Endoplasmic reticulum stress (ERS) is a crucial mechanism for maintaining cellular homeostasis and plays a significant role in the pathogenesis of hypertrophic cardiomyopathy (HCM). Identifying ERS-associated feature genes related to HCM and ERS can provide new perspectives and potential targets for future treatments of heart diseases. Multiple datasets were downloaded from the GEO database, followed by differential analysis and weighted gene co-expression network analysis (WGCNA) to screen genes most related to HCM. ERS-related genes were intersected with the previously identified genes. Feature genes were screened using machine learning methods, diagnostic efficacy, and expression validation, and a nomogram was constructed based on the feature genes. The CIBERSORT algorithm was used to calculate the proportion of immune cell infiltration, and single-cell RNA-sequencing analysis was used to characterize the expression of the feature gene across different cell types. Finally, the expression of the feature gene was verified by real-time quantitative PCR (RT-qPCR). FKBP5, MYH6 and PROS1 were regarded as feature genes. The transcriptomic classification nomogram model constructed based on these genes shows favorable internal apparent performance (AUC = 0.974). Immune analysis revealed that 8 types of immune cells in HCM were significantly changed, and monocytes were significantly correlated with FKBP5 and PROS1. Single‑cell RNA‑sequencing analysis revealed that PROS1 was localized to cardiomyocytes within HCM myocardial specimens. RT-qPCR experiment verified that the expression trends of MYH6 and PROS1 in the Ang II-induced cardiomyocyte hypertrophy model were consistent with the results of bioinformatics analysis. This study identified three ERS‑associated candidate feature genes in HCM that showed a significant correlation with immune cells. The findings provide a theoretical basis and reference value for exploring the relationship between HCM, ER stress, and immunity.
CHARGE syndrome (CS) is a recognizable genetic syndrome with known pattern of multiple organ malformations and the association between SEMA3E and CS is not well understood. In a one-year-old boy who suspected diagnosis with CS, all his clinical data were collected and trio whole exome sequencing (WES) and Sanger sequencing were performed in the patient and his parents for genetic tests. The patient had global developmental delay, microcephaly, hypertelorism, low-set ears, auricular malformation, bilateral renal hypoplasia, and gastroesophageal reflux. A de novo heterozygous SEMA3E c.1983_1984insCTTT was found in the patient and it was not documented in SNP databases. Bioinformatics analysis predicts that this frameshift mutation terminates at codon 668, resulting in the production of a truncated protein with approximately 88 amino acid residues missing at the C-terminus; this truncation occurs within the disulfide-bond domain (residues 654–729) of the SEMA3E protein. In addition, exploratory non-targeted metabolomics analysis (Supplementary Materials) revealed differences in the patient’s metabolic profile compared to that of his parents; however, due to limitations related to the single-subject nature of the study and confounding factors, these results cannot yet serve as mechanistic evidence supporting a SEMA3E-related pathogenesis and should be considered only as a hypothesis for future research. A novel, likely pathogenic variant identified in this study provides evidence that SEMA3E is a candidate gene for CHARGE syndrome.
Human cytomegalovirus (HCMV) encodes multiple microRNAs that modulate viral replication, immune evasion, and latency. Among them, miR-UL112-3p, miR-UL22A-3p, and miR-US25-1-3p have been implicated in regulating host antiviral responses and viral persistence. This study evaluated the expression of three HCMV-encoded miRNAs and their corresponding viral transcripts in solid organ transplant recipients with plasma HCMV DNA loads > 10,000 copies/mL or no detectable plasma HCMV DNA, compared with healthy controls. Thirty solid organ transplant recipients with plasma HCMV DNA loads > 10,000 copies/mL (active group), 30 recipients with no detectable plasma HCMV DNA (inactive group), and 30 healthy controls were enrolled. HCMV DNA was quantified by TaqMan real-time PCR, and expression of UL112, UL22A, US25, miR-UL112-3p, miR-UL22A-3p, and miR-US25-1-3p was assessed by SYBR Green real-time RT-PCR. The expression levels of HCMV UL112, UL22A, and US25 transcripts and their corresponding miRNAs were evaluated in blood samples using quantitative real-time PCR. UL112, UL22A, and US25 transcript expression was higher in recipients with plasma HCMV DNA loads > 10,000 copies/mL than in recipients with no detectable plasma HCMV DNA and healthy controls. miR-UL112-3p was also elevated in active infection. Spearman correlation analysis demonstrated a moderate inverse association between UL112 mRNA and miR-UL112-3p in the inactive HCMV group (ρ = −0.46, P = 0.009), whereas no significant association was observed in the active group (ρ = 0.129, P = 0.49). UL22A and miR-UL22A-3p were not significantly correlated in either group. In contrast, US25 mRNA and miR-US25-1-3p showed strong positive correlations in both inactive (ρ = 0.92, P < 0.0001) and active HCMV infection (ρ = 0.90, P < 0.0001). HCMV-encoded miRNAs and their corresponding viral transcripts exhibit locus-specific relationships that differ between groups defined by plasma HCMV DNA status. The inverse association between UL112 and miR-UL112-3p in inactive infection, absence of correlation for UL22A/miR-UL22A-3p, and strong concordance between US25 and miR-US25-1-3p indicate that viral miRNA-transcript relationships are not uniform across HCMV loci.
Leber congenital amaurosis encompasses a genetically heterogeneous spectrum of inherited retinal disorders, typically following an autosomal recessive inheritance pattern, with onset in infancy or early childhood. Several genes have been associated with LCA, including GUCY2D, which encodes retinal membrane guanylyl cyclase involved in phototransduction. Whole-exome sequencing was employed to identify candidate variants in an Iranian proband from a consanguineous family presenting with severe visual impairment consistent with LCA. Variant interpretation was performed using database resources and in silico prediction tools. Sanger sequencing was performed in the proband, both parents, and the clinically unaffected sister to confirm the variant and assess its segregation within the family. Structural modeling of wild-type and p.(Gly982Ser) GUCY2D proteins was performed, followed by molecular docking analysis with GTP using AutoDock Vina. Docking results were evaluated to explore potential structural differences between the wild-type and mutant proteins. A homozygous missense variant in GUCY2D, NM_000180.4:c.2944G > A, p.(Gly982Ser), was identified in the proband and confirmed by Sanger sequencing. This variant has previously been reported in a Kurdish child with LCA1, and the present study represents an independent observation in a Kurdish family. Exploratory molecular docking revealed model-dependent differences in GTP pose sampling between wild-type and G982S GUCY2D. However, the observed Vina-score differences were small relative to the uncertainty of the scoring function, and no retained pose simultaneously satisfied all predefined catalytic-geometry criteria. This study reports an independent observation of the homozygous GUCY2D p.(Gly982Ser) variant in a patient with a clinical presentation consistent with LCA. The genetic findings and computational analyses provide additional context for understanding the potential molecular effects of this variant. However, further experimental functional studies are required to determine its biological consequences and contribution to LCA1 pathogenesis.
Alzheimer’s disease (AD) and progressive supranuclear palsy (PSP) are neurodegenerative tauopathies characterized by accumulation of insoluble hyperphosphorylated tau protein in the brain. Despite neuropathological similarities, these conditions are also differentiated by distinct pathological and clinical features informed by complex molecular mechanisms. Therefore, exploring the molecular mechanisms that are conserved or distinct in AD and PSP is crucial for understanding the pathogenic risk factors contributing to these conditions and will ultimately advance preventive strategies in precision medicine. Although alternative splicing is among the most important forms of transcriptional regulation in the brain, its role in distinguishing or unifying mechanisms between these conditions remains incompletely understood. Transcriptomic analysis was performed on RNA-seq data from the temporal cortex of 82 AD patients, 84 PSP patients, and 78 controls in the discovery set, and 96 AD and 96 PSP patients in the replication set. Differential exon skipping events in AD and PSP were identified, followed by functional enrichment analysis. Whole-genome sequencing (WGS) data from ascertained samples were further integrated to investigate their cis-genetic regulators as splicing quantitative trait loci (sQTLs). Relative to the controls, a total of 472 and 366 genes with exon skipping events (ES genes) were differentially spliced in AD and PSP, respectively. Among these, 325 ES genes were specific to AD and 219 to PSP, while 147 ES genes were common to both conditions. These shared ES genes were enriched in pathways related to neural activity and tau metabolism such as myelination, Schwann cell development, and neurotrophin production. ES genes exclusive to AD were enriched for synaptic activity, lipid metabolism, and amyloid beta processing, and those exclusive to PSP for the vesicle-mediated transport pathway. Further integration of WGS data identified three independent splicing quantitative trait loci (cis-sQTLs) that were associated with three ES events in the genes c7orf63, DEF8, and HAP1. This study highlights common and distinct molecular mechanisms mediated through exon skipping that underlie the tauopathies, AD and PSP. These findings enhance our understanding of the complex neuropathogenic mechanisms associated with these conditions and ultimately support further research aimed at identifying and developing effective treatment strategies.
