Host genetic factors, particularly human leukocyte antigen (HLA) polymorphisms, play a crucial role in determining the immunological outcome of Hepatitis C virus (HCV) infection. These genetic variations may influence susceptibility, disease progression, and the development of severe liver complications such as liver cirrhosis and hepatocellular carcinoma (HCC). Given the predominance of HCV genotype 4 in Egypt, this study was designed to investigate the association between specific HLA Class I and Class II alleles and clinical outcomes in an Egyptian cohort of patients with chronic HCV infection. A total of 223 participants were enrolled and stratified into four clinically defined categories: healthy controls (n = 55), patients with chronic hepatitis C (n = 86), liver cirrhotic patients (n = 42), and individuals with HCV-related HCC (n = 40). HLA genotyping was performed using the LABType® sequence-specific oligonucleotide (SSO) probe assay. Clinical and demographic data were collected between 2014 and 2016, before the implementation of Egypt's nationwide HCV eradication program, launched in 2018. This study identified differential HLA allele frequencies across HCV-related liver disease outcomes in an Egyptian cohort. HLA-DQB1*03, HLA-B*27, HLA-C*05, and HLA-A*26 showed high frequencies in healthy controls, whereas HLA-DRB1*14, HLA-A*69, HLA-B*52, HLA-B*58, and HLA-DRB1*15 appeared enriched in disease groups. Only HLA-DQB1*03 remained significant after Bonferroni correction. However, given the small sample size and lack of comprehensive national HLA data for direct comparison, these observations should be interpreted with caution. Alleles, such as HLA-B*44 and HLA-C*05, appeared absent in the HCC subgroup, though the significance of this finding remains uncertain due to low background frequencies in the control group. These preliminary data suggest possible associations of certain HLA alleles and HCV-related liver disease progression in Egyptian patients, possibly serving as candidate biomarkers for risk stratification. Nonetheless, larger, population-based studies with confirmed HCV genotyping, comprehensive treatment data, and updated controls reflecting current Egyptian HLA allele frequencies are needed to substantiate these associations.

The immune microenvironment plays a critical role in breast cancer progression and therapeutic response. Among circulating immune cells, B cells and natural killer (NK) cells have shown distinct and potentially opposing roles in tumor dynamics. However, whether dynamic changes in peripheral blood B- and NK-cell percentages are associated with treatment response across molecular subtypes of breast cancer remains unclear. Peripheral blood samples were collected from breast cancer patients before and after standardized subtype-guided therapy (chemotherapy, anti-HER2 therapy, and/or endocrine therapy according to clinical guidelines). Flow cytometry was performed to quantify the percentage of B cells and NK cells within CD45 + lymphocytes. Spearman's rank correlation was used to evaluate associations between B and NK cell percentages. Paired comparisons were performed using the Wilcoxon signed-rank test. Treatment efficacy was defined as objective response rate (complete response + partial response, CR + PR) according to RECIST criteria. Patients with decreased B cell percentage and increased NK cell percentage after treatment were defined as the B-NK group. Subtype A (-- group) exhibited significantly higher B cells percentages and lower NK cells percentages compared with other subtypes (P < 0.05). A significant negative correlation between B and NK cell percentages was observed in overall cohort and in most subtypes. In subtype A, B cell percentage significantly decreased after treatment (P < 0.05), whereas NK cell percentage showed variable changes. Patients classified into the B-NK group tended to have a higher objective response rate compared with non-B-NK patients across subtypes. Peripheral blood B and NK cell percentages exhibit dynamic and partially coordinated changes during treatment. A pattern characterized by decreased B cell percentage and increased NK cell percentage may be associated with treatment response. These findings are exploratory and hypothesis-generating, and further validation in prospective, well characterized cohorts is required before potential clinical application.

Pancreatic adenocarcinoma (PAAD) is a highly lethal malignancy characterized by a dismal prognosis and limited treatment options. Protein tyrosine kinase 6 (PTK6) is abnormally activated in multiple cancer types. Understanding the function of PTK6 in pancreatic cancer could reveal novel therapeutic targets and prognostic markers. We extracted the expression profiles and clinical data of patients with pancreatic adenocarcinoma from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO). The Wilcoxon rank-sum test was used to compare PTK6 gene expression levels between tumor tissues and normal pancreatic tissues. Functional enrichment analysis, Cox regression analysis, and Kaplan-Meier plot analysis were employed to determine the prognostic value of PTK6. Single-sample gene set enrichment analysis (ssGSEA) and TIMER3.0 were used to evaluate immune cell infiltration. Finally, CCK-8, RT‒qPCR, colony formation, and western blot assays were performed in vitro cellular experiments to verify the biological effects of PTK6 on the malignant behaviour of pancreatic cancer. PTK6 is highly expressed in pancreatic adenocarcinoma, with its upregulation validated in both TCGA (AUC = 0.935) and GEO (AUC = 0.796) cohorts. Elevated PTK6 expression was correlated with shorter overall survival (HR = 2.08, 95% CI: 1.36-3.19, p < 0.001), and multivariate analysis revealed that N1 stage was an independent prognostic factor for OS. In addition, PTK6 expression was negatively correlated with immune cell infiltration and key immune checkpoint genes, with the strongest correlation observed for TIGIT (ρ = -0.316, p < 0.001). In vitro experiments revealed that downregulating PTK6 expression via siRNA inhibited cell proliferation and colony formation, concomitant with increased LC3-II/LC3-I ratio, suggesting a potential role in autophagy regulation. PTK6 could serve as a potential diagnostic and prognostic biomarker for PAAD and may participate in immune modulation and autophagy regulation. These findings, supported by multi-cohort bioinformatic analysis and preliminary experimental validation, highlight its prospective clinical value in prognosis evaluation, immunotherapy, and targeted therapy for PAAD.

Colorectal cancer (CRC) is one of the most common malignancies of the gastrointestinal tract and remains a leading cause of cancer-related mortality. Hypoxia and pyroptosis are closely linked to malignant progression and may shape tumour biology and the immune microenvironment. Pyroptosis- and hypoxia-related genes were obtained from GeneCards and MSigDB, and PHRGs were defined as the intersection of pyroptosis- and hypoxia-associated gene sets for subsequent consensus clustering. Transcriptomic profiles and survival information for CRC were retrieved from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Pyroptosis- and hypoxia-related molecular subtypes were identified using consensus clustering. Least absolute shrinkage and selection operator and Cox regression analyses were applied to construct a prognostic signature. Key signature genes were preliminarily assessed at the expression level in paired CRC and adjacent normal tissues using qRT-PCR (n = 10 pairs) and western blotting (n = 6 pairs). Given the limited sample size and lack of stage- or subtype-stratified validation, these experiments provide preliminary expression-level support rather than functional confirmation. Patients were classified into four PHRG-related molecular subtypes with distinct prognoses and biological characteristics, with PHRGcluster C2 showing the worst outcome. A total of 1578 pyroptosis- and hypoxia-related differentially expressed genes (PHRDEGs) were identified across the four PHRGclusters. A six-gene prognostic signature (PHRDEGscore) was then established in the TCGA cohort. Patients with a high PHRDEGscore had shorter survival, which was validated in external cohorts (GSE39582 and GSE17536). The low-PHRDEGscore group exhibited higher immune infiltration (e.g., activated CD8+ and CD4+ T cells), and an exploratory association with immunotherapy response was observed in a non-CRC immunotherapy-treated cohort (IMvigor210). Experimental analyses indicated higher mRNA levels of HES4, SHROOM2, and OXCT1 in CRC tissues than in adjacent normal tissues (qRT-PCR, n = 10 pairs), and increased protein expression of HES4 and SHROOM2 (western blotting, n = 6 pairs). This study identified hypoxia- and pyroptosis-related molecular patterns and a six-gene prognostic signature in CRC. The observed associations are correlative and hypothesis-generating. Prospective validation in independent, well-annotated clinical cohorts is needed before any clinical application can be considered.