Current Issues in Molecular Biology

16 pages, 12952 KB

Open AccessArticle

Astrocyte Subtype-Specific Expression of the Sodium-Coupled Citrate Transporter SLC13A5 and Citrate Metabolism Genes Across Alzheimer’s Disease Pseudoprogression: A Single-Nucleus RNA Sequencing Analysis of the Human Middle Temporal Gyrus

by Patricia Fernanda Schuck, Gustavo da Costa Ferreira and Hércules Rezende Freitas

Curr. Issues Mol. Biol. 2026, 48(7), 691; https://doi.org/10.3390/cimb48070691 - 5 Jul 2026

Abstract

The sodium-coupled citrate transporter NaCT (SLC13A5) imports extracellular citrate into cells. In the CNS, SLC13A5 is described to be expressed predominantly in neurons. Cytosolic citrate levels rely on citrate generated in mitochondria and imported from other CNS cells, regulating intermediary metabolism [...] Read more.

The sodium-coupled citrate transporter NaCT (SLC13A5) imports extracellular citrate into cells. In the CNS, SLC13A5 is described to be expressed predominantly in neurons. Cytosolic citrate levels rely on citrate generated in mitochondria and imported from other CNS cells, regulating intermediary metabolism and supplying acetyl-CoA for lipid synthesis and histone acetylation. Despite evidence for NaCT’s role in neurometabolic homeostasis, its transcriptional behavior across Alzheimer’s disease (AD) progression and across astrocyte subtypes remains uncharacterized at single-cell resolution. We analyzed single-nucleus RNA sequencing data from 1,378,211 nuclei across 84 donors in the Seattle Alzheimer’s Disease Brain Cell Atlas (SEA-AD) Middle Temporal Gyrus dataset to profile SLC13A5 and seven citrate metabolism genes across a continuous AD pseudoprogression score. SLC13A5 expression was restricted to astrocytes (~20% prevalence) and concentrated in the Astro 2 supertype (24.0%), a homeostatic subtype characterized by low C3 (1.6%) and CD44 (5.5%), which expanded with pseudoprogression (Spearman rho = +0.345, FDR < 0.001). The A1-reactive Astro 3 supertype, where SLC13A5 prevalence was 0.87%, declined concordantly (rho = −0.393). Opposing compositional and transcriptional forces produced apparent stability in overall SLC13A5 prevalence. SLC13A3 and ACO1 showed progressive donor-level declines correlating with Braak stage and Thal phase (rho range: −0.307 to −0.349, FDR < 0.01). APOE4 carriers exhibited lower SLC13A5 prevalence specifically within Astro 2 nuclei (median 17.6% vs. 25.9%; Wilcoxon p = 0.025), though this association did not survive multivariate regression. No difference in Astro 2 SLC13A5 expression was detected between cognitively resilient and expected-AD donors with equivalent high Braak burden (p = 0.888). Contrary to the prevailing description of NaCT as a neuronal transporter, SLC13A5 transcript in the SEA-AD MTG dataset was detected almost exclusively in astrocyte nuclei, concentrated in the homeostatic Astro 2 subtype, and maintained as this subtype expanded with advancing AD pathology. Because these are nuclear transcript measurements, they delimit where SLC13A5 mRNA is detectable rather than establishing the cellular site of NaCT protein or activity, which requires in situ validation. Full article

(This article belongs to the Special Issue Molecular Dialogues: Signaling Networks of the Aging Nervous System)

13 pages, 545 KB

Open AccessArticle

Alpha-Lipoic Acid Modulates Melanoma Survival Networks via ER Stress Induction, Mitochondrial Apoptosis, and Kinase Pathway Suppression in B16F10 Cells

by Ömer Kokaçya, Percin Pazarci and Halil Mahir Kaplan

Curr. Issues Mol. Biol. 2026, 48(7), 690; https://doi.org/10.3390/cimb48070690 - 3 Jul 2026

Abstract

Background/Objectives: Malignant melanoma is characterized by constitutive PI3K/Akt/mTOR and MAPK activation, driving aggressive behavior and therapeutic resistance. Alpha-lipoic acid (αLA), a naturally occurring dithiol compound with an established clinical safety profile, has shown anticancer potential; however, its integrated molecular mechanisms in melanoma remain [...] Read more.

Background/Objectives: Malignant melanoma is characterized by constitutive PI3K/Akt/mTOR and MAPK activation, driving aggressive behavior and therapeutic resistance. Alpha-lipoic acid (αLA), a naturally occurring dithiol compound with an established clinical safety profile, has shown anticancer potential; however, its integrated molecular mechanisms in melanoma remain poorly defined. This study aimed to comprehensively evaluate the cytotoxic and mechanistic effects of αLA in B16F10 murine melanoma cells. Methods: Antiproliferative effects were assessed by MTT assay at four concentrations (250, 500, 750, 1000 µM) over 48 h. Protein levels of apoptotic markers (Bax, Bcl-2, Caspase-3, AIF), kinase signaling components (p-Akt, p-mTOR, p-ERK, p-JNK), ER stress markers (GRP78, GADD153/CHOP), and cell cycle regulator Wee1 were quantified by ELISA at a specifically selected sub-lethal concentration of 750 µM (inducing ~38% growth inhibition). Results: αLA dose-dependently inhibited B16F10 proliferation. At 750 µM, it triggered robust intrinsic apoptotic signaling, evidenced by a nearly 10-fold shift in the Bax/Bcl-2 ratio and greater than 9-fold Caspase-3 activation. Elevated AIF suggested profound mitochondrial stress and the potential priming of concurrent caspase-independent cell death mechanisms. αLA suppressed survival signaling by reducing p-Akt (44%), p-mTOR, p-ERK, and p-JNK. Treatment triggered lethal ER stress via GRP78 and GADD153/CHOP upregulation and upregulated Wee1, suggesting the induction of stress-responsive checkpoint signaling. The simultaneous CHOP upregulation and p-Akt suppression highlight a concurrent dysregulation of stress and survival pathways, suggesting a potential pro-apoptotic interplay. Conclusions: αLA exerts potent multi-target anticancer effects by inducing a broad spectrum of associated molecular changes, including the suppression of PI3K/Akt/mTOR and MAPK networks, induction of ER stress, engagement of cell cycle checkpoints, and activation of the mitochondrial Bax/Bcl-2/Caspase-3 axis. Importantly, these correlative findings do not establish proven pathway dependencies. Nevertheless, this concurrent dysregulation positions αLA as a potential disruptor of inter-pathway resilience underlying drug resistance. Full article

(This article belongs to the Section Molecular Pharmacology)

► Show Figures

Graphical abstract

18 pages, 1072 KB

Open AccessArticle

5-ALA Photodynamic Therapy Induces Competing Death and Survival Pathways in Glioblastoma Cells

by Julia Inglot, Dorota Bartusik-Aebisher, Joanna Katarzyna Strzelczyk, Angelika Myśliwiec, Klaudia Dynarowicz, Dorota Hudy, Oliwia Trzaskoś, Jacek Tabarkiewicz, Aleksandra Kawczyk-Krupka, Magdalena Moś and David Aebisher

Curr. Issues Mol. Biol. 2026, 48(7), 689; https://doi.org/10.3390/cimb48070689 - 3 Jul 2026

Abstract

Glioblastoma multiforme (GBM), isocitrate dehydrogenase (IDH)-wildtype, is the most aggressive primary malignant tumor of the central nervous system, characterized by poor prognosis and high recurrence rates despite standard multimodal treatment. This study investigates the molecular response of glioblastoma cells to 5-aminolevulinic acid (5-ALA)-based [...] Read more.

Glioblastoma multiforme (GBM), isocitrate dehydrogenase (IDH)-wildtype, is the most aggressive primary malignant tumor of the central nervous system, characterized by poor prognosis and high recurrence rates despite standard multimodal treatment. This study investigates the molecular response of glioblastoma cells to 5-aminolevulinic acid (5-ALA)-based photodynamic therapy (PDT), focusing on gene expression changes associated with apoptosis, ferroptosis, and oxidative stress. Human glioblastoma T98G cells were treated with 5-ALA followed by light irradiation, and gene expression was analyzed using RT-qPCR. PDT induced moderate upregulation of pro-apoptotic genes (BAX, CASP3, FAS) alongside increased expression of the anti-apoptotic gene BCL2, indicating simultaneous activation of cell death and survival pathways. Ferroptosis-related genes showed mixed responses, with slight upregulation of ACSL4 and downregulation of GPX4, suggesting increased susceptibility to lipid peroxidation. The most significant change was observed in GCH1 expression, reflecting activation of oxidative stress response mechanisms. However, none of the observed changes reached statistical significance, likely due to the limited sample size. These findings demonstrate that PDT induces a complex and dual biological response in glioblastoma cells, involving both cytotoxic and adaptive mechanisms. This may limit therapeutic efficacy and contribute to treatment resistance. The results support the rationale for combining PDT with targeted molecular therapies aimed at inhibiting antioxidant defenses and anti-apoptotic pathways. Additionally, personalized therapeutic strategies based on tumor molecular profiles may enhance treatment outcomes. Further studies with larger sample sizes and functional validation are required to confirm these preliminary observations. Full article

(This article belongs to the Special Issue Cancer-Associated Remodeling of Functional Molecular Pathways)

24 pages, 1544 KB

Open AccessArticle

Optimized Preparation of Gastrodiae elata Extract Enhances Antiepileptic Effects by Regulating Neuroinflammation, Oxidative Stress, and Neuronal Apoptosis in Rats

by He Wang, Shiyi Lun, Hu Ding, Zhimeng Li, Xian Wu, Huiyang Yuan, Bo Yang, Guoxin Ji, Huan Wang and Shumin Wang

Curr. Issues Mol. Biol. 2026, 48(7), 688; https://doi.org/10.3390/cimb48070688 - 3 Jul 2026

Abstract

Epilepsy is a common chronic neurological disorder characterized by recurrent seizures. Gastrodia elata, the dried tuber of G. elata Bl. (Orchidaceae), is a valuable medicinal and edible botanical resource. This study optimized the preparation of Yellow Rice Wine-Processed G. elata (YPGE) and [...] Read more.

Epilepsy is a common chronic neurological disorder characterized by recurrent seizures. Gastrodia elata, the dried tuber of G. elata Bl. (Orchidaceae), is a valuable medicinal and edible botanical resource. This study optimized the preparation of Yellow Rice Wine-Processed G. elata (YPGE) and investigated its antiepileptic effects and underlying mechanisms in a pentylenetetrazol (PTZ)-kindled rat model. Processing parameters were optimized using single-factor experiments combined with an analytic hierarchy process (AHP)-entropy weight method (EWM) weighting strategy and Box–Behnken design–response surface methodology. The optimal parameters were determined as 18% alcohol by volume, 72 °C drying temperature, and 32 h drying time. Compared with unprocessed G. elata (GE), YPGE exhibited 0.54-, 0.13-, 1.87-, and 3.58-fold increases in the contents of gastrodin (GAS), G. elata polysaccharides (GEPs), p-hydroxybenzyl alcohol (p-HBA), and total parishins (TP), respectively, and demonstrated significantly enhanced in vitro antioxidant activity (IC50 values of 2.604, 2.719, and 4.046 mg/mL for DPPH, ABTS, and hydroxyl radicals). In vivo, both GE and YPGE significantly reduced seizure severity, decreased inflammatory cytokines (TNF-α, IL-1β), alleviated oxidative stress (increased SOD and GSH-Px, decreased MDA), and modulated neurotransmitter balance (reduced Glu, increased GABA) in brain tissues. YPGE also upregulated P-glycoprotein expression and reduced neuronal apoptosis in the hippocampal CA1 region by upregulating Bcl-2 and downregulating Bax. These findings suggest that YPGE exerts multi-target antiepileptic effects through synergistic anti-inflammatory, antioxidant, and anti-apoptotic actions, providing experimental evidence for the development of novel antiepileptic therapies based on processed G. elata. Full article

(This article belongs to the Section Bioorganic Chemistry and Medicinal Chemistry)

45 pages, 1558 KB

Open AccessReview

Liver Macrophages in the Pathogenesis of Viral Hepatitis

by Ioannis Tsomidis, Angeliki Tsakou, Argyro Voumvouraki and Elias Kouroumalis

Curr. Issues Mol. Biol. 2026, 48(7), 687; https://doi.org/10.3390/cimb48070687 - 3 Jul 2026

Abstract

Chronic hepatitis B virus (HBV) and hepatitis C virus (HCV) infection remain a world health problem leading to fibrosis and cirrhosis. Liver damage is primarily mediated by the innate and adaptive immune responses since HBV and HCV are not directly cytotoxic. Kupffer cells [...] Read more.

Chronic hepatitis B virus (HBV) and hepatitis C virus (HCV) infection remain a world health problem leading to fibrosis and cirrhosis. Liver damage is primarily mediated by the innate and adaptive immune responses since HBV and HCV are not directly cytotoxic. Kupffer cells and liver-recruited macrophages are heavily implicated in both viral elimination and progression of the disease. HBV and HCV proteins polarize macrophages into either an M1 pro-inflammatory phenotype, promoting hepatocyte damage or into an M2 immunosuppressive phenotype, leading to viral persistence and fibrogenesis via cytokines such as interleukin-10 (IL-10) and transforming growth factor-beta (TGF-β). In this review a brief overview of the heterogeneity of liver macrophages in health and during chronic viral infection is presented. Recognition of viruses by macrophages and the modulation of macrophages by viral proteins in the pathogenesis of liver inflammation and injury are discussed in detail. Most importantly, the mechanisms that HBV and HCV are using to manipulate macrophages and escape elimination are also presented. The role of macrophages in the evolution of acute-on-chronic liver failure is analyzed. Finally, a concise presentation of the emerging, but not yet clinically used, therapeutic strategies targeting macrophages to control chronic HBV infection and restore the dysregulated immune response is discussed. In conclusion, this integrated review of liver macrophage implication summarizes the pathophysiology and pathogenesis of HBV and HCV including acute-on-chronic- liver failure and viral cirrhosis. Full article

(This article belongs to the Special Issue Advances in Molecular Biology Methods in Hepatology Research, 2nd Edition)

► Show Figures

Figure 1

13 pages, 11926 KB

Open AccessArticle

Functional Analysis of IL-6 Genetic Variants and Their Potential Role in Lipid Homeostasis and Inflammatory Regulation in Colombian Athletes

by Diana Carolina Zambrano Ríos, Miguel Ángel Gómez, Juan Manuel Gómez, Felipe Alberto Polo, Betty Oviedo Sarria, Julián Andrés Rivera and Andrés Jenuer Matta

Curr. Issues Mol. Biol. 2026, 48(7), 686; https://doi.org/10.3390/cimb48070686 - 3 Jul 2026

Abstract

Obesity and metabolic dysregulation are closely associated with chronic low-grade inflammation, in which interleukin-6 (IL-6) plays a key regulatory role. Genetic variation in the IL-6 gene may influence inflammatory responses and metabolic homeostasis. To identify single-nucleotide variants (SNVs) in the IL-6 gene in [...] Read more.

Obesity and metabolic dysregulation are closely associated with chronic low-grade inflammation, in which interleukin-6 (IL-6) plays a key regulatory role. Genetic variation in the IL-6 gene may influence inflammatory responses and metabolic homeostasis. To identify single-nucleotide variants (SNVs) in the IL-6 gene in a cohort of Colombian high-performance athletes and to evaluate their potential functional and structural consequences using bioinformatic prediction and protein-modeling approaches. A descriptive observational study was conducted in a cohort of 23 high-performance Colombian athletes from Valle del Cauca representing cycling, karate, and weightlifting disciplines. Genomic Deoxyribonucleic Acid (DNA) extracted from peripheral blood samples was analyzed using Next-Generation Sequencing (NGS). Identified variants were evaluated using several in silico prediction tools, including Basic Local Alignment Search Tool (BLAST version 2.16.0), Expert Protein Analysis System (ExPASy version 4.0), Open Reading Frame Finder (ORFfinder version 0.4.3), and population databases such as Genome Aggregation Database (gnomAD version 4.0). Structural modeling was used to explore the potential impact of amino-acid substitutions on IL-6 protein stability. Eight single-nucleotide variants were identified in the IL-6 gene. Among them, the rs1524107 variant generated a missense substitution predicted to modify the amino-acid sequence of the IL-6 protein. Structural modeling suggested a potential alteration in protein stability associated with this variant. The rs1524107 variant may influence IL-6 protein structure according to computational predictions. These findings provide preliminary hypothesis-generating evidence regarding the potential role of IL-6 genetic variation in inflammatory regulation; however, functional validation and larger cohort studies are required to determine their biological significance. Full article

(This article belongs to the Special Issue Mechanisms and Pathophysiology of Obesity)

► Show Figures

Figure 1

2 pages, 158 KB

Open AccessCorrection

Correction: Yuan et al. Molecular Diversity, Structure–Function Relationship, Mechanism of Action, and Transformative Potential of Black Soldier Fly Antimicrobial Peptides Against Multidrug-Resistant Pathogens. Curr. Issues Mol. Biol. 2026, 48, 62

by Ru-Xi Yuan, Xiao-Yang Ma, Yang Lv and Hong-Bin Si

Curr. Issues Mol. Biol. 2026, 48(7), 685; https://doi.org/10.3390/cimb48070685 - 3 Jul 2026

Abstract

Figure Legend [...] Full article

(This article belongs to the Section Bioorganic Chemistry and Medicinal Chemistry)

14 pages, 283 KB

Open AccessReview

Research Progress on the Regulatory Mechanisms of Salt-Stress Response and Functional Genes in Populus

by Peiyang He and Hanyang Cai

Curr. Issues Mol. Biol. 2026, 48(7), 684; https://doi.org/10.3390/cimb48070684 - 3 Jul 2026

Abstract

Soil salinization represents one of the most severe abiotic constraints on global forest productivity. Populus, the most widely cultivated fast-growing timber tree and a premier model woody plant, exhibits striking intrageneric variation in salt tolerance—from the extremely halophytic Populus euphratica to highly [...] Read more.

Soil salinization represents one of the most severe abiotic constraints on global forest productivity. Populus, the most widely cultivated fast-growing timber tree and a premier model woody plant, exhibits striking intrageneric variation in salt tolerance—from the extremely halophytic Populus euphratica to highly salt-sensitive cultivated clones. Understanding the molecular basis of this variation has profound implications for saline–alkali land reclamation and salt-tolerant variety breeding. This review systematically synthesizes current knowledge on Populus salt-stress responses, covering three primary injury mechanisms (osmotic stress, ionic toxicity, and oxidative damage) and the corresponding physiological countermeasures. We further survey functional genes across four major categories: ion transporters, osmotic-adjustment enzymes, antioxidant-defense components, and transcription factors. Crucially, we extend beyond the herbaceous-plant paradigm by examining salt-tolerance strategies that are specific to the woody architecture of Populus: long-distance radial and axial Na⁺ transport through tall stems, salt sequestration in senescent bark and wood parenchyma, and deep-root ion exclusion strategies. Comparative insights from other woody genera are incorporated to highlight convergent and divergent mechanisms. On this basis, we propose an integrated multi-level regulatory model in which Na⁺ compartmentalization/efflux serves as the core, ROS homeostasis as the key regulatory axis, and osmotic adjustment as the auxiliary strategy. Outstanding challenges—including unresolved primary salt-signal perception, insufficient pathway integration, and limited in planta gene-function verification—are critically assessed, and future research priorities encompassing multi-omics integration, CRISPR-based gene editing, and natural-population genomics are outlined. Full article

(This article belongs to the Special Issue Molecular Mechanisms and Omics Approaches in Plant Stress Tolerance)

20 pages, 1991 KB

Open AccessArticle

Phenotypic Biodiversity and Niche-Associated Functional Traits in Lactiplantibacillus plantarum

by Gianluca Paventi, Mariantonietta Succi, Katia Maglieri, Catello Di Martino, Maria Virginia Soldovieri and Massimo Iorizzo

Curr. Issues Mol. Biol. 2026, 48(7), 683; https://doi.org/10.3390/cimb48070683 - 2 Jul 2026

Abstract

Lactiplantibacillus plantarum is a highly versatile lactic acid bacterium, widely distributed across diverse ecological niches. Although often described as a nomadic species, increasing evidence suggests that strains from different habitats may retain niche-associated functional traits. This study investigated the phenotypic biodiversity of forty [...] Read more.

Lactiplantibacillus plantarum is a highly versatile lactic acid bacterium, widely distributed across diverse ecological niches. Although often described as a nomadic species, increasing evidence suggests that strains from different habitats may retain niche-associated functional traits. This study investigated the phenotypic biodiversity of forty L. plantarum strains isolated from four ecologically distinct environments: wine, honeybee gut, trout intestine, and pre-weaning infant feces. Growth performance at different temperatures and on various carbon sources, acidification capacity, and β-glucosidase activity were evaluated and integrated using multivariate statistical analyses. Significant differences in β-glucosidase activity were observed among ecological groups (Kruskal–Wallis, p = 0.001), with wine-associated strains exhibiting the highest enzymatic activities and trout-derived isolates the lowest. Growth and acidification traits showed more limited variation among habitats, indicating that these physiological characteristics are largely conserved within the species. Heatmap visualization, principal component analysis (PCA), and hierarchical clustering revealed substantial phenotypic heterogeneity among strains. PCA indicated that growth performance and acidification traits contributed primarily to the first principal component, whereas β-glucosidase activity and differential fructose utilization were major contributors to the second component. Permutational multivariate analysis of variance (PERMANOVA) confirmed a significant effect of ecological origin on the overall phenotypic structure (p = 0.006), although habitat explained only 15.3% of the total variance (R² = 0.153). Overall, the results show that ecological origin contributes to the phenotypic diversification of L. plantarum populations while preserving the extensive functional versatility characteristic of this species. β-Glucosidase activity emerged as the most discriminating phenotypic trait among ecological groups and represented the principal niche-associated functional signature identified in this study. Full article

(This article belongs to the Collection Feature Papers Collection in Molecular Microbiology)

► Show Figures

Figure 1

23 pages, 15923 KB

Open AccessArticle

Lysimachiae Herba Modulates FXR to Alleviate Cholestatic Liver Injury: Insights from Serum Pharmacochemistry and Experimental Validation

by Wei Zhao, Bao Yu, Chengli Li, Jingjing Li, Haijun Huang and Weiguo Cao

Curr. Issues Mol. Biol. 2026, 48(7), 682; https://doi.org/10.3390/cimb48070682 - 2 Jul 2026

Abstract

Cholestatic liver injury (CLI) is a complex condition for which current treatment options remain limited. Lysimachiae Herba (LH), a traditional Chinese medicine with hepatoprotective properties, has not yet been fully characterized in terms of its active constituents and underlying mechanisms in CLI. This [...] Read more.

Cholestatic liver injury (CLI) is a complex condition for which current treatment options remain limited. Lysimachiae Herba (LH), a traditional Chinese medicine with hepatoprotective properties, has not yet been fully characterized in terms of its active constituents and underlying mechanisms in CLI. This study was designed to systematically determine the chemical composition of LH, characterize its absorbed constituents in vivo, and elucidate its therapeutic mechanisms against CLI. UPLC-Q-TOF-MS/MS was employed to analyze the chemical composition of LH and its absorbed components in rat serum. Key targets and signaling pathways were predicted using network pharmacology and molecular docking, followed by experimental validation in an ANIT-induced CLI mouse model and LCA-treated HepG₂ cells through biochemical assays, histological examination, transcriptomic analysis, qRT-PCR, Western blotting, and immunofluorescence analysis. A total of 129 compounds were tentatively identified in LH, among which 26 were detected in the bloodstream. Network analysis and molecular docking suggested that LH regulates bile acid homeostasis predominantly by the FXR signaling pathway. Both in vivo and in vitro experiments provided convergent evidence that LH modulates the FXR-related bile acid regulatory network, enhances bile acid efflux transporter expression, and alleviates CLI. In conclusion, this study systematically elucidates the chemical composition, absorbed constituents, and pharmacological mechanisms of LH in CLI, highlighting the involvement of FXR-related bile acid regulation as an important mechanism and providing a scientific basis for the potential development of LH for cholestatic liver injury. Full article

(This article belongs to the Special Issue Medicinal Plants and Phytochemicals for the Prevention and Intervention of Metabolic Disease and Its Complications)

► Show Figures

Figure 1

18 pages, 1974 KB

Open AccessReview

Gene-Edited Stem Cells for Ischemic Vascular Disease: Current Advances and Future Perspectives

by Seongho Han and Sung-Whan Kim

Curr. Issues Mol. Biol. 2026, 48(7), 681; https://doi.org/10.3390/cimb48070681 - 2 Jul 2026

Abstract

Ischemic vascular diseases remain a leading cause of morbidity and mortality worldwide and are frequently associated with irreversible tissue damage. Although stem cell-based therapies have shown promise for vascular regeneration, their clinical translation has been limited by poor survival, insufficient engraftment, functional heterogeneity, [...] Read more.

Ischemic vascular diseases remain a leading cause of morbidity and mortality worldwide and are frequently associated with irreversible tissue damage. Although stem cell-based therapies have shown promise for vascular regeneration, their clinical translation has been limited by poor survival, insufficient engraftment, functional heterogeneity, and immune rejection. Recent advances in genome-editing technologies, including CRISPR/Cas9, base editing, and prime editing, have provided powerful tools for overcoming these limitations through precise genetic modification of stem cells. Gene editing can enhance angiogenic potential, improve resistance to ischemic stress, augment paracrine activity, promote endothelial maturation, and reduce immunogenicity. In this review, we outline the current genome-editing toolbox and its application to stem cell engineering for vascular regeneration in ischemic disease. We also examine emerging therapeutic concepts, universal donor cell platforms, and key issues in safety and ethics, with a focus on translational pathways. Taken together, advances at the interface of genome editing and stem cell biology are likely to accelerate the development of regenerative therapies that deliver more durable vascular repair in ischemic vascular disease. Full article

(This article belongs to the Special Issue Stem Cell-Derived Cells and Bioengineering: Molecular Bases in Regeneration and Disease Research)

► Show Figures

Figure 1

29 pages, 1485 KB

Open AccessReview

Advancements and Clinical Applications Prospects of Epigenetic Biomarkers in Liquid Biopsy for Oral Squamous Cell Carcinoma

by Yuan Li, Yao Liu, Yuyi Cong, Juan Liu, Wen Pan, Xiaobing Guan and Jiaqi Wang

Curr. Issues Mol. Biol. 2026, 48(7), 680; https://doi.org/10.3390/cimb48070680 - 1 Jul 2026

Abstract

Oral squamous-cell carcinoma (OSCC) is a prevalent malignancy of the head and neck region. A delay in the diagnosis of OSCC often results in a high metastatic tendency, which is the main reason for the high patient mortality. Dynamic monitoring and management of [...] Read more.

Oral squamous-cell carcinoma (OSCC) is a prevalent malignancy of the head and neck region. A delay in the diagnosis of OSCC often results in a high metastatic tendency, which is the main reason for the high patient mortality. Dynamic monitoring and management of the onset and progression of OSCC are critical for improving patient survival rates. Liquid biopsy technology—characterized by its non-invasive nature, procedural convenience, and capacity for longitudinal monitoring—is a promising adjunct to histopathological examination for the early diagnosis of OSCC. Epigenetic alterations, characterized by reversibility and long-term stability in physiological fluids, are critical enablers of liquid biopsy and its clinical utility. Advances in detection technologies, including quantitative polymerase chain reaction (qPCR), digital droplet PCR (ddPCR), next-generation sequencing (NGS), and electrochemical biosensors, have significantly facilitated the research and clinical translation of epigenetic biomarkers in oral liquid biopsies. However, translating epigenetic biomarkers from research discovery to clinical practice for OSCC remains hindered by several critical challenges: the scarcity of large-scale, rigorously designed cohort studies, limited multicenter validation, inconsistent preprocessing protocols, and a lack of harmonized analytical platforms. Finally, we propose a conceptual framework to outline potential clinical application models for these biomarkers. Full article

(This article belongs to the Special Issue Oral Cancer: Prophylaxis, Etiopathogenesis and Treatment, 2nd Edition)

► Show Figures

Graphical abstract

16 pages, 1003 KB

Open AccessArticle

A Scalable Lentiviral Workflow for Laboratory-Scale Generation of BCMA/GPRC5D Co-Transduced CAR-T Cells in Multiple Myeloma

by Ewa Nowak, Emilia Morawiec, Adam Pudełko, Agnieszka Polak, Mateusz Broncel, Daria Matczyńska, Dawid Zamojski, Michał Czerwinski and Anna Bednarska-Czerwińska

Curr. Issues Mol. Biol. 2026, 48(7), 679; https://doi.org/10.3390/cimb48070679 - 30 Jun 2026

Abstract

Efficient and reproducible lentiviral vector production and T-cell transduction remain important technical challenges in CAR-T (Chimeric Antigen Receptor T-cell) cell manufacturing. In this study, we optimized HEK293T transfection and primary T-cell transduction parameters for lentiviral CAR constructs targeting BCMA (B-cell maturation antigen) and [...] Read more.

Efficient and reproducible lentiviral vector production and T-cell transduction remain important technical challenges in CAR-T (Chimeric Antigen Receptor T-cell) cell manufacturing. In this study, we optimized HEK293T transfection and primary T-cell transduction parameters for lentiviral CAR constructs targeting BCMA (B-cell maturation antigen) and GPRC5D (G-protein coupled receptor family C group 5 member D). Lipofectamine 3000 and TurboFectin 8.0 were compared across different seeding densities and reagent-to-DNA ratios, with vector yields quantified by qPCR (Quantitative Polymerase Chain Reaction) and p24 ELISA (Enzyme-linked Immunosorbent Assay). Lipofectamine 3000 consistently generated higher viral titers and transduction efficiencies, as reflected by a greater proportion of GFP-positive (Green Fluorescent Protein) cells than TurboFectin 8.0, reaching peak titers of 9.65 × 10⁸ copies/mL for the anti-GPRC5D and 5.33 × 10⁸ copies/mL for the anti-BCMA vectors. Under optimized conditions, transduction efficiencies reached 43.8% GFP⁺ cells for BCMA-CAR and approximately 13–14% GFP-positive transduced cells for the GPRC5D construct within the tested TU/mL range. Co-transduction experiments yielded approximately 62–66% GFP⁺ cells with detectable BCMA-binding and presumptive GPRC5D-CAR-expressing subpopulations identified based on GFP reporter expression. Immunophenotypic analysis demonstrated a relatively stable CD4/CD8 distribution (~65/35), enrichment of effector memory CD8⁺ cells, and expression of activation-associated markers. Collectively, these findings describe an optimized lentiviral transfection and transduction workflow that may support the further development of dual-targeting BCMA/GPRC5D CAR-T manufacturing strategies in research and early translational settings. Full article

(This article belongs to the Section Molecular Medicine)

► Show Figures

Graphical abstract

25 pages, 29699 KB

Open AccessArticle

Unraveling the Skeletal Growth-Promoting Mechanism of the Seahorse Hippocampus erectus: From Active Fraction Screening to Signaling Pathway Regulation

by Lianghua Huang, Zhaoji Pan, Meng Bai, Jiyan Guo, Jian Xiao and Chenghai Gao

Curr. Issues Mol. Biol. 2026, 48(7), 678; https://doi.org/10.3390/cimb48070678 - 30 Jun 2026

Abstract

As a traditional element of Chinese medicine, Hippocampus erectus is well known for promoting adolescent growth, yet its active fractions and underlying molecular mechanisms remain unclear. In this study, the aqueous extract of H. erectus was subjected to in vitro simulated gastrointestinal digestion [...] Read more.

As a traditional element of Chinese medicine, Hippocampus erectus is well known for promoting adolescent growth, yet its active fractions and underlying molecular mechanisms remain unclear. In this study, the aqueous extract of H. erectus was subjected to in vitro simulated gastrointestinal digestion and ultrafiltration to separate three molecular weight fractions (<10 kDa, 10–30 kDa, >30 kDa). Their chemical profiles were characterized, and osteogenic activities were systematically evaluated using cell assays, a juvenile rat model, and integrated transcriptomics and data-independent acquisition (DIA) proteomics. Results revealed that chemical profiling showed the >30 kDa fraction was mainly composed of hemocyanin subunits, and the 10–30 kDa fraction was enriched in growth-related amino acids and steroid derivatives; functionally, the 10–30 kDa fraction promoted preosteoblast proliferation and early differentiation via enhanced alkaline phosphatase (ALP) activity, while the >30 kDa fraction dominated late osteoblast maturation and mineralization. Both fractions significantly increased rat body and bone length by expanding growth plate proliferative zones and elevating serum insulin-like growth factor-1 (IGF-1)/bone morphogenetic protein-2 (BMP-2) levels. Transcriptomic and proteomic analyses identified vascular endothelial growth factor (VEGF), Wingless-related integration site (Wnt), phosphatidylinositol 3-kinase-protein kinase B (PI3K-Akt), and extracellular matrix (ECM)–receptor interaction as potential core regulatory pathways. Integrated multi-omics analysis further confirmed Frizzled-related protein B (Frzb) and AKT1 substrate 1 (Akt1s1) as candidate key regulatory targets enriched in the Wnt and adenosine monophosphate-activated protein kinase (AMPK) signaling pathways. These findings elucidate the multi-fraction, multi-pathway mechanism of H. erectus in promoting skeletal development, providing scientific evidence for its traditional use and a theoretical basis for growth-promoting functional food development. Full article

(This article belongs to the Special Issue Natural Products in Biomedicine and Pharmacotherapy, 2nd Edition)

10 pages, 1255 KB

Open AccessArticle

Multiplex PCR Fluorescence Method for Detection of Genetically Modified Maize Strains

by Wenxiu Yin, Wenxin Zhang, Quan Zhang, Zhengping Ying, Shan Wu, Huizhen Yu and Mingzhe Zhang

Curr. Issues Mol. Biol. 2026, 48(7), 677; https://doi.org/10.3390/cimb48070677 - 30 Jun 2026

Abstract

The rapid proliferation of genetically modified (GM) crops and the uncontrolled distribution of GM-based food and feed have become a growing global concern, posing new challenges for regulatory oversight and traceability. The traditional PCR detection method cannot simultaneously meet the needs of high-throughput, [...] Read more.

The rapid proliferation of genetically modified (GM) crops and the uncontrolled distribution of GM-based food and feed have become a growing global concern, posing new challenges for regulatory oversight and traceability. The traditional PCR detection method cannot simultaneously meet the needs of high-throughput, high-specificity and high-sensitivity detection of transgenic organisms. In this study, a multiplex fluorescence PCR-capillary electrophoresis platform was developed by labeling primers of endogenous and exogenous genes with different fluorescent groups. The system enabled the simultaneous detection of 27 GM-related genes and events in a single analytical workflow. The results demonstrated accurate identification of all seven GM maize events, with correct detection achieved for each individual strain. In addition, the method enabled precise discrimination of a mixed sample containing five GM maize varieties. The assay also achieved a detection sensitivity of 0.1% in gradient mixtures with different GM contents. Our platform integrates a larger number of targets into a single PCR reaction, thereby simplifying the detection workflow while maintaining high analytical performance. Furthermore, the combination of multicolor fluorescence labeling and capillary electrophoresis provides high-resolution fragment discrimination and robust multiplex detection capability. This platform provides a novel and effective tool for rapid detection in food safety of transgenic crops and related areas, and can be applied in import/export inspection, quarantine, and biosafety surveillance. Full article

(This article belongs to the Section Molecular Plant Sciences)

► Show Figures

Figure 1

20 pages, 2111 KB

Open AccessReview

Elevated CO₂ as a Biostimulatory Approach to Enhance the Nutraceutical Potential of Ginseng

by Hamad Hussain, Nooral Amin, Imran Ali, Abdul Wakeel Umar and Naveed Ahmad

Curr. Issues Mol. Biol. 2026, 48(7), 676; https://doi.org/10.3390/cimb48070676 - 30 Jun 2026

Abstract

The continued rise in atmospheric carbon dioxide (CO₂) concentrations presents a strategic opportunity to harness climate change variables within the framework of precision agriculture. Despite the well-established role of elevated CO₂ (eCO₂) in enhancing biomass accumulation, its largely [...] Read more.

The continued rise in atmospheric carbon dioxide (CO₂) concentrations presents a strategic opportunity to harness climate change variables within the framework of precision agriculture. Despite the well-established role of elevated CO₂ (eCO₂) in enhancing biomass accumulation, its largely underexplored potential to drive the biosynthesis of secondary metabolites represents a more significant and promising avenue of investigation. This review appraises the physiological and molecular mechanisms through which eCO₂ enrichment redirects metabolic flux toward secondary metabolite biosynthesis, with far-reaching implications for plant productivity and resilience. Special emphasis is placed on critically evaluating the scientific literature to explore how CO₂-mediated modulation of the carbon–nutrient balance (CNB) can be strategically leveraged to enhance secondary metabolite yields. Moving from observation to application, integrated strategies are proposed to exploit CO₂ enrichment in advanced bioreactor systems and controlled-environment greenhouses as a means of maximizing bioactive compound production in ginseng. Pinpointing the regulatory sweet spots at which carbon saturation elicits maximum ginsenoside expression opens a promising avenue for engineering ginseng cultivation systems with sustainable potency and superior bioactivity. Though the full molecular architecture of these pathways in Panax awaits elucidation, converging evidence from related plant systems furnishes a credible mechanistic scaffold for future research. Full article

(This article belongs to the Special Issue Molecular Mechanisms of Plant Stress Responses and Development)

► Show Figures

Figure 1

20 pages, 1703 KB

Open AccessArticle

Differential Anti-Inflammatory Effects of Semaglutide and Tirzepatide in Experimental Diabetes Mellitus

by Roxana-Cristina Dobriceanu, Ianis Kevyn Stefan Boboc, Liliana Mititelu Tartau, Andreea Daniela Meca, Carmen Nicoleta Oancea, Maria-Alexandra Paceana, Marius-Mihai Pastiu, Adina Turcu-Stiolica and Maria Bogdan

Curr. Issues Mol. Biol. 2026, 48(7), 675; https://doi.org/10.3390/cimb48070675 - 30 Jun 2026

Abstract

Background: Type 2 diabetes mellitus is associated with chronic low-grade inflammation contributing to endothelial dysfunction, metabolic imbalance, and cardiovascular complications. Although semaglutide (SEM) and tirzepatide (TIR) provide important metabolic and cardioprotective benefits, their early anti-inflammatory effects and potential sex-dependent differences remain incompletely [...] Read more.

Background: Type 2 diabetes mellitus is associated with chronic low-grade inflammation contributing to endothelial dysfunction, metabolic imbalance, and cardiovascular complications. Although semaglutide (SEM) and tirzepatide (TIR) provide important metabolic and cardioprotective benefits, their early anti-inflammatory effects and potential sex-dependent differences remain incompletely understood. This study comparatively evaluated the effects of SEM and TIR on systemic inflammatory biomarkers in a murine model of streptozotocin-induced diabetes mellitus. Methods: Thirty BALB/c mice were allocated into six experimental groups according to sex and treatment: control, SEM, and TIR groups (n = 5/group). Diabetes was induced by intraperitoneal streptozotocin administration, followed by treatment with SEM or TIR. Circulating interleukin-1β (IL-1β) and pentraxin-3 (PTX-3) levels were measured at baseline, one week after streptozotocin administration, and after six weeks of treatment. Results: Control groups exhibited progressive increases in IL-1β and PTX-3 levels, indicating sustained inflammatory activation. In contrast, SEM- and TIR-treated animals showed attenuated inflammatory responses characterized by transient or stabilized biomarker profiles. Differential inflammatory responses were observed between treatments and sexes. Male SEM and Male TIR groups demonstrated stable IL-1β levels, whereas female treated groups showed persistent elevations, particularly Female TIR animals. PTX-3 responses also displayed differential sex-dependent patterns, with Female SEM animals exhibiting the most stable inflammatory profile. Conclusions: These findings suggest differential early immunomodulatory effects of the two modern antidiabetic drugs, characterized by distinct biomarker responses according to sex and inflammatory marker profile. IL-1β and PTX-3 may represent complementary biomarkers for the assessment of early inflammatory activation associated with diabetes mellitus and its cardiometabolic complications. Full article

(This article belongs to the Special Issue Novel Pharmacological Strategies and Molecular Mechanisms in Nonclinical Research)

► Show Figures

Figure 1

14 pages, 2563 KB

Open AccessArticle

Cytokine Profiles as Molecular Markers Associated with Physical Exercise and Insulin Therapy in Patients with Type 2 Diabetes Mellitus

by Danielle Cristina Honorio França, Alan Cardec Barbosa, Elton Brito Ribeiro, Anibal Monteiro de Magalhães Neto, Emanuelle Carolina Honorio França, Maraisa Delmult Borges, Patrícia Gelli Feres de Marchi, Adenilda Cristina Honorio-França and Eduardo Luzía França

Curr. Issues Mol. Biol. 2026, 48(7), 674; https://doi.org/10.3390/cimb48070674 - 30 Jun 2026

Abstract

Background: Type 2 diabetes mellitus is characterized by chronic low-grade inflammation, immune dysregulation, and metabolic impairment. This study investigated cytokine profiles associated with physical exercise and insulin therapy in patients with Type 2 diabetes mellitus. Methods: Blood samples were collected from 51 volunteers [...] Read more.

Background: Type 2 diabetes mellitus is characterized by chronic low-grade inflammation, immune dysregulation, and metabolic impairment. This study investigated cytokine profiles associated with physical exercise and insulin therapy in patients with Type 2 diabetes mellitus. Methods: Blood samples were collected from 51 volunteers to evaluate metabolic parameters and cytokine concentrations. According to glycemic status and insulin use, participants were classified into non-diabetic, non-insulin-dependent Type 2 Diabetes Mellitus, and insulin-dependent Type 2 Diabetes Mellitus groups. Results: Physically active individuals with non-insulin-dependent Type 2 Diabetes Mellitus exhibited increased IL-4, IL-6, and IL-10 levels, suggesting enhanced immunoregulatory and anti-inflammatory responses. Physically active patients with insulin-dependent Type 2 Diabetes Mellitus showed elevated IL-17 concentrations. In contrast, sedentary individuals with insulin-dependent Type 2 Diabetes Mellitus exhibited higher TNF-α levels, indicating a more pronounced proinflammatory profile. IFN-γ concentrations were significantly higher in patients with insulin-dependent Type 2 Diabetes Mellitus, regardless of exercise status. Correlation analyses demonstrated distinct cytokinemetabolic interaction patterns according to metabolic condition and physical exercise. Conclusion: Cytokines can serve as molecular markers of immunometabolic responses associated with physical exercise and insulin therapy in Type 2 Diabetes Mellitus, reflecting alterations in systemic inflammatory regulation and immune–metabolic crosstalk related to glycemic adaptation. Full article

(This article belongs to the Special Issue Molecular Research on Metabolic Disease)

► Show Figures

Figure 1

18 pages, 1070 KB

Open AccessReview

Selected Chemokines as Prognostic Biomarkers and Therapeutic Targets in Ovarian Cancer

by Anna Długaszek, Jacek Kabut, Małgorzata Domagała-Haduch, Anita Gorzelak-Magiera, Joanna Sadurska, Maria-Laura Morawiec, Aleksandra Mielczarek-Palacz and Iwona Gisterek-Grocholska

Curr. Issues Mol. Biol. 2026, 48(7), 673; https://doi.org/10.3390/cimb48070673 - 30 Jun 2026

Abstract

Ovarian cancer, particularly high-grade serous ovarian cancer (HGSOC), remains one of the most lethal gynecological malignancies due to late diagnosis and the development of chemoresistance. The tumor microenvironment (TME) plays an important role in disease progression, with chemokines influencing cell recruitment, angiogenesis, metastasis, [...] Read more.

Ovarian cancer, particularly high-grade serous ovarian cancer (HGSOC), remains one of the most lethal gynecological malignancies due to late diagnosis and the development of chemoresistance. The tumor microenvironment (TME) plays an important role in disease progression, with chemokines influencing cell recruitment, angiogenesis, metastasis, and immune modification. This review synthesizes current evidence on key chemokine axes in ovarian cancer, highlighting their dual roles as prognostic biomarkers and therapeutic targets. The most important axes include CXCL12/CXCR4 (which drives tumor proliferation, angiogenesis and chemoresistance via epithelial–mesenchymal transition), CCL2/CCR2 (promoting immunosuppressive tumor-associated macrophages and resistance), and CCL5/CCR5 (enhancing pro-oncogenic signaling and Treg/MDSC infiltration). Pro-angiogenic ELR+CXC chemokines like CXCL8 induce vascularization and inflammation. On the contrary, effector chemokines (CXCL9/10/11/13) correlate with “hot” immune subtypes and improved survival in several studies. High expression of immunosuppressive chemokines predicts poorer prognosis and therapy resistance, while immune-attracting profiles associate with better outcomes and chemotherapy responsiveness. Therapeutically, inhibitors like plerixafor (CXCR4), PF-04136309 (CCR2), and maraviroc (CCR5) show preclinical promise, synergizing with chemotherapy, anti-VEGF, and checkpoint inhibitors. Chemokines also represent actionable molecular targets to overcome ovarian cancer’s “cold” immune phenotype. Future research should validate multi-chemokine signatures for patient stratification and advanced clinical trials toward personalized therapies. Full article

(This article belongs to the Section Molecular Medicine)

► Show Figures

Figure 1

Journal Description

Current Issues in Molecular Biology

Latest Articles

Journal Menu

Journal Browser

Highly Accessed Articles

Latest Books

E-Mail Alert

News

Topics

Conferences

Special Issues

Topical Collections

Further Information

Guidelines

MDPI Initiatives

Follow MDPI