Cells

18 pages, 4668 KB

Open AccessArticle

Streptozotocin Causes Blood–Brain Barrier and Astrocytic Dysfunction In Vitro

by Sarah A. Habib, Mohamed M. Kamal, Mohamed H. Aly, Heba R. Ghaiad, Sherine M. Rizk, William A. Banks and Michelle A. Erickson

Cells 2025, 14(21), 1745; https://doi.org/10.3390/cells14211745 - 6 Nov 2025

Viewed by 1033

Abstract

Streptozotocin (STZ) is an alkylating agent that has neurotoxic effects when injected into the cerebral ventricles (ICV) and also models many other features of Alzheimer’s disease. However, the mechanisms of STZ neurotoxicity are not well understood. In this study, we hypothesized that some [...] Read more.

Streptozotocin (STZ) is an alkylating agent that has neurotoxic effects when injected into the cerebral ventricles (ICV) and also models many other features of Alzheimer’s disease. However, the mechanisms of STZ neurotoxicity are not well understood. In this study, we hypothesized that some of the neurotoxic effects of STZ could be due to direct activities on brain endothelial cells and astrocytes, which are key in forming and supporting the functions of the blood–brain barrier (BBB), respectively. To test this hypothesis, we characterized the changes induced by STZ either in cultures of human-induced pluripotent stem cell (iPSC)-derived brain endothelial-like cells (iBECs), which form an in vitro BBB model, or in primary human astrocytes. We found that STZ at a dosage of 5 mM caused a delayed reduction in the transendothelial electrical resistance (TEER) of iBECs at 7–11 days post-treatment, indicating induction of BBB leakage. Additionally, we observed significant increases in albumin leakage across the monolayer, altered iBEC morphology, and reductions in tight junction proteins, suggesting that STZ causes BBB disruption. We further found that the BBB glucose transporter GLUT-1 was reduced in iBECs, as was the total number of iBECs. In astrocytes, the 5 mM dose of STZ reduced the GFAP signal and total number of cells, suggesting that STZ has anti-proliferative and/or toxic effects on astrocytes. Together, these data support that STZ’s neurotoxic effects could be due, in part, to its direct toxic activities on brain endothelial cells and astrocytes. Full article

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29 pages, 5933 KB

Open AccessArticle

Gap Junctional Communication Required for the Establishment of Long-Term Robust Ca²⁺ Oscillations Across Human Neuronal Spheroids and Extended 2D Cultures

by Jasmin Kormann, Eike Cöllen, Ayla Aksoy-Aksel, Jana Schneider, Yaroslav Tanaskov, Kevin Wulkesch, Marcel Leist and Udo Kraushaar

Cells 2025, 14(21), 1744; https://doi.org/10.3390/cells14211744 - 6 Nov 2025

Viewed by 999

Abstract

Synchronized oscillatory fluctuations in intracellular calcium concentration across extended neuronal networks represent a functional indicator of connectivity and signal coordination. In this study, a model of human immature neurons (differentiated from LUHMES precursors) has been used to establish a robust protocol for generating [...] Read more.

Synchronized oscillatory fluctuations in intracellular calcium concentration across extended neuronal networks represent a functional indicator of connectivity and signal coordination. In this study, a model of human immature neurons (differentiated from LUHMES precursors) has been used to establish a robust protocol for generating reproducible intracellular Ca²⁺ oscillations in both two-dimensional monolayers and three-dimensional spheroids. Oscillatory activity was induced by defined ionic conditions in combination with potassium channel blockade. It was characterized by stable frequencies of approximately 0.2 Hz and high synchronization indices across millimeter-scale cultures. These properties were consistently reproduced in independent experiments and across laboratories. Single-cell imaging confirmed that oscillations were coordinated throughout large cell populations. Pharmacological interventions demonstrated that neither excitatory nor inhibitory chemical synaptic transmission influenced oscillatory dynamics. Gap junction blockers completely disrupted synchronization, while leaving individual cell activity unaffected. Functional dye-transfer assays provided additional evidence for electrical coupling. This was further supported by connexin-43 expression profiles and immunostaining. Collectively, these findings indicate that synchronized Ca²⁺ oscillations in LUHMES cultures are mediated by gap junctional communication rather than by conventional synaptic mechanisms. This system offers a practical platform for studying fundamental principles of network coordination and for evaluating pharmacological or toxicological modulators of intercellular coupling. Moreover, it may provide a relevant human-based model to explore aspects of neuronal maturation and to assess compounds with potential neurodevelopmental toxicity. Full article

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17 pages, 1520 KB

Open AccessReview

O-GlcNAcylation: A Nutrient-Sensitive Metabolic Rheostat in Antiviral Immunity and Viral Pathogenesis

by Thomas I. Odo and Maya Saleh

Cells 2025, 14(21), 1743; https://doi.org/10.3390/cells14211743 - 6 Nov 2025

Viewed by 1086

Abstract

Viruses account for the most abundant biological entities in the biosphere and can be either symbiotic or pathogenic. While pathogenic viruses have developed strategies to evade immunity, the host immune system has evolved overlapping and redundant defenses to sense and fight viral infections. [...] Read more.

Viruses account for the most abundant biological entities in the biosphere and can be either symbiotic or pathogenic. While pathogenic viruses have developed strategies to evade immunity, the host immune system has evolved overlapping and redundant defenses to sense and fight viral infections. Nutrition and metabolic needs sculpt viral–host interactions and determine the course and outcomes of the infection. In this review, we focus on the hexosamine biosynthesis pathway (HBP), a nutrient-sensing pathway that controls immune responses and host–viral interactions. The HBP converges on O-GlcNAcylation, a dynamic post-translational modification of cellular proteins, that emerged as a critical effector of immune cell development, differentiation, and effector functions. We present a broad overview of uncovered O-GlcNAc substrates identified in the context of viral infections and with a functional impact on antiviral immunity and viral restriction, or conversely on exacerbating viral-induced pathologic inflammation or viral oncogenesis. We discuss the clinical implications of these findings, current limitations, and future perspectives to harness this pathway for therapeutic purposes. Full article

(This article belongs to the Special Issue The Cellular and Molecular Mechanisms of Immunity to Infectious Viruses)

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23 pages, 5199 KB

Open AccessArticle

Butyrate Enhances Antimicrobial Defence in Chicken Macrophages Through Reactive Oxygen Species Generation and Autophagy Activation

by James R. G. Adams, Faisal R. Anjum, Jai W. Mehat, Roberto M. La Ragione and Shahriar Behboudi

Cells 2025, 14(21), 1742; https://doi.org/10.3390/cells14211742 - 6 Nov 2025

Viewed by 928

Abstract

Sodium butyrate has been documented to support gut function and help control pathogens in the gastrointestinal tract. However, the precise mechanisms of dietary sodium butyrate’s control over enteric pathogens in chickens remain unclear. Our study demonstrated that priming chicken bone marrow-derived macrophages (BMDMs) [...] Read more.

Sodium butyrate has been documented to support gut function and help control pathogens in the gastrointestinal tract. However, the precise mechanisms of dietary sodium butyrate’s control over enteric pathogens in chickens remain unclear. Our study demonstrated that priming chicken bone marrow-derived macrophages (BMDMs) or the HD11 cell line with 1 mM sodium butyrate significantly enhanced their antimicrobial capacity against key bacterial pathogens (Escherichia coli, Salmonella Typhimurium, Pseudomonas aeruginosa, and Staphylococcus aureus) in gentamicin protection assays (p < 0.05; ≥1 log reduction in CFU/mL). This in vitro enhancement was associated with increased production of reactive oxygen species (ROS), as detected by DCFH-DA assays, showing approximately a 30% increase in HD11 cells and a 12% increase in BMDMs. Butyrate priming was observed to result in autophagy activation, potentially through mTOR pathway inhibition, evidenced by changes in related gene expression using RT-qPCR assay and a 2.5-fold increase in GFP-LC3B accumulation. Supporting this, pharmacological inhibition of ROS using the ROS scavenger N-acetyl-L-cystine (NAC) or autophagy with chloroquine reduced the butyrate-enhanced bacterial clearance. Furthermore, the mTOR inhibitor rapamycin synergized with butyrate priming, whereas the mTOR activator L-leucine counteracted enhanced antimicrobial activity. These findings offer crucial insights for improving host defence against bacterial infections and developing novel therapeutic strategies in chickens. Full article

(This article belongs to the Section Cellular Immunology)

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20 pages, 1589 KB

Open AccessArticle

Polo-like Kinase 1 Activation Regulates Angiotensin II-Induced Contraction in Pudendal and Small Mesenteric Arteries from Mice

by Raiana Anjos Moraes, Olufunke O. Arishe, James Pratt, Stephanie Wilczynski, Rinaldo Rodrigues dos Passos, Diana L. Silva-Velasco, Tiago Tomazini Gonçalves, Tianxin Zhang, Darizy Flavia Silva, R. Clinton Webb and Fernanda Priviero

Cells 2025, 14(21), 1741; https://doi.org/10.3390/cells14211741 - 6 Nov 2025

Cited by 1 | Viewed by 652

Abstract

Polo-like kinase 1 (PLK1) is a serine/threonine protein kinase expressed in smooth muscle cells (SMCs), with emerging roles in regulating contraction. We hypothesize that PLK1 contributes to smooth muscle contractility in pudendal arteries (PA), small mesenteric arteries (SMA), and the corpus cavernosum (CC). [...] Read more.

Polo-like kinase 1 (PLK1) is a serine/threonine protein kinase expressed in smooth muscle cells (SMCs), with emerging roles in regulating contraction. We hypothesize that PLK1 contributes to smooth muscle contractility in pudendal arteries (PA), small mesenteric arteries (SMA), and the corpus cavernosum (CC). Using male C57BL/6J mice, we assessed mRNA and protein expression of PLK1 in these tissues. In addition, the arteries and CC were mounted in myographs for isometric force measurement. We then investigated whether PLK1 regulates SMC contractility induced by phenylephrine (PE), U46619, and angiotensin II (Ang II) in arteries, and by PE, serotonin (5-HT), and electrical field stimulation (EFS; 1–16 Hz) in the CC, both in the presence and absence of the PLK1 inhibitor volasertib. PLK1 expression was confirmed in the SMA, PA, and CC by RT-qPCR or Western blotting. Notably, PLK1 inhibition significantly reduced Ang II-induced contraction in the PA and SMA and attenuated EFS-induced contraction at 2 and 4 Hz in the CC. In contrast, responses to PE, U46619, and 5-HT were unaffected by PLK1 inhibition. These results suggest that PLK1 selectively mediates contraction in response to Ang II and neurogenic stimuli. PLK1 may therefore represent a novel, stimulus-specific regulator of vascular and erectile smooth muscle contractility. Full article

(This article belongs to the Special Issue Molecular Pathways and Potential Therapeutic Targets of Vascular Dysfunction)

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17 pages, 4415 KB

Open AccessArticle

Distinct Tumor-Associated Macrophage Signatures Shape the Immune Microenvironment and Patient Prognosis in Renal Cell Carcinoma

by Youngsoo Han, Aidan Shen, Cheng-chi Chao, Lucas Yeung, Aliesha Garrett, Jianming Zeng, Satoru Kawakita, Jesse Wang, Zhaohui Wang, Alireza Hassani, Xiling Shen and Chongming Jiang

Cells 2025, 14(21), 1740; https://doi.org/10.3390/cells14211740 - 6 Nov 2025

Viewed by 1113

Abstract

Renal cell carcinoma (RCC) accounts for 90% of adult renal cancer cases and is characterized by significant heterogeneity within its tumor microenvironment. This study tests the hypothesis that tumor-associated macrophages (TAMs) influence RCC progression and patient response to treatment by investigating the prognostic [...] Read more.

Renal cell carcinoma (RCC) accounts for 90% of adult renal cancer cases and is characterized by significant heterogeneity within its tumor microenvironment. This study tests the hypothesis that tumor-associated macrophages (TAMs) influence RCC progression and patient response to treatment by investigating the prognostic implications of TAM signatures. Utilizing independent single-cell RNA sequencing data from RCC patients, we developed eight distinct TAM signatures reflective of TAM presence. A LASSO Cox regression model was constructed to predict survival outcomes, evaluated using the TCGA dataset, and validated across independent RCC cohorts. Model performance was assessed through Kaplan–Meier survival plots, receiver operating characteristic (ROC) curves, and principal component analysis. Survival analysis demonstrated that specific TAM signature gene expressions serve as significant prognostic markers, identifying TAM signatures positively correlated with patient survival and macrophage infiltration. A 27-gene TAM risk model was established, successfully stratifying patients into risk categories, with low-risk patients showing improved overall survival. These findings provide insights into the role of TAMs in modulating the RCC tumor immune microenvironment and their impact on patient prognosis, suggesting that TAM-based signatures may serve as useful prognostic markers and potential targets to enhance RCC treatment strategies. Full article

(This article belongs to the Section Cell Microenvironment)

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19 pages, 1144 KB

Open AccessReview

Glymphatic Dysfunction in Neuro-Pulmonary Complications Following Subarachnoid Hemorrhage: A New Perspective on Brain–Lung Axis Disruption

by Eun Chae Lee and Jae Sang Oh

Cells 2025, 14(21), 1739; https://doi.org/10.3390/cells14211739 - 5 Nov 2025

Viewed by 1849

Abstract

Subarachnoid hemorrhage (SAH), often resulting from aneurysmal rupture, remains a life-threatening cerebrovascular disorder with high morbidity and mortality. While previous research has focused primarily on cerebral damage and neurological outcomes, growing evidence suggests that SAH also causes systemic complications, including pulmonary dysfunction. The [...] Read more.

Subarachnoid hemorrhage (SAH), often resulting from aneurysmal rupture, remains a life-threatening cerebrovascular disorder with high morbidity and mortality. While previous research has focused primarily on cerebral damage and neurological outcomes, growing evidence suggests that SAH also causes systemic complications, including pulmonary dysfunction. The underlying mechanisms linking SAH to lung injury, however, are not fully understood. The glymphatic system, a perivascular network that facilitates the clearance of cerebrospinal fluid (CSF) and interstitial waste from the brain, plays a critical role in maintaining central nervous system (CNS) homeostasis. Aquaporin-4 (AQP4) water channels, predominantly expressed in astrocytic end feet, are essential for efficient glymphatic flow. Emerging studies have shown that SAH impairs glymphatic function by disrupting AQP4 polarity and CSF circulation, resulting in the accumulation of neurotoxic substances and neuroinflammation. Recent findings further suggest that glymphatic dysfunction may exert systemic effects beyond the CNS, contributing to a breakdown of the brain–lung axis. The release of pro-inflammatory cytokines, blood degradation products, and damage-associated molecular patterns (DAMPs) into systemic circulation can promote pulmonary endothelial injury and trigger immune responses in the lungs. This phenomenon is exacerbated by impaired clearance via the glymphatic system, amplifying systemic inflammation and increasing the risk of acute lung injury (ALI) or neurogenic pulmonary edema (NPE). This review proposes a novel perspective linking glymphatic impairment with pulmonary complications after SAH. Understanding this connection could open new therapeutic avenues—such as targeting AQP4 function, enhancing CSF circulation, or modulating the inflammatory response—to mitigate both neurological and respiratory sequelae in SAH patients. Full article

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20 pages, 1368 KB

Open AccessReview

Targeting Mitochondrial Dynamics via EV Delivery in Regenerative Cardiology: Mechanistic and Therapeutic Perspectives

by Dhienda C. Shahannaz, Tadahisa Sugiura, Brandon E. Ferrell and Taizo Yoshida

Cells 2025, 14(21), 1738; https://doi.org/10.3390/cells14211738 - 5 Nov 2025

Cited by 2 | Viewed by 1718

Abstract

Mitochondrial dysfunction is a key contributor to cardiac injury and heart failure, and extracellular vesicles (EVs) have emerged as promising therapeutic agents due to their ability to deliver mitochondrial-targeted cargo. This review systematically maps the evidence on how EVs modulate mitochondrial dynamics—including fusion, [...] Read more.

Mitochondrial dysfunction is a key contributor to cardiac injury and heart failure, and extracellular vesicles (EVs) have emerged as promising therapeutic agents due to their ability to deliver mitochondrial-targeted cargo. This review systematically maps the evidence on how EVs modulate mitochondrial dynamics—including fusion, fission, mitophagy, and biogenesis—in regenerative cardiology. We comprehensively searched PubMed, Scopus, and Web of Science up to September 2025 for original studies. A total of 48 studies were included, with most utilizing EVs from mesenchymal stem cells, induced pluripotent stem cells, or cardiac progenitors. The review found that EV cargo influences key pathways such as DRP1 and MFN2, restores mitochondrial membrane potential, reduces ROS accumulation, and improves cardiomyocyte survival. While engineered EVs showed enhanced specificity, a lack of standardized preparation and quantitative assessment methods remains a significant challenge. We conclude that EV-mediated mitochondrial modulation is a promising strategy for cardiac repair, but the field needs harmonized protocols, deeper mechanistic understanding, and improved translational readiness to advance beyond preclinical research. The future of this research lies in integrating systems biology and precision targeting. Full article

(This article belongs to the Special Issue Advances in Cardiomyocyte and Stem Cell Biology in Heart Disease)

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16 pages, 1750 KB

Open AccessArticle

Adaptive Gene Expression Induced by a Combination of IL-1β and LPS in Primary Cultures of Mouse Astrocytes

by Thierry Coppola, Gwénola Poupon, Hélène Rangone, Stéphane Martin and Patricia Lebrun

Cells 2025, 14(21), 1737; https://doi.org/10.3390/cells14211737 - 5 Nov 2025

Viewed by 965

Abstract

Astrocytes are vital cells within the central nervous system (CNS), as they perform a critical role in supporting neurons by providing nutrients, such as lactate for energy, and safeguarding them against the toxicity of excessive neurotransmitters, such as glutamate. This study investigates astrocyte [...] Read more.

Astrocytes are vital cells within the central nervous system (CNS), as they perform a critical role in supporting neurons by providing nutrients, such as lactate for energy, and safeguarding them against the toxicity of excessive neurotransmitters, such as glutamate. This study investigates astrocyte adaptive mechanisms in response to chronic inflammation. The primary aim is to assess the long-term effects of an inflammation-induced environment using a combination of lipopolysaccharide (LPS) and interleukin-1β (IL-1β), on the expression of key genes involved in essential metabolic pathways for astrocyte function, including glutamate metabolism and clearance, lactate synthesis and transport, and glucose metabolism. We observed an upregulation of the glutamate transporter eaat2 (but not eaat1), leading to glutamate accumulation and altered glutamate-glutamine cycling, as well as increased glycolytic activity and lactate production/export via hexokinases (hk1 and hk2) and the mct4 lactate transporter. Interestingly, these mechanisms are reversible, indicating a precisely controlled adaptive system. This investigation facilitated the identification of the signaling pathways involved in astrocyte adaptive responses to stress. This will further guide our investigations towards the more complex domain of resistance and adaptation of CNS in pathophysiological conditions. Full article

(This article belongs to the Section Cellular Neuroscience)

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23 pages, 3576 KB

Open AccessArticle

The BUD31 Homologous Gene in Schizosaccharomyces pombe Is Evolutionarily Conserved and Can Be Linked to Cellular Processes Regulated by the TOR Pathway

by Ildikó Vig, Lajos Acs-Szabo, Zsigmond Benkő, Silvia Bagelova Polakova, László Attila Papp, Juraj Gregan and Ida Miklós

Cells 2025, 14(21), 1736; https://doi.org/10.3390/cells14211736 - 5 Nov 2025

Viewed by 737

Abstract

The human BUD31 gene has been associated with various processes including cancer. To better understand its function, we used genetic methods to study Schizosaccharomyces pombe cells lacking the BUD31 homologous gene (cwf14) and performed sequence analysis using bioinformatics methods. Mutant cells [...] Read more.

The human BUD31 gene has been associated with various processes including cancer. To better understand its function, we used genetic methods to study Schizosaccharomyces pombe cells lacking the BUD31 homologous gene (cwf14) and performed sequence analysis using bioinformatics methods. Mutant cells lacking the cwf14 gene showed cell size and division defects, altered stress response, rapamycin sensitivity, enhanced chronological aging, and increased sporulation tendency. These processes are known to be regulated by the TOR pathway. The cwf14-TOR link was also supported by further experiments. We demonstrated that most protein-coding genes affected by cwf14 deletion are upregulated, encode hydrolases, oxidoreductases, and are often involved in transport. GO enrichment drew our attention to genes related to nitrogen transport, while additional data pointed to a nutrient/nitrogen (N) sensing problem. Although Cwf14 protein is associated with spliceosome complex, most genes affected by the absence of cwf14 do not contain introns, suggesting that they are influenced indirectly by the cwf14 gene. In silico experiments have revealed that BUD31 orthologous genes are found from yeast to humans, are evolutionarily conserved with a high degree of sequence identity, conserved motifs, and structures. Since the human gene partially complemented the mutant phenotype of S. pombe cells, indicating functional homology, our data can help better understand pathological mechanisms observed in human cancer cells. Full article

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17 pages, 1413 KB

Open AccessArticle

Secondhand Smoke Exposure Timing Triggers Distinct Placental Responses in Mouse Pregnancy

by Archarlie Chou, Ethan Frank, Matt Reall, Olivia Hiatt, Logan Beck, Paul R. Reynolds, Brett E. Pickett and Juan A. Arroyo

Cells 2025, 14(21), 1735; https://doi.org/10.3390/cells14211735 - 5 Nov 2025

Viewed by 724

Abstract

Secondhand smoke (SHS), found in about 57.6% of global public areas as a widespread environmental hazard, has been associated with negative effects during pregnancy, such as preeclampsia (PE) and intrauterine growth restriction (IUGR). Our research investigated the impact of SHS on placental issues [...] Read more.

Secondhand smoke (SHS), found in about 57.6% of global public areas as a widespread environmental hazard, has been associated with negative effects during pregnancy, such as preeclampsia (PE) and intrauterine growth restriction (IUGR). Our research investigated the impact of SHS on placental issues in a C57BL/6 model that simulates PE and IUGR in mice. We administered SHS to pregnant mice through a nose-only delivery method, beginning either on embryonic day 12.5 (prior to spiral artery (SA) invasion; labeled SHS-6D) or day 14.5 (following SA invasion; labeled SHS-4D), continuing up to E18.5. Control animals received only ambient air. We employed bulk RNA sequencing to assess and describe changes in placental gene expression patterns. For the SHS-4D group, which mimicked IUGR, compared to untreated controls, results showed elevated levels of inflammation-related genes (IL11RA, CHI3L1) alongside likely interference in pathways for antibody-triggered complement activation, marked by reduced expression of C1QA, C1QB, and C1QC. Immune profiling also indicated decreased macrophage activity in the placentas of the SHS-4D group relative to those from normal pregnancies at term. In contrast, the SHS-6D versus control analysis revealed lowered expression of collagen-related genes (COL1A1, COL4A5, COL4A6, COL17A1). Additionally, SHS-6D exhibited higher levels of genes associated with cell-based lysis processes compared to SHS-4D. An evaluation of the existing literature revealed that nearly every differentially expressed gene (DEG) identified in our work has been reported in studies associated with SHS exposure. Yet, few of these DEGs are discussed alongside PE or IUGR in prior reports, highlighting gaps in knowledge about how SHS triggers these conditions. Overall, we determined that the timing of SHS exposure in pregnant mice results in unique patterns of gene regulation and involvement in biological pathways. Full article

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40 pages, 1578 KB

Open AccessReview

Does Amyotrophic Lateral Sclerosis (ALS) Have Metabolic Causes from Human Evolution?

by Michael Spedding

Cells 2025, 14(21), 1734; https://doi.org/10.3390/cells14211734 - 5 Nov 2025

Viewed by 2237

Abstract

As so many drugs have failed in ALS a new approach is needed. The author proposes that recent human genetic variants may play major roles in the disease, changing metabolism. Evolution of hominins was accelerated 3–2.5 Mya, by cytidine monophospho-N-acetylneuraminic acid hydroxylase (CMAH) [...] Read more.

As so many drugs have failed in ALS a new approach is needed. The author proposes that recent human genetic variants may play major roles in the disease, changing metabolism. Evolution of hominins was accelerated 3–2.5 Mya, by cytidine monophospho-N-acetylneuraminic acid hydroxylase (CMAH) becoming a unitary pseudogene after a pathogenic infection, changing the sialome, and hence metabolism, brain development and neuromuscular junctions (NMJs). This was when hominins evolved to run in Africa and develop bigger brains. Deletion of CMAH in mice allows them to run for longer (~50%). The enzyme CMAH is critical for the sialome, particularly the neurotrophin GM1, a critical hub for viral infection and for NMJ stability, but which is lost from NMJs at the beginning of denervation, probably due a 10-fold increase in spinal cord glucosylceramidases (non-lysosomal GBA2). A GBA2 inhibitor, ambroxol, is currently in phase II for ALS. Human-specific GM1 may be critical for human evolution, lactate metabolism and ALS. Lipid/lactate metabolism changed to support these evolutionary changes and lactate is a major body/brain fuel, but compromised in ALS patients and a marker of disease progression. Recent progress in sports science involving lactate metabolism and human performance may also be relevant to ALS therapies, and incidence. Full article

(This article belongs to the Special Issue Pathology and Treatments of Amyotrophic Lateral Sclerosis (ALS))

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16 pages, 467 KB

Open AccessReview

Obesity-Related Inflammatory Biomarkers in the Elderly Population

by Georgia Vamvakou, Nikolaos Theodorakis, Dimitris Anagnostou, Magdalini Kreouzi, Loukianos S. Rallidis, Vasiliki Katsi, Effie Simou, Stefanos Archontakis, George Skalis, Christos Hitas, Konstantinos Toutouzas and Maria Nikolaou

Cells 2025, 14(21), 1733; https://doi.org/10.3390/cells14211733 - 4 Nov 2025

Viewed by 1148

Abstract

Obesity in elderly individuals is associated with increased levels of inflammatory biomarkers, indicating a state of chronic low-grade inflammation, which has been recently termed as adipaging. Several studies have demonstrated this relationship: overweight and obese middle-aged and elderly individuals show elevated levels of [...] Read more.

Obesity in elderly individuals is associated with increased levels of inflammatory biomarkers, indicating a state of chronic low-grade inflammation, which has been recently termed as adipaging. Several studies have demonstrated this relationship: overweight and obese middle-aged and elderly individuals show elevated levels of inflammatory markers like CXCL-16, IL-6, and adipokines compared to normal weight counterparts. These markers positively correlate with anthropometric parameters indicating increased cardiovascular risk. C-reactive protein (CRP) and fibrinogen levels increase progressively with higher obesity classes in the general population, including the elderly. For instance, CRP levels nearly double with each increase in weight class compared to normal weight individuals. Additionally, the presence of obesity-related comorbidities like hypertension or diabetes further elevates these inflammatory markers. In conclusion, obesity in the elderly is characterized by elevated levels of various inflammatory biomarkers, reflecting a state of chronic low-grade inflammation. This inflammatory state may contribute to the development of obesity-related comorbidities. The clarification of the complementary or independent role of these biomarkers in aging and obesity could lead to targeted therapeutic interventions in this vulnerable population group. Full article

(This article belongs to the Special Issue Integrated Approaches Between Metabolomics and Cellular Aging)

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25 pages, 2860 KB

Open AccessArticle

Alterations in Circulating T-Cell Subsets with Gut-Homing/Residency Phenotypes Associated with HIV-1 Status and Subclinical Atherosclerosis

by Etiene Moreira Gabriel, Jonathan Dias, Abdelali Filali-Mouhim, Ramon Edwin Caballero, Tomas Raul Wiche Salinas, Manon Nayrac, Carl Chartrand-Lefebvre, Jean-Pierre Routy, Madeleine Durand, Mohamed El-Far, Cécile Tremblay and Petronela Ancuta

Cells 2025, 14(21), 1732; https://doi.org/10.3390/cells14211732 - 4 Nov 2025

Viewed by 717

Abstract

Antiretroviral therapy (ART) controls HIV-1 replication in people with HIV-1 (PWH), but intestinal integrity impairment persists and fuels microbial translocation and chronic immune activation, thus heightening the cardiovascular disease (CVD) risk. Here, we sought to identify novel immunological correlates of the HIV and [...] Read more.

Antiretroviral therapy (ART) controls HIV-1 replication in people with HIV-1 (PWH), but intestinal integrity impairment persists and fuels microbial translocation and chronic immune activation, thus heightening the cardiovascular disease (CVD) risk. Here, we sought to identify novel immunological correlates of the HIV and CVD status in ART-treated PWH (HIV⁺; n = 42) and uninfected participants (HIV⁻; n = 40) of the Canadian HIV and Aging Cohort Study (CHACS), with/without subclinical coronary atherosclerotic plaques, measured by Coronary Computed Tomography Angiography as total plaque volume (TPV, mm³). PBMCs were analyzed by flow cytometry for the expression of T-cell lineage (CD45, CD3, CD4, CD8αα, CD8αβ, TCRαβ, TCRγδ), epithelial cell (EpCAM/CD326), activation (HLA-DR), and gut-homing/residency markers (CD69, CD196/CCR6, CD199/CCR9, CD49d/Itgα4, CD103/ItgαE, Itgβ7). Alterations in the CD3⁺ T-cell pool, such as increased frequencies of CD8⁺TCRαβ⁺ and TCRγδ⁺ cells, to the detriment of CD4⁺TCRαβ⁺ subsets, were associated with the HIV status. Also, CD4⁺ T-cells with CD326⁺CD69⁺CCR6⁺ItgαE⁺ and CCR6⁺Itgβ7⁻ phenotypes were increased in frequency in HIV⁺ vs. HIV⁻ participants, together with a decreased frequency of CD8⁺ T-cells with an intraepithelial lymphocyte (IEL)-like CD3⁺CD4⁻TCRαβ⁺TCRγδ⁻CD8αα⁺CD8αβ⁻ phenotype. Finally, multivariate logistic regression identified the frequency of ItgαE⁺CD8⁺, ItgαE⁻CD8⁺, CCR6⁺CD4⁺, and CCR6⁺Itgβ7⁻CD4⁺ T-cells as strong positive correlates of HIV status and atherosclerotic plaque in ART-treated PWH. Full article

(This article belongs to the Special Issue Human Immunodeficiency Virus/Simian Immunodeficiency Virus and Host Interactions)

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19 pages, 770 KB

Open AccessReview

Biological Mechanisms Involved in Muscle Dysfunction in COPD: An Integrative Damage–Regeneration–Remodeling Framework

by Joaquim Gea, Mauricio Orozco-Levi, Sergi Pascual-Guàrdia, Carme Casadevall, César Jessé Enríquez-Rodríguez, Ramon Camps-Ubach and Esther Barreiro

Cells 2025, 14(21), 1731; https://doi.org/10.3390/cells14211731 - 4 Nov 2025

Cited by 1 | Viewed by 1865

Abstract

Skeletal muscle dysfunction is a major systemic manifestation of COPD that shapes symptoms, exercise tolerance and mortality. Current evidence can be integrated within a Damage–Regeneration–Remodeling framework linking mechanics and biology to clinical phenotypes. Pulmonary hyperinflation and chest wall geometry chronically load the diaphragm [...] Read more.

Skeletal muscle dysfunction is a major systemic manifestation of COPD that shapes symptoms, exercise tolerance and mortality. Current evidence can be integrated within a Damage–Regeneration–Remodeling framework linking mechanics and biology to clinical phenotypes. Pulmonary hyperinflation and chest wall geometry chronically load the diaphragm and other respiratory muscles in COPD, whereas inactivity and exacerbation-related disuse underload locomotor muscles. Across muscle compartments, oxidative/nitrosative stress, activation of proteolytic pathways, mitochondrial and endoplasmic reticulum stress, microvascular limitations, neuromuscular junction instability, and myosteatosis degrade muscle quality. The diaphragm adapts with a fast-to-slow fiber shift, greater oxidative capacity, and sarcomere foreshortening, improving endurance, whereas limb muscles show atrophy, a glycolytic shift, reduced oxidative enzymes, extracellular matrix accrual, and fat infiltration. Translational levers that address these mechanisms include: (I) Reduce damage: bronchodilation, lung-volume reduction, oxygen, non-invasive ventilation, early mobilization, pulmonary rehabilitation, neuromuscular stimulation, and corticosteroid stewardship; (II) Enable regeneration: progressive resistance plus high-intensity/heavy-load endurance training; adequate protein and vitamin-D intake, and endocrine correction; and (III) Steer remodeling: increase physical activity (with/without coaching/telecoaching), functional assessment and CT or MRI monitoring, inspiratory-muscle training, and phenotype-guided adjuncts in selected cases. This framework clarifies why lung deflation strategies benefit inspiratory mechanics, whereas limb recovery requires behavioral and metabolic interventions layered onto systemic optimization. Full article

(This article belongs to the Special Issue Plasticity of Skeletal, Cardiac and Smooth Muscles in COPD: Repair and Remodeling)

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31 pages, 1151 KB

Open AccessReview

Aging and Corneal Nerve Health: Mechanisms of Degeneration and Emerging Therapies for the Cornea

by Hanieh Niktinat, Melinda Alviar, Marziyeh Kashani, Hamed Massoumi, Ali R. Djalilian and Elmira Jalilian

Cells 2025, 14(21), 1730; https://doi.org/10.3390/cells14211730 - 4 Nov 2025

Viewed by 2099

Abstract

Corneal nerves play a crucial role in maintaining ocular surface homeostasis by supporting the functional integrity of corneal epithelial, stromal, and endothelial cells; modulating tear secretion; and facilitating sensory responses essential for overall ocular health. With advancing age, these highly specialized peripheral sensory [...] Read more.

Corneal nerves play a crucial role in maintaining ocular surface homeostasis by supporting the functional integrity of corneal epithelial, stromal, and endothelial cells; modulating tear secretion; and facilitating sensory responses essential for overall ocular health. With advancing age, these highly specialized peripheral sensory fibers undergo progressive attrition and morphologic distortion driven by the canonical hallmarks of aging including genomic instability, impaired proteostasis, mitochondrial dysfunction, and chronic low-grade inflammation. The resulting neuro-immune dysregulation reduces trophic support, delays wound healing, and predisposes older adults to dry-eye disease, neurotrophic keratopathy, and postsurgical hypoesthesia. Age-exacerbating cofactors including diabetes, dyslipidemia, neurodegenerative disorders, topical preservatives, chronic contact-lens wear, herpes zoster ophthalmicus, and ocular-surface hypoxia further accelerate sub-basal nerve rarefaction and functional decline. This review provides an overview of age-related physiological alterations in ocular surface nerves, with a particular emphasis on corneal innervation. It also discusses risk factors that speed up these changes. Given the inherently limited regenerative capacity of corneal nerves and their inability to fully restore to baseline conditions following injury or degeneration, it is critical to identify and develop effective strategies aimed at mitigating or delaying physiological nerve degeneration and promoting nerve regeneration. This review also brings up emerging therapeutic strategies, including regenerative medicine, neuroprotective agents, and lifestyle interventions aimed at mitigating age-related corneal nerve degeneration. Full article

(This article belongs to the Special Issue Understanding Aging Mechanisms to Prevent Age-Related Diseases: 2nd Edition)

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22 pages, 2496 KB

Open AccessArticle

Advanced Glycation End Products Mediate Epigenetic Alteration of H3K27me3 in Renal Proximal Tubular Cells: Potential Role in Metabolic Memory

by Lore Ludewig, Tzvetanka Bondeva, Marita Liebisch, Jonas Ihle, Ivonne Loeffler and Gunter Wolf

Cells 2025, 14(21), 1729; https://doi.org/10.3390/cells14211729 - 4 Nov 2025

Viewed by 854

Abstract

The accumulation of advanced glycation end products (AGEs) is a hallmark of prolonged high glucose levels in diabetes mellitus. We have previously reported that hypoxia and AGEs cause epigenetic modification of the repressive mark H3K27me3 in podocytes by downregulation of enhancer of zeste [...] Read more.

The accumulation of advanced glycation end products (AGEs) is a hallmark of prolonged high glucose levels in diabetes mellitus. We have previously reported that hypoxia and AGEs cause epigenetic modification of the repressive mark H3K27me3 in podocytes by downregulation of enhancer of zeste homolog 2 (EZH2) and nuclear inhibitor of protein phosphatase 1 (NIPP1). However, their impact on proximal tubular cells remains unclear. The aim of this study was to investigate the role of AGEs and diabetes on the epigenetic modifications of EZH2 and H3K27me3 in proximal tubular cells and in diabetic (db/db) mice. Our results show that AGEs reduced EZH2 expression in TKPTS cells, thereby decreasing the tri-methylation of H3K27. qRT-PCR analysis revealed upregulation of genes known to contribute to diabetic nephropathy and kidney injury as Ctgf, Snai1, and p27^Kip1. Consistently, immunofluorescent staining of renal sections from db/db mice confirmed the reduction in H3K27me3 levels in proximal tubules compared to non-diabetic controls. In summary, we show that AGEs induce epigenetic changes in proximal tubular cells by suppressing EZH2, thereby facilitating the transcription of genes involved in progression of diabetic nephropathy. These results provide new insights into metabolic memory, a process in which prior poor glucose control triggers ongoing renal damage despite current normoglycemia. Full article

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20 pages, 2139 KB

Open AccessArticle

Inhalation of Ultrafine Carbon Black-Induced Mitochondrial Dysfunction in Mouse Heart Through Changes in Acetylation

by Rahatul Islam, Jackson E. Stewart, William E. Mullen, Dena Lin, Salik Hussain and Dharendra Thapa

Cells 2025, 14(21), 1728; https://doi.org/10.3390/cells14211728 - 4 Nov 2025

Viewed by 764

Abstract

Air pollution, particularly from fine and ultrafine particulate matter (PM), has been increasingly associated with cardiovascular diseases. Ultrafine carbon, a component of ultrafine PM widely used in industrial settings, is both an environmental and occupational hazard. But the cardiac toxicity of repeated inhalation [...] Read more.

Air pollution, particularly from fine and ultrafine particulate matter (PM), has been increasingly associated with cardiovascular diseases. Ultrafine carbon, a component of ultrafine PM widely used in industrial settings, is both an environmental and occupational hazard. But the cardiac toxicity of repeated inhalation exposure to ultrafine carbon black (CB) remains unclear. In this study, we investigated how repeated inhalation of CB affects cardiac mitochondrial function, focusing on metabolic pathways and regulatory mechanisms involved in energy production. Male C57BL/6J mice were exposed to either filtered air or CB aerosols (10 mg/m³) for four consecutive days. Cardiac tissues were collected and analyzed to assess changes in metabolic enzyme activity, protein expression, and mitochondrial function using Western blotting, enzymatic assays, and immunoprecipitation. Despite there being few changes in overall protein expression levels, we observed significant impairments in fatty acid oxidation, increased glucose oxidation, and disrupted electron transport chain (ETC) supercomplex assembly, particularly in Complexes III and IV. These changes were accompanied by increased hyperacetylation of mitochondrial proteins and elevated levels of GCN5L1, a mitochondrial acetyltransferase. We also found increased lipid peroxidation and hyperacetylation of antioxidant enzyme SOD2 at the K-122 site, which reflects reduced enzymatic activity contributing to oxidative stress. Our findings suggest that repeated CB inhalation leads to mitochondrial dysfunction in the heart by dysregulating substrate utilization, impairing ETC activities, and weakening antioxidant defenses primarily through lysine acetylation. These findings reveal a potential role of key post-translational mechanisms in environmental particulate exposure to mitochondrial impairment and provide a potential therapeutic target for CB-induced cardiotoxicity. Full article

(This article belongs to the Special Issue Cellular Mechanisms in Mitochondrial Function and Calcium Signaling)

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31 pages, 1693 KB

Open AccessReview

Therapeutic and Clinical Potential of Adipose-Derived Stem Cell Secretome for Skin Regeneration

by Anna Sendera, Hubert Kubis, Anna Pałka and Agnieszka Banaś-Ząbczyk

Cells 2025, 14(21), 1727; https://doi.org/10.3390/cells14211727 - 4 Nov 2025

Viewed by 4456

Abstract

Adipose-derived stem cells (AT-MSCs) exhibit great potential for application in stem cell therapy, primarily due to their unique pro-regenerative capabilities, which include supporting skin regeneration. AT-MSCs secrete a variety of biomolecules with immunomodulatory, re-epithelializing, antifibrotic, antiapoptotic, proangiogenic, and neurotrophic activity as well as [...] Read more.

Adipose-derived stem cells (AT-MSCs) exhibit great potential for application in stem cell therapy, primarily due to their unique pro-regenerative capabilities, which include supporting skin regeneration. AT-MSCs secrete a variety of biomolecules with immunomodulatory, re-epithelializing, antifibrotic, antiapoptotic, proangiogenic, and neurotrophic activity as well as the ability to promote proliferation and migration of skin cells. Recently, therapy using AT-MSC secretome alone has garnered increasing attention due to its potentially safer and more effective application ability than the use of whole AT-MSCs. This review provides a comprehensive summary of the current state of knowledge regarding the potential use of AT-MSC secretome as a promising cell-free therapeutic strategy for wound healing, alopecia, skin rejuvenation, and skin inflammatory diseases. We critically analyze and discuss findings from in vitro, in vivo, and clinical studies. Moreover, we briefly discuss possible approaches to enhance the secretion of AT-MSC biomolecules, such as AT-MSC preconditioning with low-frequency electromagnetic fields and hypoxia. In conclusion, the evidence presented in this review paper underscores that the AT-MSC secretome alone can be a highly effective approach as a stem cell-free therapy for skin repair, with significant translational potential. Full article

(This article belongs to the Special Issue Adipose-Derived Stem Cells for Tissue Regeneration)

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22 pages, 3025 KB

Open AccessArticle

Osmotic Fragility in Leukodepleted Stored Red Blood Cells: Implications for Neurocritical Care Transfusion Strategies

by Marta Peris, Maria A. Poca, Ana Ortuño, Verónica Pons, Nuria Rodríguez-Borrero, Desiree Jurado, Rafael Parra-López, Marina Rierola and Juan Sahuquillo

Cells 2025, 14(21), 1726; https://doi.org/10.3390/cells14211726 - 3 Nov 2025

Viewed by 1335

Abstract

Background: Anemia is frequent in critically ill patients with traumatic brain injury (TBI) and worsens neurological outcomes. Red blood cell (RBC) transfusion is a cornerstone of management, but storage-related biochemical and structural changes may impair oxygen delivery. This study examined the effect [...] Read more.

Background: Anemia is frequent in critically ill patients with traumatic brain injury (TBI) and worsens neurological outcomes. Red blood cell (RBC) transfusion is a cornerstone of management, but storage-related biochemical and structural changes may impair oxygen delivery. This study examined the effect of storage duration on osmotic fragility (OF) and free hemoglobin (fHb) in leukodepleted packed RBCs (pRBCs) as indicators of membrane stability and hemolysis. Methods: Twenty-four leukodepleted pRBC units in SAGM (saline, adenine, glucose, and mannitol) solution were analyzed from Day 3 to Day 42. OF was assessed by Beutler’s method with H50 values derived from logistic models, and fHb was quantified spectrophotometrically. Flow cytometry with phosphate-buffered saline (PBS)-induced osmotic stress provided complementary OF data. Results: OF increased significantly beyond 28 days, with Week 6 H₅₀ values exceeding those at Weeks 2 and 4 (p < 0.0001). fHb rose progressively with storage: 7.3 ± 4.3 µmol/L (Week 2), 14.6 ± 7.9 (Week 4), and 25.7 ± 12.1 (Week 6) (p < 0.0001). Hemolysis remained below the 0.8% threshold but increased from 0.09% to 0.29% (p < 0.0001). Conclusions: pRBC storage beyond 28 days leads to greater OF and fHb release, reflecting reduced membrane stability. These changes may compromise transfusion efficacy and oxygen delivery in neurocritical care. Full article

(This article belongs to the Special Issue Highlights in Red Blood Cell Research)

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18 pages, 12830 KB

Open AccessArticle

Desloratadine Induces TP53-Dependent Apoptosis in MCF-7 Breast Cancer Cells

by Syed Rashel Kabir, Taufique Abdullah, Gausul Azam, Tamzid Hossain Molla, Hasan Ali, Mojnu Miah, Mohammad Taufiq Alam and Sayem Miah

Cells 2025, 14(21), 1725; https://doi.org/10.3390/cells14211725 - 3 Nov 2025

Viewed by 1188

Abstract

Breast cancer remains a leading cause of mortality among women despite advances in early detection and targeted therapies, underscoring the need for safer and more effective treatment options. Drug repurposing offers a promising strategy by leveraging existing pharmacological agents with established safety profiles. [...] Read more.

Breast cancer remains a leading cause of mortality among women despite advances in early detection and targeted therapies, underscoring the need for safer and more effective treatment options. Drug repurposing offers a promising strategy by leveraging existing pharmacological agents with established safety profiles. Desloratadine, a second-generation H1-histamine receptor antagonist widely prescribed for allergic conditions, has attracted interest in oncology because histamine signaling influences proliferation, angiogenesis, and immune responses, yet its anticancer potential remains poorly understood. In this study, we investigated its effects in MCF-7 breast cancer cells, which harbor wild-type TP53. Desloratadine inhibited cell viability in a dose-dependent manner, with an IC₅₀ of 14.2 µg/mL. Mechanistic analyses revealed that growth inhibition was primarily mediated through apoptosis, confirmed by Hoechst 33342 staining, ROS generation, annexin V/PI staining, and caspase-dependent pathways. Gene expression profiling demonstrated upregulation of TP53, FAS, and BAX, alongside reduced PARP-1 and NF-κB expression, with no detectable STAT3 or BCL2 expression. Flow cytometry indicated accumulation of cells in the sub-G₁ phase and G₂/M arrest, consistent with apoptosis induction. Molecular docking further supported these findings, showing that Desloratadine binds with high affinity to p53 (−7.0 kcal/mol), FAS (−6.8 kcal/mol), and NF-κB (−6.5 kcal/mol), forming stabilizing hydrogen bonds and hydrophobic interactions aligned with the observed gene expression changes. To confirm the functional role of TP53, we generated CRISPR-Cas9 knockout MCF-7 cells. Compared with wild-type cells, these knockout cells displayed markedly reduced sensitivity to Desloratadine, with the IC₅₀ shifting from 14.2 µg/mL to 36.4 µg/mL, demonstrating that p53 is a key mediator of the drug’s cytotoxic effect. Collectively, these findings identify Desloratadine as a potential repurposed drug candidate for breast cancer therapy, acting at least in part through a p53-dependent apoptotic pathway. Full article

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27 pages, 1583 KB

Open AccessReview

Dysferlin and the Regulation of Ca²⁺ Release in Skeletal Muscle

by Robert J. Bloch, Joaquin Muriel and Valeriy Lukyanenko

Cells 2025, 14(21), 1724; https://doi.org/10.3390/cells14211724 - 3 Nov 2025

Viewed by 1363

Abstract

Dysferlin is a large transmembrane protein that is mutated or absent in Limb Girdle Muscular Dystrophy Type R2 (LGMD R2). Although it may have several functions in healthy skeletal muscle, most research on dysferlin has addressed its roles in repair of the sarcolemma [...] Read more.

Dysferlin is a large transmembrane protein that is mutated or absent in Limb Girdle Muscular Dystrophy Type R2 (LGMD R2). Although it may have several functions in healthy skeletal muscle, most research on dysferlin has addressed its roles in repair of the sarcolemma and in maintaining proper control of Ca²⁺ homeostasis at the triad junction, where it concentrates. Here, we review the literature on the role of dysferlin in both membrane repair and in Ca²⁺ homeostasis, with a focus on the latter. We propose that pathophysiology in LGMD R2 is in part the result of increased leak of Ca²⁺ at the triad junction, which in turn reduces the amplitude of Ca²⁺ transients and, by activating Ca²⁺-induced Ca²⁺ release, or CICR, at the triad junction, induces Ca²⁺ waves. We discuss the mechanisms that regulate Ca²⁺ leak and Ca²⁺ levels at the triad junction under physiological and pathophysiological conditions. Our results suggest that suppression of abnormal leak and CICR may be therapeutic for LGMD R2 and other diseases of muscle linked to dysregulation of Ca²⁺ homeostasis. Full article

(This article belongs to the Special Issue Advances in Muscle Research in Health and Disease—2nd Edition)

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23 pages, 3285 KB

Open AccessArticle

Kv7 Channels as an Important Contributor to Alcohol-Induced Modulation of Neuronal Excitability in Neonatal Rat Superior Cervical Ganglion

by Da-Jeong Jeong, Jin-Nyeong Woo, Tery Yun, Myungin Baek and Byung-Chang Suh

Cells 2025, 14(21), 1723; https://doi.org/10.3390/cells14211723 - 1 Nov 2025

Viewed by 699

Abstract

Normal alcohols (n-alcohols) exhibit contrasting effects on neuronal excitability; specifically, ethanol enhances neuronal firing, while hexanol suppresses it. Both compounds are known to inhibit sodium currents, yet the mechanisms behind their differing effects remain unclear. Our previous studies demonstrated that Kv7 [...] Read more.

Normal alcohols (n-alcohols) exhibit contrasting effects on neuronal excitability; specifically, ethanol enhances neuronal firing, while hexanol suppresses it. Both compounds are known to inhibit sodium currents, yet the mechanisms behind their differing effects remain unclear. Our previous studies demonstrated that Kv7 channels are modulated differently by alcohol chain length, prompting investigation into their role in these contrasting effects. We conducted whole-cell patch clamp recordings on neonatal (P5-P7) rat superior cervical ganglion neurons to assess alcohol impacts on action potential firing and ionic currents, utilizing tetrodotoxin (TTX), XE991, and retigabine (RTG). Ethanol (100 mM) increased action potential frequency, whereas hexanol (3 mM) decreased it, despite both inhibiting sodium currents by 12% and 45%, respectively. Notably, ethanol inhibited Kv7 currents by 16%, while hexanol enhanced them by 29%. TTX alone did not affect firing frequency until sodium current inhibition exceeded 76%, indicating moderate sodium channel blockade cannot fully explain the effects of alcohol. XE991 increased firing frequency and depolarized the resting membrane potential, while retigabine produced opposite effects. The combination of TTX with Kv7 modulators replicated the effects observed with each alcohol. These findings suggest Kv7 channel modulation plays an important role in the chain length-dependent effects of alcohol on neuronal excitability. Full article

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12 pages, 2080 KB

Open AccessArticle

The Molecular Mechanism of PDE1 Regulation

by Jacob Nielsen, Morten Langgård, Josefine Fussing Tengberg and Jan Kehler

Cells 2025, 14(21), 1722; https://doi.org/10.3390/cells14211722 - 1 Nov 2025

Viewed by 703

Abstract

The phosphodiesterase 1 genes PDE1A, PDE1B, and PDE1C encode calcium-regulated cyclic nucleotide phosphodiesterases that mediate the interplay between calcium and cyclic nucleotide signaling in the brain, heart, and vasculature. While an inhibitory domain and a calmodulin-binding domain have been identified in PDE1, the [...] Read more.

The phosphodiesterase 1 genes PDE1A, PDE1B, and PDE1C encode calcium-regulated cyclic nucleotide phosphodiesterases that mediate the interplay between calcium and cyclic nucleotide signaling in the brain, heart, and vasculature. While an inhibitory domain and a calmodulin-binding domain have been identified in PDE1, the mechanism of regulation is not understood. In this study, we investigated the regulatory mechanism through a series of experiments. The experimental data, supported by AlphaFold structure predictions, consistently point to the following model of PDE1 regulation: In the absence of calcium, the inhibitory domain of PDE1 binds to and blocks the catalytic site via molecular interactions that closely resemble those observed in autoinhibited PDE4. Upon calcium/calmodulin binding to PDE1’s calmodulin-binding domain, steric constraints prevent the inhibitory domain from reaching the catalytic site, thereby activating PDE1. Understanding this mode of PDE1 regulation may open new avenues for pharmacological intervention. Moreover, it establishes PDE1 and PDE4 as a second mechanistic class of phosphodiesterase regulation in addition to the GAF-domain-mediated regulation known to control the activity of several other PDEs. Full article

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29 pages, 4096 KB

Open AccessArticle

Acute Myeloid Leukemia: A Key Role of DGKα and DGKζ in Cell Viability

by Elisa Gorla, Marco Cristiano Cartella, Edoardo Borghetti, Ginevra Lovati, Luisa Racca, Teresa Gravina, Giorgio Biazzo, Gabriele Bonello, Valeria Malacarne, Veronica De Giorgis, Davide Corà, Marcello Manfredi, Alberto Massarotti, Andrea Graziani and Gianluca Baldanzi

Cells 2025, 14(21), 1721; https://doi.org/10.3390/cells14211721 - 1 Nov 2025

Viewed by 1063

Abstract

Acute myeloid leukemia (AML) is a heterogeneous disease with an unmet need for novel therapeutic drugs. Previous studies have reported the upregulation of diacylglycerol kinases (DGKs) in AML. This study investigated the effects of ritanserin, a DGKα-specific inhibitor, and DGKζ-IN4 or BAY 2965501, [...] Read more.

Acute myeloid leukemia (AML) is a heterogeneous disease with an unmet need for novel therapeutic drugs. Previous studies have reported the upregulation of diacylglycerol kinases (DGKs) in AML. This study investigated the effects of ritanserin, a DGKα-specific inhibitor, and DGKζ-IN4 or BAY 2965501, DGKζ-selective inhibitors, on a panel of AML cell lines. Ritanserin induced apoptotic cell death across all tested models, whereas DGKζ inhibitors triggered both apoptosis and necrosis to variable extents, with HL-60 cells being the most responsive to both compounds. Drug sensitivity did not correlate with DGKα or DGKζ expression levels, indicating that additional factors may influence cellular susceptibility. THP-1 proteomic profiling revealed that ritanserin broadly downregulated proteins involved in antigen presentation, cell cycle and metabolism, while BAY 2965501 affected a smaller and distinct but functionally similar protein subset, implying different mechanisms of action. Gene silencing confirmed AML cell line-specific dependence on DGK isoforms: HEL cells were sensitive to DGKα knockdown, HL-60 to DGKζ silencing, whereas K562 and THP-1 were resistant to both. These findings indicate that DGKs targeting can effectively reduce AML cell viability. However, AML heterogeneity and the limited selectivity of current inhibitors underscore the need for predictive biomarkers and combinatorial strategies to translate DGK inhibition into effective therapy. Full article

(This article belongs to the Special Issue Molecular Mechanisms of Leukemias)

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29 pages, 1422 KB

Open AccessReview

Functions of TIP60/NuA4 Complex Subunits in Cell Differentiation

by Fatemeh Hashemi, Aida Nourozi, Mojtaba Shaban Loushab and Karl Riabowol

Cells 2025, 14(21), 1720; https://doi.org/10.3390/cells14211720 - 1 Nov 2025

Viewed by 1262

Abstract

The TIP60/NuA4 complex is a large, multifunctional histone acetyltransferase assembly of ~1.7 megadaltons, composed of 17–20 subunits, which plays a central role in epigenetic regulation. Through recognition of H3K4me3 by the ING3 reader, TIP60/NuA4 is recruited to sites of active transcription, where it [...] Read more.

The TIP60/NuA4 complex is a large, multifunctional histone acetyltransferase assembly of ~1.7 megadaltons, composed of 17–20 subunits, which plays a central role in epigenetic regulation. Through recognition of H3K4me3 by the ING3 reader, TIP60/NuA4 is recruited to sites of active transcription, where it remodels chromatin to regulate gene expression. Its activities include histone acetylation, histone variant exchange, transcriptional co-activation, and regulation of the cell cycle and apoptosis. In this review, we examine how altered subunit levels or mutations impact the chromatin structure and transcriptional activity, and how these changes influence differentiation across diverse cell types. We emphasize the molecular mechanisms by which TIP60/NuA4 shapes lineage specification, including histone H2A and H4 acetylation by the KAT5 catalytic subunit, H2A.Z incorporation by EP400, and interactions with transcription factors such as MyoD, PPARγ, and Myc. By integrating mechanistic and functional insights, we highlight how TIP60/NuA4 acts as a central epigenetic hub in differentiation and contributes to proper developmental transitions. Full article

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24 pages, 4341 KB

Open AccessArticle

EGFR mRNA-Engineered Mesenchymal Stem Cells (MSCs) Demonstrate Radioresistance to Moderate Dose of Simulated Cosmic Radiation

by Fay Ghani, Peng Huang, Cuiping Zhang and Abba C. Zubair

Cells 2025, 14(21), 1719; https://doi.org/10.3390/cells14211719 - 1 Nov 2025

Viewed by 901

Abstract

Galactic cosmic ray (GCR) radiation is a major barrier to human space exploration beyond Earth’s magnetic field protection. Mesenchymal stem cells (MSCs) are found in all organs and play a critical role in repair and regeneration of tissue. We engineered bone marrow-derived MSCs [...] Read more.

Galactic cosmic ray (GCR) radiation is a major barrier to human space exploration beyond Earth’s magnetic field protection. Mesenchymal stem cells (MSCs) are found in all organs and play a critical role in repair and regeneration of tissue. We engineered bone marrow-derived MSCs and evaluated their response to ionizing radiation exposure. Epidermal growth factor receptor (EGFR) expression by certain types of cancers has been shown to induce radioresistance. In this study, we tested the feasibility of transfecting MSCs to overexpress EGFR (eMSC-EGFR) and their capacity to tolerate and recover from X-ray exposure. Quantitative real-time PCR (qRT-PCR) and immunoblotting results confirmed the efficient transfection of EGFR into MSCs and EGFR protein production. eMSC-EGFR maintained characteristics of human MSCs as outlined by the International Society for Cell & Gene Therapy. Then, engineered MSCs were exposed to various dose rates of X-ray (1–20 Gy) to assess the potential radioprotective role of EGFR overexpression in MSCs. Post-irradiation analysis included evaluation of morphology, cell proliferation, viability, tumorigenic potential, and DNA damage. eMSC-EGFR showed signs of radioresistance compared to naïve MSCs when assessing relative proliferation one week following exposure to 1–8 Gy X-rays, and significantly lower DNA damage content 24 h after exposure to 4 Gy. We establish for the first time the efficient generation of EGFR overexpressing MSCs as a model for enhancing the human body to tolerate and recover from moderate dose radiation injury in long-term manned space travel. Full article

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19 pages, 1312 KB

Open AccessReview

Beyond Tumor Suppression: The Multifaceted Functions of HOPX in Tissue Differentiation, Metabolism, and Immunity

by Fabian Munzert, Miljana Nenkov, Alexander Berndt, Tim Sandhaus, Susanne Lang, Nikolaus Gaßler and Yuan Chen

Cells 2025, 14(21), 1718; https://doi.org/10.3390/cells14211718 - 1 Nov 2025

Viewed by 954

Abstract

The transcription factor homeodomain-only protein X (HOPX) is the smallest member of the homeodomain protein family. Lacking a DNA-binding domain, it acts as a co-effector, interacting with other transcription factors such as serum response factor (SRF) and GATA-binding factor 6 (GATA6) to regulate [...] Read more.

The transcription factor homeodomain-only protein X (HOPX) is the smallest member of the homeodomain protein family. Lacking a DNA-binding domain, it acts as a co-effector, interacting with other transcription factors such as serum response factor (SRF) and GATA-binding factor 6 (GATA6) to regulate the differentiation and development of the heart and lung. HOPX exerts a tumor-suppressive function in various types of epithelial-derived carcinoma, while it promotes oncogenic effects in mesenchymal-derived sarcoma, indicating a distinct role of HOPX in the two major types of the malignancy. In addition, accumulating evidence shows that HOPX is expressed in the immune system and involved in the differentiation of immune cells. Recently, the emerging role of HOPX in metabolism has gained attention. This review describes the identification of HOPX in various tissues and discusses its role in carcinogenesis, as well as its functions in tissue differentiation, lipid metabolism, immunity, and the tumor microenvironment. The participation of HOPX in carcinogenesis and immunity implies that it may serve as a potential enhancer in tumor immunotherapy. Full article

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18 pages, 5999 KB

Open AccessArticle

Effectiveness of Graphene Oxide (GO) in Activating the Mitochondrial Pathway of Oxidative Stress-Induced Apoptosis in Breast Cancer Cells

by Rafał Krętowski, Beata Szynaka, Małgorzata Borzym-Kluczyk, Natalia Tyszka, Agata Jabłońska-Trypuć, Maciej Gil and Marzanna Cechowska-Pasko

Cells 2025, 14(21), 1717; https://doi.org/10.3390/cells14211717 - 1 Nov 2025

Cited by 1 | Viewed by 828

Abstract

Due to its unique physicochemical properties, graphene oxide (GO) is used in nanomedicine. Many studies have examined the effects of GO on cancer cells. However, there are no data on the mechanisms of action of GO in breast cancer. The aim of this [...] Read more.

Due to its unique physicochemical properties, graphene oxide (GO) is used in nanomedicine. Many studies have examined the effects of GO on cancer cells. However, there are no data on the mechanisms of action of GO in breast cancer. The aim of this study was to analyze the cytotoxic effect and mechanisms of action of GO on MDA-MB-231 and ZR-75-1 cell lines. Our findings show that GO induced cytotoxicity in MDA-MB-231 but not in ZR-75-1 cells. The cytotoxic effect of GO on fibroblasts was negligible. Cytotoxicity was associated with ROS synthesis, decreased mitochondrial membrane potential, and apoptosis/necrosis in MDA-MB-231 cells. In addition, we observed cell cycle arrest and increased P21 protein expression in MDA-MB-231 cells. Furthermore, we observed increased levels of proapoptotic proteins, decreased levels of antiapoptotic proteins, and activation of caspase-9 and caspase-3/7 in MDA-MB-231 cells. This study elucidates the possible mechanisms of action of GO in breast cancer cells. Full article

(This article belongs to the Special Issue Focus on Machinery of Cell Death)

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13 pages, 14281 KB

Open AccessArticle

Exenatide Is Neuroprotective in a New Rabbit Model of Hypoxia-Ischemia

by Eridan Rocha-Ferreira, Malin Carlsson, Pernilla Svedin, Kerstin Ebefors, Owen Herrock, Anna-Lena Leverin and Henrik Hagberg

Cells 2025, 14(21), 1715; https://doi.org/10.3390/cells14211715 - 1 Nov 2025

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Abstract

Hypoxia-ischemia is a serious perinatal complication affecting neonates globally. Animal models have increased the understanding of its pathophysiology and have been used to investigate potential therapies. Exenatide, clinically used for the treatment of type 2 diabetes mellitus, also protects the rodent brain from [...] Read more.

Hypoxia-ischemia is a serious perinatal complication affecting neonates globally. Animal models have increased the understanding of its pathophysiology and have been used to investigate potential therapies. Exenatide, clinically used for the treatment of type 2 diabetes mellitus, also protects the rodent brain from hypoxia-ischemia. The rabbit brain has an earlier neurodevelopmental maturation than rodents, as well as similar postnatal maturation to humans. We hereby introduce a new, reproducible hypoxia-ischemia model in rabbit kits at postnatal day (P) 3–4. Following hypoxia-ischemia, rabbit kits received different exenatide concentrations: 170 μg/g (2-dose) or 500 μg/g (1- or 2-dose), or vehicle. The brains were collected seven days later for histological assessment showing that 500 μg/g exenatide, either as a 1- or 2-dose regimen, reduced brain tissue loss by 90% in hypoxia-ischemia experiments both at P3 and P4. A second cohort received a 1-dose 500 μg/g of exenatide or vehicle, and were sacrificed at different early time-points for glucose, ketone bodies, body weight, and temperature measurements. Our results showed a transient 2-fold increase in ketone bodies (0.6 to 1.3 mmol/L) at 6 h. Exenatide did not affect glucose, body temperature or weight gain and appears to be safe and well tolerated in the rabbit model of hypoxia-ischemia. Full article

(This article belongs to the Special Issue Perinatal Brain Injury—from Pathophysiology to Therapy)

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