Cells

17 pages, 730 KB

Open AccessEditor’s ChoiceReview

Exploring the Muco-Microbiotic Interface as a Hub for Microbial Metabolites and Immune Regulation in Gastroenteric Health and Disease

by Adelaide Carista, Melania Ionelia Gratie, Enrico Tornatore, Salvatore Accomando, Giovanni Tomasello, Domiziana Picone, Stefano Burgio and Francesco Cappello

Cells 2026, 15(1), 45; https://doi.org/10.3390/cells15010045 - 25 Dec 2025

Cited by 3 | Viewed by 2097

Abstract

The mucus layer covering the gastrointestinal tract forms a specialised interface where mucins, microbes, and extracellular vesicles create a dynamic, self-regulating ecosystem. Here, we introduce the concept of the muco-microbiotic layer as an integrated eco-physiological system that maintains mucosal homeostasis through coordinated structural, [...] Read more.

The mucus layer covering the gastrointestinal tract forms a specialised interface where mucins, microbes, and extracellular vesicles create a dynamic, self-regulating ecosystem. Here, we introduce the concept of the muco-microbiotic layer as an integrated eco-physiological system that maintains mucosal homeostasis through coordinated structural, metabolic, and immune functions. The MuMi layer varies regionally in its biochemical composition, microbial inhabitants, and environmental parameters—from the acidic stomach to the anaerobic colon—thereby generating distinct niches for microbial colonisation and metabolite production. We summarise current evidence on how mucin glycans, mucus-associated microbiota, and vesicle-mediated signalling sustain barrier integrity, nutrient flux, and immune tolerance. Perturbations in any of these components lead to barrier failure, microbial encroachment, and inflammation, contributing to a broad spectrum of disorders, including gastritis, inflammatory bowel disease, colorectal cancer, and metabolic syndrome. Methodological advances such as organoid and mucus-on-chip models, spatial multi-omics, and vesiculomics are now enabling site-specific analyses of this complex system. Conceptually, defining the mucus, microbiota, and vesicular compartments as a single MuMi layer provides a new framework for understanding mucosal physiology and pathophysiology, emphasising the interdependence between structure and function. Integrating this perspective into experimental and clinical research may open new avenues for diagnostics and therapies targeting mucosal health. Full article

(This article belongs to the Special Issue Gut Microbiota, Metabolites, and Immune Regulation in Gastrointestinal Diseases)

► Show Figures

Figure 1

28 pages, 869 KB

Open AccessEditor’s ChoiceReview

Cell-Free Nucleic Acids in Cardiovascular Disease: From Biomarkers to Mechanistic Drivers and Therapeutic Opportunities

by Hannah Morgan, Keara Little, Suchandrima Dutta, Sophie Chen, Jiantao Gong, Siddu Koduri, Asma Raja, Wendy Lin, Kanishka Saini, Riya Bhullar and Wei Huang

Cells 2026, 15(1), 33; https://doi.org/10.3390/cells15010033 - 23 Dec 2025

Cited by 6 | Viewed by 2903

Abstract

Cardiovascular disease (CVD) remains the leading cause of morbidity and mortality worldwide, with heart failure (HF) representing a major contributor to hospitalizations, healthcare costs, and death. Effective management of HF is hindered by the limitations of current biomarkers and diagnostic tools. Conventional biomarkers, [...] Read more.

Cardiovascular disease (CVD) remains the leading cause of morbidity and mortality worldwide, with heart failure (HF) representing a major contributor to hospitalizations, healthcare costs, and death. Effective management of HF is hindered by the limitations of current biomarkers and diagnostic tools. Conventional biomarkers, such as natriuretic peptides, primarily reflect downstream hemodynamic stress and often lack specificity, particularly in HF with preserved ejection fraction or multiple comorbidities. While imaging provides valuable structural and functional information, it is resource-intensive, costly, and unsuitable for frequent longitudinal monitoring. As a result, these conventional approaches are inadequate to capture the dynamic and heterogeneous nature of HF pathophysiology. Circulating cell-free nucleic acids (cfNAs), including cell-free DNA (cfDNA) and RNA (cfRNA), have emerged as promising noninvasive liquid biopsy biomarkers capable of providing real-time insight into upstream pathological events, such as cardiomyocyte injury, immune activation, inflammation, and maladaptive remodeling. Importantly, cfNAs also act as active mediators of CVD pathology. When released under stress or injury, cfNAs interact with pattern recognition receptors (PRRs) that trigger sterile inflammation, cardiovascular cell dysfunction, and adverse cardiac remodeling. This review summarizes the origins, mechanistic roles, and clinical significance of cfNAs in HF and related CVD, highlighting their dual roles as diagnostic biomarkers and mechanistic effectors of disease. Finally, we discuss emerging cfNA-targeted therapeutic strategies, challenges, and future opportunities for precision medicine in HF and HF-associated CVD. Full article

(This article belongs to the Special Issue Molecular Pathogenesis of Cardiovascular Diseases)

► Show Figures

Figure 1

22 pages, 3828 KB

Open AccessEditor’s ChoiceArticle

Potential Role of Serum Cytokines and Chemokines as Biomarkers of Injury Severity and Functional Outcomes Following Pediatric Traumatic Brain Injury

by Kathryn Swaby, Alexander J. Skirvin, Natalie Machado, Maria Mateo Chavez, Julia Alexis Bernal, Ana Fuentes, Charlene P. Pringle, Kourtney Guthrie, Jennifer Coto, Rajderkar Dhanashree, Joslyn Gober, Paula Karina Perez, Juan P. Solano, Heather J. McCrea, Ricardo Loor-Torres, Joyce Kaufman, Ayham Alkhachroum, Kristine H. O’Phelan, Firas Kobeissy, Robert W. Keane, Kevin K. Wang, W. Dalton Dietrich, Juan Pablo de Rivero Vaccari and Jennifer C. Munoz Pareja Show full author list Hide full author list

Cells 2026, 15(1), 19; https://doi.org/10.3390/cells15010019 - 22 Dec 2025

Viewed by 1896

Abstract

Traumatic brain injury (TBI) is one of the leading causes of death and neurological disability worldwide. The search for biomarkers that indicate TBI severity and prognosis with greater accuracy is ongoing. This study aimed to evaluate the significance of several neuroinflammatory cytokines and [...] Read more.

Traumatic brain injury (TBI) is one of the leading causes of death and neurological disability worldwide. The search for biomarkers that indicate TBI severity and prognosis with greater accuracy is ongoing. This study aimed to evaluate the significance of several neuroinflammatory cytokines and chemokines, assessing their potential as biomarkers in pediatric TBI (pTBI). This was an exploratory analysis of inflammatory cytokines and chemokines measured in a subset of 26 children aged 0–18 years with TBI and 21 controls. TBI severity was determined by GCS. The functional outcome was measured via the GOS-E score at 6 weeks and 3, 6, 9, and 12 months post-injury. Serum samples were analyzed for ICAM-1, VCAM-1, SAA, CRP, IFN-g, IL-10, IL-12p70, IL-13, IL-1b, IL-2, IL-4, IL-6, IL-8, TNF-a, TNF-b, eotaxin, eotaxin-3, IP-10, MCP-1, MCP-4, MDC, MIP-1a, MIP-1b and TARC. Levels of IL-6, IL-10, IL-13, IL-16, MDC, and GM-CSF were increased, and IFN-γ, IL-5, IL-8, and eotaxin-3 were decreased at enrollment when compared with controls. Elevated IL-6 and IL-10 at enrollment were associated with severe TBI (AUC of 1, p = 0.0002 and p = <0.0001, respectively). IL-6, IL-10, IL-16, and TNF-β at enrollment and IL-5 at 24 h were elevated in children with unfavorable outcomes, with an AUC > 0.8, suggesting biomarker potential. Our data indicate that several cytokines and chemokines measured after TBI may aid in the assessment of pTBI severity and prognosis. IL-6, IL-10, and IL-16 may show potential as biomarkers for pTBI severity and outcomes. Full article

(This article belongs to the Special Issue Cellular and Molecular Mechanisms in Neuropsychiatric and Neurological Disorders)

► Show Figures

Figure 1

26 pages, 1500 KB

Open AccessEditor’s ChoiceReview

Platelet Subpopulations in Health and Disease: Heterogeneity, Clinical Associations, and Therapeutic Targeting

by Deepa Gautam, Giovanni Goggi and Elisabeth M. Battinelli

Cells 2026, 15(1), 11; https://doi.org/10.3390/cells15010011 - 20 Dec 2025

Cited by 4 | Viewed by 2239

Abstract

Platelets are often described in simple terms as small anucleate cells that mediate hemostasis, but studies over more than half a century have shown that circulating platelets are heterogeneous in size, density, age, and functional responses. These subtypes not only contribute to normal [...] Read more.

Platelets are often described in simple terms as small anucleate cells that mediate hemostasis, but studies over more than half a century have shown that circulating platelets are heterogeneous in size, density, age, and functional responses. These subtypes not only contribute to normal hemostasis but also play pivotal roles in the pathogenesis of diverse diseases, including cardiovascular, inflammatory, and malignant disorders. Accumulating evidence indicates that alterations in specific platelet subtypes are closely linked to disease onset, progression, and severity, underscoring their importance as both biomarkers and potential therapeutic targets. Current clinical assessments of platelet status rely primarily on platelet count and mean platelet volume (MPV) as part of routine complete blood count analysis. However, these global measures often fail to capture differences in platelet subtypes, which may remain undetected despite their significant contribution to disease pathology. This gap highlights the necessity of moving beyond conventional metrics toward a more nuanced understanding of platelet heterogeneity and its clinical implications. In this review, we discuss the diversity of platelet subpopulations and their roles in health and disease, emphasizing how specific subsets contribute to divergent pathological mechanisms. We also highlight emerging strategies that target defined platelet subpopulations, illustrating how this knowledge could pave the way for more precise diagnostic and therapeutic approaches. Full article

(This article belongs to the Special Issue Molecular and Cellular Insights into Platelet Function, 2nd Edition)

► Show Figures

Figure 1

19 pages, 7200 KB

Open AccessEditor’s ChoiceArticle

Cardiomyocyte Glucocorticoid Receptors Exacerbate Stress Effects in Myocardial Ischemia Injury in Mice

by Analilia Cardenas-Garza, Lilly A. Kamberov, Hemangini A. Dhaibar, Tanja Dudenbostel, Gopi Krishna Kolluru, Christopher G. Kevil, Robert H. Oakley, John A. Cidlowski, Luca Cucullo and Diana Cruz-Topete

Cells 2025, 14(24), 2017; https://doi.org/10.3390/cells14242017 - 18 Dec 2025

Viewed by 1782

Abstract

An increase in mental stress is a recognized risk factor for cardiovascular disease (CVD). The present study investigated the relationships between stress, glucocorticoid receptors (GR), and ischemia/reperfusion (I/R) injury. We subjected male and female mice lacking cardiomyocyte GR (CardioGRKO) and their respective controls [...] Read more.

An increase in mental stress is a recognized risk factor for cardiovascular disease (CVD). The present study investigated the relationships between stress, glucocorticoid receptors (GR), and ischemia/reperfusion (I/R) injury. We subjected male and female mice lacking cardiomyocyte GR (CardioGRKO) and their respective controls to a murine model of mental stress (restraint stress). Following stress exposure, mice from both experimental and control groups underwent I/R injury via surgical ligation of the left anterior descending coronary artery. Our findings suggest that the absence of cardiomyocyte GR mitigates the detrimental effects of restraint stress on infarct size and improves post-I/R survival rates in female mice. We found that cardiomyocyte GR deficiency protects the female heart from stress-induced damage by reducing oxidative stress (superoxide and lipid peroxide production). This study is the first to test the impact of systemic stress on cardiomyocyte GR activation, linking it to redox stress in the heart during I/R injury. Our findings provide proof of concept that stress exacerbates cardiomyocyte GR-mediated responses to myocardial infarction (MI) in the female heart. These insights may contribute to the development of sex-specific treatments and therapies tailored for women. Full article

(This article belongs to the Special Issue Sex Differences in Cardiovascular Structure and Function—from Mechanisms to Therapeutics)

► Show Figures

Graphical abstract

15 pages, 1474 KB

Open AccessEditor’s ChoiceArticle

Piezo1 Mechanosensor Expression in Rare Hematopoietic Cells Controls Systemic Inflammatory Response in Mice

by Shiv Vardan Singh, Anastasia Iris Karkempetzaki, Nasi Huang, Vipul C. Chitalia, Saravanan Subramaniam and Katya Ravid

Cells 2025, 14(24), 1999; https://doi.org/10.3390/cells14241999 - 16 Dec 2025

Viewed by 967

Abstract

Mutations in the Piezo1 mechanosensor are associated with blood cell anomalies. The objective of our study was to explore the role of Piezo1 in the development and function of the megakaryocyte (MK) lineage. To this end, PF4-Cre mice, bearing Cre recombinase under the [...] Read more.

Mutations in the Piezo1 mechanosensor are associated with blood cell anomalies. The objective of our study was to explore the role of Piezo1 in the development and function of the megakaryocyte (MK) lineage. To this end, PF4-Cre mice, bearing Cre recombinase under the control of the Pf4 gene promoter—which drives expression to hematopoietic progenitors and to the MK/platelet lineage—were crossbred with Piezo1-floxed mice to generate Piezo1 knockout (KO) mice. In our results, the hematopoietic stem cell (HSC) count—including Multipotent Progenitors 2 (MPP2) progenitors that give rise to MKs—tended to be augmented in KO mice, while the level of MPP3 progenitors that give rise to white blood cells (WBCs) tended to be reduced, as compared to matching controls. The level of circulating WBCs was significantly reduced in the KO mice compared to controls. In addition, while platelet count was modestly elevated, platelet activation response was reduced in Piezo1 KO mice compared to controls. MK levels and ploidy were similar in both groups. Baseline serum pro-and anti-inflammatory cytokine profiles were also similar in the two experimental groups. However, upon LPS challenge, there was a significant reduction in IL-6 and INF-γ levels in the sera of Piezo1 KO mice compared to controls. Our findings point to an immunoregulatory and thrombotic potential of Piezo1 in relatively rare bone marrow cells, along with an ability to modulate WBC count. Full article

(This article belongs to the Section Cell Microenvironment)

► Show Figures

Graphical abstract

27 pages, 3096 KB

Open AccessEditor’s ChoiceArticle

B Cells Can Trigger the T-Cell-Mediated Autoimmune Response Against Melanocytes in Psoriasis

by Mengwen He, Melissa Bernhardt, Akiko Arakawa, Song-Min Kim, Sigrid Vollmer, Burkard Summer, Yukiyasu Arakawa, Tatsushi Ishimoto, Andreas Schlosser and Jörg Christoph Prinz

Cells 2025, 14(24), 2002; https://doi.org/10.3390/cells14242002 - 16 Dec 2025

Cited by 3 | Viewed by 1689

Abstract

Psoriasis vulgaris is a T-cell-mediated skin disease that may involve an autoimmune response against melanocytes. It develops through still unexplained pathomechanisms. Streptococcal tonsillopharyngitis is a major trigger of psoriasis onset and relapses. HLA-C*06:02 is the main psoriasis risk gene. Here we find that [...] Read more.

Psoriasis vulgaris is a T-cell-mediated skin disease that may involve an autoimmune response against melanocytes. It develops through still unexplained pathomechanisms. Streptococcal tonsillopharyngitis is a major trigger of psoriasis onset and relapses. HLA-C*06:02 is the main psoriasis risk gene. Here we find that B cells isolated from streptococci-infected tonsils or peripheral blood of HLA-C*06:02⁺ psoriasis patients stimulate an HLA-C*06:02-restricted melanocyte-reactive Vα3S1/Vβ13S1 T-cell receptor (TCR) from a lesional psoriatic CD8⁺ T cell clone in an IFN-γ-enhanced manner. Patients’ B cells furthermore induce proliferation of autologous blood CD8⁺ T cells. We identify several HLA-C*06:02-presented self-peptides in the immunopeptidomes we had isolated from four HLA-C*06:02 homozygous B-cell lines that stimulate the Vα3S1/Vβ13S1 TCR and differ from the melanocyte autoantigen recognized by this TCR. These data suggest that the proinflammatory environment of streptococcal tonsillopharyngitis may enable B cells to activate autoreactive CD8⁺ T cells that, owing to the polyspecificity of T-cell receptors, recognize several B-cell self-peptides presented by HLA-C*06:02 and subsequently cross-react against melanocytes in the skin, thereby triggering psoriasis. The capacity of B cells to stimulate a cross-reactive autoimmune response through HLA class I-presented B-cell peptides is a previously unknown mechanism in the induction of autoimmunity that could explain psoriasis onset and persistence. Full article

(This article belongs to the Special Issue Genetic and Cellular Basis of Autoimmune Diseases)

► Show Figures

Graphical abstract

15 pages, 3187 KB

Open AccessEditor’s ChoiceArticle

PGE1 Suppresses the Expression of M2 Markers on Macrophages Through Prostaglandin Receptors

by Hiroyuki Tsuchiya, Takehiko Hanaki, Jun Yoshida, Mayu Obora, Yoshiyuki Fujiwara and Daisuke Nanba

Cells 2025, 14(24), 1992; https://doi.org/10.3390/cells14241992 - 15 Dec 2025

Cited by 2 | Viewed by 998

Abstract

M2-like tumor-associated macrophages (TAMs) are a promising target for cancer immunotherapy, particularly for cancer patients who are refractory to current immune checkpoint inhibitors (ICIs). Previously, we showed that prostaglandin E1 (PGE1) enhances the expression of M1 markers, including HLA-DR, on macrophages and induces [...] Read more.

M2-like tumor-associated macrophages (TAMs) are a promising target for cancer immunotherapy, particularly for cancer patients who are refractory to current immune checkpoint inhibitors (ICIs). Previously, we showed that prostaglandin E1 (PGE1) enhances the expression of M1 markers, including HLA-DR, on macrophages and induces the M1 polarization of TAMs in vivo. This study investigated the pharmacological mechanisms by which PGE1 and its derivatives suppress the expression of M2 markers, including TREM2 and CXCR2. Macrophages were cultured in ultralow attachment dishes either alone or in combination with liver cancer cell lines to generate homospheroids or heterospheroids. Cell surface marker expression was assessed by flow cytometry. Compared with homospheroids, M2 marker expression on macrophages in heterospheroids was significantly increased, suggesting that heterospheroid culture promotes M2 polarization. PGE1 decreased M2 marker expression in heterospheroids more effectively compared with PGE2, PGE3, misoprostol, and 13,14-dihydro-15-keto-PGE1, whereas the suppressive effects of 15-keto- and 13,14-dihydro-PGE1s, and lubiprostone were comparable to that of PGE1. Pharmacological inhibition of prostaglandin receptors revealed that EP2 and EP4 receptors are involved in the PGE1-induced reprogramming of M2-like macrophages to M1 macrophages. In summary, PGE1 and its derivatives are promising TAM-targeting immunotherapeutics. Full article

(This article belongs to the Special Issue Macrophages in Cancer: Immunosuppression, Immunoregulation, and Immunotherapy)

► Show Figures

Figure 1

24 pages, 1966 KB

Open AccessEditor’s ChoiceReview

The Expanding Role of HLA-E in Host Defense: A Target for Broadly Applicable Vaccines and Immunotherapies

by Mahsa Rafieiyan, Marco Pio La Manna, Francesco Dieli, Nadia Caccamo and Giusto Davide Badami

Cells 2025, 14(24), 1983; https://doi.org/10.3390/cells14241983 - 14 Dec 2025

Cited by 1 | Viewed by 1651

Abstract

Human leukocyte antigen (HLA)-E, a non-classical class I molecule with limited polymorphism, bridges innate and adaptive immunity. Traditionally, the role of HLA-E had been associated with regulating natural killer (NK) cell activity via CD94/NKG2 receptors, by presenting self-peptides derived from the leader sequence [...] Read more.

Human leukocyte antigen (HLA)-E, a non-classical class I molecule with limited polymorphism, bridges innate and adaptive immunity. Traditionally, the role of HLA-E had been associated with regulating natural killer (NK) cell activity via CD94/NKG2 receptors, by presenting self-peptides derived from the leader sequence of HLA-I. Recent findings reveal its ability to present pathogen-derived peptides to CD8⁺ T cells, eliciting unconventional cytotoxic responses. This review examines the expanding role of HLA-E-restricted T cells in viral and bacterial infections and their capacity to recognize diverse microbial peptides and enhance immune response when classical HLA pathways are impaired. We also highlight key advances in immunotherapy and vaccine development, including CMV-vectored platforms, donor-unrestricted TCR-based strategies, and peptide prediction algorithms. The minimal polymorphism of HLA-E, its resistance to viral immune evasion, and its ability to present conserved pathogen peptides position it as a promising target for universal vaccines and next-generation immunotherapies. Understanding these unconventional roles may pave the way for broadly applicable immunotherapies and vaccines against infectious diseases. Full article

(This article belongs to the Section Cellular Immunology)

► Show Figures

Figure 1

19 pages, 1107 KB

Open AccessEditor’s ChoiceReview

Distinct Roles of Monocyte Subsets in Cancer

by Maria Amparo Sahagun Cortez, Wolf Eilenberg, Christoph Neumayer and Christine Brostjan

Cells 2025, 14(24), 1982; https://doi.org/10.3390/cells14241982 - 13 Dec 2025

Cited by 2 | Viewed by 1703

Abstract

While the distinct roles of lymphocyte populations are well characterized in adaptive immunity, the phenotypic and functional diversity of innate immune cells is less explored. In recent years, subsets of monocytes have gained attention, as prominent shifts in population frequencies have been observed [...] Read more.

While the distinct roles of lymphocyte populations are well characterized in adaptive immunity, the phenotypic and functional diversity of innate immune cells is less explored. In recent years, subsets of monocytes have gained attention, as prominent shifts in population frequencies have been observed in disease states such as cancer. This narrative review summarizes current knowledge of the distribution and functional differences among the three major monocyte subsets (classical, intermediate, non-classical) in tumor settings. It includes rare populations, such as neutrophil-like, CD56+, and Tie2-expressing monocytes. Scientific evidence indicates that the phenotypical and functional heterogeneity of monocyte subsets determines their roles in either preventing cancer development or supporting the progression of disease through a remarkable diversity of mechanisms. Of note, alterations in the distribution of monocyte subsets and their functional reprogramming have been identified as drivers of cancer progression. While changes in monocyte frequencies have limited diagnostic biomarker potential for cancer detection, they may reflect the progression of disease and response to therapy. Based on subset-specific properties, distinct monocyte populations are increasingly recognized as promising targets of cancer immunotherapy. Yet novel strategies targeting monocyte populations must consider the risk of treatment reversal given the high plasticity of these cells. Full article

► Show Figures

Figure 1

21 pages, 2914 KB

Open AccessEditor’s ChoiceArticle

Hyperactivity and Differential Gene Expression in lbx1a⁽^−/⁻⁾ Zebrafish Larvae

by Carsten Drepper, Laura Kettenstock, Simon Stöckl, Anna Elsenbach, Carina Lechermeier, Wonhyeok Lee, Susanne Kneitz, Klaus-Peter Lesch, Marcel Romanos and Christina Lillesaar

Cells 2025, 14(24), 1980; https://doi.org/10.3390/cells14241980 - 12 Dec 2025

Viewed by 1259

Abstract

Lbx1 plays important roles in different processes, including the development of sensory pathways, neuronal cell fate regulation, and muscle cell precursor migration. Genetic variation in the LBX1 locus has been associated with several human disease conditions, such as idiopathic scoliosis, congenital limb malformation, [...] Read more.

Lbx1 plays important roles in different processes, including the development of sensory pathways, neuronal cell fate regulation, and muscle cell precursor migration. Genetic variation in the LBX1 locus has been associated with several human disease conditions, such as idiopathic scoliosis, congenital limb malformation, and neuropsychiatric illness, including attention-deficit/hyperactivity disorder (ADHD) and anxiety disorders. Zebrafish (Danio rerio) were used to investigate the behavioral consequences of the loss of function of the two orthologs to the human LBX1 gene, zebrafish lbx1a and lbx1b. We observed a consistent locomotor hyperactivity phenotype induced by a novel environment in lbx1a mutants. Repeated dark stimuli provoked similar responses in both mutant lines, including the novelty-induced hyperactivity. We performed RNAseq on total RNA isolated from the head region of mutant and wildtype larvae. Several differentially expressed genes were identified, giving more insights into Lbx1 target genes and pathways, which could be relevant regarding the evaluation of zebrafish lbx1a or lbx1b as a human disease model. Furthermore, the analysis was complemented with a comparison to the expression profile of human LBX1 overexpression in cell culture, revealing a convergence on just two commonly regulated genes, namely alpha-Internexin (INA) and Fibrillin-3 (FBN3). In conclusion, our findings might further elucidate the multitude of functions of Lbx1 and its involvement in various human disease conditions. Full article

(This article belongs to the Special Issue Advances in Zebrafish Disease Models)

► Show Figures

Figure 1

23 pages, 1268 KB

Open AccessEditor’s ChoiceReview

CD74 and Proteases: Impact of Location on Immune and Cellular Functions

by Iztok Dolenc

Cells 2025, 14(24), 1960; https://doi.org/10.3390/cells14241960 - 10 Dec 2025

Cited by 3 | Viewed by 2250

Abstract

Proteases represent a diverse family of enzymes that catalyze the hydrolysis of peptide bonds, modulating numerous biological processes. Among their substrates, CD74—also known as the invariant chain—has received increased research attention due to its multifunctional roles in both innate and adaptive immunity. This [...] Read more.

Proteases represent a diverse family of enzymes that catalyze the hydrolysis of peptide bonds, modulating numerous biological processes. Among their substrates, CD74—also known as the invariant chain—has received increased research attention due to its multifunctional roles in both innate and adaptive immunity. This review provides an overview of current knowledge on protease-mediated interactions with CD74. The protein was originally identified as a chaperone for major histocompatibility complex class II (MHC-II) molecules. Proteolytic cleavage of CD74, most notably by cathepsin S, is essential for the release of MHC-II and the initiation of antigen presentation. However, CD74 has since emerged as a central regulator of processes extending well beyond antigen presentation. More recent findings reveal that CD74, acting as a receptor of macrophage migration inhibitory factor, also participates in signaling pathways in non-immune cells, independent of its classical chaperone function. Proteolytic processing of CD74 can trigger signaling cascades that modulate gene expression, underscoring its multifunctionality. Dysregulation of CD74 cleavage and its interaction with proteases has been linked to diverse pathological conditions, including cancer and autoimmune diseases, where aberrant protease activity disrupts CD74 function and promotes disease progression. Full article

► Show Figures

Figure 1

19 pages, 1532 KB

Open AccessEditor’s ChoiceReview

The Stress Response Is Adaptive in a Context- and State-Dependent Manner

by Harmen J. Krugers and Marian Joëls

Cells 2025, 14(24), 1957; https://doi.org/10.3390/cells14241957 - 9 Dec 2025

Cited by 5 | Viewed by 2880

Abstract

A long-standing question in stress physiology is when and how the adaptive stress response can become maladaptive. In this narrative review primarily focusing on experimental studies in (male) animals, we argue that this is not merely a semantic matter but that—within limits—the organism’s [...] Read more.

A long-standing question in stress physiology is when and how the adaptive stress response can become maladaptive. In this narrative review primarily focusing on experimental studies in (male) animals, we argue that this is not merely a semantic matter but that—within limits—the organism’s response to an acute or chronic stressor and the interpretation thereof as adaptive or maladaptive very much depends on the context and the circumstances under which the response is studied, including the task configuration and the state of the organism, in relation to its recent and past stress history. We substantiate this by providing examples at the behavioral level. The behavioral findings can be understood from neurophysiological studies, which show that the recent and distant (stress) history of an organism shapes its brain function and plasticity. Consequently, renewed exposure to a challenging situation evokes a different cellular and circuit response than it would without this history, pointing to metaplastic changes. Overall, we argue that many of the stress-induced effects that are currently interpreted as maladaptive are, in fact, adaptive as long as the environment aligns with the circumstances predicted by the individual’s experience. Adaptation to novel circumstances will require (behavioral) flexibility. Understanding the relationship between metaplastic or state-dependent cellular responses on the one hand and flexible behavioral outcomes that foster adaptation on the other hand may contribute to developing interventions or preventing stress-related mental disorders. Full article

► Show Figures

Figure 1

25 pages, 6468 KB

Open AccessEditor’s ChoiceReview

Plant-Derived Antioxidants as Modulators of Redox Signaling and Epigenetic Reprogramming in Cancer

by Thi Thuy Truong, Alka Ashok Singh, Soonhyuk Tak, Sungsoo Na, Jaeyeop Choi, Junghwan Oh and Sudip Mondal

Cells 2025, 14(24), 1948; https://doi.org/10.3390/cells14241948 - 8 Dec 2025

Cited by 14 | Viewed by 1666

Abstract

Redox imbalance and epigenetic dysregulation, which both contribute to tumor initiation, survival, and resistance to therapy, are intimately linked to the progression of cancer. Reactive oxygen species (ROS) have two contrasting effects: at moderate concentrations, they promote angiogenesis and oncogenic signaling, whereas at [...] Read more.

Redox imbalance and epigenetic dysregulation, which both contribute to tumor initiation, survival, and resistance to therapy, are intimately linked to the progression of cancer. Reactive oxygen species (ROS) have two contrasting effects: at moderate concentrations, they promote angiogenesis and oncogenic signaling, whereas at high concentrations, they trigger apoptosis. Oxidative stress alters histone modifications, DNA methylation, and non-coding RNA (ncRNA) expression, reshaping the epigenetic landscape and supporting malignant phenotypes. Plant-derived antioxidants, including flavonoids, polyphenols, alkaloids, and terpenoids, act as dual modulators of cancer biology. They scavenge or regulate reactive oxygen species (ROS), restore redox balance, activate tumor suppressor pathways, inhibit oncogenic mechanisms, and reverse abnormal epigenetic marks. Compounds such as resveratrol, curcumin, epigallocatechin gallate (EGCG), quercetin, and sulforaphane modulate DNA methyltransferases (DNMTs), histone deacetylases (HDACs), and non-coding RNA networks, and can enhance chemotherapy and radiation therapy. Despite promising mechanisms, challenges remain in translational efficacy, optimal dosing, and bioavailability. This review emphasizes the potential of plant-derived antioxidants as precision oncology adjuncts and highlights the need for biomarker-guided strategies, nano-delivery systems, and clinical validation to fully realize their therapeutic benefits. Plant-derived antioxidants mitigate ROS-induced oncogenic signaling, as evidenced by in vitro and clinical models. Full article

► Show Figures

Figure 1

21 pages, 2532 KB

Open AccessEditor’s ChoiceArticle

Electrophysiological Phenotyping of hiPSC-Derived Atrial Cardiomyocytes Using Automated Patch-Clamp: A Platform for Studying Atrial Inherited Arrhythmias

by Verónica Jiménez-Sábado, Hosna Babini, Peter C. Ruben, Eric A. Accili, Thomas W. Claydon, Leif Hove-Madsen and Glen F. Tibbits

Cells 2025, 14(24), 1941; https://doi.org/10.3390/cells14241941 - 6 Dec 2025

Cited by 2 | Viewed by 1716

Abstract

Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) represent a robust platform for modelling inherited cardiac disorders. Comparative analysis of ion channel activity in patient-specific and isogenic control lines provides critical insights into the molecular mechanisms underlying channelopathies and arrhythmias. Atrial-specific hiPSC-CMs (hiPSC-aCMs) exhibit distinct [...] Read more.

Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) represent a robust platform for modelling inherited cardiac disorders. Comparative analysis of ion channel activity in patient-specific and isogenic control lines provides critical insights into the molecular mechanisms underlying channelopathies and arrhythmias. Atrial-specific hiPSC-CMs (hiPSC-aCMs) exhibit distinct electrophysiological properties governed by unique ion channel expression profiles, underscoring the need for optimized methodologies to record atrial ionic currents accurately. Here, we characterized the electrophysiological features of hiPSC-aCMs using the Nanion Patchliner automated patch-clamp system. An optimized cell dissociation protocol was developed to enhance cell integrity and seal formation, while tailored intra- and extracellular solutions were employed to isolate specific ionic currents. Using this approach, we reliably recorded major atrial currents, including the sodium current (I_Na), L-type calcium current (I_CaL), transient outward potassium current (I_to), ultrarapid component of the delayed rectifier current (I_Kur), small-conductance calcium-activated potassium current (I_SK), and pacemaker funny current (I_f). The resulting current profiles were reproducible and consistent with those observed in native atrial cardiomyocytes. These findings establish the feasibility of the automated electrophysiological characterization of ion channels in hiPSC-aCMs. This platform enables more efficient investigation of pathogenic variants and facilitates the development of targeted therapeutics for atrial arrhythmias and related channelopathies. Full article

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

► Show Figures

Graphical abstract

25 pages, 1719 KB

Open AccessEditor’s ChoiceReview

Astrocyte-Mediated Plasticity: Multi-Scale Mechanisms Linking Synaptic Dynamics to Learning and Memory

by Masaya Yamamoto and Tetsuya Takano

Cells 2025, 14(24), 1936; https://doi.org/10.3390/cells14241936 - 5 Dec 2025

Cited by 8 | Viewed by 5115

Abstract

Astrocytes play a pivotal role in shaping synaptic function and in learning, memory, and emotion. Recent studies show that perisynaptic astrocytic processes form structured interactions with pre- and postsynaptic elements, which extends synaptic diversity beyond neuron–neuron connections. Accumulating evidence indicates that astrocytic Ca [...] Read more.

Astrocytes play a pivotal role in shaping synaptic function and in learning, memory, and emotion. Recent studies show that perisynaptic astrocytic processes form structured interactions with pre- and postsynaptic elements, which extends synaptic diversity beyond neuron–neuron connections. Accumulating evidence indicates that astrocytic Ca²⁺ signaling, gliotransmission, and local translation modulate synaptic efficacy and contribute to the formation and stabilization of memory traces. It is therefore essential to define how astrocytic microdomains, multisynaptic leaflet domains, and network-level ensembles cooperate to regulate circuit computation across space and time. Advances in super-resolution and volumetric in vivo imaging and spatial transcriptomics now enable detailed, cell-type- and compartment-specific analysis of astrocyte–synapse interactions in vivo. In this review, we highlight these approaches and synthesize classical and emerging mechanisms by which astrocytes read neuronal activity, write to synapses, and coordinate network states. We also discuss theoretical frameworks such as neuron–astrocyte associative memory models that formalize astrocytic calcium states as distributed substrates for storage and control. This integrated view provides new insight into the multicellular logic of memory and suggests paths toward understanding and treating neurological and psychiatric disorders. Full article

(This article belongs to the Special Issue Synaptic Plasticity and the Neurobiology of Learning and Memory)

► Show Figures

Figure 1

26 pages, 5299 KB

Open AccessEditor’s ChoiceArticle

PeriTox-M, a Cell-Based Assay for Peripheral Neurotoxicity with Improved Sensitivity to Mitochondrial Inhibitors

by Anna-Katharina Holzer, Mira Dürr, Selina Multrus, Laura Dangel, Viktoria Magel and Marcel Leist

Cells 2025, 14(23), 1929; https://doi.org/10.3390/cells14231929 - 4 Dec 2025

Cited by 1 | Viewed by 1264

Abstract

Human cell-based assays for neurotoxicity (NT) and developmental neurotoxicity (DNT) have reached a high level of readiness, but some tests require improvements in the specificity and sensitivity at which mitochondrial toxicants are detected. This study aimed to optimize the PeriTox assay, which uses [...] Read more.

Human cell-based assays for neurotoxicity (NT) and developmental neurotoxicity (DNT) have reached a high level of readiness, but some tests require improvements in the specificity and sensitivity at which mitochondrial toxicants are detected. This study aimed to optimize the PeriTox assay, which uses peripheral neurons (PNs) and predicts the potential of chemicals to trigger peripheral neuropathies. By introducing a glucose-to-galactose switch in the medium composition, cells were forced to rely on mitochondrial respiration. Using pre-differentiated PNs cultured in either glucose (Glc) or galactose (Gal), we observed no major differences in baseline phenotype, gene expression, neurite outgrowth, or total ATP content. However, a marked metabolic shift was confirmed by the increased oxygen consumption in Gal conditions. Based on measurements of neurite growth and ATP levels, Gal-adapted neurons showed a heightened sensitivity, up to 7500-fold, to a range of mitochondrial respiratory chain (MRC) inhibitors. The sensitivity shift was high for inhibitors of MRC complexes I and III and modest or absent for unrelated compounds such as proteasome inhibitors or cytoskeletal poisons. For complex I-III inhibitors, the enhanced detection of mitochondrial neurotoxicants was coupled with a more accurate distinction between cytotoxic and neurite-specific effects, i.e., an improved assay specificity. In conclusion, our study on 39 compounds suggests that running the PeriTox assay in galactose increases its sensitivity and specificity for several mitochondrial toxicants, while no general disadvantages or shortcomings were observed. The modified version (PeriTox-M) may increase the performance of in vitro test batteries for scientific and regulatory applications. Full article

(This article belongs to the Special Issue Stress Signaling and Cell Fate Determination: The Role of Mitochondria)

► Show Figures

Graphical abstract

16 pages, 536 KB

Open AccessEditor’s ChoiceReview

Ferroptosis and Radiotherapy in Lung Cancer

by Prem C. Patel and Eva M. Galvan

Cells 2025, 14(23), 1927; https://doi.org/10.3390/cells14231927 - 4 Dec 2025

Cited by 2 | Viewed by 2833

Abstract

Background: Lung cancer (LC) is a leading cause of cancer mortality worldwide. While radiotherapy (RT) has been a lasting cornerstone of LC management, there are concerns due to tumor radioresistance and unintended damage to surrounding healthy tissue. Ferroptosis is a recently described mechanism [...] Read more.

Background: Lung cancer (LC) is a leading cause of cancer mortality worldwide. While radiotherapy (RT) has been a lasting cornerstone of LC management, there are concerns due to tumor radioresistance and unintended damage to surrounding healthy tissue. Ferroptosis is a recently described mechanism of programmed cell death which has potential to serve as a complementary adjunct to facilitate RT-based LC treatment. Objectives: This review is a comprehensive overview of ferroptosis in the broader context of synergism with RT for LC. Summary: Ferroptosis is essentially driven by intracellular iron overload, which drives the formation of reactive oxygen species, ultimately resulting in membrane instability and cell death. LC lines have been shown to exhibit a heterogeneous mix of pro- and anti-ferroptotic changes. RT shows promise as a potential ferroptosis inducer, especially when complemented with pharmacologic agents such as erastin. Conclusions: Ferroptosis represents a promising modern adjunct to a traditional therapeutic strategy. Future work should focus on rigorous dosage standards to avoid unintended toxicity, repurposing of currently available drugs into ferroptosis inducers, and establishment of safety protocols to begin the pathway towards clinical studies. Full article

(This article belongs to the Special Issue Lung Cancer: Molecular Mechanisms and Therapeutic Opportunities)

► Show Figures

Figure 1

42 pages, 1600 KB

Open AccessEditor’s ChoiceReview

Bridging the Translational Gap: Rethinking Smooth Muscle Cell Plasticity in Atherosclerosis Through Human-Relevant In Vitro Models

by Liliana Som and Nicola Smart

Cells 2025, 14(23), 1913; https://doi.org/10.3390/cells14231913 - 2 Dec 2025

Cited by 5 | Viewed by 3364

Abstract

While vascular smooth muscle cell (SMC) plasticity is increasingly recognised as a critical driver of atherosclerosis progression, most mechanistic insights derive from murine models that fail to fully capture the diversity and complexity of human SMC phenotypes. This creates a translational gap in [...] Read more.

While vascular smooth muscle cell (SMC) plasticity is increasingly recognised as a critical driver of atherosclerosis progression, most mechanistic insights derive from murine models that fail to fully capture the diversity and complexity of human SMC phenotypes. This creates a translational gap in our understanding of disease-relevant cell states. Human single-cell and genetic studies reveal a broader spectrum of SMC phenotypes, many of which remain uncaptured by existing experimental models. In this review, we argue that better human in vitro models, when critically assessed and integrated with omics data from human disease, can help bridge this gap. We examine how different in vitro systems, from simple monocultures to advanced co-culture and 3D platforms, can model human SMC plasticity, and how benchmarking against human single-cell and multi-omics data can guide model selection, validation, and refinement. Full article

(This article belongs to the Special Issue Emerging Topics in Smooth Muscle Cell Fate and Plasticity in Atherosclerosis)

► Show Figures

Figure 1

22 pages, 2395 KB

Open AccessEditor’s ChoiceArticle

Characterization of Stimulated γδ T Cells: Phenotypic Analysis and Implications for Allogeneic Cellular Immunotherapy

by Anna Bold, Heike Gross, Marco Bardenbacher, Elisabeth Holzmann, Stefan Knop and Martin Wilhelm

Cells 2025, 14(23), 1917; https://doi.org/10.3390/cells14231917 - 2 Dec 2025

Cited by 1 | Viewed by 1813

Abstract

Due to their anti-tumor activity and non-major histocompatibility complex (MHC) binding T cell receptor, γδ T cells are suitable candidates for allogeneic cellular immunotherapy in cancer. Recently, we developed a new protocol called Ko-Op for stimulation of γδ T cells (specifically Vy9Vδ2 T [...] Read more.

Due to their anti-tumor activity and non-major histocompatibility complex (MHC) binding T cell receptor, γδ T cells are suitable candidates for allogeneic cellular immunotherapy in cancer. Recently, we developed a new protocol called Ko-Op for stimulation of γδ T cells (specifically Vy9Vδ2 T cells) that generates a cell product consisting mainly of γδ T cells with preserved anti-tumor activity targeted for clinical-grade application. In this study, we investigated the phenotype of stimulated γδ T cells and correlated this with results of functional assays to obtain a deeper understanding of the characteristics of stimulated γδ T cells. Additionally, an intensive analysis of surface molecules of unstimulated and stimulated γδ T cells is presented. Since heterogeneous results regarding the response to therapy with γδ T cells observed in earlier clinical trials could be a consequence of various extents of γδ T cell adhesion and migration ability, we addressed surface molecules associated with cellular activity and adhesion and migration functions as well. By investigating correlations between the phenotype of unstimulated γδ T cells and cellular cytotoxicity, as well as the degranulation ability of stimulated γδ T cells, we could draw conclusions about optimal donors for further allogeneic cellular therapies. Finally, we demonstrated that the phenotype varies over the time of culture and is clearly modifiable by changing the stimulation protocol. Full article

(This article belongs to the Special Issue Unconventional T Cells in Health and Disease)

► Show Figures

Figure 1

16 pages, 1469 KB

Open AccessEditor’s ChoiceArticle

New Biomarkers in the Diagnosis and Prognosis of Dilated Cardiomyopathy: Pro-Resolving Lipids and miRNAs

by Rafael I. Jaén, Sergio Sánchez-García, María Fernández-Velasco, Irene Cuadrado, Beatriz de las Heras, Lisardo Boscá and Patricia Prieto

Cells 2025, 14(23), 1916; https://doi.org/10.3390/cells14231916 - 2 Dec 2025

Cited by 1 | Viewed by 1545

Abstract

Dilated cardiomyopathy is a major cause of heart failure and is one of the most common forms of cardiomyopathy worldwide. Although there has been significant progress in its clinical management, early diagnosis and precise prognosis remain challenging due to the lack of specificity [...] Read more.

Dilated cardiomyopathy is a major cause of heart failure and is one of the most common forms of cardiomyopathy worldwide. Although there has been significant progress in its clinical management, early diagnosis and precise prognosis remain challenging due to the lack of specificity in current biomarkers. As inflammation plays a key role in DCM, we determined the levels of systemic inflammatory markers and specific pro-resolving lipid mediators (SPMs) in a cohort of DCM patients. Our data show that the levels of lipoxin A4 significantly increased in DCM patients (343 + 75.1 pg/mL in controls vs. 482.2 ± 159.1 pg/mL in DCM patients), whereas the opposite was observed for resolving D1 (57.18 ± 32.68 pg/mL in controls vs. 38.55 ± 25.13 pg/mL in DCM patients). These results may indicate that SPMs could be considered new biomarkers related to the progression of this pathology. Moreover, since microRNAs (miRNAs) are also considered potential biomarkers at the molecular level, we conducted comprehensive miRNA expression profiling using a high-throughput array platform in our cohort. Of the differentially expressed miRNAs identified, we chose to focus on two that were significantly upregulated (miR378-3p and miR486-5p; more than two-folds) or downregulated (miR142-3p and miR328-3p < 20% and 40% vs. the control, respectively) in DCM patients, all of them strongly associated with inflammatory pathways. The selected miRNAs showed considerable potential as biomarkers, exhibiting statistical significance after ROC analysis. In fact, improved performance was observed when combining both miR142-3p and miR328-3p, using a LASSO regression model. However, we found no correlation between miRNAs and traditional inflammatory markers or SPMs ruling out the possibility to proposing them as combined biomarkers in this case. The heterogeneity of DCM leads to the need to identify new biomarkers that, either individually or in combination, may improve the prognosis of affected individuals. In our study, we have identified that some of the main SPMs can provide valuable information about disease progression, in addition to the combination of certain circulating miRNAs, which show promising prognostic values in our cohort. Thus, we have identified novel biomarkers that integrate inflammatory profiles with specific circulating miRNA expression patterns is an important step towards more targeted patient stratification in DCM. This approach can improve DCM diagnosis and prognosis, supporting the development of personalized treatments through a multi-parameter panel of biomarkers that can be measured in peripheral blood and used in routine clinical practice. Such a strategy can enable earlier treatment, resulting in better patient outcomes and quality of life. Full article

► Show Figures

Graphical abstract

17 pages, 12009 KB

Open AccessEditor’s ChoiceArticle

Endometrial Stromal Cells from Endometriosis Patients Reflect Lesion-Type-Specific Heterogeneity

by Daniel Rodriguez Gutierrez, Marianne R. Spalinger, Alina Astourian, Olivera Evrova, Lucie Berclaz, Monique Hartmann, Ioannis Dedes, Patrick Imesch, Julian M. Metzler, Isabelle Witzel, Mohaned Shilaih, Valentina Vongrad and Brigitte Leeners

Cells 2025, 14(23), 1891; https://doi.org/10.3390/cells14231891 - 28 Nov 2025

Cited by 4 | Viewed by 2599

Abstract

Endometriosis, a disease affecting about one out of ten women, is characterized by the growth of endometrial-like tissue outside the uterine cavity. There is significant disease heterogeneity, but the pathophysiological mechanisms underlying differences in clinical presentation are poorly understood. Here, we investigated endometrial [...] Read more.

Endometriosis, a disease affecting about one out of ten women, is characterized by the growth of endometrial-like tissue outside the uterine cavity. There is significant disease heterogeneity, but the pathophysiological mechanisms underlying differences in clinical presentation are poorly understood. Here, we investigated endometrial stromal cells (ESCs) from different types of endometrial lesions (endometrioma, superficial, and deep endometrial lesions), which revealed distinct differences in proliferation, migration, and contractility among different lesion types and when compared to ESC from normal (eutopic) endometrium. In particular, ESCs from endometriotic lesions showed reduced proliferation but increased migratory capacity, an effect most pronounced in endometrioma ESCs but also evident in ESCs from superficial and deep lesions. ESCs from superficial and deep lesions—but not those from endometrioma—showed increased contractility, a feature involved in tissue scarring and pain perception. Transcriptomics and proteomics revealed changes in genes and proteins involved in cell division, proliferation, extracellular matrix organization, and migration in endometriosis vs. healthy ESCs. Overall, our results demonstrate that stromal cells from different endometriotic lesions show distinct in vitro phenotypes that might explain differences in clinical presentation. Further, these cells represent an excellent in vitro model for studying patho-mechanisms involved in endometriosis heterogeneity. Full article

► Show Figures

Figure 1

28 pages, 586 KB

Open AccessEditor’s ChoiceReview

A New Look at the Role of Radiation-Related Epigenetic Mechanisms in Diagnosis and Anticancer Therapies

by Adam Jan Olichwier, Magdalena Bruzgo-Grzybko, Izabela Suwda Kalita, Natalia Bielicka, Ewa Chabielska and Anna Gromotowicz-Poplawska

Cells 2025, 14(23), 1885; https://doi.org/10.3390/cells14231885 - 27 Nov 2025

Cited by 2 | Viewed by 2000

Abstract

Epigenetics encompasses heritable but reversible modifications of gene expression that occur without changes in the DNA sequence and involve mechanisms such as DNA and RNA methylation and histone modifications. These mechanisms modulate chromatin architecture, genome stability, and cellular responses to environmental stressors, and [...] Read more.

Epigenetics encompasses heritable but reversible modifications of gene expression that occur without changes in the DNA sequence and involve mechanisms such as DNA and RNA methylation and histone modifications. These mechanisms modulate chromatin architecture, genome stability, and cellular responses to environmental stressors, and their dysregulation contributes to oncogenesis and cancer progression. In parallel, radiotherapy remains a cornerstone of cancer treatment; furthermore, ionizing radiation induces epigenetic modifications alongside direct DNA double-strand breaks and oxidative damage. Radiation-induced epigenetic changes, including global or locus-specific DNA methylation shifts (e.g., genes promoter CpG islets), histone acetylation and methylation imbalances, are increasingly recognized as key contributors to molecular radioresistance. These adaptive responses may enhance tumor cell survival, affect therapeutic efficacy, and promote metastasis. Understanding the interplay between radiation exposure and epigenetic remodeling opens new perspectives for precision oncology and diagnostics. Epigenetic biomarkers hold potential for predicting treatment response and prognosis, while epigenetic modifiers may sensitize tumors to radiation. This review summarizes current evidence on radiation-induced epigenetic mechanisms and evaluates their diagnostic, prognostic, and therapeutic implications in cancer management. Full article

(This article belongs to the Topic Overview of Cancer Metabolism)

► Show Figures

Figure 1

17 pages, 2618 KB

Open AccessEditor’s ChoiceArticle

Alterations in Resident Immune Cells in Prenatal Trisomy 21 Lungs

by Andrew Frauenpreis, Soumyaroop Bhattacharya, Randa Belgacemi, Pauline Sokolskiy, Gail Deutsch, Nicholas Jendzjowsky, Ian A. Glass, Thomas J. Mariani, Denise Al Alam and Soula Danopoulos

Cells 2025, 14(23), 1866; https://doi.org/10.3390/cells14231866 - 26 Nov 2025

Viewed by 945

Abstract

Respiratory tract infections (RTIs) are amongst the leading causes of hospitalizations in children with Down syndrome (DS). Their elevated susceptibility likely stems from structural differences in the airways and immune system abnormalities. The aim of this study was to characterize immune cells in [...] Read more.

Respiratory tract infections (RTIs) are amongst the leading causes of hospitalizations in children with Down syndrome (DS). Their elevated susceptibility likely stems from structural differences in the airways and immune system abnormalities. The aim of this study was to characterize immune cells in prenatal Trisomy 21 (T21) lungs, potentially explaining vulnerability to RTIs. Single-cell RNA sequencing was used to profile immune cells in prenatal T21 (n = 5) and non-T21 (n = 4) prenatal lungs. Spatial phenotypes were assessed via fluorescent in situ hybridization and immunofluorescent staining on prenatal lung tissue sections. Gene expression analysis was also performed on isolated immune cells from lung single-cell suspensions. Several major immune cell populations were identified. A total of 84 DEGs were identified in at least 1 of the 14 different clusters. A significant decrease in the percentage of B cells was observed in T21 lungs (FDR = 0.0037, * p < 0.05). Furthermore, qRT-PCR demonstrated B cell markers were significantly decreased in T21, including those associated with B cell maturation (* p < 0.05 and ** p < 0.01). Several of these markers were also decreased at the protein level (i.e., CD20 and CD38; * p < 0.05 and ** p < 0.01). Our data demonstrate changes in the T21 pulmonary immune system in utero, primarily within the B cell population, which may contribute to the increased susceptibility to RTIs observed in children with DS. Full article

(This article belongs to the Special Issue Cellular and Molecular Mechanisms in Immune Regulation—Second Edition)

► Show Figures

Figure 1

34 pages, 3961 KB

Open AccessEditor’s ChoiceReview

Cellular and Molecular Mechanisms of Wound Repair: From Biology to Therapeutic Innovation

by Caijun Jin, Yongxun Jin, Zhiyuan Ding, Kong Srey Nuch, Mira Han, JungHee Shim, Pham Ngoc Chien and Chan Yeong Heo

Cells 2025, 14(23), 1850; https://doi.org/10.3390/cells14231850 - 24 Nov 2025

Cited by 60 | Viewed by 12622

Abstract

Wound repair preserves tissue integrity through four overlapping phases—hemostasis, inflammation, proliferation, and remodeling—coordinated by platelets, neutrophils, macrophages, fibroblasts, keratinocytes, endothelial cells, and stem/progenitor cells acting with growth factors, chemokines, extracellular matrix, and intracellular signaling. Disruption of these programs results in chronic non-healing wounds [...] Read more.

Wound repair preserves tissue integrity through four overlapping phases—hemostasis, inflammation, proliferation, and remodeling—coordinated by platelets, neutrophils, macrophages, fibroblasts, keratinocytes, endothelial cells, and stem/progenitor cells acting with growth factors, chemokines, extracellular matrix, and intracellular signaling. Disruption of these programs results in chronic non-healing wounds or fibrotic scarring. Recent work delineates microbial influences, epigenetic and transcriptomic regulation, and cellular heterogeneity resolved by single-cell and spatial omics. Concurrent advances in biomaterials, engineered scaffolds, stem cell-derived products, and genome-targeted approaches are enabling mechanism-based therapies. Persistent challenges include wound heterogeneity, systemic modifiers such as diabetes and aging, and safe, effective delivery of biologics. This review summarizes cellular and molecular mechanisms of cutaneous repair, outlines deviations that underlie pathological healing, and evaluates emerging concepts and translational strategies. Integrating classical models with contemporary insights supports the development of precision wound medicine and personalized interventions to improve outcomes and quality of life. Full article

(This article belongs to the Special Issue Cellular and Molecular Mechanisms of Wound Repair)

► Show Figures

Figure 1

24 pages, 2412 KB

Open AccessEditor’s ChoiceReview

Innate Immune Pairing: Eosinophils as Hidden Architects of T Cell Immunity

by Kriti Gupta, Natalie A. Falta and Lisa A. Spencer

Cells 2025, 14(22), 1826; https://doi.org/10.3390/cells14221826 - 20 Nov 2025

Cited by 5 | Viewed by 2291

Abstract

Eosinophils, once primarily considered strictly end-stage effector cells in parasitic infections and allergic inflammation, are now emerging as vital immunoregulatory cells. This review focuses on eosinophil contributions to cell-mediated adaptive immunity by exploring the multifaceted interactions between eosinophils and T cells that underlie [...] Read more.

Eosinophils, once primarily considered strictly end-stage effector cells in parasitic infections and allergic inflammation, are now emerging as vital immunoregulatory cells. This review focuses on eosinophil contributions to cell-mediated adaptive immunity by exploring the multifaceted interactions between eosinophils and T cells that underlie their unique contributions to immune modulation in allergic diseases. We begin by reviewing key features of eosinophil immunobiology within the context of their relevance to the development, differentiation, and function of CD4⁺ and CD8⁺ T cells in homeostasis and immunity. Building on this framework, we review recent literature revealing new roles for eosinophils in homeostatic immunosuppression, adaptive immune initiation, and immunomodulation within the context of an active immune response. We further explore the significance of eosinophil functionality impacting the structure and function of primary and secondary lymphoid organs, including thymic involution and regeneration, on cell-mediated immunity. This review presents an evolving paradigm that positions eosinophils as essential players in shaping multiple layers of the immune landscape in allergic diseases and beyond. Full article

(This article belongs to the Special Issue Eosinophils and Their Role in Allergy and Related Diseases)

► Show Figures

Figure 1

35 pages, 3906 KB

Open AccessEditor’s ChoiceReview

A Comprehensive Review of Modern Cancer Therapies Utilizing Oncolytic Viruses

by Michał Sułek and Agnieszka Szuster-Ciesielska

Cells 2025, 14(22), 1825; https://doi.org/10.3390/cells14221825 - 20 Nov 2025

Cited by 4 | Viewed by 7154

Abstract

Oncolytic viruses (OVs) are gaining traction as advanced tools in cancer therapy. They are distinguished by their ability to destroy malignant cells while sparing normal tissue specifically. In addition to their direct tumor-lysing properties, an essential benefit of oncolytic virus therapy is its [...] Read more.

Oncolytic viruses (OVs) are gaining traction as advanced tools in cancer therapy. They are distinguished by their ability to destroy malignant cells while sparing normal tissue specifically. In addition to their direct tumor-lysing properties, an essential benefit of oncolytic virus therapy is its capacity to activate both the innate and adaptive immune systems. To enhance these therapeutic actions, many OVs have been genetically engineered to encode immune-modulating factors that reestablish or strengthen antitumor immune responses. Recent studies show that combining OVs with other forms of immunotherapy—such as immune checkpoint inhibitors, CAR-T cells, specific T-cell receptor therapies, or autologous tumor-infiltrating lymphocytes—offers significant advances in cancer treatment. This article reviews how OVs work, discusses strategies to enhance their immunogenicity further, and presents the latest rational combinations of oncolytic viruses with other immunotherapies based on current preclinical and clinical research. Full article

(This article belongs to the Special Issue How Can We Optimise Cancer Therapy? Tumour Microenvironment and Immune Evasion Strategies)

► Show Figures

Figure 1

21 pages, 983 KB

Open AccessEditor’s ChoiceReview

Valeric Acid: A Gut-Derived Metabolite as a Potential Epigenetic Modulator of Neuroinflammation in the Gut–Brain Axis

by Chiara Paciolla, Michele Manganelli, Mariagiovanna Di Chiano, Francesca Montenegro, Anna Gallone, Fabio Sallustio and Gabriella Guida

Cells 2025, 14(22), 1823; https://doi.org/10.3390/cells14221823 - 20 Nov 2025

Cited by 25 | Viewed by 3282

Abstract

The gut–brain axis (GBA) is a critical area of research for understanding the pathogenesis of neuroinflammatory and neurodegenerative diseases. Metabolites produced by the gut microbiota, particularly short-chain fatty acids (SCFAs), act as key mediators in this bidirectional communication. While the roles of acetate, [...] Read more.

The gut–brain axis (GBA) is a critical area of research for understanding the pathogenesis of neuroinflammatory and neurodegenerative diseases. Metabolites produced by the gut microbiota, particularly short-chain fatty acids (SCFAs), act as key mediators in this bidirectional communication. While the roles of acetate, propionate, and butyrate are well-established, valeric acid (VA), a five-carbon SCFA, is poorly understood. This comprehensive review explores VA as a gut-derived physiological epigenetic modulator, examining its microbial biosynthesis and systemic effects. This review discusses how VA acts as a selective histone deacetylase inhibitor (HDACi), particularly targeting Class I HDACs, to modulate gene expression and exert neuroprotective and anti-inflammatory effects. The analysis compares VA with its pharmacological analog, valproic acid (VPA), a well-known but non-selective HDACi. This comparison highlights how VA’s physiological nature may offer a more targeted and safer intervention. In conclusion, elucidating VA’s role as a microbiome-derived epigenetic regulator would open promising avenues for therapeutic strategies that directly connect gut and CNS health within the GBA. Full article

(This article belongs to the Special Issue Molecular and Cellular Advances in Gut-Brain Axis)

► Show Figures

Figure 1

61 pages, 5195 KB

Open AccessEditor’s ChoiceReview

Precision Oncology: Current Landscape, Emerging Trends, Challenges, and Future Perspectives

by Diane Qiao, Richard C. Wang and Zhixiang Wang

Cells 2025, 14(22), 1804; https://doi.org/10.3390/cells14221804 - 17 Nov 2025

Cited by 16 | Viewed by 9943

Abstract

Precision oncology is broadly defined as cancer prevention, diagnosis, and treatment specifically tailored to the patient based on his/her genetics and molecular profile. In simple terms, the goal of precision medicine is to deliver the right cancer treatment to the right patient, at [...] Read more.

Precision oncology is broadly defined as cancer prevention, diagnosis, and treatment specifically tailored to the patient based on his/her genetics and molecular profile. In simple terms, the goal of precision medicine is to deliver the right cancer treatment to the right patient, at the right dose, at the right time. Precision oncology is the most studied and widely applied subarea of precision medicine. Now, precision oncology has expanded to include modern technology (big data, single-cell spatial multiomics, molecular imaging, liquid biopsy, CRISPR gene editing, stem cells, organoids), a deeper understanding of cancer biology (driver cancer genes, single nucleotide polymorphism, cancer initiation, intratumor heterogeneity, tumor microenvironment ecosystem, pan-cancer), cancer stratification (subtyping of traditionally defined cancer types and pan-cancer re-classification based on shared properties across traditionally defined cancer types), clinical applications (cancer prevention, early detection, diagnosis, targeted therapy, minimal residual disease monitoring, managing drug resistance), lifestyle changes (physical activity, smoking, alcohol consumption, sunscreen), cost management, public policy, and more. Despite being the most developed area in precision medicine, precision oncology is still in its early stages and faces multiple challenges that need to be overcome for its successful implementation. In this review, we examine the history, development, and future directions of precision oncology by focusing on emerging technology, novel concepts and principles, molecular cancer stratification, and clinical applications. Full article

► Show Figures

Figure 1

20 pages, 695 KB

Open AccessEditor’s ChoiceReview

Retinal Neurovascular Coupling: From Mechanisms to a Diagnostic Window into Brain Disorders

by Wen Shen

Cells 2025, 14(22), 1798; https://doi.org/10.3390/cells14221798 - 16 Nov 2025

Cited by 15 | Viewed by 3011

Abstract

Retinal neurovascular coupling reflects the precise coordination between neuronal activity, glial support, and vascular responses, mirroring key neurovascular mechanisms in the brain. This review emphasizes the cellular and molecular processes underlying retinal neurovascular coupling and positions the retina as a sensitive and accessible [...] Read more.

Retinal neurovascular coupling reflects the precise coordination between neuronal activity, glial support, and vascular responses, mirroring key neurovascular mechanisms in the brain. This review emphasizes the cellular and molecular processes underlying retinal neurovascular coupling and positions the retina as a sensitive and accessible model for investigating neurovascular function in the brain. It highlights how parallel neurovascular degeneration in the brain and retina provides critical insights into the pathophysiology of neurodegenerative and vascular disorders. Advances in retinal imaging, including functional optical coherence tomography (fOCT), OCT angiography (OCTA), and functional electrophysiology, offer unprecedented opportunities to detect early neuronal and vascular dysfunction, establishing the retina as a non-invasive biomarker for early detection, disease monitoring, and therapeutic evaluation in Alzheimer’s, Parkinson’s and Huntington’s disease, and stroke. By integrating structural, functional, and mechanistic approaches, the review emphasizes the retina’s potential as a translational platform bridging basic science and clinical applications in neurovascular research. Full article

(This article belongs to the Special Issue Cell Stress and Intervention in Neurological Disease: In Honor of Dr. Jang-Yen Wu for His Long and Distinguished Career in Neuroscience)

► Show Figures

Figure 1

24 pages, 4730 KB

Open AccessEditor’s ChoiceArticle

NeoPAIR-T: Functional Mapping of Neoantigen–TCR Pairs Using a CRISPR-Engineered Jurkat Reporter System

by Koji Nagaoka, Yukari Kobayashi and Kazuhiro Kakimi

Cells 2025, 14(22), 1789; https://doi.org/10.3390/cells14221789 - 14 Nov 2025

Viewed by 2133

Abstract

Targeting mutation-derived neoantigens is a promising strategy for personalized immunotherapies. However, identifying true neoantigens and cognate T cell receptors (TCRs) remains challenging because computational prediction of neoantigen peptides is uncertain and most tumor-infiltrating lymphocytes are bystanders rather than tumor-reactive, necessitating functional validation. Here, [...] Read more.

Targeting mutation-derived neoantigens is a promising strategy for personalized immunotherapies. However, identifying true neoantigens and cognate T cell receptors (TCRs) remains challenging because computational prediction of neoantigen peptides is uncertain and most tumor-infiltrating lymphocytes are bystanders rather than tumor-reactive, necessitating functional validation. Here, we developed NeoPAIR-T (Neoantigen–TCR Pairing Assay using reporter T cells), a functional assay based on co-culture of TCR–T reporter cells and autologous antigen-presenting cells (APCs) to screen neoantigen–TCR pairs. Reporter T cells are Jurkat-derived cells engineered to express a luciferase/eGFP dual reporter, providing quantitative readouts of TCR activation, while APCs are immortalized autologous cells transfected with tandem minigenes (TMGs) encoding predicted neoantigens, bypassing peptide synthesis. NeoPAIR-T also includes TCRα-knockout with targeted knock-in of candidate TCRs at the TCRβ locus to prevent mispairing and enables parallel testing of multiple reporter T cell clones co-cultured with the same APCs for efficient identification of functional pairs. Using lung cancer samples, whole-exome and RNA sequencing predicted 63 candidate peptides assembled into three TMGs. Single-cell RNA/TCR sequencing identified eight TCR clonotypes, introduced into reporter T cells and tested in parallel. Co-culture with TMG-expressing APCs revealed two functional neoantigen–TCR pairs validated by peptide assays (EC₅₀: 10^−9.2–10^−6.7 M). Collectively, NeoPAIR-T streamlines neoantigen–TCR identification for vaccine and TCR-T applications. Full article

(This article belongs to the Section Cellular Immunology)

► Show Figures

Graphical abstract

22 pages, 8835 KB

Open AccessEditor’s ChoiceArticle

High-Frequency Irreversible Electroporation Alters Proteomic Profiles and Tropism of Small Tumor-Derived Extracellular Vesicles to Promote Immune Cell Infiltration

by Kelsey R. Murphy, Kenneth N. Aycock, Spencer Marsh, Liping Yang, Jonathan Hinckley, Aubrie Selmek, Robert Gourdie, Shay Bracha, Rafael V. Davalos, John H. Rossmeisl and Nikolaos G. Dervisis

Cells 2025, 14(22), 1782; https://doi.org/10.3390/cells14221782 - 13 Nov 2025

Cited by 2 | Viewed by 1421

Abstract

High-frequency irreversible electroporation (H-FIRE) is a nonthermal tumor ablation technique that disrupts the blood–brain barrier (BBB) in a focal and reversible manner. However, the mechanisms underlying this disruption remain poorly understood, particularly the role of small tumor-derived extracellular vesicles (sTDEVs) released from ablated [...] Read more.

High-frequency irreversible electroporation (H-FIRE) is a nonthermal tumor ablation technique that disrupts the blood–brain barrier (BBB) in a focal and reversible manner. However, the mechanisms underlying this disruption remain poorly understood, particularly the role of small tumor-derived extracellular vesicles (sTDEVs) released from ablated tumor cells. In this study, we investigate the proteomic and functional alterations of sTDEVs released from F98 glioma and LL/2 Lewis lung carcinoma cells following H-FIRE ablation. Mass spectrometry analysis revealed 108 unique proteins in sTDEVs derived from ablative doses of H-FIRE, which are capable of disrupting the BBB in an in vitro model. Proteomic analysis of TDEVs highlights key changes in pathways related to integrin signaling, Platelet-derived growth factor receptor (PDGFR) signaling, and ubiquitination, which may underline their interactions with brain endothelial cells. These “disruptive” sTDEVs exhibit enhanced tropism for cerebral endothelial cells both in vitro and in vivo, where they persist in the brain longer than sTDEVs released after non-ablative H-FIRE doses. Notably, when introduced into a healthy Fischer rat model, disruptive sTDEVs are associated with increased recruitment of Iba1+ immune cells, suggesting a potential role in modulating post-ablation immune responses. However, despite their altered protein composition, these vesicles do not directly increase BBB permeability in vivo. This study is the first to demonstrate that electroporation-based tumor ablation significantly alters the composition and functionality of tumor-derived extracellular vesicles, potentially influencing the tumor microenvironment post-ablation. These findings have important implications for developing multimodal treatment strategies that combine H-FIRE with systemic therapies to enhance efficacy while managing the peritumoral microenvironment. Full article

(This article belongs to the Special Issue Immune Cell Effect on the Endothelium)

► Show Figures

Figure 1

26 pages, 14048 KB

Open AccessEditor’s ChoiceReview

The Gut Microbiome as a Biomarker and Therapeutic Target of Immune Checkpoint Inhibitors: A Review for Oncologists

by Thiti Susiriwatananont, Panuch Eiamprapaporn, Maria Vazquez Roque, Francis A. Farraye, Adam Perlman and Saranya Chumsri

Cells 2025, 14(22), 1779; https://doi.org/10.3390/cells14221779 - 12 Nov 2025

Cited by 13 | Viewed by 5617

Abstract

Immune checkpoint inhibitors (ICIs) have transformed cancer therapy, yet their benefits remain limited to a subset of patients, underscoring the need for more reliable biomarkers and novel therapeutic strategies. The gut microbiome has emerged as a critical modulator of systemic immunity and a [...] Read more.

Immune checkpoint inhibitors (ICIs) have transformed cancer therapy, yet their benefits remain limited to a subset of patients, underscoring the need for more reliable biomarkers and novel therapeutic strategies. The gut microbiome has emerged as a critical modulator of systemic immunity and a promising determinant of ICI response. Evidence links specific microbial features, taxa, and bioactive metabolites to enhanced antitumor immunity, whereas disruptions, such as antibiotic exposure, are associated with poorer outcomes. Advances in sequencing and multi-omics technologies have provided more profound insights into microbiome-immune crosstalk, though methodological heterogeneity continues to challenge reproducibility. Translational studies demonstrate that microbiome-based intervention, including fecal microbiota transplantation (FMT), biotics supplementation, and engineered microbial strains, can enhance ICI efficacy or mitigate immune-related toxicities. Despite encouraging early clinical signals, broader implementation requires methodological rigor, standardized protocols, and innovative trial designs that account for host and environmental factors. For clinicians, the most immediate strategies involve prudent antibiotic stewardship and patient enrollment in microbiome-focused clinical trials. Overall, the gut microbiome is a promising biomarker and a therapeutic target, representing a new frontier for personalizing immunotherapy and improving patient outcomes in oncology. Full article

(This article belongs to the Special Issue Cellular Mechanisms of Anti-Cancer Therapies)

► Show Figures

Graphical abstract

25 pages, 4735 KB

Open AccessEditor’s ChoiceArticle

Role of Nitric Oxide and Nrf2 to Counteract Vascular Endothelial Dysfunction Induced by Periodontal Pathogens Using HUVECs

by Gunaraj Dhungana, Chethan Sampath, Vineeta Sharma, Olga Korolkova and Pandu R. Gangula

Cells 2025, 14(22), 1777; https://doi.org/10.3390/cells14221777 - 12 Nov 2025

Cited by 4 | Viewed by 2548

Abstract

Background: Polybacterial infections associated with periodontitis are increasingly linked to systemic vascular complications, yet the underlying endothelial mechanisms remain unclear. This study investigated how a consortium of red-complex bacteria (Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola) and orange complex ( [...] Read more.

Background: Polybacterial infections associated with periodontitis are increasingly linked to systemic vascular complications, yet the underlying endothelial mechanisms remain unclear. This study investigated how a consortium of red-complex bacteria (Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola) and orange complex (Fusobacterium nucleatum) affects oxidative stress, inflammation, metabolism, and apoptosis in endothelial cells, and whether L-Sepiapterin [a tetrahydrobiopterin (BH4) precursor via salvage pathway] or bardoxolone methyl (CDDO-Me) [a potent nuclear factor erythroid 2-related factor 2 (Nrf2) activator)] could provide protection. Methods: Human umbilical vein endothelial cells (HUVECs) were infected for 12–72 h and treated with L-Sepiapterin or CDDO-Me. Nitric oxide (NO), BH4, and reactive oxygen species (ROS) levels were quantified, and mRNA expression of key genes regulating nitric oxide synthase activity, antioxidant defense, inflammation (TLR4/NF-κB, cytokines), metabolism (PI3K-AKT-PEA-15), and apoptosis (FAS–caspase pathway) was analyzed. Results: Infection markedly reduced NO and BH4, elevated ROS, activated TLR4/NF-κB and proinflammatory cytokines, disrupted PI3K/AKT signaling, and triggered endothelial apoptosis. Treatments with L-Sepiapterin and CDDO-Me restored NO bioavailability, reduced oxidative and inflammatory responses, normalized metabolic gene expression, and attenuated apoptosis, with CDDO-Me showing more promising effects. This study provides the mechanistic insight linking periodontal polybacterial infection to endothelial dysfunction and metabolic impairment such as diabetes, suggesting that redox-modulating strategies such as L-Sepiapterin and CDDO-Me may help prevent vascular damage associated with periodontal disease. Full article

(This article belongs to the Special Issue Redox Regulation by Nrf2 in Health and Disease)

► Show Figures

Graphical abstract

16 pages, 9263 KB

Open AccessEditor’s ChoiceArticle

Immunohistochemical Characterization and CT-Derived Volume of Epicardial Adipose Tissue in Patients with Coronary Artery Disease

by Matija Furtula, Igor Zivkovic, Slobodan Micovic, Zoran Tabakovic, Gorica Vidovic, Zelimir Antonic, Jelica Vukmirovic, David Savic, Milovan Bojic, Branko Beleslin, Milan Dobric and Jelena Rakocevic

Cells 2025, 14(22), 1760; https://doi.org/10.3390/cells14221760 - 11 Nov 2025

Cited by 3 | Viewed by 3135

Abstract

Background: Epicardial adipose tissue (EAT) is a visceral fat depot surrounding the myocardium. It contributes to coronary artery disease (CAD) through local inflammation, while its metabolic activity, including the expression of uncoupling protein-1 (UCP-1) and incretin receptors (GLP-1R, GIPR), may exert protective effects. [...] Read more.

Background: Epicardial adipose tissue (EAT) is a visceral fat depot surrounding the myocardium. It contributes to coronary artery disease (CAD) through local inflammation, while its metabolic activity, including the expression of uncoupling protein-1 (UCP-1) and incretin receptors (GLP-1R, GIPR), may exert protective effects. The relationship between EAT immunohistochemical features and imaging-derived volume remains unclear. Methods: We prospectively studied 50 patients undergoing cardiac surgery: 25 with CAD undergoing coronary artery bypass grafting and 25 without CAD undergoing valve replacement. EAT samples were immunohistochemically stained for CD3, CD68, MPO, UCP-1, GLP-1R, and GIPR. Preoperative CT was used to quantify EAT volume. Results: Patients with CAD more frequently had higher CD3 immunopositivity compared to the control group (84.0 vs. 58.3%, p = 0.047), with no difference in MPO and CD68 immunoexpression. UCP-1 expression was elevated in CAD patients (p = 0.004), whereas GLP-1R and GIPR immunopositivity were similar. EAT volume did not differ between CAD and non-CAD patients (102.87 cm³ vs. 99.38 cm³, p = 0.964) but correlated modestly with BMI (r_s = 0.325, p = 0.021). UCP-1 and GLP-1R immunopositivity, as well as larger LVEDD (left ventricular end-diastolic diameter), were positively associated with greater EAT volume. Conclusions: EAT in CAD exhibits increased T-cell infiltration and elevated UCP-1 expression, indicating an inflammatory yet metabolically active profile. Larger EAT volume was associated with UCP-1 and GLP-1R expression, underscoring the immunometabolic role of EAT in CAD. Full article

(This article belongs to the Special Issue Ischemic Heart Disease: From Cellular Level to Clinical Approaches)

► Show Figures

Figure 1

23 pages, 1693 KB

Open AccessEditor’s ChoiceArticle

A Bispecific Antibody Blocking Both TSLP and IL-4Rα for the Treatment of Allergic Inflammatory Diseases

by Mingcan Yu, Peng Chen, Ying Jin, Sheng Huang, Hao Jiang, Fulai Zhou, Mark L. Chiu and Di Zhang

Cells 2025, 14(22), 1747; https://doi.org/10.3390/cells14221747 - 7 Nov 2025

Cited by 5 | Viewed by 5475

Abstract

Thymic stromal lymphopoietin (TSLP) works synergistically with Th2 cytokines to regulate infection, inflammation, and metabolic homeostasis. However, their aberrant activities lead to the onset and sustaining of many types of allergic inflammatory diseases. While biologics drug molecules blocking either TSLP or IL-4/IL-13 show [...] Read more.

Thymic stromal lymphopoietin (TSLP) works synergistically with Th2 cytokines to regulate infection, inflammation, and metabolic homeostasis. However, their aberrant activities lead to the onset and sustaining of many types of allergic inflammatory diseases. While biologics drug molecules blocking either TSLP or IL-4/IL-13 show clinical efficacies, the broader effect of simultaneously targeting these cytokines remains to be explored. We generated a bispecific antibody (BsAb) targeting both TSLP and IL-4Rα, which effectively blocked the signaling cascades driven by TSLP, IL-4, and IL-13. The BsAb also neutralized TSLP-driven CD4⁺ T cell proliferation as well as IL-4 and IL-13-driven TF-1 cell proliferation. The BsAb reduced CCL17 release from CD14⁺ monocytes activated by LPS, TSLP, and IL-4 and reduced allergen-induced CCL26 and IL-5 release from co-cultures of PBMC, MRC-5, and A549 cells. In a TSLP/OVA-induced asthma model with transgenic human TSLP, TSLP receptor, IL-4, and IL-4Rα mice, the BsAb reduced every single allergic/inflammatory hallmark, while the single target blockade antibody failed to have such comprehensive effects. Our data suggested that simultaneous blocking of TSLP, IL-4, and IL-13 could offer broader control of allergic inflammation, which could translate to a more effective treatment of related disorders. Full article

(This article belongs to the Section Cellular Immunology)

► Show Figures

Figure 1

18 pages, 5999 KB

Open AccessEditor’s ChoiceArticle

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 7 | Viewed by 1421

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)

► Show Figures

Figure 1

27 pages, 20279 KB

Open AccessEditor’s ChoiceArticle

Novel Taxol-Derivative, STO-1, Induces Selective Anti-Tumor Immunity and Sustained Remission of Glioblastoma Without Triggering Autoimmune Reactions

by Shubhasmita Mohapatra, Adrian Guerrero, Neha Rahman, Khondoker Takia Zaman, Jing Wu, Callistus Onyeagba, Chanyue Hu, Matteo Pellegrini, Jayaram Vankudoth, Seiya Kitamura, Lauren O’Donnell, Youssef Zaim Wadghiri and Probal Banerjee

Cells 2025, 14(21), 1703; https://doi.org/10.3390/cells14211703 - 30 Oct 2025

Cited by 5 | Viewed by 2397

Abstract

Reprogramming of macrophages into the inflammatory state (also known as M1) is currently considered as an effective way of eliminating cancer cells, but systemic deployment of this strategy is likely to induce dangerous autoimmune reactions. Consequently, converting immunosuppressive M2-type macrophages into M1 systemically [...] Read more.

Reprogramming of macrophages into the inflammatory state (also known as M1) is currently considered as an effective way of eliminating cancer cells, but systemic deployment of this strategy is likely to induce dangerous autoimmune reactions. Consequently, converting immunosuppressive M2-type macrophages into M1 systemically is not a safe and effective therapeutic approach against cancer. Through cleavable covalent linking of curcumin to the chemotherapeutic agent Paclitaxel (Taxol), we have created a novel prodrug (STO-1) that, upon intravenous delivery, selectively reprograms tumor-associated microglia and macrophages (TAMs) and eliminates glioblastoma (GBM) without triggering autoimmunity. Demonstrating its therapeutic efficacy, prolonged treatment of six orthotopic GBM-bearing mice with STO-1 resulted in 67% long-term survival, with three surviving mice exhibiting complete tumor clearance and one displaying minimal residual disease, as confirmed by high-resolution ex vivo T2-weighted MRI 85 days after tumor inoculation. In contrast, the vehicle-treated mice displayed extensive intracranial tumors with edema and hemorrhage. Mechanistically, scRNA-seq analysis indicated induction of multiple M1-associated transcripts (ccrl2, cxcl9, ccr2, ccl5) consistent with robust TAMs reprogramming. In striking contrast to the M2⟶M1 reprogramming of TAMs, M1-type macrophages were suppressed in the spleens of STO-1-treated cancer-free mice. Therefore, STO-1 induces selective anti-tumor immunity and GBM elimination without triggering systemic autoimmune reactions. Full article

(This article belongs to the Special Issue Decoding the Tumor-Immune Intersection: Opportunities for Breakthroughs in Cancer Therapies)

► Show Figures

Figure 1

54 pages, 9515 KB

Open AccessEditor’s ChoiceReview

Impact of the ECM on the Mechanical Memory of Cancer Cells

by Claudia Tanja Mierke

Cells 2025, 14(21), 1707; https://doi.org/10.3390/cells14211707 - 30 Oct 2025

Cited by 9 | Viewed by 5394

Abstract

Besides genomic and proteomic analyses of bulk and individual cancer cells, cancer research focuses on the mechanical analysis of cancers, such as cancer cells. Throughout the oncogenic evolution of cancer, mechanical inputs are stored as epigenetic memory, which ensures versatile coding of malignant [...] Read more.

Besides genomic and proteomic analyses of bulk and individual cancer cells, cancer research focuses on the mechanical analysis of cancers, such as cancer cells. Throughout the oncogenic evolution of cancer, mechanical inputs are stored as epigenetic memory, which ensures versatile coding of malignant characteristics and a quicker response to external environmental influences in comparison to solely mutation-based clonal evolutionary mechanisms. Cancer’s mechanical memory is a proposed mechanism for how complex details such as metastatic phenotypes, treatment resistance, and the interaction of cancers with their environment could be stored at multiple levels. The mechanism appears to be similar to the formation of memories in the brain and immune system like epigenetic alterations in individual cells and scattered state changes in groups of cells. Carcinogenesis could therefore be the outcome of physiological multistage feedback mechanisms triggered by specific heritable oncogenic alterations, resulting in a tumor-specific disruption of the integration of the target site/tissue into the overall organism. This review highlights and discusses the impact of the ECM on cancer cells’ mechanical memory during their metastatic spread. Additionally, it demonstrates how the emergence of a mechanical memory of cancer can give rise to new degrees of individuality within the host organism, and a connection to the cancer entity is established by discussing a connection to the metastasis cascade. The aim is to identify common mechanical memory mechanisms of different types of cancer. Finally, it is emphasized that efforts to identify the malignant potency of tumors should go way beyond sequencing approaches and include a functional diagnosis of cancer physiology and a dynamic mechanical assessment of cancer cells. Full article

(This article belongs to the Special Issue Physics of Cancer: How Mechanobiology Drives Cancer Progression)

► Show Figures

Graphical abstract

16 pages, 324 KB

Open AccessEditor’s ChoiceSystematic Review

The Influence of Carbon Nanotubes and Graphene on Immune Cells

by Davide Frumento and Ştefan Ţălu

Cells 2025, 14(21), 1700; https://doi.org/10.3390/cells14211700 - 30 Oct 2025

Cited by 14 | Viewed by 1755

Abstract

Recent studies propose that nanomaterials, either independently or coupled with biomolecular conjugates, have the ability to influence immune activity directly, creating new opportunities for advancing immunotherapies targeting infections and cancer. This review highlights current findings on how functionalized carbon nanotubes (f-CNTs), graphene, and [...] Read more.

Recent studies propose that nanomaterials, either independently or coupled with biomolecular conjugates, have the ability to influence immune activity directly, creating new opportunities for advancing immunotherapies targeting infections and cancer. This review highlights current findings on how functionalized carbon nanotubes (f-CNTs), graphene, and carbon nanohorns interact with immune cells. Among these, f-CNTs have been the most thoroughly explored, though research interest in graphene has been rising steadily. Analysis of published work shows that macrophages are the most frequently studied immune cells (56%), followed by lymphocytes (30%), particularly T cells (22%). Investigations into monocytes and dendritic cells represent 7%, mixed populations such as peripheral blood mononuclear cells make up 6%, and studies on B cells and natural killer (NK) cells remain minimal (1%). Much of the available research has focused on assessing cytotoxicity and compatibility rather than uncovering precise mechanisms of immune modulation. Nonetheless, recent large-scale gene expression profiling has revealed novel immunomodulatory properties of f-CNTs, including stimulation of certain inflammatory signaling pathways. Research on graphene’s immune interactions is still developing. Overall, this review consolidates evidence on the immunological potential of biocompatible f-CNTs and graphene, offering groundwork for their future application in immunology and medicine. Full article

15 pages, 1801 KB

Open AccessEditor’s ChoiceArticle

Impact of Tobacco Smoke Exposure on Male Fertility: An In Vivo Study Using Drosophila melanogaster

by Natasha Gomes de Miranda, Ana Gajeiro, Ana Martins-Bessa and Isabel Gaivão

Cells 2025, 14(21), 1689; https://doi.org/10.3390/cells14211689 - 28 Oct 2025

Cited by 1 | Viewed by 2928

Abstract

Tobacco smoke has been consistently associated with impaired spermatozoa quality in men, including decreased concentration, motility, and increased morphological abnormalities. Key tobacco-related toxins such as nicotine and cadmium induce oxidative stress, leading to DNA damage in germ cells. This study aimed to evaluate [...] Read more.

Tobacco smoke has been consistently associated with impaired spermatozoa quality in men, including decreased concentration, motility, and increased morphological abnormalities. Key tobacco-related toxins such as nicotine and cadmium induce oxidative stress, leading to DNA damage in germ cells. This study aimed to evaluate the effects of tobacco smoke exposure on male fertility using Drosophila melanogaster as an in vivo model. Fertility, fecundity, parental toxicity, lifespan, and spermatozoa morphology were assessed in two strains: the wild-type Oregon K and the DNA repair-deficient mus308. Males were exposed to whole-tobacco smoke in a controlled environment for periods of 0, 1 and 7 min. Tobacco smoke exposure reduced fertility, fecundity, and offspring longevity in both strains. Additionally, spermatozoa from mus308 males exhibited a higher frequency of morphological abnormalities. These findings demonstrate the detrimental impact of tobacco smoke on male reproductive function and suggest increased vulnerability in organisms with impaired DNA repair capacity. Full article

(This article belongs to the Special Issue Drosophila as a Model for Understanding Human Disease)

► Show Figures

Figure 1

26 pages, 12785 KB

Open AccessEditor’s ChoiceArticle

Altered DNA Methylation Pattern Contributes to Differential Epigenetic Immune Signaling in the Upper Respiratory Airway of Unvaccinated COVID-19 Patients

by Melissa Govender, Jyotirmoy Das, Francis R. Hopkins, Cecilia Svanberg, Johan Nordgren, Marie Hagbom, Jonas Klingström, Åsa Nilsdotter-Augustinsson, Yean K. Yong, Vijayakumar Velu, Sivadoss Raju, Johanna Sjöwall, Esaki M. Shankar, Sofia Nyström and Marie Larsson

Cells 2025, 14(21), 1673; https://doi.org/10.3390/cells14211673 - 27 Oct 2025

Cited by 2 | Viewed by 2987

Abstract

SARS-CoV-2 infection remains a global health concern, with its impact on host immune responses not fully understood. In a case–control study, we examined how COVID-19 affects DNA methylation patterns in the upper respiratory airway of hospitalized individuals. DNA methylation arrays were performed on [...] Read more.

SARS-CoV-2 infection remains a global health concern, with its impact on host immune responses not fully understood. In a case–control study, we examined how COVID-19 affects DNA methylation patterns in the upper respiratory airway of hospitalized individuals. DNA methylation arrays were performed on nasopharyngeal samples at inclusion/hospitalization and 6 weeks post-inclusion. We found a distinct DNA methylation pattern in COVID-19 patients compared to healthy controls, identifying 510,099 differentially methylated CpGs. Within the transcription start sites (TSSs) and gene body, COVID-19 patients displayed a higher number of genes/CpGs with elevated methylation levels. Enrichment analysis of TSS-methylated genes revealed effects of SARS-CoV-2 on genes associated with type I interferons, anti-viral and inflammatory responses, and immune functions. Some CpG methylations were transient, and normalized at group level by 6 weeks post-inclusion. Several IFN-regulated genes, including OAS1, OAS3, IFIT3, and MX1, were identified. Among the top regulators were IL17A and ERK1/2, both involved in inflammatory processes. Networks nodes included IGF1 and EGF, associated with processes including tissue repair and activation of immune responses. Overall, our data suggests that COVID-19 can impact the upper airway by modifying gene methylation patterns. This could have implications for conditioning of the airways, how individuals respond to future airway infections, and therapeutic interventions. Full article

(This article belongs to the Special Issue DNA Methylation in Human Health and Disease: From Molecular Mechanisms to Clinical Applications)

► Show Figures

Figure 1

28 pages, 680 KB

Open AccessEditor’s ChoiceReview

Cells Versus Cell-Derived Signals in Cardiac Regenerative Therapy: A Comparative Analysis of Mechanisms and Clinical Evidence

by Julia Soczyńska, Wiktor Gawełczyk, Krzysztof Majcherczyk, Julia Rydzek, Adrian Muzyka, Mateusz Żołyniak and Sławomir Woźniak

Cells 2025, 14(21), 1674; https://doi.org/10.3390/cells14211674 - 27 Oct 2025

Cited by 4 | Viewed by 2600

Abstract

Heart failure (HF) and other cardiac pathologies represent leading causes of hospitalization and mortality worldwide, underscoring the urgent need for effective regenerative therapies. In recent years, considerable research has focused on developing cell-based therapeutic strategies, with stem cells receiving particular attention. Approaches that [...] Read more.

Heart failure (HF) and other cardiac pathologies represent leading causes of hospitalization and mortality worldwide, underscoring the urgent need for effective regenerative therapies. In recent years, considerable research has focused on developing cell-based therapeutic strategies, with stem cells receiving particular attention. Approaches that harness cellular signaling pathways have also been investigated. Experimental studies conducted in both animal models and human subjects have demonstrated that cell-based therapies hold remarkable potential, showing efficacy through improvements in cardiac function, patient quality of life, and overall safety. Clinical data concerning therapies based on cellular signals, while sometimes inconclusive, often yield outcomes comparable to or even superior to those of cell-based interventions. Nonetheless, both approaches face substantial challenges, including the need to ensure reproducibility of results, standardization of therapeutic product preparation, and addressing ethical and regulatory considerations. To translate these promising strategies into clinical practice, a greater number of large-scale, multicenter, and diverse clinical trials will be required. Full article

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

► Show Figures

Figure 1

16 pages, 1716 KB

Open AccessEditor’s ChoiceReview

Alternative Lengthening of Telomeres: A Prognostic Paradox in Cancer

by Ji-Yong Sung

Cells 2025, 14(20), 1613; https://doi.org/10.3390/cells14201613 - 17 Oct 2025

Cited by 6 | Viewed by 4069

Abstract

Telomere maintenance enables unlimited cell proliferation by counteracting telomere erosion. While the majority of tumors activate telomerase, a significant subset—approximately 10–15%—utilizes alternative lengthening of telomeres (ALT), a recombination-based mechanism. ALT-positive cancers are classically associated with genomic instability, anaphase bridges, chromosomal rearrangements, and resistance [...] Read more.

Telomere maintenance enables unlimited cell proliferation by counteracting telomere erosion. While the majority of tumors activate telomerase, a significant subset—approximately 10–15%—utilizes alternative lengthening of telomeres (ALT), a recombination-based mechanism. ALT-positive cancers are classically associated with genomic instability, anaphase bridges, chromosomal rearrangements, and resistance to DNA-damaging therapies. This process is closely associated with genetic instability, which contributes to chromosomal rearrangements and tumor evolution. Consequently, ALT has traditionally been considered an adverse prognostic marker in aggressive malignancies such as osteosarcoma, pancreatic neuroendocrine tumors, and high-grade sarcomas. Paradoxically, recent evidence demonstrates that ALT positivity correlates with improved survival in glioblastoma (GBM) and chondrosarcoma, two tumor types that have historically been regarded as immune-cold and therapeutically intractable. This favorable outcome likely reflects a convergence of factors, including replication stress and DNA damage that impose a fitness cost in slow-growing or metabolically constrained tumors. Loss of ATRX/DAXX, while enabling ALT, further amplifies chromatin fragility, and ALT-mediated instability may paradoxically enhance immunogenicity within immune-quiescent microenvironments. Moreover, ALT-positive cells exhibit unique therapeutic vulnerabilities, particularly to ATR and PARP inhibitors. Together, these observations support a context-dependent model in which ALT functions as a double-edged sword, acting as a driver of malignant aggressiveness in rapidly proliferating cancers while serving as a relative liability in slower-growing, immune-cold tumors. Understanding this duality not only refines prognostic stratification but also opens opportunities for precision oncology. By integrating ALT-specific biomarkers into clinical workflows and exploiting ALT-related DNA repair dependencies, clinicians may transform a once uniformly negative prognostic factor into an actionable therapeutic target. Full article

(This article belongs to the Special Issue Telomere Maintenance and Metabolic Reprogramming in the Tumor Microenvironment)

► Show Figures

Figure 1

19 pages, 2243 KB

Open AccessEditor’s ChoiceReview

Pyruvate Kinase M2 Role in Cardiovascular Repair

by Mohd Rihan, Lior Zangi and Ajit Magadum

Cells 2025, 14(20), 1623; https://doi.org/10.3390/cells14201623 - 17 Oct 2025

Cited by 7 | Viewed by 2877

Abstract

Adult cardiomyocytes (CMs) lose their proliferative capacity shortly after birth, posing a major challenge for cardiac repair following injury such as myocardial infarction (MI). Despite significant advances over the past decade, many strategies for promoting cardiac regeneration have faced limitations, underscoring the need [...] Read more.

Adult cardiomyocytes (CMs) lose their proliferative capacity shortly after birth, posing a major challenge for cardiac repair following injury such as myocardial infarction (MI). Despite significant advances over the past decade, many strategies for promoting cardiac regeneration have faced limitations, underscoring the need to identify novel molecular pathways and targets. Pyruvate kinase muscle isoform 2 (PKM2), a key metabolic enzyme, has emerged as a compelling candidate in this context due to its multifaceted roles in cellular metabolism, proliferation, redox balance, angiogenesis, and master gene regulator in repair. Recent studies highlight the critical function of PKM2 in cardiac repair and regeneration. PKM2 not only promotes the proliferation of CMs but also protects the heart from oxidative stress by redirecting glycolytic intermediates toward the pentose phosphate pathway (PPP), thereby increasing nicotinamide adenine dinucleotide phosphate (NADPH) levels, reducing reactive oxygen species (ROS), and minimizing DNA damage. Moreover, PKM2 interacts with key signaling molecules, including β-catenin, hypoxia-inducible factor 1α (HIF-1a), and checkpoint kinase 1 (CHK1), to promote CM cell cycle reentry, angiogenesis, and enhanced cell survival. Collectively, these multifaceted actions highlight PKM2 as both a metabolic and signaling hub in cardiac repair by promoting myocardial remuscularization, protection, and revascularization and position PKM2 as a promising therapeutic. This review explores the diverse roles of PKM2 in myocardial repair and discusses its potential as a novel avenue for advancing regenerative therapies in cardiovascular medicine. Full article

(This article belongs to the Special Issue Cellular and Molecular Mechanisms of Heart Diseases)

► Show Figures

Figure 1

19 pages, 4150 KB

Open AccessEditor’s ChoiceArticle

Cisplatin-Loaded M1 Macrophage-Derived Vesicles Have Anti-Cancer Activity in Osteosarcoma

by Namrata Anand, Joseph Robert McCorkle, David S. Schweer, Lan Li, Kristen S. Hill, Melissa A. Fath, Derek B. Allison, Christopher L. Richards and Jill M. Kolesar

Cells 2025, 14(20), 1616; https://doi.org/10.3390/cells14201616 - 17 Oct 2025

Cited by 9 | Viewed by 3639

Abstract

Osteosarcoma (OS) is a relatively rare bone malignancy that primarily affects children and young adults and is associated with significant morbidity and mortality. Cisplatin is a mainstay of treatment, but its efficacy is limited by off-target toxicities. Immunotherapy is not effective due to [...] Read more.

Osteosarcoma (OS) is a relatively rare bone malignancy that primarily affects children and young adults and is associated with significant morbidity and mortality. Cisplatin is a mainstay of treatment, but its efficacy is limited by off-target toxicities. Immunotherapy is not effective due to a poor antigenic tumor microenvironment. Here, we address these challenges by using manufactured M1 macrophage-derived vesicles (MVs) loaded with cisplatin. Human blood and mouse RAW 264.7 M1 macrophages were used to prepare empty (E-MVs) and cisplatin-loaded MVs (C-MVs). Human OS cell lines were used in vitro and in a tibia xenograft mouse model to evaluate the anti-cancer and immune-stimulating abilities of MVs. C-MVs had lower IC50s but equivalent DNA damage in OS cell lines when compared with free cisplatin. E-MVs and C-MVs were observed to accumulate in the tumor in OS tumor-bearing mice. C-MVs significantly reduced tumor burden and prolonged survival in a mouse model of OS. Animals dosed with free cisplatin experienced weight loss and renal and hepatic toxicity, while equivalent doses of C-MVs did not cause these effects. In addition, both E-MVs and C-MVs showed immunomodulation of the tumor microenvironment with a significant increase in the M1/M2 macrophages ratio (7-fold and 22-fold, respectively) and increased levels of TNF-α in serum (1.8-fold and 2.1-fold, respectively) compared to control mice. Collectively, these experiments support further development of C-MVs for the treatment of OS. Full article

(This article belongs to the Special Issue Advances in Smart and Innovative Nanoparticles for Precision Tumor Therapy: Design, Delivery, Immune Modulation, and Clinical Translation)

► Show Figures

Graphical abstract

17 pages, 23793 KB

Open AccessEditor’s ChoiceArticle

Dental Pulp Stem Cell-Derived Organoids: Advancing the Development of 3D Structures

by Loreto Lancia, Fanny Pulcini, Emanuela Mari, Luca Piccoli, Leda Assunta Biordi, Luciano Mutti, Claudio Festuccia, Giovanni Luca Gravina, Vincenzo Mattei, Annunziata Mauro, Valentina Notarstefano and Simona Delle Monache

Cells 2025, 14(20), 1603; https://doi.org/10.3390/cells14201603 - 15 Oct 2025

Cited by 3 | Viewed by 1884

Abstract

Two-dimensional cell cultures are crucial research tools, and they have been widely used, although they are not completely representative of biological processes in vivo due to the lack of tissue architecture and complexity. Recent advances in organoid technology have addressed these limitations and [...] Read more.

Two-dimensional cell cultures are crucial research tools, and they have been widely used, although they are not completely representative of biological processes in vivo due to the lack of tissue architecture and complexity. Recent advances in organoid technology have addressed these limitations and are revolutionizing the tools available for in vitro culture. Although there are no unified protocols for generating organoids, they can be obtained with various techniques, leading to cell aggregation by promoting cell adhesion. This work aims to generate and characterise organoid models of dental pulp from dental pulp stem cells (DPSCs), a type of mesenchymal stem/stromal cells known for their high regenerative potential and ease of accessibility, to establish a model for translational studies. The organoids were subjected to osteogenic differentiation conditions. Cell viability was evaluated using a CCK-8 assay, while osteogenic morphology and mineralization were confirmed by Alizarin red analysis, Raman microspectroscopy, and by immunofluorescence for the lineage markers expression. The Alizarin red analysis indicated a higher presence of calcium phosphate deposits in the differentiated organoids than in the control group (CTR). These results were confirmed by spectral profiles obtained using Raman microspectroscopy, which were attributable to a hydroxyapatite-based biomaterial. Immunofluorescence analysis also revealed increased expression of odonto/osteogenic markers (RUNX and OSX), alongside reduced expression of stemness markers. In conclusion, the organoids appeared to have successfully differentiated into an osteogenic lineage, forming a mineralized matrix containing hydroxyapatite and showing increased expression of relevant lineage markers. Full article

(This article belongs to the Special Issue 3D Cultures and Organ-on-a-Chip in Cell and Tissue Cultures)

► Show Figures

Figure 1

19 pages, 694 KB

Open AccessEditor’s ChoiceReview

Evolutionary Perspective of Nonclassical MHC Class I and Innate-like T Cells Relevance in Immune Surveillance

by Jacques Robert and Elnaz Najafi-Majd

Cells 2025, 14(20), 1592; https://doi.org/10.3390/cells14201592 - 14 Oct 2025

Cited by 4 | Viewed by 2242

Abstract

Unlike conventional T cells, which express a highly diverse repertoire of dimeric αβ T-cell receptors (TCRs) restricted by classical, polymorphic MHC class I molecules (MHC-Ia), a distinct group of T cells—collectively termed “innate-like T (iT) cells”—exhibits limited TCR diversity and depends instead on [...] Read more.

Unlike conventional T cells, which express a highly diverse repertoire of dimeric αβ T-cell receptors (TCRs) restricted by classical, polymorphic MHC class I molecules (MHC-Ia), a distinct group of T cells—collectively termed “innate-like T (iT) cells”—exhibits limited TCR diversity and depends instead on nonclassical, nonpolymorphic MHC class I molecules (MHC-Ib) for their development and function. While mounting evidence supports the role of iT cells as pivotal regulators and effectors in both innate and adaptive immune responses, many aspects of their biology remain incompletely understood. In humans, iT cells represent a significant fraction of the total T cell population, and evolutionarily conserved subsets have also been identified in other mammals and amphibians. Moreover, the expanding catalog of nonpolymorphic MHC-Ib genes and lineages—distinct from polymorphic MHC-Ia genes—across jawed vertebrate genomes suggests a broader and potentially more integral role for MHC-Ib molecules in T cell function and immune surveillance. In this review, we explore the immunological significance of MHC-Ib molecules and iT cells through an evolutionary lens, highlighting recent advances that shed light on their contributions to immune homeostasis and defense. Full article

(This article belongs to the Special Issue T Cells in Inflammation and Cancer)

► Show Figures

Graphical abstract

26 pages, 4528 KB

Open AccessEditor’s ChoiceArticle

House Dust Mite Nebulization Drives Alarmin and Complement Activation in a Murine Tracheal Air–Liquid Interface Culture System

by Janti Haj Ahmad, Philip Einwohlt, Mareike Ohms, Doris Wilflingseder and Jörg Köhl

Cells 2025, 14(20), 1598; https://doi.org/10.3390/cells14201598 - 14 Oct 2025

Cited by 3 | Viewed by 1772

Abstract

Air–liquid interface (ALI) cultures offer a physiologically relevant in vitro model of the airway epithelium (AE), capable of recapitulating key structural and functional features observed in vivo. In this study, we established and validated a murine ALI culture system comprising pseudostratified epithelia with [...] Read more.

Air–liquid interface (ALI) cultures offer a physiologically relevant in vitro model of the airway epithelium (AE), capable of recapitulating key structural and functional features observed in vivo. In this study, we established and validated a murine ALI culture system comprising pseudostratified epithelia with functional tight junctions, ciliated cells and goblet cells. To assess their innate immune functions, we designed and 3D-printed an autoclavable aerosol deposition chamber, which allowed us to expose differentiated AE cultures to house dust mite (HDM) allergen. Upon HDM exposure, AE cells mounted a time-dependent innate immune response characterized by the secretion of complement component C3, the generation of its active cleavage products C3a and increased expression of C3aR and C5aR1. This was associated with increased intracellular TSLP and IL-25 production and TSLP release in AE cells. Progressive loss of tight junction integrity and reduced transepithelial electrical resistance (TEER) demonstrated epithelial susceptibility to allergen protease-induced cell damage. Together, we established a murine ALI system preserving airway epithelial architecture and a nebulization system to study innate immune activation of AE cells in response to HDM mimicking the initial phase of allergen sensitization. More generally, we described a powerful and accessible platform for studying epithelial-driven mechanisms in murine airway immune responses. Full article

(This article belongs to the Special Issue Novel Insights into Molecular Mechanisms and Therapy of Asthma)

► Show Figures

Figure 1

34 pages, 8250 KB

Open AccessEditor’s ChoiceReview

From Cytokines to Biomarkers: Mapping the Immunopathology of Inflammatory Bowel Disease

by Sarah Baum, Kamron Hamedi, Caroline Loftus, Gannett Loftus, Emily-Rose Zhou and Sergio Arce

Cells 2025, 14(20), 1589; https://doi.org/10.3390/cells14201589 - 13 Oct 2025

Cited by 16 | Viewed by 5574

Abstract

Inflammatory bowel disease (IBD) is a chronic immune-mediated condition of the gastrointestinal tract, characterized by dysregulated inflammatory responses throughout the gastrointestinal tract. It includes two major phenotypes, Crohn’s disease (CD) and ulcerative colitis (UC), which present with varying gastrointestinal and systemic symptoms. The [...] Read more.

Inflammatory bowel disease (IBD) is a chronic immune-mediated condition of the gastrointestinal tract, characterized by dysregulated inflammatory responses throughout the gastrointestinal tract. It includes two major phenotypes, Crohn’s disease (CD) and ulcerative colitis (UC), which present with varying gastrointestinal and systemic symptoms. The pathophysiology of IBD is multifactorial including genetic predisposition, mucosal and epithelial dysfunction, environmental injury, and both innate and adaptive immune response abnormalities. Several predisposing genetic factors have been associated with IBD explaining the strong hereditary risk for both CD and UC. For example, Caspase Recruitment Domain 9 (CARD9) variant rs10781499 increases risk for IBD, while other variants are specific to either CD or UC. CD is related to loss-of-function mutations in the nucleotide oligomerization domain containing the protein 2 (NOD2) gene and Autophagy-Related 16-like 1 (ATG16L1) gene. UC risk is increased particularly in Chinese populations by the A-1661G polymorphism of the Cytotoxic T-lymphocyte antigen 4 (CTLA-4) gene. This abnormal CTLA-4 interferes with B- and T-cell responses causing predisposition to autoimmune conditions. Previous studies suggested that IBD results from breakdown of the adaptive immune system, primarily of T-cells. However, new evidence suggests that a primary breakdown of the innate immune system in both CD and UC increases susceptibility to invasion by viruses and bacteria, with a compensatory overactivation of the adaptive immune system as a result. When this viral and microbial invasion continues, further damage is incurred, resulting in a downward cycle of further cytokine activation and epithelial damage. Released biomarkers also affect the permeability of the epithelial membrane, including lactoferrin, nitric oxide (NO), myeloperoxidase (MPO) and its activation of hypochlorous acid, matrix metalloproteinases (MMPs), especially MMP-9, omentin-1, and others. Increased macrophage and dendritic cell dysfunction, increased neutrophil activity, increased numbers of innate lymphoid cells, increased T-cells with decreased regulatory T-cells (Tregs), and changes in B-cell populations and immunoglobulin (Ig) functions are all associated with IBD. Finally, treatment of IBD has typically consisted of medical management (e.g., aminosalicylates and corticosteroids) and lifestyle modification, and surgical intervention in extreme cases. New classes of medications with more favorable side effect profiles include anti-integrin antibodies, vedolizumab, etrolizumab, and carotegrast methyl. Additionally, fecal microbiota transplant (FMT) is a newer area of research for treatment of IBD along with TNF-blockers, JAK inhibitors, and S1PR modulators. However, expense and long preparation time have limited the usefulness of FMT. Full article

(This article belongs to the Special Issue Molecular and Signaling Pathways in Pathological Changes of Esophageal, Gastric and Intestinal Epithelial Cells and Methods of Cell Protective Treatment)

► Show Figures

Figure 1

Journal Menu

Journal Browser

Editor’s Choice Articles

Further Information

Guidelines

MDPI Initiatives

Follow MDPI