Cells

12 pages, 2080 KB

Open AccessArticle

The Molecular Mechanism of PDE1 Regulation

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

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

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

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

► Show Figures

Figure 1

29 pages, 4096 KB

Open AccessArticle

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

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

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

Abstract

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

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

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

► Show Figures

Figure 1

29 pages, 1422 KB

Open AccessReview

Functions of TIP60/NuA4 Complex Subunits in Cell Differentiation

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

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

Abstract

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

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

► Show Figures

Figure 1

24 pages, 4341 KB

Open AccessArticle

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

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

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

Abstract

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

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

► Show Figures

Figure 1

19 pages, 1312 KB

Open AccessReview

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

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

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

Abstract

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

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

► Show Figures

Figure 1

18 pages, 5999 KB

Open AccessArticle

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

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

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

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

13 pages, 14281 KB

Open AccessArticle

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

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

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

Abstract

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

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

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

► Show Figures

Figure 1

18 pages, 4533 KB

Open AccessArticle

Acute Kidney Injury Induces Lung Damage via Mitochondrial DAMPs by Activating TREM-1 and cGAS-STING Pathways

by Zhi Tian, Runze Ni, Nadezhda N. Zheleznova, Diane Allen-Gipson, Lei Wang, Vijay Subramanian, Kiran Dhanireddy, Sarah Y. Yuan, Nohely Hernandez Soto, Jose D. Herazo-Maya, Kristof Williams, Isabella Lozonschi, Andrew Bedard, Gabrielle Morrison and Ruisheng Liu

Cells 2025, 14(21), 1716; https://doi.org/10.3390/cells14211716 (registering DOI) - 31 Oct 2025

Abstract

Acute kidney injury (AKI) is a leading cause of distant organ dysfunction among critically ill patients. Mitochondrial dysfunction is considered a key factor driving the damage after renal ischemia–reperfusion (IR) injury. Damaged mitochondria release mitochondrial damage-associated molecular patterns (mtDAMPs) into the cytosol, which [...] Read more.

Acute kidney injury (AKI) is a leading cause of distant organ dysfunction among critically ill patients. Mitochondrial dysfunction is considered a key factor driving the damage after renal ischemia–reperfusion (IR) injury. Damaged mitochondria release mitochondrial damage-associated molecular patterns (mtDAMPs) into the cytosol, which initiate a systemic inflammatory response. To better understand the underlying mechanism, mice were challenged with 30 min of bilateral renal ischemia followed by 24 h of reperfusion. The cytokine profiling in mouse lung tissues revealed that TREM-1 was significantly increased. Western Blot (WB) analysis demonstrated that the cGAS and STING pathway was increased in AKI mice. Transmission electron microscopy (TEM) images indicated that the mtDAMPs were released from damaged kidney mitochondria. Injection of mtDAMPs into mice induced an inflammatory response in the lungs similar to that induced by AKI. Mouse macrophages and lung epithelial cells were utilized to verify if inhibition of the TREM-1 and cGAS-STING pathways reduces mtDAMP-induced lung injury. Electric Cell-substrate Impedance Sensing (ECIS) results demonstrated that inhibiting the TREM-1 and cGAS-STING pathways significantly increased cell proliferation and migration while reducing mtDAMP-induced cytotoxicity. In conclusion, our findings suggest that targeting TREM-1 and cGAS-STING has the potential to attenuate acute lung injury in IR-AKI. Full article

(This article belongs to the Section Tissues and Organs)

21 pages, 2847 KB

Open AccessArticle

Evaluation of Rat Testicular Cell Populations in Experimental Condition of Diabetes Induced in Early Postnatal Life

by Ekaterina Pavlova, Rosen Ivanov, Desislava Abadjieva, Yordanka Gluhcheva, Emilia Petrova, Ivelin Vladov, Emilia Lakova and Nina Atanassova

Cells 2025, 14(21), 1714; https://doi.org/10.3390/cells14211714 (registering DOI) - 31 Oct 2025

Abstract

Diabetes mellitus (DM) causes male infertility through the suppression of spermatogenesis and testosterone biosynthesis. The impact of DM on male reproduction has mainly been investigated in adulthood, therefore we aimed to study the developmental effects of DM, induced in early life, on testicular [...] Read more.

Diabetes mellitus (DM) causes male infertility through the suppression of spermatogenesis and testosterone biosynthesis. The impact of DM on male reproduction has mainly been investigated in adulthood, therefore we aimed to study the developmental effects of DM, induced in early life, on testicular cell population and fertility. Neonatal (NDM) and prepubertal DM (PDM) were induced in immature rats by streptozotocin administration on day 1 or day 10, respectively. Germ (GCs) and somatic cells (Sertoli—SCs and Leydig cells—LCs) were counted in pubertal (25 day) and post-pubertal (45 day) rats in tandem with the measurement of serum testosterone levels and the protein expression of androgen receptor. Glucose levels were higher in PDM than in NDM. Incomplete spermatogenesis and reduced GC number were found in PDM but not in NDM. LC number, testosterone, and luteinizing hormone (LH) levels were differently altered by both types of DM with a pronounced negative impact of PDM. Protein expression of androgen receptor in SCs was altered only in PDM. Reduced sperm concentration and motility was found in both groups. Thus, our results provide new insights into different mechanisms of action of PDM and NDM on developing germ cells that involved disturbances in androgen production by Leydig cells and androgen action in Sertoli cells. Full article

(This article belongs to the Section Reproductive Cells and Development)

14 pages, 1555 KB

Open AccessArticle

Ladarixin Potential over the Effects of IL-8 and of Serum from Patients with Abdominal Aortic Aneurysm on Human Aortic Cells

by Lucia Spartano, Maria Lombardi, Vincenzo Ardita, Roberto Chiesa, Andrea Aramini, Marcello Allegretti, Domenico Baccellieri, Lidia De Filippis and Chiara Foglieni

Cells 2025, 14(21), 1713; https://doi.org/10.3390/cells14211713 (registering DOI) - 31 Oct 2025

Abstract

Early cellular alterations in abdominal aortic aneurysm (AAA) are scarcely investigated. Aortic remodeling inflammation-related suggested the CXCR2/CXCL1/IL-8 axis as a therapeutic target. This study investigates CXCR1/CXCR2 antagonism in primary human aortic endothelial (HAOEC) and smooth muscle cells (HAOSMC) conditioned with IL-8 or serum [...] Read more.

Early cellular alterations in abdominal aortic aneurysm (AAA) are scarcely investigated. Aortic remodeling inflammation-related suggested the CXCR2/CXCL1/IL-8 axis as a therapeutic target. This study investigates CXCR1/CXCR2 antagonism in primary human aortic endothelial (HAOEC) and smooth muscle cells (HAOSMC) conditioned with IL-8 or serum from patients with AAA (sPT). Ladarixin (10 μM Lad or 25 μM) served as an inhibitor. Readouts included RT-qPCR for CXCL1, CXCL8, CXCR2, MMP9, NFKB1, and VEGF-A; zymography for MMP9 activity confocal microscopy for F-actin and mitochondria; NADPH/NADH diaphorase histochemistry for redox activity; and ATP assay. In HAOEC, IL-8 downregulated CXCR2, increased MMP9 activity, and induced cytoskeletal and mitochondria disorganization without altering NADH/NADPH diaphorases but increasing ATP release. At concentration of 10 μM Lad rescued cell organization and gene expression. sPT upregulated CXCL8, CXCR2, and MMP9, decreased NADH/NADPH diaphorases, and altered cytoskeleton and mitochondria organization in HAOEC. At concentration of 10 μM Lad (partially) and 25 μM Lad reverted gene upregulation and mitochondria distribution; both doses increased diaphorase and released ATP. HAOSMC were scantily susceptible to IL-8 and weakly responsive to sPT, slightly upregulating CXCR2 and VEGF-A but increasing proMMP9 gelatinolysis. Ladarixin recovered proMMP9 activity and modulated CXCL1. AAA-like vascular cell alterations involve multiple inflammatory factors and are modulable by inhibition of IL-8 receptors. The results underline careful dose calibration. Full article

(This article belongs to the Special Issue Advances in the Cellular and Molecular Mechanisms of Cardiovascular Diseases)

► Show Figures

Figure 1

19 pages, 3032 KB

Open AccessArticle

Dual ROCK1/2–MYLK4 Kinase Inhibition Preserves Visual Function in a Rat Model of Neuromyelitis Optica Spectrum Disorder Optic Neuritis

by Chin-Te Huang, Monir Hossen, Tu-Wen Chen, Chih-Wei Fu, Yi-Hsun Chen, Tzu-Lun Huang and Rong-Kung Tsai

Cells 2025, 14(21), 1712; https://doi.org/10.3390/cells14211712 (registering DOI) - 31 Oct 2025

Abstract

Background: Neuromyelitis optica spectrum disorder (NMOSD) causes severe optic nerve (ON) inflammation and vision loss. Current treatments remain limited, prompting exploration of new therapeutic strategies. This study evaluated the efficacy of ITRI-E-(S)4046 (ITRI-ES), a dual ROCK1/2 and MYLK4 kinase inhibitor, in a [...] Read more.

Background: Neuromyelitis optica spectrum disorder (NMOSD) causes severe optic nerve (ON) inflammation and vision loss. Current treatments remain limited, prompting exploration of new therapeutic strategies. This study evaluated the efficacy of ITRI-E-(S)4046 (ITRI-ES), a dual ROCK1/2 and MYLK4 kinase inhibitor, in a rat model of NMOSD optic neuritis. Methods: NMOSD-like optic neuritis was induced in rats by applying NMOSD patient serum-soaked sponges around the ON. Rats received intravitreal injections of either 0.2% ITRI-ES, phosphate-buffered saline (PBS), or intraperitoneal methylprednisolone (MP). Visual function was assessed using flash visual-evoked potentials (fVEP). Retinal ganglion cell (RGC) survival and apoptosis were quantified using FluoroGold retrograde labeling and TUNEL assay. ON inflammation and demyelination were evaluated via immunohistochemistry and Western blot analysis of aquaporin-4 (AQP4), myelin basic protein (MBP), glial fibrillary acidic protein (GFAP), and inflammatory markers. Results: ITRI-ES significantly preserved visual function, restoring fVEP amplitudes (~36 μV vs. ~21 μV in PBS-treated, p < 0.05) and RGC density (~85% of normal vs. ~37% PBS). RGC apoptosis was reduced (~2.3-fold lower vs. PBS, p < 0.05). PBS-treated rats showed decreased AQP4 and MBP (2.5–2.8-fold vs. sham) and increased GFAP (2.8-fold). ITRI-ES maintained higher AQP4 (~3.5-fold) and MBP (~1.5-fold) levels, suppressed GFAP (~5.5-fold vs. PBS), reduced NF-κB, IL-1β, TNF-α, microglia activation, and macrophage infiltration, and increased anti-inflammatory Arg1 and CD206 markers (~3-fold vs. PBS). Conclusions: ITRI-ES alleviates optic nerve inflammation, preserves retinal integrity, and maintains visual function in NMOSD-associated optic neuritis, underscoring kinase inhibition as a promising therapeutic strategy. Full article

(This article belongs to the Section Cells of the Nervous System)

► Show Figures

Figure 1

41 pages, 3718 KB

Open AccessArticle

Behavioral Balance in Tryptophan Turmoil: Regional Metabolic Rewiring in Kynurenine Aminotransferase II Knockout Mice

by Ágnes Szabó, Zsolt Galla, Eleonóra Spekker, Diána Martos, Mónika Szűcs, Annamária Fejes-Szabó, Ágnes Fehér, Keiko Takeda, Kinuyo Ozaki, Hiromi Inoue, Sayo Yamamoto, Péter Monostori, József Toldi, Etsuro Ono, László Vécsei and Masaru Tanaka

Cells 2025, 14(21), 1711; https://doi.org/10.3390/cells14211711 (registering DOI) - 31 Oct 2025

Abstract

Background: Cognitive, emotional, and social impairments are pervasive across neuropsychiatric conditions, where alterations in the tryptophan (Trp)–kynurenine pathway and its product kynurenic acid (KYNA) from kynurenine aminotransferases (KATs) have been linked to Alzheimer’s disease, Parkinson’s disease, depression, and post-traumatic stress disorder. In novel [...] Read more.

Background: Cognitive, emotional, and social impairments are pervasive across neuropsychiatric conditions, where alterations in the tryptophan (Trp)–kynurenine pathway and its product kynurenic acid (KYNA) from kynurenine aminotransferases (KATs) have been linked to Alzheimer’s disease, Parkinson’s disease, depression, and post-traumatic stress disorder. In novel CRISPR/Cas9-engineered KAT II knockout (aadat^−/− also known as kat2^−/−) mice, we observed despair-linked depression-like behavior with peripheral excitotoxicity and oxidative stress. KAT II’s role and its crosstalk with serotonin, indole-pyruvate, and tyrosine–dopamine remain unclear. It is unknown whether deficits extend to cognitive, emotional, motor, and social domains or whether brain tissues mirror peripheral stress. Objectives: Delineate domain-wide behaviors, brain oxidative/excitotoxic profiles, and pathway interactions attributable to KAT II. Results: Behavior was unchanged across strains. kat2^−/− deletion remodeled Trp metabolic pathways: 3-hydroxykynurenine increased, xanthurenic acid decreased, KYNA fell in cortex and hippocampus but rose in striatum, quinaldic acid decreased in cerebellum and brainstem. These region-specific changes indicate metabolic stress across the brain and align with higher oxidative load and signs of excitotoxic pressure. Conclusions: Here, we show that KAT II deletion reshapes regional Trp metabolism and amplifies oxidative and excitotoxic imbalance. Although domain-wide behavioral measures, spanning cognition, sociability, and motor coordination, remained largely unchanged, these neurochemical alterations signify a latent emotional bias rather than overt depressive-like behavior. This work, therefore, refines prior findings by delineating KAT II–linked biochemical vulnerability as a potential substrate for stress-reactive affective dysregulation. Full article

(This article belongs to the Special Issue Kynurenic Acid: Unveiling Its Impact on the Brain’s Neurotransmission and Neurological Disorders)

24 pages, 7280 KB

Open AccessArticle

Cellular and Molecular Effects of Targeting the CBP/β-Catenin Interaction with PRI-724 in Melanoma Cells, Drug-Naïve and Resistant to Inhibitors of BRAF^V600 and MEK1/2

by Anna Gajos-Michniewicz, Michal Wozniak, Katarzyna Anna Kluszczynska and Malgorzata Czyz

Cells 2025, 14(21), 1710; https://doi.org/10.3390/cells14211710 (registering DOI) - 31 Oct 2025

Abstract

Targeted therapies, including treatment with inhibitors of BRAF^V600 and MEK kinases, have improved outcomes in advanced melanoma. However, most patients relapse due to acquired resistance, underscoring the need for new drug targets. This study evaluated PRI-724, a CBP/β-catenin inhibitor, in patient-derived drug-naïve [...] Read more.

Targeted therapies, including treatment with inhibitors of BRAF^V600 and MEK kinases, have improved outcomes in advanced melanoma. However, most patients relapse due to acquired resistance, underscoring the need for new drug targets. This study evaluated PRI-724, a CBP/β-catenin inhibitor, in patient-derived drug-naïve melanoma cells and their trametinib- or vemurafenib-resistant counterparts. While PRI-724 has demonstrated efficacy in preclinical models and clinical trials in different cancer types, its potential in melanoma has not been previously assessed. We found that treatment with PRI-724 downregulated survivin and other CBP/β-catenin target proteins, reduced invasiveness, and induced apoptosis in drug-naïve and trametinib- and vemurafenib-resistant cells. Trametinib-resistant melanoma cells showed the greatest sensitivity to PRI-724, indicating that CBP/β-catenin transcriptional activity may represent a new therapeutic vulnerability. Transcriptomic and immunoblotting analyses revealed the highest survivin levels in vemurafenib-resistant cells, which may underlie their reduced responsiveness to PRI-724. Bioinformatic analyses (TCGA and GSE50509) confirmed that a high survivin level predicts poor prognosis and reduced response to treatment. The results of the study point to the potential of PRI-724 as a chemotherapeutic agent for the treatment of melanoma. Its efficacy might depend on CBP/β-catenin transcriptional activity in melanoma cells, and further evaluation of this signaling with survivin as a biomarker is therefore warranted. Full article

23 pages, 673 KB

Open AccessReview

Calcium Dynamics in Astrocyte-Neuron Communication from Intracellular to Extracellular Signaling

by Agnieszka Nowacka, Maciej Śniegocki and Ewa A. Ziółkowska

Cells 2025, 14(21), 1709; https://doi.org/10.3390/cells14211709 (registering DOI) - 31 Oct 2025

Abstract

Astrocytic calcium signaling is a central mechanism of neuron-glia communication that operates across multiple spatial and temporal scales. Traditionally, research has focused on intracellular Ca²⁺ oscillations that regulate gliotransmitter release, ion homeostasis, and metabolic support. Recent evidence, however, reveals that extracellular calcium [...] Read more.

Astrocytic calcium signaling is a central mechanism of neuron-glia communication that operates across multiple spatial and temporal scales. Traditionally, research has focused on intracellular Ca²⁺ oscillations that regulate gliotransmitter release, ion homeostasis, and metabolic support. Recent evidence, however, reveals that extracellular calcium ([Ca²⁺]o) is not a passive reservoir but a dynamic signaling mediator capable of influencing neuronal excitability within milliseconds. Through mechanisms such as calcium-sensing receptor (CaSR) activation, ion channel modulation, surface charge effects, and ephaptic coupling, astrocytes emerge as active partners in both slow and rapid modes of communication. This dual perspective reshapes our understanding of brain physiology and disease. Disrupted Ca²⁺ signaling contributes to network instability in epilepsy, synaptic dysfunction in Alzheimer’s and Parkinson’s disease, and impaired maturation in neurodevelopmental disorders. Methodological advances, including Ca²⁺-selective microelectrodes, genetically encoded extracellular indicators, and computational modeling, are beginning to uncover the richness of extracellular Ca²⁺ dynamics, though challenges remain in achieving sufficient spatial and temporal resolution. By integrating classical intracellular pathways with emerging insights into extracellular signaling, this review highlights astrocytes as central architects of the ionic landscape. Recognizing calcium as both an intracellular messenger and an extracellular signaling mediator provides a unifying framework for neuron–glia interactions and opens new avenues for therapeutic intervention. Full article

► Show Figures

Figure 1

25 pages, 5654 KB

Open AccessArticle

Six1-Eya1 Axis Governs Myofiber Remodeling and Fibrosis in Extraocular Myopathy: Insights from Single-Cell RNA Sequencing and Mesenchymal Stem Cell Therapy in Thyroid Eye Disease

by Hyun-Ah Shin, Mira Park, Hey Jin Lee, Jong Hyun Moon, Jasvinder Paul Banga and Helen Lew

Cells 2025, 14(21), 1708; https://doi.org/10.3390/cells14211708 (registering DOI) - 31 Oct 2025

Abstract

Thyroid eye disease (TED) is an autoinflammatory condition characterized by fibrosis in orbital fat and extraocular muscles, primarily driven by TSH receptor antibodies and inflammatory cytokines. While research has predominantly focused on the involvement of fat tissue, the understanding of myopathy in TED [...] Read more.

Thyroid eye disease (TED) is an autoinflammatory condition characterized by fibrosis in orbital fat and extraocular muscles, primarily driven by TSH receptor antibodies and inflammatory cytokines. While research has predominantly focused on the involvement of fat tissue, the understanding of myopathy in TED remains limited. This study developed a TED mouse model and isolated myoblasts from both control individuals and TED patients for analysis. Single-cell RNA sequencing was used to investigate myofiber type changes in TED and their alterations following treatment with human-derived mesenchymal stem cells. Key regulatory genes involved in myofiber differentiation and fibrosis in myofibroblasts were identified, and their expression balance was validated in myoblasts derived from both normal individuals and TED patients. Our analysis revealed a disease-associated shift in myofiber types and identified Six1 and Eya1 as central regulators of myofiber differentiation and fibrosis suppression. These regulatory effects were validated in primary myoblasts isolated from both control and TED patients. Collectively, our findings uncover a novel role for the Six1/Eya1 axis in modulating muscle remodeling and fibrosis in TED and provide a foundation for the development of targeted therapies for TED-associated myopathy. Full article

► Show Figures

Figure 1

54 pages, 9515 KB

Open AccessReview

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

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

Figure 1

13 pages, 3727 KB

Open AccessArticle

Pulsatilla Saponin D Suppresses Proliferation and Induces Apoptosis in Human Prostatic Cells

by Yuzhong Chen, Ping Zhou, Yangtao Jin, Dongyan Huang, Xin Su, Congcong Shao, Juan Jiang, Rongfu Yang and Jianhui Wu

Cells 2025, 14(21), 1706; https://doi.org/10.3390/cells14211706 - 30 Oct 2025

Abstract

The growing global aging population is contributing to an increasing burden of benign prostatic hyperplasia (BPH), highlighting the need for novel, highly effective and low-toxicity therapies. In light of its well-documented anti-inflammatory and anti-tumor properties, we investigated the potential of the natural product [...] Read more.

The growing global aging population is contributing to an increasing burden of benign prostatic hyperplasia (BPH), highlighting the need for novel, highly effective and low-toxicity therapies. In light of its well-documented anti-inflammatory and anti-tumor properties, we investigated the potential of the natural product Pulsatilla saponin D (PSD) in treating BPH. For the first time, we demonstrate that PSD significantly inhibits the proliferation of and induces apoptosis in the immortalized human normal prostatic stromal cell line, human prostate fibroblasts, and the human benign prostatic hyperplasia epithelial cell line. Mechanistic studies involving transcriptome analysis and RT-qPCR validation revealed that PSD likely exerts its effects by downregulating the expression of the androgen receptor and by modulating multiple signaling pathways synergistically, including the Phosphatidylinositol 3-kinase/Protein Kinase B, Tumor Necrosis Factor, Hypoxia-Inducible Factor-1 and Interleukin-17 pathways. Full article

► Show Figures

Figure 1

17 pages, 1045 KB

Open AccessReview

SERBP1: A Multifunctional RNA-Binding Protein Linking Gene Expression, Cellular Metabolism, and Diseases

by Zezhao Ji and Abduxukur Ablimit

Cells 2025, 14(21), 1705; https://doi.org/10.3390/cells14211705 - 30 Oct 2025

Abstract

SERBP1 (SERPINE1 mRNA-Binding Protein 1), as an RNA-binding protein with multiple biological functions, has become a research hotspot in the field of life sciences in recent years. Its unique molecular structure, such as the presence of RG/RGG repeat sequences and the absence of [...] Read more.

SERBP1 (SERPINE1 mRNA-Binding Protein 1), as an RNA-binding protein with multiple biological functions, has become a research hotspot in the field of life sciences in recent years. Its unique molecular structure, such as the presence of RG/RGG repeat sequences and the absence of typical RNA-binding domains, enables it to exert diverse roles in cells. This article systematically reviews the research progress of SERBP1 in various fields including cellular stress response, tumorigenesis and development, reproductive system regulation, nervous system function, and viral infection, elaborates on its mechanism of action in detail (including newly supplemented content on cell cycle regulation, interaction with PARP1, and ribosome biogenesis), and outlines future research directions. It aims to provide a reference for in-depth understanding of the biological functions of SERBP1 and the diagnosis and treatment of related diseases. Full article

► Show Figures

Figure 1

21 pages, 41265 KB

Open AccessArticle

Preventive Effect of Fisetin on Follicular Granulosa Cells Senescence via Attenuating Oxidative Stress and Upregulating the Wnt/β-Catenin Signaling Pathway

by Juan Dong, Zhaoyu Yang, Qiongyu Yuan, Weidong Zeng, Yuling Mi and Caiqiao Zhang

Cells 2025, 14(21), 1704; https://doi.org/10.3390/cells14211704 - 30 Oct 2025

Abstract

Oxidative stress-mediated dysfunction of granulosa cells (GCs) is recognized as a pivotal driver of prehierarchical follicular atresia in poultry, contributing substantially to the reduced egg production in aged laying hens. Here, we investigated the protective effects of the natural flavonol, fisetin, on aged [...] Read more.

Oxidative stress-mediated dysfunction of granulosa cells (GCs) is recognized as a pivotal driver of prehierarchical follicular atresia in poultry, contributing substantially to the reduced egg production in aged laying hens. Here, we investigated the protective effects of the natural flavonol, fisetin, on aged chicken follicular GCs. A D-galactose (D-gal)-induced aging model of GCs was established to evaluate the protective role of fisetin against cellular senescence. Small yellow follicles (SYFs) from 580-day-old hens were cultured with fisetin for 72 h to verify its ameliorative effect on naturally aged follicles. Fisetin reduced the typical characteristic of senescence in D-gal-induced GCs, as reflected by decreased senescence-associated β-galactosidase (SA-β-gal) activity and increased expression of proliferation-related proteins, including cyclin D1 (CCND1), cyclin-dependent kinase 2 (CDK2), cyclin-dependent kinase 1 (CDK1), and Cyclin B1. Furthermore, fisetin enhanced the activity of antioxidant enzymes by activating the Nrf2/HO-1 signaling pathways, while attenuating mitochondrial dysfunction and promoting ATP production in senescent GCs. Additionally, fisetin significantly promoted nuclear translocation of β-catenin, and suppressed the expression of senescence marker proteins p53 and p21, thereby alleviating cell cycle arrest in D-gal-induced senescent GCs. Simultaneous inhibition of Nrf2/HO-1 and β-catenin signaling also abolished the beneficial effects of fisetin on oxidative stress and cell proliferation in naturally senescent follicles. These findings indicate that fisetin prevents follicular atresia by suppressing GCs oxidative damage and improving cell cycle arrest via activating the Nrf2/HO-1 and Wnt/β-catenin signaling pathways. Full article

► Show Figures

Figure 1

Journal Description

Cells

Latest Articles

Journal Menu

Journal Browser

Highly Accessed Articles

Latest Books

E-Mail Alert

News

Topics

Conferences

Special Issues

Topical Collections

Further Information

Guidelines

MDPI Initiatives

Follow MDPI