a potential contributor to antimicrobial resistance. Although their antimicrobial properties are encouraging, current evidence remains predominantly preclinical, and clinically effective and safe antimicrobial dosing has not been established. At present, their antimicrobial use should be regarded as proof-of-concept rather than a clinically validated therapeutic strategy. Further pharmacokinetic, pharmacodynamic, animal, and clinical studies are required before routine antimicrobial application can be considered. Full article

Background: Benzo[a]pyrene (BaP), a common food contaminant, is a recognized gastric carcinogen. This study aimed to identify therapeutic targets and repurposed drugs for gastric cancer (GC) using BaP as a network toxicology query. Methods: An integrated strategy combining network toxicology, multi-omics, machine learning (Random Forest, LASSO, SVM-RFE), and experimental validation was applied. Results: By intersecting GC-associated genes with BaP-related targets and machine learning, we identified three hub genes. The logistic regression model further revealed KCNE2 as a protective factor (OR = 0.515, 95% CI: 0.383–0.692), while SULF1 (OR = 2.940, 95% CI: 1.399–6.179) and TIMP1 (OR = 5.351, 95% CI: 2.020–16.743) were identified as potential risk factors. Survival analysis confirmed their prognostic significance. Single-cell transcriptomics descriptively showed TIMP1 and SULF1 enrichment in malignant/stromal cells and fibroblasts, respectively, whereas KCNE2 was restricted to normal epithelial cells and silenced in tumors. GSVA implicated epigenetic regulation, ECM remodeling, and TGF-β signaling. Molecular docking and dynamics simulations suggested that BaP can form stable complexes with DNMT1 and DNMT3A. Accordingly, drug enrichment analysis identified DNMT inhibitor 5-azacytidine as a top candidate. Cellular experiments confirmed that 5-azacytidine selectively inhibited GC cells and was associated with modulation of the DNMT3A–KCNE2 axis. Conclusions: Our findings provide a novel molecular target and a repurposed drug for GC from the perspective of a food contaminant. Full article

Background/Objectives: Allergic rhinitis (AR) is a highly prevalent chronic inflammatory disease of the upper airway characterized by immune dysregulation. This study aimed to systematically characterize the immunomodulatory landscape of AR and identify exploratory molecular and cellular features associated with disease-related immune remodeling. Methods: Public bulk transcriptomic datasets and single-cell RNA sequencing data were integrated to identify immunomodulatory-related differentially expressed genes (IMRDEGs), construct an immunomodulatory score (IM.Score), evaluate immune cell infiltration, and characterize cell-type composition and intercellular communication. Machine learning was used to derive an exploratory molecular signature, and L1000CDS2-based drug repositioning analysis was performed to identify in silico candidate compounds predicted to oppose AR-associated transcriptional signatures. Results: A total of 12 IMRDEGs were identified and used to construct the IM.Score, which was lower in AR samples relative to control samples. Immune infiltration analysis demonstrated that IM.Score stratification was associated with distinct immune microenvironmental profiles. Single-cell analysis revealed potential reductions in CD4⁺ central memory T cells, which represent an immune cellular alteration requiring further experimental verification, alongside attenuated intercellular communication involving this cell population. A six-gene exploratory molecular signature comprising NFKBIA, PDCD1, MYC, IFNG, FOXP3, and CD274 showed favorable performance in the training cohort (AUC = 0.992, 95% CI: 0.974–1.000) but failed to generalize in the external validation cohort (AUC = 0.500, 95% CI: 0.245–0.755), precluding clinical diagnostic interpretation at this stage. Drug repositioning analysis identified candidate compounds, including narciclasine and BRD-K91370081, with the potential to reverse AR-associated transcriptional alterations. Conclusions: This integrative multi-omics analysis identifies coordinated molecular, cellular, and communication-level immune alterations in AR. The IM.Score, altered CD4⁺ central memory T-cell abundance, and six-gene exploratory molecular signature may be regarded as hypothesis-generating candidate clues pending further experimental and clinical validation. Full article