Current Issues in Molecular Biology最新文献

Genome-wide association studies (GWAS) have transformed the study of chronic kidney disease (CKD) by identifying hundreds of genetic loci associated with multiple aspects of kidney function, including albuminuria and CKD risk factors, in diverse populations. A major challenge is translating statistically significant signals into causal genes and mechanisms, as most CKD-associated variants lie in non-coding regulatory regions and often act in a cell type- and context-specific manner. In this review, we provide an overview of the current strategies for moving from GWAS signals toward the identification of causal genes for CKD. We discuss advances in four areas: statistical and functional fine-mapping, molecular quantitative trait locus (QTL) mapping, colocalization, and transcriptome-wide associations, highlighting the advantages and disadvantages of each. We further examined how emerging kidney-specific single-cell, single-nucleus, and spatial transcriptomic atlases have enabled the mapping of genetic risk to specific renal cell types and microanatomical niches. By combining these approaches with chromatin interaction data, multi-omics analytics, and clustered regularly interspaced short palindromic repeats (CRISPR)-based studies, the process of generating causal relationships and mechanistic understanding has been further refined. Importantly, this review provides a unifying framework that synthesizes cross-sectional and longitudinal GWAS with kidney-specific functional genomics to distinguish genetic determinants of CKD susceptibility from modifiers of disease progression, thereby highlighting how regulatory variation and disease trajectories inform precision nephrology. As a result, we can provide insights into the role of genetically informed gene prioritization for experimentation, therapeutic target discovery, and the development of a framework for precision nephrology. Together, these advancements highlight how human genetics, in conjunction with functional genomics and experimental biology, can link an association signal to a clinically relevant interpretation of CKD.

Significant advances in coronavirus immunoprophylaxis have enabled the control of the SARS-CoV-2 pandemic. However, the continued emergence of SARS-CoV-2 variants with immune escape potential highlights the need for effective direct-acting antivirals targeting conserved viral enzymes. The SARS-CoV-2 main protease (M^pro) remains one of the most promising antiviral drug targets due to its essential role in viral replication and the high conservation of its active site across coronavirus variants. Building upon the established GC373 scaffold, we designed, synthesized, and biochemically evaluated two novel GC373-like peptidomimetic inhibitors incorporated modified glutamine-mimic residues. These analogs were designed to enhance solubility and metabolic resilience while retaining key recognition features within the M^pro active site. Both compounds demonstrated micromolar inhibitory activity in enzymatic assays, supported by molecular docking and MM-PBSA analyses consistent with stable binding. The proposed inhibitors represent viable scaffolds for further optimization of electrophilic warheads and S1/S2 residue interactions. These findings contribute to the rational design of next-generation M^pro inhibitors and align with ongoing efforts to expand the chemical space of SARS-CoV-2 antiviral agents.

C-C motif chemokine ligand 2 (CCL2) may reflect subtle metabolic-inflammatory changes in pregnancy. This study evaluated CCL2 concentrations and their peripartum changes in women with uncomplicated term pregnancies, focusing on associations with maternal metabolic status. Serum CCL2 was measured before delivery and 48 h postpartum; urinary CCL2 was assessed postpartum. Peripartum serum change (ΔCCL2) was calculated. BMI was recorded pre-pregnancy (or early pregnancy), at delivery, and 48 h postpartum; total BMI change (ΔBMI) was derived. Participants were stratified into two groups (ΔBMI > 1 kg/m² vs. ≤1 kg/m²). Peripartum serum CCL2 changes differed significantly between ΔBMI groups. In the total cohort, CCL2 correlated with HbA1c and selected body composition indices, including fat tissue index, lean tissue index, and body cell mass. In women with ΔBMI > 1 kg/m², additional associations were found with BMI, peripartum BMI change, HbA1c, ferritin, creatinine, and total body water. Among women with ΔBMI ≤ 1 kg/m², significant relationships were observed with uric acid and triglycerides. Peripartum CCL2 dynamics appear to reflect maternal metabolic status, even in metabolically "normal" pregnancies, but these findings are exploratory and should be interpreted cautiously. CCL2 is a promising marker of subtle metabolic alterations in late pregnancy and the early postpartum period, but further validation is required before clinical application.

Background: Lung adenocarcinoma (LUAD) remains a leading cause of cancer-related mortality. The prognostic significance and functional role of sodium overload-induced necrosis (a novel form of regulated cell death driven by disrupted sodium homeostasis, hereafter abbreviated as NECSO) in LUAD are largely unexplored. Methods: A prognostic model was constructed utilizing the NECSO key gene TRPM4 and analyzed through Cox, LASSO, and multivariate Cox regression analyses. LUAD patients were stratified into high- and low-risk groups. The model's predictive performance was evaluated using time-dependent ROC curves and nomograms. Functional enrichment analysis elucidated underlying biological disparities. The tumor immune microenvironment was characterized using ESTIMATE, ssGSEA, CIBERSORTx, and TIDE algorithms, with results corrected for multiple testing. Drug sensitivity to chemotherapeutic and targeted agents was predicted. The functional role of a key gene, DENND1C, was validated in vitro. Its association with immunotherapy survival outcomes was assessed in a real-world cohort. Results: The NECSO-based prognostic signature demonstrated robust performance in risk stratification across training and independent validation cohorts. Patients in the high-risk group exhibited significantly shorter overall survival. Functional enrichment revealed associations with processes related to plasma membrane integrity, cell death, metabolism, and immune response. Multi-algorithm immunogenomic analyses consistently identified an immunosuppressive microenvironment in high-risk patients. The risk score was predictive of differential sensitivity to therapeutics, including taxanes and EGFR inhibitors. In vitro experiments confirmed DENND1C as a tumor suppressor, inhibiting LUAD cell proliferation, invasion, and migration. Furthermore, high DENND1C expression was associated with improved survival in patients receiving immunotherapy. Conclusions: This study establishes and validates a novel NECSO-based prognostic model for LUAD. DENND1C is identified as a key tumor suppressor and a potential biomarker for immunotherapy, offering insights for personalized treatment strategies in LUAD.

Gitelman syndrome (GS) is a rare, autosomal recessive salt-losing tubulopathy caused by mutations in the SLC12A3 gene. It involves dysfunction of the sodium-chloride cotransporter positioned on the apical membranes of the distal convoluted tubule cells, causing sodium shortage and mimicking the use of thiazide diuretics. Hyperaldosteronism secondary to sodium depletion and hypovolemia causes hypokalaemia and metabolic alkalosis. This is associated with inhibition of the Transient Receptor Potential Cation Channel, Subfamily M, Member 6 -TRPM6 channel, which leads to urinary magnesium leakage and hypomagnesemia, subsequently stopping PTH secretion and resulting in hypocalcemia and hypocalciuria. Gitelman syndrome frequently presents later in life, as the symptoms are usually not very threatening. However, early identification, diagnosis, and urgent intervention are essential to improve patient prognosis and quality of life. Importantly, both hypomagnesemia and hypokalaemia can impair insulin secretion and sensitivity. Furthermore, hyperaldosteronism caused by the secondary activation of the R-A-A system can also lead to these disorders. Glucose metabolism problems have been shown to prevail amongst GS patients and manifest more frequently in comparison to the general population. When it comes to the treatment used to reduce hyperglycemia in GS-related T2DM, we consider which of the available drugs are the best for those patients. The article analyses the association of Gitelman syndrome with diabetes mellitus based on the available medical literature-as there are no clinical trials or meta-analyses available for this group, it is presented as a narrative review.

Neurodegenerative diseases, including Alzheimer's disease, are marked by cholinergic dysfunction, oxidative stress, and reduced neurotrophic support, which drives the quest for multifunctional therapeutic agents. This pilot study presents four novel monoterpene-aminoadamantane conjugates (MACs 1-4) designed to combine the antioxidant and neuromodulatory characteristics of monoterpenes with the neuroprotective properties of aminoadamantane derivatives. Their physicochemical characteristics, blood-brain barrier permeability, and binding affinity to human acetylcholinesterase (AChE) were evaluated using molecular docking and in silico descriptor analysis. In vivo, the neuroprotective efficacy of the MACs was investigated in a scopolamine-induced dementia model in rats, employing behavioral tests. Biochemical assays conducted in the hippocampus and prefrontal cortex assessed AChE activity, antioxidant enzyme performance, lipid peroxidation levels, total glutathione content, and BDNF concentrations. The findings indicate that MAC1, MAC3, and MAC4 demonstrate favorable calculated blood-brain barrier permeability, strong predicted affinity for AChE, and significant in vivo alleviation of scopolamine-induced memory deficits, in conjunction with improvement of key markers of oxidative stress and cholinergic function. These results show that the structural hybridization of myrtenal with aminoadamantane frameworks produces promising multifunctional ligands that are relevant for Alzheimer's-type neurodegeneration.

The Zinc Finger X-Linked Duplicate B (ZXDB) gene is one of a pair of replicated zinc finger genes on chromosome Xp11.21. The homologous gene of ZXDB in mice is Zxdb. Recent studies have found that Zxdb plays a role in the spermatogenic process of mice; however, its impact on the female reproductive system has not yet been explored. In our study, we found, for the first time, that the loss of function of Zxdb leads to reduced decidualization rates and a decrease in litter size in female mice. Secondly, we found that maternal loss of Zxdb is the determinant of these phenotypes. Thirdly, the transcriptional and proteomic differential expression genes in the uterine tissues of wild-type (WT) and Zxdb knockout (Zxdb-KO) mice were significantly enriched in signaling pathways such as adhesion molecules. Finally, we demonstrated that the disorder of expression and uneven distribution of adhesion molecules in mouse uterine tissue may be the main reason for the decline in embryo implantation rate. In conclusion, we have established for the first time a link between the Zxdb gene and reduced female fertility. This study will help provide guidance and genetic counseling for future common clinical complications such as Recurrent Spontaneous Abortion (RSA) or Recurrent Implantation Failure (RIF).

The incidence of primary liver cancer is increasing annually, with extremely high mortality and suboptimal therapeutic outcomes. The inefficient presentation of tumor antigens and low infiltration of specific cytotoxic T lymphocytes (CTLs) result in insufficient immunogenicity, which limits the efficacy of immunotherapy. Despite the popularity of immune checkpoint inhibitors (ICIs), insufficient immune activation means only a small subset of hepatocellular carcinoma (HCC) patients exhibit clinical responses to ICIs, showing significant inter-individual variability. The activation of the cyclic GMP-AMP synthase(cGAS)- stimulator of interferon genes(STING) pathway initiates the expression of type I interferons (IFNs) and inflammatory cytokines, promoting the formation of a pro-inflammatory environment at the tumor site. This pathway enhances anti-tumor immune responses by facilitating antigen processing and presentation, T cell priming and activation, and remodeling of the immunosuppressive microenvironment. Our research found that cucurbitacin B (CuB), a natural component derived from traditional Chinese medicine, had significant anti-hepatocellular carcinoma properties and exerted anti-tumor effects through the cGAS-STING pathway. Specifically, CuB regulated ferroptosis by down-regulating the expression of Solute Carrier Family 7 Member 11 (SLC7A11) and Glutathione Peroxidase 4 (GPX4) and upregulating the expression of Transferrin Receptor Protein 1 (TFR1) and Long-chain Acyl-CoA Synthetase 4 (ACSL4). These actions involved lipid substrates, iron ion homeostasis, and antioxidant defense systems. The release of mitochondrial DNA (mtDNA) triggered by ferroptosis activated the cGAS-STING immune signaling pathway, leading to the up-regulation of cGAS, phosphorylated STING (p-STING), phosphorylated TANK-binding kinase 1 (TBK1), phosphorylated Interferon regulatory factor3 (IRF3), and Interferon-β (IFN-β). This cascade activation pattern provides new insights into the drug treatment of tumors.

The patch-clamp technique is widely regarded as the gold standard in cellular electrophysiology and can be applied in several configurations. In the cell-attached (C-A) mode, it enables the recording of single-channel currents, whereas the whole-cell (W-C) mode allows for the measurement of macroscopic currents, representing the collective activity of many channels. When the recording configuration was switched from C-A to W-C on the same cell, the current amplitude increased dramatically, while action currents (ACs) were completely abolished, indicating a profound alteration in the cell's electrophysiological response under the new setup. In excitable cells, the occurrence of ACs, representing propagated action potentials, can interfere with C-A single-channel recordings. To address this, a high-K⁺ solution is typically applied to the bath to suppress the ACs. The inwardly rectifying K⁺ (Kir), ATP-sensitive K⁺ (K_ATP) and large-conductance Ca²⁺-activated K⁺ (BK_Ca) channels are crucial members of the K⁺ channel family that facilitate the efflux of K⁺ ions, driven by the K⁺ electrochemical gradient. These channels are primarily distinguished by their rectification properties and gating kinetics. For instance, K_ATP channels exhibit a bursting kinetic pattern with inward rectifying property, while BK_Ca channels display strong outward rectification. Mitoxantrone, which belongs to a class of drugs called anthracenediones, can suppress the activity of Kir channels in differentiated RAW 264.7 cells, with no change in single-channel conductance. The respiratory stimulator GAL-021 acts as a BK_Ca channel inhibitor, and it suppresses channel activity and shifts the activation curve to the right, suggesting a voltage-dependent blockade that stabilizes the channel in a closed state. GAL-021 does not change the single-channel conductance, indicating it is a gating modifier rather than an open-pore blocker. The functional roles of ion channels are fundamentally important. Correspondingly, the field is transitioning to artificial intelligence for automated single-cell patch-clamp experiments, though brain slice recordings still require manual techniques.

Approximately 50% of individuals diagnosed with Stage I liver cancer live beyond four years; however, a small subset of Stage I patients die within the first year. A prognostic biomarker panel that can identify high-risk Stage I patients may be extremely valuable. In this study, we used the Long-Evans Cinnamon (LEC) rat model of Wilson's Disease and hepatocellular carcinoma (HCC), along with data from The Cancer Genome Atlas (TCGA) human database, to create a novel biomarker panel. We generated and analyzed a rat microarray gene expression profile by comparing liver tumor tissues with adjacent normal tissues from the same animals, covering approximately 30,000 genes. The microarray results were translated into a five-gene panel associated with 1-year survival in Stage I liver cancer patients based on TCGA data, in combination with machine learning and bioinformatics approaches. The panel was internally validated following the "REporting recommendations for Tumor MARKer prognostic studies (REMARK)" guidelines. With no existing Stage-I-specific prognostic tools, a biomarker panel associated with 1-year survival in patients with Stage I liver cancer is a potential candidate for rigorous external validation.