Cells doi: 10.3390/cells15121077

Authors: Ja Young Cho A Young Jeon Hyun Tae Kim Jung-Ha Kang Jae Hun Cheong Jae Hoon Choi

Spermatogonial stem cells (SSCs) are pivotal in surrogate broodstock technology. However, species-specific protocols for the efficient enrichment and long-term preservation of SSCs in olive flounder (Paralichthys olivaceus) are not yet fully established. In this study, we evaluated and optimized methods for the isolation and cryopreservation of P. olivaceus SSCs. First, we compared two enrichment methods, including Percoll density gradient centrifugation (PDGC) and differential plating (DP). Although SSCs enriched by both methods showed increased expression of SSC-specific marker genes, PDGC resulted in significantly greater enrichment than DP. A combination of PDGC and DP did not further improve enrichment efficiency, suggesting that PDGC alone is sufficient in P. olivaceus. Second, we optimized cryopreservation conditions according to various cryoprotectants. Among the conditions, SSCs cryopreservation using 1.3 M propylene glycol (PG) as a permeating agent and 0.2 M raffinose (Raf) as a non-permeating cryoprotectant provided the highest cell viability (56.1%), demonstrating a synergistic protective effect. Finally, preliminary in vivo migration and localization ability of the cryopreserved SSCs was confirmed through xenotransplantation into zebrafish (Danio rerio) larvae. PKH26-labeled donor cells exhibited successful initial localization and short-term persistence within the presumptive gonadal ridge of the recipients at 5 days post-transplantation. These findings provide an optimized protocol for the handling and preservation of P. olivaceus germline resources, contributing to the technical advancement of surrogate reproduction strategies in this species.

Cells doi: 10.3390/cells15121076

Authors: Dominika Przywara Wiktor Babiuch Alicja Petniak Małgorzata Wasilewska Jarosław Krzyżanowski Monika Czuba Arkadiusz Krzyżanowski Adrianna Kondracka Janusz Kocki Paulina Gil-Kulik

Umbilical cord mesenchymal stromal cells (UC-MSCs) are a key element of regenerative medicine due to their ability to secrete growth factors that stimulate proliferation and angiogenesis, and modulate the inflammatory response. Despite their widespread use, the influence of the perinatal microenvironment on their biological properties remains poorly understood. The aim of this study was to assess the influence of pH and blood gas parameters in umbilical cord blood on the global transcriptomic profile of UC-MSCs and to analyze the correlation between the metabolic status of the newborn and the expression of key trophic factors: EGF, FGF2, FGFR1, FGFR3, GDNF, HGF, IGF1, NES, NGF, and PGF. Methods: The study was conducted in two stages. In the first phase, transcriptomic screening was performed using Affymetrix HuGene 2.0 ST microarray on cells isolated from three environmental groups defined by cord blood pH: acidic (pH < 7.35), physiological (7.35–7.39), and alkaline (pH ≥ 7.4). In the second phase, the results were validated using qPCR on an expanded study group (N = 50). Gene expression levels (RQ) were related to blood gas parameters (pH, pCO2, pO2, cHCO3) and the presence of clinical features of threatened neonatal asphyxia. Results: Microarray analysis revealed that environmental pH acts as a molecular phenotypic switch. Under low pH conditions (<7.35), a shift in cell profile from proliferative to structural–migratory was observed. Significant overexpression of genes responsible for extracellular matrix (ECM) organization and adhesion (e.g., COMP, DCN, LUM, FMOD) was observed, while pathways related to cell cycle and cell division (↓CDK1, AURKA, TOP2A) were downregulated. qPCR validation confirmed these observations, demonstrating a strong positive correlation between blood pH and the expression of regenerative mediators: FGFR1 (r = 0.28), EGF (r = 0.30), NGF (r = 0.39), and IGF1 (r = 0.30). A negative correlation was also found between carbon dioxide pressure (pCO2) and the expression of NGF, FGFR1, and EGF. A significant clinical finding was that in newborns diagnosed with threatened asphyxia, EGF, FGFR1, and NGF gene expression was significantly reduced, indicating impaired trophic potential of the cells in response to metabolic stress. Conclusions: These results indicate that cord blood gas parameters are critical regulators of the genetic activity of UC-MSCs. Metabolic and respiratory acidosis not only inhibit the cells’ proliferative potential but also force them into a matrix remodeling mode, permanently modifying their transcriptomic profile. This suggests that the neonatal acid–base status may serve as an objective indicator of the “biological quality” of isolated stromal cells, which has significant implications for their future applications in cell therapies.

Cells doi: 10.3390/cells15121075

Authors: Andrey Kostin Anton Saevskiy Md Aftab Alam Yiqun Jiang Natalia Suntsova Md Noor Alam

Aging disrupts sleep, but how these changes are structured across circadian time, vigilance states, and sex remains poorly understood, because most prior studies used single-sex cohorts and few days of recordings. We continuously recorded 14 days of EEG/EMG in 24 C57BL/6J mice using a balanced 2 × 2 design (young vs. old; male vs. female; n = 6/group). A comprehensive multiscale analysis of the extended dataset enabled detailed reconstruction of 24 h sleep–wake architecture, better characterization of natural day-to-day variability including across multiple estrous cycles, and detection of rare bouts and transition events. Across seven levels of analysis, from circadian profiles to EEG spectral parameterization, the strongest aging effect was a dark-phase-specific 17–18% loss of theta-dominant active wake (TDW) in both sexes, with reciprocal increases in quiet wake (nTDW) and NREM sleep. We also identified a recurring N-shaped structural motif at the dark-to-light transition, where age-related and several sex-associated differences were most apparent. Broadly, old mice exhibited (i) shorter TDW bouts; (ii) a shift in NREM exit kinetics toward wakefulness; (iii) more brief and poorly consolidated “out-block” NREM episodes; and (iv) a slowing of waking theta and higher low-frequency TDW power. Variance decomposition indicated that statistical power depends more on sample size than on recording length. Together, aging reflects a coordinated, circadian-phase-specific reorganization of sleep–wake architecture. Sex-related and interaction findings should be interpreted as hypothesis-generating pending larger cohorts.

Cells doi: 10.3390/cells15121074

Authors: Gianluca Aguiari Nicoletta Bianchi Ornella Franzese

Colorectal cancer (CRC) remains a leading cause of cancer-related morbidity and mortality, with substantial heterogeneity that is not fully explained by genetic alterations alone. Emerging evidence positions metabolic reprogramming as a central driver of tumor behavior, integrating glycolysis, mitochondrial function, lipid and amino acid metabolism, and autophagy into coordinated networks that extend beyond cancer cells to the tumor microenvironment. Tumor–immune metabolic competition and metabolite-mediated signaling shape immune responses, often promoting immunosuppression and resistance to immunotherapy, particularly in microsatellite-stable (MSS) CRC. Systemic factors, including obesity, insulin resistance, and the diet–microbiota axis, further modulate tumor metabolism and immune function, reinforcing disease progression. Metabolic biomarkers reflecting these multi-level interactions, spanning tumor-intrinsic pathways, immune contexture, and host metabolism, offer promising opportunities for improved patient stratification and therapeutic targeting, although clinical validation remains limited. Current treatments, including chemotherapy, targeted agents, and immune checkpoint inhibitors, are effective in selected subgroups but are constrained by resistance mechanisms. In this review, we propose an integrative immunometabolic framework in which tumor, immune, and systemic metabolic processes co-evolve, defining CRC progression and treatment response. Targeting this interconnected network through combinatorial and metabolism-oriented strategies may enable precision therapies, particularly for immunotherapy-resistant MSS CRC.

Cells doi: 10.3390/cells15121073

Authors: Lining Wang Siyu Zhong Jianan Zhao Ligang Liu Changyong Li

Lactylation is an emerging lactate-derived post-translational modification that may link tumour metabolic reprogramming, epigenetic regulation and DNA damage repair. Enhanced glycolysis and lactate accumulation are common in many tumours, and lactate has been reported to induce histone and non-histone lactylation in specific experimental contexts. Recent studies suggest that lactylation is associated with several DNA repair pathways, including base excision repair/single-strand break repair, nucleotide excision repair, homologous recombination and non-homologous end joining, and may contribute to therapy resistance in selected cancer models. Specifically, XRCC1 lactylation has been reported to promote nuclear translocation and repair activity in glioblastoma models; H4K12 lactylation has been linked to PARP inhibitor resistance through RAD23A activation in ovarian cancer models; and BLM lactylation has been associated with enhanced homologous recombination repair in bladder cancer models. Lactylation of NBS1, RAD51 and XLF has also been implicated in DNA repair regulation in specific experimental systems, although some mechanistic links are inferred from pathway activation or functional rescue experiments rather than directly demonstrated across multiple tumour types. These findings suggest that lactylation may modulate DNA repair and therapeutic response in a context-dependent manner. Targeting lactate metabolism, transport and lactylation regulators, including LDHA, MCT1/4, ACAT1, AARS1 and GCN5, or using site-specific lactylation-inhibiting peptides may improve chemotherapy and PARP inhibitor efficacy, but clinical translation remains limited by heterogeneity, metabolic plasticity, toxicity and insufficient validation.

Cells doi: 10.3390/cells15121072

Authors: Poorvi Subramanian Loganayaki Periyasamy Sreenidhi Mohanvelu Sheeja Aravindan Natarajan Aravindan

Neuroblastoma (NB), the most common extracranial solid tumor of childhood, exemplifies one of the most formidable paradigms of tumor immune evasion (TIME) in pediatric oncology. Despite significant advances in multimodal therapy and the clinical integration of immunotherapeutic strategies, high-risk NB (HR-NB) remains largely refractory to durable immune control. This failure reflects not an absence of immune engagement, but the presence of a highly evolved and developmentally wired immune escape architecture. In this review, we synthesize emerging insights from single-cell, multi-omics, and functional studies to define how developmental lineage, cellular plasticity, metabolic rewiring, epigenetic regulation, and therapy-induced adaptation converge to engineer immune blindness in NB. We discuss how NB’s neural crest origin establishes a baseline of low immunogenicity, which is subsequently reinforced through coordinated suppression of antigen presentation, dominance of immune checkpoint signaling, and profound dysfunction of cytotoxic T and natural killer cells within an immunosuppressive tumor microenvironment. Central to this process is tumor-intrinsic plasticity, whereby lineage instability and dedifferentiation, exacerbated by therapeutic pressure, embed immune silence as a stable tumor state. We highlight evidence positioning RD3 as a master upstream regulator linking cellular identity to immune visibility, governing antigen presentation, innate immune sensing, checkpoint expression, and cytotoxic lymphocyte engagement. Beyond tumor-intrinsic mechanisms, we examine the roles of immunosuppressive myeloid populations, tumor-derived exosomes, metabolic stress, hypoxia, and ferroptosis-associated pathways in reinforcing immune paralysis. Finally, we outline emerging therapeutic strategies aimed at dismantling this architecture, including combinatorial checkpoint blockade, metabolic and epigenetic reprogramming, exosome-targeted interventions, and next-generation immune engineering platforms. Together, this review reframes TIME in NB as a programmable, developmentally rooted process and provides a mechanistic roadmap for restoring immune competence and therapeutic susceptibility in HR disease.

Cells doi: 10.3390/cells15121071

Authors: Shuyue Cheng Zeyue Mu Feng Zhang Jianyou Song Jiapeng Shao Yunqi Yan Anastasios A. Daskalakis Yunjie Wang Bin Zhang Yashuang Jiang Le Wang Fang Liu

Background: Mitochondrial dysfunction is a central event in the pathogenesis of ischemic stroke. The roles of specific mitochondrial complex subunits, such as NDUFA4 and NDUFB3, in cerebral ischemia–reperfusion injury remain poorly defined. This study aims to investigate the dynamic expressions and functional impact of NDUFA4 and NDUFB3 in ischemic stroke. Methods: A transient middle cerebral artery occlusion (MCAO) model was established in male C57BL/6J mice. Label-free quantitative proteomics and Western blotting were employed to analyze protein expression in the ischemic penumbra. Highly differentiated PC12 cells were subjected to oxygen-glucose deprivation/reperfusion (OGD/R) or glutamate excitotoxicity to mimic ischemic injury in vitro. The functional consequences of NDUFB3 knockdown and overexpression were assessed by measuring ATP levels, reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm), and apoptosis. The involvement of the JNK-mediated mitochondrial apoptotic pathway was also examined. Results: Proteomic analysis revealed a significant upregulation of NDUFA4 and NDUFB3 in the ischemic penumbra of MCAO mice, as verified by western blot. In highly differentiated PC12 cells, both OGD/R and glutamate exposure induced a time-dependent increase in these proteins in mitochondrial fractions. Functional studies demonstrated that NDUFB3 knockdown significantly rescued OGD/R-induced mitochondrial dysfunction, as indicated by restored ATP production, reduced ROS generation, and stabilized ΔΨm. Furthermore, NDUFB3 silencing attenuated apoptosis by inhibiting JNK phosphorylation and decreasing BAX levels. Conversely, overexpression of NDUFB3 alone was sufficient to induce mitochondrial abnormalities, including loss of ΔΨm and elevated oxidative stress in highly differentiated PC12 cells. Conclusions: Ischemic injury triggers the upregulation of mitochondrial complex subunits NDUFA4 and NDUFB3. While this may initially act as a compensatory response, our findings identify NDUFB3 as a critical mediator of ischemic stroke pathology, whose overexpression drives mitochondrial dysfunction and apoptosis. In contrast, the suppression of NDUFB3 provides protection against ischemic injury. Therefore, NDUFB3 may be a potential candidate therapeutic target for reducing mitochondrial damage in ischemic stroke, but this role requires further validation in additional experimental and translational models.

Cells doi: 10.3390/cells15121070

Authors: Zhongwei Zhang Huiran Li Zhonglan Shi Xuan Bai Peipei Yin Lingguang Yang

Tyrosinase is a pivotal therapeutic target for hyperpigmentation disorders, yet current inhibitors frequently exhibit limited potency and suboptimal safety. Here, we employed structure-based virtual screening of an FDA-approved drug library against a refined human tyrosinase homology model, identifying methotrexate and selinexor as potent anti-melanogenic candidates. Both compounds markedly suppressed cellular tyrosinase activity and melanin synthesis (IC50 < 1 µM) in MNT-1 melanoma cells. Mechanistically, they orchestrate a multi-pronged downregulation of microphthalmia-associated transcription factor (MITF) by attenuating cAMP/PKA/CREB signaling, promoting β-catenin degradation, and accelerating MITF proteolysis via AKT/ERK activation. Additionally, they bolster the intracellular antioxidant defense system. These findings unveil a sophisticated regulatory network and suggest that with strict control of systemic exposure through optimized topical formulations, these FDA-approved agents could be further investigated as potential localized treatments for pigmentary disorders.

Cells doi: 10.3390/cells15121069

Authors: Akvilė Žilytė Vilma Petrikaitė

In this study, we evaluated the impact of different in vitro 3D culture modelling methods on the activity of doxorubicin (DOX) and 5-fluorouracil (5-FU) in human melanoma spheroids. Human melanoma A375 and IGR39 spheroids were generated using the hanging drop and non-adhesive surface methods. Spheroid growth dynamics were assessed by measuring changes in spheroid diameter. To compare the effects of anticancer drugs in spheroids of different sizes, spheroids of approximately 200 and 400 µm were formed. Drug activity was evaluated based on spheroid growth and cell viability using the MTT assay. A375 spheroids formed using the non-adhesive surface method were more sensitive to DOX than spheroids formed using the hanging drop method. In smaller A375 spheroids, 10 µM 5-FU reduced cell viability more effectively in spheroids formed using the hanging drop method. In contrast, IGR39 spheroids formed by the hanging drop method were more resistant than those formed on a non-adhesive surface. However, in IGR39 spheroids, the effects of DOX and 5-FU on growth and viability did not significantly differ between formation methods. In conclusion, A375 spheroid growth was not significantly influenced by the formation method, whereas IGR39 spheroid growth depended on the method used. A375 spheroids formed on non-adhesive surfaces were more sensitive to DOX, whereas 5-FU activity depended on drug concentration and spheroid size. In IGR39 spheroids, the effects of DOX and 5-FU on growth and viability were largely independent of the spheroid formation method.

Cells doi: 10.3390/cells15121068

Authors: Helena Branco Joana Carreira Inês Soure Cristina P. R. Xavier Andreia Rosário Maria Amorim Hugo Osório José E. Guimarães Ana Bela Sarmento-Ribeiro Manuel A. Sobrinho-Simões Hugo R. Caires M. Helena Vasconcelos

Measurable residual disease (MRD) in acute myeloid leukemia (AML) is monitored through detection of leukemia-associated phenotypic protein markers (LAPMs) in bone marrow aspirates, hindering disease real-time monitoring. We explored peripheral blood (PB), extracellular vesicle (EV)-based methods for MRD monitoring. To confirm that LAPMs are present in AML-derived EVs, EVs were isolated from OCI-AML3 cells by differential centrifugation and characterized according to their size (nanoparticle tracking analysis), morphology (transmission electron microscopy) and protein cargo (proteomic analysis and Western blot). CD14 and CD33 were detected in OCI-AML3 cells and their released EVs. To select a method to isolate EVs from the PB of AML patients, three techniques were tested: size exclusion chromatography followed by ultrafiltration (SEC-UF), Total Exosome Isolation Kit (Invitrogen) and Exo-spin™ Exosome Purification Kit (Cell Guidance Systems). SEC-UF allowed EV isolation with higher purity and less aggregates than the other techniques. LAPMs were detected in those EVs, but their presence depended on the isolation method. Finally, EVs from seven AML patients’ plasma were isolated by SEC-UF. LAPMs were identified in paired samples at diagnosis and remission, with differential expression throughout disease evolution. This proof-of-concept study highlights the possibility of real-time MRD monitoring through LAPMs’ analysis in AML patient’s circulating EVs.

Cells doi: 10.3390/cells15121067

Authors: Junmyeong Park Seungye Kang Soojin Kim Donghyun Kim Borami Shin Ji Young Mun Yurim Park Johnny Kim Steven A. Goldman Kee-Pyo Kim

Oligodendrocytes (OLs) are essential for myelin formation in the central nervous system, and their loss or dysfunction is a hallmark of various demyelinating and neurodegenerative disorders. Although oligodendrocyte precursor cells (OPCs) represent a promising cell source for remyelination therapies, existing protocols for generating OPCs from human-induced pluripotent stem cells (iPSCs) are often limited by prolonged culture duration, low efficiency, and cellular heterogeneity. Here, we report an efficient and reproducible platform for generating OPCs from iPSC-derived neural progenitor cells (iNPCs) through stage-specific modulation of developmental signaling pathways. Directed differentiation of iNPCs recapitulated key developmental transitions, progressing through OLIG2+/NKX2.2+ progenitors to CD140a+/O4+ OPCs within a significantly shortened timeframe compared to conventional approaches. Notably, iNPC-derived spheres functioned as a progenitor-like niche, enabling sustained OPC production through serial replating. Purified OPCs could differentiate into MBP+ oligodendrocytes and demonstrated myelination capacity both in vitro, via nanofiber ensheathment and in vivo following transplantation into shiverer (shi/shi) mice, where they formed myelin sheaths around host axons. Despite these advances, OPC differentiation and maturation efficiencies remained suboptimal, highlighting the need for further optimization. Collectively, our findings establish a scalable and time-efficient strategy for iPSC-derived OPC generation and underscore their potential for disease modeling and cell-based remyelination therapies.

Cells doi: 10.3390/cells15121066

Authors: Zhi-Wei Ye Leilei Zhang Xuhong Zhang John Culpepper Eduardo N. Maldonado Kenneth D. Tew Jie Zhang Danyelle M. Townsend

Melanoma is a highly aggressive and metabolically adaptable cancer that often resists conventional therapies. Targeting core bioenergetic pathways may, therefore, represent an effective strategy to improve therapeutic responses, particularly in tumors dependent on mitochondrial function. SC18 is an imidazolidine-2,4-dione compound that binds the NADH-binding pocket of voltage-dependent anion channels (VDACs), inducing mitochondrial dysfunction. VDAC expression is increased in melanoma and strongly associated with advanced disease stage and poor prognosis. In this study, we evaluated the effects of SC18 in melanoma cell lines with distinct pigmentation states, including melanin-rich melanotic human MNT-1 and mouse B16-F1, as well as low/amelanotic human SKMel28 and mouse YUMM cells. VDAC1, VDAC2 and VDAC3 were highly expressed across these melanoma lines, all of which relied on both glycolysis and mitochondrial oxidative phosphorylation for ATP production. SC18 reduced mitochondrial membrane potential and oxygen consumption rates, accompanied by declines in intracellular ATP levels and TCA cycle substrate utilization. SC18 also increased reactive oxygen species, mitochondrial superoxide, and lipid peroxidation, indicating enhanced oxidative stress. These metabolic and redox disturbances were associated with reduced cell viability and significantly impaired migration in multiple melanoma cell lines, supporting a potential anti-metastatic effect. In addition, SC18 showed synergistic cytotoxicity when combined with other chemotherapeutic agents. Overall, SC18 disrupted mitochondrial metabolism, induced oxidative stress, and impaired survival and motility pathways, with more pronounced effects in low/amelanotic than in melanotic melanoma cells. Together, these findings support the further development of SC18 as a mitochondrial metabolic disruptor that targets redox vulnerabilities in melanoma.

Cells doi: 10.3390/cells15121065

Authors: Cynthia Van Rompay Kevin Tabury Emil Rehnberg Zoë Janssen Sarah Baatout Marianne S. Carlon Xavier Casadevall i Solvas Bjorn Baselet

Three-dimensional (3D) cardiac models, including spheroids, organoids, and organ-on-chips, are advanced systems for studying human physiology, disease, and drug responses with greater biological relevance than 2D models. As their use expands in biomedical research, tissue engineering, and regenerative medicine, reliable preservation methods are needed. However, cryopreservation often fails to protect 3D systems due to limited cryoprotectant penetration, ice formation, and mechanical stress during freezing and thawing. Room-temperature (RT) preservation has emerged as a promising alternative for short-term transport. This study evaluated a RT-based transport medium (CellShip®) for preserving cardiac organoids for up to seven days, compared with conventional cryopreservation using slow-freezing in Cryostor®CS10. Viability and functionality were assessed using apoptosis, ATP levels, beating activity, proliferation, and size. During maturation, organoids showed increased size, ATP levels, and beating capacity. Cryopreservation reduced size, proliferation, ATP levels, and altered beating, while increasing apoptosis. In contrast, RT preservation maintained stable viability and functionality after recovery. These findings demonstrate that RT preservation effectively maintains cardiac organoid integrity and function, offering a promising alternative for short-term storage and transport, with potential terrestrial and nonterrestrial applications.

Cells doi: 10.3390/cells15121064

Authors: Jürgen Becker Jörg Wilting

There are a few molecules that are regularly used as markers for lymphatic endothelial cells (LECs) such as the adhesion molecule CD31/PEACAM1, the transcription factor PROX1, the Vascular Endothelial Growth Factor Receptor-3 (VEGFR3/FLT4), the glycoprotein podoplanin, and the hyaluronan receptor LYVE1. However, none of the molecules are exclusively expressed in LECs, and there is molecular and functional heterogeneity of LECs in initial lymphatics, lymphatic collectors and lymph nodes. Therefore, a combination of markers must be applied to identify lymphatics. This is particularly true for the characterization of conditions such as lymphatic malformations or cancers, in which the molecular profile of vessels may be variable or abnormal. Here we present two molecules that can help distinguish between endothelial cells of blood and lymphatic vessels: the scaffold protein liprin β-1 (PPFIBP1) and the intermediate filament synemin. We collected own data on the RNA and protein expression of the two molecules in humans, and studied publicly available databases. PPFIBP1 appears to be a suitable marker of LECs in initial lymphatics, collectors and lymph nodes, while synemin appears to be more restricted to initial lymphatics. We hope this will stimulate monoclonal antibody development and help expand the range of LEC markers in health and disease.

Cells doi: 10.3390/cells15121063

Authors: Sara Marcoccia Giulia Chiacchierini Patrizia Campolongo

The entorhinal cortex (EC) is a central structure of the medial temporal lobe, functioning as the main cortical gateway to the hippocampus (HPC) and playing a crucial role in memory, spatial navigation, and temporal representation. This review outlines the distinct yet complementary contributions of its two main subdivisions, the medial (MEC) and lateral (LEC) entorhinal cortices. Despite being historically viewed as functionally segregated, they operate instead in close coordination to support the encoding and retrieval of multidimensional experiences. While the MEC is prominently involved in mapping spatial relationships and movement through specialized cell populations, and the LEC in processing object-related and contextual information, growing evidence shows substantial integration between these domains, challenging strict dichotomies. The MEC encodes elapsed time through persistent firing and time cell sequences, while the LEC signals temporal context via rate remapping; their convergent projections to the hippocampus enable the formation of temporally structured episodic memories. The review assesses recent findings on memory, navigation, and time processing, and highlights how the EC supports each through its layered architecture, local microcircuitry, and widespread interactions with HPC, cortical, and subcortical networks. Moreover, alterations in EC activity patterns emerge as the earliest signs of pathologies such as Alzheimer’s disease and temporal lobe epilepsy. Altogether, this review offers an up-to-date view of the EC not as a set of parallel modules, but as a highly interactive and dynamic system essential for structuring experience across space, time, and context.

Cells doi: 10.3390/cells15121062

Authors: Daniel Alberto Carrillo-Vázquez Beatriz Alcalá-Carmona Jennifer Tiaré Balderas Miranda Yatzil Reyna-Juárez María José Ostos-Prado Fabiola Cassiano-Quezada Samuel Govea-Peláez Nancy R. Mejía-Domínguez Guillermo Juárez-Vega Karina Santana-De Anda Jiram Torres-Ruiz Diana Gómez-Martín

Background: Neutrophils play a role in idiopathic inflammatory myopathies (IIMs), especially in vasculopathic manifestations. While neutrophil extracellular traps (NETs) and low-density granulocytes (LDGs) have been described, the functional relevance of other subsets—such as naïve neutrophils, reverse transendothelial migration neutrophils (rTEM), myeloid-derived suppressor cells (MDSCs), and regulatory neutrophils—and their association with vasculopathy remains unknown. Objective: We aimed to characterize the phenotypic and functional profile of neutrophil subsets and their association with vasculopathic manifestations in IIM, adjusting for disease activity. Methods: We conducted a cross-sectional, single-center study including 59 IIM patients diagnosed by muscle biopsy and fulfilling 2017 ACR/EULAR criteria. Flow cytometry was used to immunophenotype myeloid subsets, and functional assays assessed phagocytosis and respiratory burst. Patients were stratified by clinical activity and presence of vasculopathy. Results: Vasculopathic patients showed expansion of LDGs (p = 0.0092), granulocytic and monocytic MDSCs expressing Arginase-1 (p = 0.0078, p = 0.0003) and PD-L1 (p = 0.0258, p = 0.0087), and rTEM neutrophils (p = 0.0775). In contrast, they exhibited a reduction in naïve neutrophils (p = 0.0004), phagocytosis (p < 0.0001) and respiratory burst (p = 0.0006). Multivariate analysis identified naïve neutrophils and activated CD177+ neutrophils as independent predictors of vasculopathy. A positive correlation between activated and naïve neutrophils were observed in patients with vasculopathic features (r = 0.43; p < 0.05). Conclusions: IIM patients with vasculopathic features display a distinct immune profile characterized by an imbalance between proinflammatory and regulatory neutrophil subsets, alongside persistent functional impairment.

Cells doi: 10.3390/cells15121061

Authors: Binglin Yue Wen Hu Shuo Zhu Du’an Chen Huanyu Guan Zhuoying Zhao Hui Wang Jiabo Wang Jincheng Zhong Haitao Shi

Skeletal myogenesis is an extremely complex process that mononuclear myoblasts undergo proliferation, differentiation, and fusion to form multinucleated contractile muscle fibers, involving a balance between synthesis and degradation metabolism. Skeletal muscle requires an effective mechanism to balance rapid proliferation by degrading supernumerary or damaged organelles/proteins, or by activating cellular signals to regulate subsequent muscle differentiation. In recent years, three important cellular processes—apoptosis, ubiquitin–proteasome system (UPS), and autophagy—have received extensive attention in skeletal myogenesis. The UPS supports the early differentiation process and initiates apoptosis, and the increase in apoptosis activates autophagy to clear damaged organelles and proteins, which in turn inhibits apoptosis, preventing excessive cell death and maintaining cellular stability. The coordination among apoptosis, UPS, and autophagy is more intricate, as they interact through a dynamic balancing mechanism, determining the balance between cell death and survival, and enabling proper muscle differentiation. Here, we explore the molecular signals that mediate apoptosis, UPS, and autophagy, with a focus on analyzing their interrelationship in skeletal myogenesis. Studying the regulatory mechanisms of these molecules will help in understanding the role of cell death in skeletal muscle development, especially how they affect muscle cell differentiation, providing new insights into mammalian skeletal myogenesis.

Cells doi: 10.3390/cells15121059

Authors: Leandro Rodrigues-Freitas Luísa Pinto Teresa Canedo

Sex differences are increasingly recognized as key determinants of vulnerability, clinical presentation, and treatment response in depression. Rather than arising from a single mechanism, these differences emerge from the interplay of multiple biological and non-biological factors. Converging evidence points to the hippocampus as a central region where these processes intersect, with adult neurogenesis and astrogliogenesis representing a potential mechanistic link between sex-specific biological factors and behavioral outcomes in depression. In this review, we integrate findings from human studies and preclinical models to examine how sex impacts depression while considering the multiple origins of sexual differentiation in the central nervous system. We discuss the importance of studying sex as a biological variable and acknowledge current limitations in the field. Finally, we highlight how cytogenic processes in the adult hippocampus are modulated in a sex-dependent manner, how their disruption may contribute to the pathophysiology of depression, and their potential role in precision psychiatry. Adult cytogenesis provides a promising target for developing therapeutic strategies aimed at promoting the integration of these cells in neural circuits, which may counterbalance the cellular impairments observed in stress-induced depression, representing a therapeutic avenue for this disorder.

Cells doi: 10.3390/cells15121060

Authors: Tohru Tezuka Wei Jie Nicholas Yang Keisuke Kitahata Aya Nohara Sun Joo Park Minsoo Kim

Ubiquitin-like proteins (UBLs) such as SUMO, NEDD8, ISG15, FAT10, and UFM1 are proteins that share structural similarities to ubiquitin. Like ubiquitin, they function as protein modifiers, catalyzing modifications through a conserved enzymatic cascade of E1 activating enzymes, E2 conjugating enzymes, and E3 ligases. In doing so, UBLs regulate a diverse set of cellular processes, including stress response, antiviral activity, nuclear transport, cancer development, and autophagy. In recent years, the roles of UBLs during pathogenic bacteria infection have gained attention, although much still remains elusive. This review describes current findings related to UBL systems in the context of pathogenic bacteria infection, focusing on NEDD8, ISG15, FAT10, and UFM1. Specifically, we look at how the host UBL system responds to bacterial infection by inducing the host’s defense system, and how pathogenic bacteria manipulate the host UBL system to ensure successful infection.

Cells doi: 10.3390/cells15121058

Authors: Leopold Eckhart Supawadee Sukseree Heinz Fischer

Sebaceous glands consist of epithelial cells, known as sebocytes, that undergo differentiation to deliver the components of sebum into the sebaceous duct and eventually to the hair and skin surface. The final step of the terminal differentiation program is called holocrine secretion because the entire cell content is converted into sebum. Holocrine secretion is a mode of programmed cell death, which involves the degradation of the nucleus and other organelles and the rupture of the cell membrane. Here, we review the current knowledge of differentiation-associated death of sebocytes and discuss open questions regarding its mechanism and functions. In vivo studies have provided evidence for degradation of nuclear and mitochondrial DNA by lysosomal deoxribonuclease 2 (DNase 2), indicating a key role of lysosomes in holocrine secretion. We discuss the influence of tight junctions on the spatial localization of holocrine secretion within glands, the regulation of holocrine cell death by autophagy and potential mediators of membrane lysis. Further studies of holocrine secretion are needed to fully uncover its molecular control and to determine potential clinical implications.

Cells doi: 10.3390/cells15121057

Authors: Nele Johanne Czaniera Wiebke Schulten Katja Nowak Diana Pschik Jonas Joneleit Barbara Kaltschmidt Christian Kaltschmidt

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterized by progressive memory impairment and cognitive decline. The APOE4 allele represents one of the most prominent genetic risk factors. In this study, we investigated the impact of APOE4 on the cholinergic neuronal development and on the neuronal inflammatory response to TNF-α stimulation. To address this, human induced pluripotent stem cells (hiPSCs) carrying a homozygous APOE4 genotype and an isogenic APOE3 control were differentiated into cholinergic-like induced neurons (iNs) by LHX8 overexpression. APOE4 was associated with accelerated early neuronal differentiation, as reflected by earlier downregulation of the progenitor marker Nestin. However, delayed expression of synaptophysin indicated impaired synaptic maturation. Functionally, APOE3 iNs exhibited a robust but temporally regulated response to TNF-α, whereas APOE4 iNs were characterized by a delayed yet sustained induction of inflammatory signaling. Moreover, APOE4 iNs displayed an enhanced stress-associated transcriptional response at early differentiation stages. Collectively, these findings suggest that APOE4 influences both neuronal development and the timing and persistence of inflammatory responses, potentially predisposing cholinergic neurons to later dysfunction in AD.

Cells doi: 10.3390/cells15121056

Authors: Vitale Miceli Mattia Emanuela Ligotti Vincenzo Raffo Silvia Lopa Viviana Ippolito Alessia Gallo Nicola Cuscino Simone Dario Scilabra Margot Lo Pinto Simone Messina Salvatore D’Arpa Matteo Moretti Laura de Girolamo Matteo Bulati Alessandra Colombini

Osteoarthritis is a chronic degenerative joint disease characterized by inflammation and cartilage degradation, for which current treatments are mainly symptomatic and unable to halt disease progression. Adipose-derived mesenchymal stem cells (ASCs) represent a promising therapeutic option due to their regenerative and immunomodulatory properties, which may be further enhanced through specific priming strategies. In this study, primary human ASCs were exposed to interleukin-1 beta (IL1β), interferon-gamma (IFNγ), or hypoxic priming, and subsequently analyzed using a multi-omics approach integrating RNA sequencing, proteomics of secretome, and exosomal miRNA profiling. Differential gene expression, protein abundance, and miRNA signatures were assessed together with functional enrichment and network analyses. IL1β priming induced marked transcriptional reprogramming of ASCs, while hypoxia and IFNγ priming produced limited changes. IL1β also profoundly reshaped the ASC secretome and exosomal miRNA cargo, revealing coordinated regulation of pathways involved in immune modulation and cartilage remodeling. In contrast, the other priming conditions showed minimal and less integrated molecular effects. Overall, IL1β priming consistently generated a multi-layered molecular signature linking immunoregulatory and regenerative pathways. These findings suggest that IL1β priming enhances the functional properties of ASCs and provides mechanistic insight supporting their potential use in osteoarthritis therapy.

Cells doi: 10.3390/cells15121054

Authors: Hossein Omidian Erma J. Gill Umadevi Kandalam

Hydrogel scaffolds have emerged as instructive microenvironments for craniofacial tissue regeneration, moving beyond passive cell carriers toward platforms that regulate cell fate, vascularization, immune remodeling, and tissue-specific architecture. This review synthesizes hydrogel-associated strategies across dental pulp, periodontal ligament, gingival, bone marrow, jawbone, endothelial, oral mucosal, induced pluripotent stem cell (iPSC), extracellular vesicle (EV), exosome, secretome, and acellular systems. The evidence indicates that craniofacial hydrogel performance is governed by reciprocal interactions among biological source, scaffold composition, matrix mechanics, spatial architecture, mineral or ionic signaling, growth factor delivery, vesicle-mediated communication, and inflammatory niche modulation. Mineralized and ion-releasing hydrogels most consistently supported osteogenesis and bone repair, whereas extracellular matrix (ECM)-mimetic, peptide, collagen, fibrin, gelatin methacryloyl (GelMA), alginate, hyaluronic acid (HA), and chitosan-based systems enabled pulp–dentin, periodontal, peri-implant, oral mucosal, and soft-tissue reconstruction. Responsive, antimicrobial, antioxidant, conductive, and immunomodulatory hydrogels further expanded the field by targeting diseased microenvironments rather than regeneration alone. Despite strong preclinical evidence, translation remains limited by heterogeneity in scaffold formulations, biological sources, analytical endpoints, defect models, and long-term functional validation. Future progress will require standardized characterization, tissue-specific design criteria, clinically relevant large-animal models, scalable cell-free technologies, and integrated assessment of regeneration, immunity, vascularization, innervation, mechanics, and safety.

Cells doi: 10.3390/cells15121055

Authors: Catalin-Bogdan Satala Alina-Mihaela Gurau Daniela Mihalache Gabriela Patrichi Roxana-Cristina Mehedinti Andy Radu Leibovici Gabriela Gurău

Gastric cancer peritoneal metastasis is not simply an extension of the primary tumor into the abdominal cavity. It represents a biologically distinct disease context shaped by interactions between disseminated tumor cells, peritoneal fluid, mesothelial surfaces, submesothelial stroma, extracellular matrix, immune populations, and malignant ascites. In this narrative review, we examine peritoneal metastasis as a transition between three related but physiologically different states: the primary gastric tumor, free-floating tumor cells or spheroids in the peritoneal fluid, and established mesothelial or submesothelial metastatic implants. We discuss how tumor cells acquire dissemination competence in the primary tumor, survive detachment and fluid-phase stress, adhere to remodeled mesothelium, recruit stromal and immune support, and adapt to ascites-mediated signaling. We also review how the peritoneal niche may contribute to biomarker discordance, immune exclusion, therapeutic resistance, and limitations of conventional response assessment. Where relevant, we distinguish evidence derived directly from gastric cancer peritoneal metastasis from preclinical data, extrapolation from other peritoneal malignancies, and hypothesis-generating interpretation. Finally, we summarize practical implications for tissue sampling, ascites and lavage analysis, biomarker interpretation, translational modeling, and peritoneal-directed therapeutic strategies. A clearer understanding of the biological divergence between the primary tumor, the fluid-phase compartment, and peritoneal implants may improve the study and clinical management of gastric cancer peritoneal metastasis.

Cells doi: 10.3390/cells15121053

Authors: Enza Maria Verde Valentina Secco Andrea Ghezzi Jessica Mandrioli Serena Carra

In order to facilitate readers’ better understanding, some language descriptions and grammar as well as the layout of some chapters have been modified [...]

Cells doi: 10.3390/cells15121052

Authors: Wei-Ping Li Karen E. Laupman Stephanie D. Beekhuis-Hoekstra Evangelia Thanou Remco V. Klaassen Patrick F. Sullivan Danielle Posthuma August B. Smit Frank Koopmans Vivi M. Heine

Astrocytes are increasingly implicated in the pathophysiology of schizophrenia (SCZ), yet how astrocytic dysfunction contributes to disease-relevant neuronal abnormalities remains unclear. Here, we used mass spectrometry–based proteomics to profile lysates (proteome) and secreted proteins (secretome) from iPSC-derived astrocytes originating from 9 SCZ patients and 8 healthy controls. Compartment-specific analyses showed that lysates were enriched for mitochondrial and nuclear pathways, whereas astrocyte-conditioned media (ACM) were enriched for extracellular matrix (ECM) and vesicle-associated proteins. Differential expression analysis revealed minimal overlap between dysregulated proteins in lysates and ACM, suggesting modality-specific effects of SCZ-associated donor background. Interestingly, ECM proteins and key secreted cues involved in synaptic development, including MFGE8 and SEMA3C, were selectively reduced in SCZ ACM, whereas RNA-processing proteins were aberrantly increased. This is in line with previously reported microRNA enrichment in extracellular vesicles (EV) derived from SCZ patients. Gene set analyses further identified the alteration in secretion and nuclear processes as well as the potential involvement of autophagy-dependent release mechanism in SCZ astrocytes. Together, these findings suggest disrupted astrocytic protein homeostasis and extracellular signalling in SCZ iPSC-derived astrocytes, providing mechanistic insight into astrocyte-mediated contributions to synaptic and circuit deficits in the disorder.

Cells doi: 10.3390/cells15121051

Authors: Gan Zhao Ziheng Chen Caifeng Yang Mingzheng Liu Weiyong Wang Chuanmao Zhang

The lamin proteins are classified into A- and B-types, and together with their associated proteins, they form the nuclear lamina, which governs diverse nuclear structures and functions, including nuclear mechanics, chromatin organization, and gene regulation. Mutations of these proteins give rise to a strikingly diverse group of tissue-specific disorders, the laminopathies, including muscular dystrophies, cardiomyopathies, lipodystrophies, neuropathies, and premature aging syndromes, despite their broad expression. Unraveling the basis of this tissue selectivity has revealed that lamins function not merely as structural elements but as active regulators. While the A-type lamins modulate nuclear stiffness, transcription, and genome integrity, the B-type lamins ensure mechanical resilience and heterochromatin tethering. Pathogenic mutations of these proteins disrupt their functions through convergent mechanisms that manifest according to tissue-specific contexts, leading to impaired nuclear mechanics, aberrant gene regulation, defective DNA repair, and cellular senescence. Advances in patient-derived cellular models and animal systems have illuminated these vulnerabilities and catalyzed therapeutic progress, ranging from farnesyltransferase inhibitors to emerging genome-editing strategies. Collectively, studies of lamin protein function reveal how the nucleus maintains its structures and functions, while studies of laminopathies demonstrate how nuclear dysfunction drives systemic disease and points toward mechanism-based therapies.

Cells doi: 10.3390/cells15121050

Authors: Md Manirujjaman Maria D. Sanchez-Pino Jasjeet Singh Farzeen Nafees Mrityunjoy Biswas Ramesh Thylur Puttalingaiah Soroor Heidari Dorota Wyczechowska Jone Garai Diana C. Polania-Villanueva Qingzhao Yu Luis Del Valle Lucio Miele Samarpan Majumder Jovanny Zabaleta Fokhrul Hossain

The mammary gland is a heterogeneous organ that modulates ductal morphogenesis and alveolar differentiation. Obesity is a significant risk factor for several cancers, including postmenopausal breast cancer. We and others have described an association between obesity and increased breast cancer growth. However, the effects of obesity on the mammary fat pad microenvironment (MFPME) remain understudied. Here, we investigated the effect of the Western Diet (WD) on immunocompetent female mice and on their MFPME. Our data suggest that the WD increased body, liver, and perigonadal white adipose tissue (pWAT) weight, as well as myeloid cell infiltration into these tissues. Interestingly, we did not find any significant change in CD4+ and CD8+ T cells in the liver, blood, and pWAT. NanoString data demonstrates that various cellular processes, including the complement system, innate immune system, phagocytic activity, immune metabolism, and NOD-like receptor (NLR) signaling, were upregulated in the MFPME of obese mice. RNA-Seq data suggest that WD significantly modulated MFPME physiology through regulation of gene expression, cellular processes, and signaling pathways. Further investigation is necessary to determine how WD-mediated changes in MFPME modulate breast cancer biology.

Cells doi: 10.3390/cells15121049

Authors: Tom Trapphoff Ursula Eichenlaub-Ritter Karoline Hohenstein Saskia Möckel Stefan Dieterle

The cryopreservation of pre-implantation embryos has become routine in medically assisted reproduction (MAR), and the proportion of frozen embryo transfers has steadily increased in recent years. Because cryopreservation through either slow-cooling protocols or ultra-rapid vitrification requires potentially cytotoxic cryoprotective agents to prevent uncontrolled and detrimental ice crystal formation, the safety of these procedures must be carefully considered. Evidence from human epidemiological studies, including retrospective and prospective controlled studies, and data from national patient registries indicate that children born after frozen embryo transfer have a higher birth weight than those born after spontaneous conception and have an increased risk of rare genomic imprinting disorders, such as Beckwith–Wiedemann, Silver–Russell, or Prader–Willi syndrome. Encompassing not only reversible DNA methylation patterns established during gametogenesis, but also the timed abundance and availability of transcripts and proteins required to establish or maintain epigenetic marks throughout development and differentiation, as well as persistent or transient post-translational histone modifications and non-coding RNAs, the epigenome may be particularly sensitive to cryopreservation. Importantly, epigenetic regulation is highly complex. Alterations of the epigenome at any developmental stage are often not monocausal, do not necessarily result in immediate disturbances in the pre-implantation embryo, and are unlikely to operate through simple all-or-nothing mechanisms; however, they may have long-lasting effects at later developmental stages. To make matters even more complex, differences between species in terms of epigenetic regulation or lineage differentiation are well known and translation from animal model systems to humans must be considered with caution. More recently, epigenetic regulation by non-coding RNAs has also come into focus, as these molecules are crucial, either directly or indirectly, for gene expression, translation, and protein biosynthesis during development. Therefore, assessing potential adverse effects of cryopreservation on the entire epigenome remains a major challenge, particularly because little is known about indirect factors, such as post-translational histone modifications and non-coding RNAs. In this review, we focus on the potential influence of the cryopreservation of pre-implantation embryos on the epigenetic profile in humans and animals. Specifically, we consider DNA methylation of imprinted genes and global DNA methylation; post-translational histone modifications; the abundance and availability of transcripts and proteins required to establish, maintain, or protect epigenetic patterns; and the presence of non-coding RNAs involved in epigenetic control.

Cells doi: 10.3390/cells15121048

Authors: Namsoo Kim Yu Jin Park Young Kyu Min Seoyoung Lim Yu Jeong Choi Seung-Tae Lee Jong Rak Choi Hongkyung Kim Saeam Shin

Background: Gene fusions play a pivotal role in the pathogenesis and classification of hematologic malignancies. RNA sequencing (RNA-seq) has emerged as a powerful tool for detecting gene fusions; however, many clinical studies have focused on targeted RNA-seq, and optimal parameters for whole transcriptome RNA-seq remain uncertain. Methods: We retrospectively analyzed whole RNA-seq data from 301 patients diagnosed with acute leukemia between October 2022 and May 2025 to characterize the landscape of pathogenic gene fusions. Fusions were identified using the Arriba algorithm, and subsampling analyses were performed on cases with recurrent fusions to determine the minimum sequencing output required for reliable detection. Results: Pathogenic gene fusions were identified in 113 of 301 patients (37.5%). Whole RNA-seq detected fusions that were not identifiable by conventional assays, including UBTF::ATXN7L3, and highlighted frequent fusion events, such as ZNF384 rearrangements. Subsampling analysis demonstrated that a sequencing output ≥ 100 million reads (moderate confidence) or ≥300 million reads (high confidence) was sufficient for 100% detection of recurrent fusions. Conclusions: Whole RNA-seq reliably detects clinically relevant gene fusions in acute leukemia, aligns well with conventional karyotyping results, and surpasses targeted RNA-seq in comprehensiveness. A sequencing output of at least 100 million reads is recommended for clinical fusion detection.

Cells doi: 10.3390/cells15121045

Authors: Mariana Pereira Nuno Vale

Drug repurposing presents as a promising strategy in oncology, particularly for urological prostate and bladder cancers, where resistance to current therapy remains a challenge. This study evaluated the anticancer potential of three antiretroviral drugs, namely ritonavir (RIT), saquinavir (SAQ), and rilpivirine (RPV), in PC-3 and UM-UC-5 cancer cell lines, using MTT, clonogenic, wound healing, toxicity assessment with fibroblast cells, and DCFDA assays; this last method included efavirenz (EFV) and etravirine (ETV) for intracellular reactive oxygen species (ROS) production. RIT and SAQ showed stronger antiproliferative effects than RPV, with lower concentration- and cell-line-dependent activity, while clonogenic assays confirmed a reduction in long-term proliferation, particularly for RIT in both cell lines and SAQ for UM-UC-5. In contrast, effects on cell migration were limited for all drugs. ROS production was cell-dependent, with EFV increasing ROS in PC-3 and SAQ and RIT in UM-UC-5 cells. Generally, all drugs showed minimal toxicity in non-malignant cells, with SAQ exhibiting some toxicity but only for concentrations higher than those required for anticancer activity. Overall, these findings suggest that antiretroviral, especially protease inhibitors, may cause anticancer effects, although these are concentration- and context-dependent, and further investigation is needed to understand the mechanisms involved.

Cells doi: 10.3390/cells15121047

Authors: Heqing Ma Abdelilah S. Gounni Ruey-Chyi Su Sam K. P. Kung

Semaphorins are a large family of proteins originally identified for their roles in axon guidance during neural development. Recent findings have established the importance of semaphorins members in modulating diverse immune responses of T cells in vitro and in vivo. Class 3 semaphorins, typified by Sema3A, signal through Neuropilin-1 and Plexin-A receptors in an activation-dependent manner, suppressing effector proliferation while promoting regulatory T cell stability and shaping cytokine profiles in autoimmunity and cancer. Sema3E and Sema3F similarly fine-tune host defense and inflammation by directing Th1/Th17 responses or restraining aberrant chemotaxis. Class 4 members, such as Sema4A and Sema4D, engage Plexin-B1, Plexin-D1, and CD72 to deliver both “forward” co-stimulatory and “reverse” signals: they amplify CD4+ and CD8+ effector functions, support T helper-B cell crosstalk, and influence tumor immunity via receptor shedding and bidirectional signaling. Finally, although less well defined, class 7 Sema7A operates indirectly—through APCs and Tregs—to regulate inflammatory recall responses and Th1/Th17 driven pathology. Together, these semaphorin-mediated pathways underscore a complex, context-dependent network that balances protective immunity against immunopathology, offering novel therapeutic targets in autoimmunity, infection, and cancer.

Cells doi: 10.3390/cells15121046

Authors: Jinrui Lv Ting Zeng Beiya Liao Ling Liu Lei Xiong Hao Xie Lin Zhu Xingzi Jiang Zhuchuyu Zhong Huaping Xie

Ezrin, expressed by the EZR gene, is a member of the ERM protein family that connects the plasma membrane to the actin cytoskeleton, participating in processes such as cell adhesion, migration, and signaling. However, its role in cardiac morphogenesis remains incompletely understood. In zebrafish (Danio rerio), two ezrin homologs, ezra and ezrb, are present. CRISPR/Cas9 gene editing technology was used to generate ezra knockout lines, and the simultaneous knockdown of ezra and ezrb was induced via morpholino oligonucleotides (MOs). To investigate the molecular mechanisms, transcriptome sequencing and bioinformatic analysis were conducted on 48 h post-fertilization (hpf) ezrin–MO embryos, with subsequent validation using a real-time quantitative polymerase chain reaction (RT-qPCR) and whole-mount in situ hybridization (WISH) experiment. The results showed that ezra−/− exhibited a compensatory upregulation of ezrb without overt developmental defects, whereas ezrin–MO embryos presented with pericardial edema, reduced cardiac chamber size, and atrioventricular valve malformations at 48 hpf. RNA-seq revealed that myocardial contraction-related genes were significantly dysregulated and apoptotic signaling pathways were activated in ezrin–MO embryos. These findings demonstrate that ezra and ezrb are functionally redundant in cardiac development and that the loss of ezrin function may lead to cardiac developmental defects and impaired myocardial contractility via the activation of apoptotic signaling pathways.

Cells doi: 10.3390/cells15121042

Authors: Jorge Manuel Vásquez-Pérez Mónica Flores-Ramos María del Pilar Ramos-Godínez Gerardo Bernabé Ramírez-Rodríguez

Major depressive disorder (MDD) is among the most common and disabling psychiatric disorders. MDD is multifactorial, influencing the central nervous, endocrine, and immune systems. Also, MDD impacts neurochemical and inflammatory pathways via shared signaling mechanisms, including metabolites, soluble factors, and extracellular vesicles (EVs, including exosomes). Here, we hypothesized that EVs from MDD patients or mice exposed to chronic unpredictable mild stress (CUMS) contain specific inflammatory signatures that may help explain the pathophysiology of that mental disorder. We included four groups: healthy female controls (n = 8), women with MDD (n = 12), healthy Balb/C female mice (n = 10), and Balb/C mice under CUMS (n = 10). We isolated and characterized exosome-enriched EVs from human and murine serum and analyzed their protein content using antibody arrays. We identified three protein sets with significant differences (p < 0.05): 36 human exosome proteins decreased in the MDD group; 18 murine exosome proteins decreased in the CUMS group; and 12 proteins showed differential expression between human and murine exosomes, mostly trending downward in the CUMS and MDD groups. We performed bioinformatic analysis to determine protein–protein interactions and gene ontology functions. We identified signaling pathways associated with MDD and chronic stress: chemokine, cytokine–cytokine receptor, JAK-STAT, pathways in cancer, Rap1, Ras, TNF signaling, and cytokine interactions. These findings highlight the importance of human and murine exosomes as critical sources for understanding depression’s molecular mechanisms.

Cells doi: 10.3390/cells15121044

Authors: Francesca Salamanna Francesca Veronesi Deyanira Contartese Giorgia Codispoti Luca Boriani Giovanni Tosini Cristiana Griffoni Alessandro Gasbarrini Gianluca Giavaresi

Adolescent idiopathic scoliosis (AIS) is a multifactorial spinal deformity with variable progression patterns, making early risk stratification challenging. Circulating and tissue biomarkers, including inflammatory, metabolic, endocrine, epigenetic, and bone-related markers, have recently been investigated as potential predictors of disease severity and progression. This systematic review evaluated the current evidence on circulating and tissue biomarkers associated with AIS severity and progression. PubMed, Scopus, and Web of Science were searched for studies published between April 2016 and April 2026. Studies assessing circulating or tissue-based inflammatory, metabolic, epigenetic, and bone-related biomarkers in AIS patients were included. Data on study design, biomarker type, analytical methods, and associations with curve severity or progression were extracted. Twenty-nine studies involving more than 4000 participants were included. Biomarkers identified included inflammatory cytokines, microRNAs, metabolic hormones, and bone metabolism markers. Most studies reported significant associations between biomarkers and curve severity, particularly for inflammatory mediators, epigenetic regulators, and bone-related markers. However, few studies evaluated longitudinal progression, and only a limited number of studies identified predictive biomarkers, including circulating miRNA panels and spermidine levels. ROBINS-I assessment showed substantial risk of bias, mainly related to confounding and selective reporting. Heterogeneity was observed across study designs and outcome definitions. Current evidence supports associations between biomarkers and AIS severity, but predictive value for progression remains limited.

Cells doi: 10.3390/cells15121043

Authors: Brigitta Ignoto Ilaria Assunta Parisi Cristin Roma Rosa Camerlingo Serena Dotolo Salvatore Tufano Monica Rosaria Maiello Nicola Normanno Alessandro Morabito Antonella De Luca Daniela Frezzetti

Mechanisms of primary and acquired resistance are responsible for treatment failure with the Epidermal Growth Factor Receptor-Tyrosine Kinase Inhibitors (EGFR-TKIs) in the majority of patients with advanced Non-Small-Cell Lung Cancer (NSCLC) carrying EGFR-activating mutations. MicroRNAs (miRNAs) are important modulators of EGFR signaling in lung cancer. Recent studies suggested the role of miR-23c as a tumor suppressor or oncogenic miRNA in different tumor types. However, the role of miR-23c in NSCLC carrying EGFR mutations and its involvement in resistance to EGFR-TKIs has not been explored yet. We found that miR-23c was strongly downregulated in H1975 and HCC827-Gefitinib-Resistant (GR) NSCLC cell lines with intrinsic and acquired resistance to gefitinib, respectively, as compared to gefitinib-sensitive cell lines. Moreover, we demonstrated that miR-23c mimic inhibited proliferation, migration, invasion, and epithelial–mesenchymal transition of resistant cells and that Interleukin-6 Receptor (IL-6R) is a direct target of miR-23c in H1975 and HCC827-GR cell lines. Importantly, miR-23c mimic re-sensitized NSCLC-resistant cells to gefitinib, whereas the combination of miR-23c mimic with a neutralizing IL-6R antibody potentiated the sensitivity to the drug. Collectively, our data demonstrated that miR-23c acts as a tumor suppressor in NSCLC cell lines carrying EGFR mutations and that the axis miR-23c/IL-6R might represent a potential target for the development of therapeutic approaches to overcome resistance to gefitinib.

Cells doi: 10.3390/cells15111041

Authors: Mario Muñoz López José Francisco López-Gil Xabier Ramírez de la piscina Viúdez Eneko Baz-Valle José Francisco Tornero Aguilera

Background: Skeletal muscle hypertrophy has traditionally been attributed to transient spikes in translational efficiency governed by the mTORC1 signaling cascade. However, contemporary molecular evidence reveals that sustained macroscopic growth is strongly associated with the physical expansion of the translational machinery itself. The activation of RNA Polymerase I and the subsequent synthesis of new ribosomes represent a critical biological correlate for long-term protein accretion. Objective: This comprehensive review critically examines ribosome biogenesis as the primary structural bottleneck shaping human skeletal muscle adaptation, differentiating acute signaling efficiency from chronic translational capacity. Synthesis: We dissect the molecular orchestration of nucleolar expansion and critically address the pervasive methodological pitfalls plaguing the current literature. Specifically, we highlight the moving denominator paradox, demonstrating how flawed bulk RNA normalization strategies systematically underestimate true ribosomal accretion in actively growing tissue. By synthesizing in vivo human evidence, we delineate how age, concurrent training, and training volume modulate this structural capacity. We further establish the high-responder phenotype as a function of successful nucleolar adaptation. Finally, we explore advanced molecular frontiers, including epigenetic chromatin remodeling, ribosomal heterogeneity as an emerging frontier, non-coding RNA regulation, and nuclear mechanotransduction via the YAP/TAZ axis. Conclusions: Acute anabolic signaling is merely permissive. Permanent hypertrophic adaptation fundamentally relies on overcoming the translational capacity bottleneck. Shifting the scientific and applied focus toward the architectural expansion of the nucleolus will fundamentally redefine practical hypertrophy programming and clinical interventions for sarcopenia.

Cells doi: 10.3390/cells15111040

Authors: Xiangyu Lu Pujun Li Lei Cao Tao Liu Yajun Ma Yue Wang Chenxi Piao Hongbin Wang

The link between neutrophil extracellular traps (NETs) and hepatocyte ferroptosis in liver ischemia–reperfusion injury (LIRI) is unclear. Adipose-derived mesenchymal stem cell exosomes (ADSCs-Exo) hold therapeutic potential for LIRI. This study employed miniature pigs to investigate the NETs’ role and ADSCs-Exo’s protection in LIRI. In vitro, established hepatocyte oxygen-glucose deprivation/reoxygenation (OGD/R) model and Transwell co-culture system with polymorphonuclear neutrophils (PMNs). In vivo, a laparoscopic minimally invasive LIRI model was constructed in miniature pigs, followed by ADSCs-Exo intervention. Results demonstrated that NETs exacerbate OGD/R-induced hepatocyte ferroptosis via myeloperoxidase. ADSCs-Exo inhibited NET formation via the NADPH/MAPK pathway, thereby mitigating ferroptosis, and ultimately improved liver histopathology and function. This study is the first to demonstrate in a large animal model that ADSCs-Exo alleviate LIRI by inhibiting NET formation via the NADPH/MAPK pathway, consequently attenuating hepatocyte ferroptosis. These findings provide novel insights into LIRI pathogenesis, support the translational potential of ADSCs-Exo as a cell-free therapeutic strategy, and highlight the value of the miniature pig model in liver research.

Cells doi: 10.3390/cells15111039

Authors: Camilla Cittadini Elisabetta Iessi Rosa Vona Paola Matarrese

Colorectal cancer (CRC) is the third most diagnosed malignancy and the second leading cause of cancer-related mortality worldwide. A consistent and epidemiologically well-documented feature of CRC is its sexual dimorphism: age-standardized incidence rates are 33–45% higher in men than in women, and mortality rates differ by 43–50%. Beyond epidemiology, biological sex influences tumor location, molecular subtype, and clinical outcome. Women more frequently develop right-sided, microsatellite-unstable tumors driven by the CpG island methylator phenotype pathway, whereas men predominantly present with left-sided, chromosomally unstable tumors harboring APC, KRAS, and TP53 mutations. Sex steroid hormones play a central modulatory role: estrogens, primarily via estrogen receptor β (ERβ), exert tumor-suppressive effects on colonic epithelium, whereas androgens promote pro-inflammatory and pro-tumorigenic signaling through androgen receptor (AR)-dependent pathways. The gut microbiome displays sex-specific compositional profiles (‘microgenderome’) and contributes to sex-specific CRC susceptibility through bidirectional interactions with sex hormones, shaping distinct immunological and metabolic microenvironments. Finally, sex influences the pharmacokinetics of fluoropyrimidines, the toxicity of targeted agents, and the response to immune checkpoint inhibitors. This review summarizes current evidence on sex-related differences in CRC epidemiology, molecular pathology, hormonal regulation, gut microbiota composition, and treatment outcomes, highlighting the need to systematically incorporate sex as a biological variable in CRC research and clinical practice.

Cells doi: 10.3390/cells15111038

Authors: Hangzhen Zhou Jiaxin Liu Lie Yang Shan Li Shuwei Li

Hair follicle (HF) stem cells are multipotent adult stem cells that play a key role in the hair follicle cycle. However, it remains poorly understood how the outer root sheath (ORS)—specifically, the stem cells in the bulge region of the hair follicle—promotes skin repair. This study aims to investigate the role of bulge stem cells in tissue growth and repair and to determine whether the ORS of transplanted hair follicles can facilitate skin repair. We further seek to elucidate the mechanisms by which bulge stem cells contribute to hair follicle development, regeneration, and skin wound healing. In this study, hair follicle samples were obtained from neonatal mice using microdissection. Hair follicle morphology was assessed by Sirius red staining, H&E staining, and transmission electron microscopy. Immunofluorescence staining was used to detect changes in CD34 and SOX9 protein expression. Additionally, microdissection-based hair follicle transplantation and Western blotting were employed to investigate protein activation and inhibition in the Wnt/β-catenin signaling pathway. The results show that the hair follicle bulge, inner root sheath, and dermal papilla all increase in size as hair follicles grow, with each structure growing relatively rapidly on day 7. Treatment with Teplinovivint effectively inhibits the expression of Wnt/β-catenin signaling pathway-related proteins and hair follicle stem cell markers. Damaged hair follicle tissues cultured in vitro are capable of self-repair. At the transplantation site, the skin gradually closes as the outer root sheath wound heals. In contrast, the central region of the outer root sheath becomes progressively filled with numerous dividing cells and extracellular matrix. The inner portion of the outer root sheath is densely populated with cells, and the markers CD34 and SOX9 are also widely distributed. This indicates that activation of the Wnt/β-catenin signaling pathway enhances the proliferation and differentiation of hair follicle stem cells, thereby promoting hair follicle growth, repair of damaged follicles, and healing of skin wounds. Furthermore, this study demonstrates the feasibility of using transplanted outer root sheath (ORS) to repair skin wounds—specifically, the potential to achieve large-scale hair regeneration from a limited number of hair follicle stem cells—providing a new approach for the clinical treatment of skin injury disorders. Nevertheless, achieving long-term, stable, and scalable clinical translation of ORS stem cells for hair follicle regeneration remains a major challenge.

Cells doi: 10.3390/cells15111037

Authors: Yan Li Yan Wang Shiying Li Kaijie Wang Jahangir Alam Shiyuan Gong Ying Zhu Jiande D. Z. Chen

The vagus nerve (innervating the gut from esophagus to proximal colon) and sacral nerve (innervating distal colon and rectum) are key parasympathetic regulators of gastrointestinal (GI) function. While vagus nerve stimulation (VNS) has shown therapeutic potential in upper GI disorders, its role in modulating distal colon and rectal function remains poorly understood. This study investigated the effects and mechanisms of VNS on distal colon transit and rectal tone in rats. Adult male Sprague Dawley rats were implanted with stimulation electrodes at the cervical or auricular vagal afferent nerve. VNS was applied with varying frequencies, pulse widths, and amplitudes. Rectal tone was assessed using a barostat device, and distal colon transit was evaluated using bead expulsion. Nitrergic and cholinergic contributions were examined using L-NAME and nNOS expression, and acetylcholine ELISA and ChAT expression, respectively. Central pathways were investigated by immunofluorescence staining of c-fos and ChAT in the nucleus tractus solitarius (NTS). Sacral efferent pathway was assessed by chemogenetic inhibition of the dorsal motor nucleus of the vagus (DMV) and Barrington nucleus (BN/PMC). VNS (5 Hz, 0.1 and 0.5 ms, 0.5 mA) significantly increased rectal volume, indicating relaxation, and accelerated distal colon transit. L-NAME abolished VNS-induced rectal relaxation, while nNOS expression in the rectum was upregulated, confirming nitrergic mediation. Distal colon transit was associated with increased acetylcholine release and ChAT expression, highlighting cholinergic involvement. VNS enhanced c-fos and ChAT-positive neurons in the NTS, suggesting central integration of vagal afferent signals. Chemogenetic inhibition of DMV and BN attenuated rectal relaxation, indicating that VNS effects are mediated via a vagal–NTS–sacral pathway. VNS modulates distal colon transit and rectal tone through coordinated nitrergic and cholinergic signaling and central vagal-to-sacral circuits. These findings reveal functional crosstalk between vagal and sacral parasympathetic pathways and provide mechanistic insight into potential VNS therapy for lower GI disorders.

Cells doi: 10.3390/cells15111036

Authors: Pawel Kordowitzki Renata Voltolini Velho Sandra Bock Jalid Sehouli Sylvia Mechsner

Background: Given the challenges in early detection and diagnosis, understanding the molecular underpinnings of endometrioid ovarian cancer (EOC) is crucial for improving patient outcomes. This multi-level study provides a new perspective on EOC, focusing on the expression of ARID1a (BAF250a) and RIF1. Methods: This study evaluates patient cohorts with EOC through semi-quantitative immunohistochemical staining of BAF250a (protein encoded by ARID1a) and RIF1 proteins alongside mutations that influence the gene expression of ARID1a and RIF1. Besides survival analyses, platinum- and taxane-based treatment responsiveness with regard to ARID1a and RIF1 expression has been analyzed using an online available database. Results: Histological and immunohistochemical analysis of clinical samples revealed a significant reciprocal alteration in protein expression, characterized by a marked reduction in the tumor suppressor BAF250a (p < 0.0001) and a concomitant elevation of RIF1 (p < 0.0001) in EOC compared to controls. Tumors harboring mutations in BRCA1 exhibited significantly (p = 2.82 × 10−4) lower ARID1a expression levels compared with corresponding wild-type tumors, whereas LAMB3-mutant tumors showed a significant (p = 5.16 × 10−3) upregulation of RIF1 mRNA expression. Conclusions: In conclusion, our study offers a new perspective, emphasizing that EOC is a distinct clinical and molecular entity. We demonstrated the expression patterns of ARID1a/BAF250a and RIF1 in EOC, establishing their potential relevance in the context of tumor biology and malignant transformation.

Cells doi: 10.3390/cells15111035

Authors: Carly M. Van Wagoner Lydia E. Kitelinger Matthew R. DeWitt Claire A. Conarroe AeRyon Kim Aaron B. Streit Richard J. Price Timothy N. J. Bullock

Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer (BrCa), owing to its lack of targetable receptors and resistance to chemical and molecularly targeted therapeutic approaches. While chemotherapy and surgical resection remain the standard of care, these interventions have significant side effects and varying patient outcomes. Thermally ablative focused ultrasound (T-FUS)—a non-invasive and non-ionizing therapy that utilizes targeted acoustic energy to debulk tumors—has displayed immunomodulatory effects in BrCa. However, T-FUS as a monotherapy has had limited clinical efficacy in TNBC due to the presence of anti-inflammatory immunosuppressive myeloid cells (isMCs). We hypothesized that the elimination of isMCs or initiating tumoricidal activity from them would lead to augmented activity of T-FUS. Thus, we interrogated the ability of myeloablative chemotherapies and antibodies; myeloid recruiting chemokine receptor blockade; and TLR agonists to remodel the tumor myeloid populations. Consistent with our previous studies, we found that while myeloablative chemotherapies decreased circulating isMCs, they had little impact on intratumoral isMCs. In contrast, antibodies targeting Ly6C and Ly6G ablated intratumoral isMCs and systemic isMCs, yet their effect was transient and was accompanied by a surprising depletion of T cells. While targeting CCR2, the dominant chemokine receptor for intratumoral isMC diminished a large subset of immunosuppressive cells within the TME; it also depleted T cells and dendritic cells. Contrary to previous studies, TLR stimulation failed to repolarize myeloid cells into a pro-inflammatory, tumoricidal phenotype but did lead to their depletion from the tumor microenvironment (TME) and mobilization of conventional dendritic cells to the draining lymph nodes. We therefore hypothesized that combining isMC depletion and TLR-driven immune activation would enhance FUS efficacy; however, this combinatorial regimen did not enhance overall survival or control tumor volume after T-FUS treatment. Thus, the BrCa TME is highly resistant to approaches intended to remodel the myeloid cell component which fail to synergize with T-FUS-mediated tumor ablation.

Cells doi: 10.3390/cells15111034

Authors: Md. Sohanur Rahman Mohammed Daira

Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) dysfunction is increasingly recognized as a key contributor to a broad spectrum of human diseases beyond classical cystic fibrosis (CF). CFTR is a cAMP-regulated chloride and bicarbonate ion channel expressed in both epithelial and non-epithelial tissues, where it regulates ion homeostasis, mucosal hydration, and cellular signaling. Both inherited CFTR mutations and acquired dysfunction resulting from environmental or inflammatory factors can disrupt these physiological processes and drive disease progression. Current evidence linking CFTR dysregulation to respiratory diseases, such as cystic fibrosis, chronic obstructive pulmonary disease (COPD), asthma, and HIV-associated airway disease, as well as cardiovascular, renal, neurological diseases, and cancer, is comprehensively discussed. Mechanistically, impaired CFTR function promotes oxidative stress, chronic inflammation, epithelial barrier dysfunction, altered mucociliary clearance, and dysregulation of signaling pathways, including NF-κB, TGF-β, PI3K/Akt, MAPK, and Wnt/β-catenin. In the context of HIV infection and cigarette smoke exposure, CFTR suppression is mediated in part by TGF-β signaling and miRNA-dependent mechanisms, resulting in compromised airway defense and increased susceptibility to pulmonary complications. Recent studies further demonstrate that CFTR dysregulation alters the expression of genes involved in fibrosis, inflammation, angiogenesis, and epithelial–mesenchymal transition (EMT). Notably, CFTR may act as either a tumor suppressor or a context-dependent oncogene, depending on tissue type and signaling milieu, highlighting its complex role in cancer biology. Advances in CFTR-targeted therapies, including potentiators, correctors, gene therapy, and combination approaches, have markedly improved outcomes in CF and may offer therapeutic potential for diseases associated with acquired CFTR dysfunction. We summarize the systemic consequences of CFTR dysregulation and the need for further mechanistic and translational research to clarify its role across diverse human diseases.

Cells doi: 10.3390/cells15111033

Authors: Alexandré Delport Ebrahim Ally Shantal Maharaj Raymond Hewer

While amyloid precursor protein (APP) overexpression in adipose tissue is a recognized consequence of high-fat diet (HFD) feeding, its role in metabolically active organs and the mechanisms linking it to systematic dysfunction remain unclear. In particular, the potential for diet-induced APP dysregulation in the other tissues and the contribution of its βC-terminal fragment (βCTF) are poorly characterized. Using a high-fat diet (HFD) mouse model to induce systematic metabolic stress, we assessed APP and βCTF levels across multiple tissues. HFD triggered a tissue-specific response, with APP levels increasing >2-fold in visceral and subcutaneous white adipose tissue (WAT) and in the kidney but remained unchanged in the liver and brain. βCTF levels were significantly elevated in the visceral WAT (3-fold) and kidney. In these responsive tissues, APP and βCTF accumulated within mitochondria, which coincided with significantly reduced complex I and IV activities. Complementary in vitro studies confirmed that APP levels can dictate mitochondrial function. Furthermore, we identified that cytokines–IL-4, IL-13, TNF-α, and IL-1β–induced APP transcription, providing a mechanistic link between diet-induced inflammation and APP dysregulation. Collectively, our findings demonstrate that APP is overexpressed in response to HFD in select peripheral tissues, which coincides with reduced mitochondrial complex enzyme activities and increased cytokine levels.

Cells doi: 10.3390/cells15111031

Authors: Asunción Espinosa-Sánchez Amancio Carnero

Cancer research has undergone a fundamental transformation in recent decades due to the integration of artificial intelligence (AI) models into the study of tumor biology. However, tumor evolution, driven by genetic and phenotypic alterations leading to heterogeneity, resistance and metastasis, remains a major challenge in oncology. To understand these processes is crucial for developing effective therapeutic strategies and improving patient outcomes. Conventional methods often fail to capture the complexity and dynamics of these processes. In contrast, AI tools have the ability to integrate and analyze large-scale multi-omics, imaging and clinical data, offering the capability to decode tumor complexity. AI-driven methods facilitate multi-modal data integration, enabling the recognition of patterns that connect molecular alterations with phenotypic outcomes. In functional genomics, AI tools predict the effects of genetic variants, identify regulatory elements and map dysregulated pathways, thus clarifying mechanisms underlying tumor development and resistance. In the imaging field, deep learning techniques improve tumor segmentation, characterization and longitudinal monitoring, providing more accurate insights into tumor progression and treatment response. Predictive modeling could allow the anticipation of tumor evolution and drug response, supporting adaptive therapeutic plans and real-time treatment adjustments. Moreover, AI supports biomarker discovery, patient stratification and decision support systems that can improve clinical trial design and accelerate the development of personalized therapies. However, these advances raise important ethical challenges, including data privacy, algorithmic bias and the preservation of patient autonomy. Addressing these concerns is essential to ensure the responsible deployment of AI in oncology.

Cells doi: 10.3390/cells15111032

Authors: Kristina Živić Marijana Pavlović Marija Ostojić Danijel Galun Aleksandar Pavić Tatjana Srdić-Rajić Jelena Grahovac

Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive cancer types with a dismal prognosis, where early metastatic dissemination and rich desmoplastic stroma limit the therapeutic efficacy. Nischarin (NISCH)/Imidazoline-1 receptor (IR-1) is a potential tumor suppressor that is involved in the regulation of cell migration, invasion, and cytoskeletal organization. Importantly, several clinically approved agonists have been shown to target this receptor. This study aimed to examine NISCH expression in PDAC and the effects of its agonists with the intent of drug repurposing. NISCH was expressed in PDAC tumor tissue, in both the epithelial and stromal compartments of tumors, and higher NISCH expression was associated with longer patient survival. Out of the three tested NISCH agonists, moxonidine, clonidine and rilmenidine, rilmenidine was the only one affecting cancer cell viability and at high doses induced cancer cell apoptosis. Transcriptome analysis of rilmenidine-treated PDAC cells revealed changes associated with cytoskeletal organization, translating into decreased adhesion and migration in vitro. In cancer-associated fibroblasts (CAFs), rilmenidine treatment decreased the expression of activation markers and limited cancer cell-CAF cytokine communication in the co-culture. Ultimately, in the in vivo tumor xenograft zebrafish model, rilmenidine reduced the metastatic spread of pancreatic cancer cells. Our results suggest that the NISCH agonist rilmenidine is a promising candidate for drug repurposing as an antimetastatic agent in PDAC, and that NISCH can be a potential target for the development of new PDAC therapeutics.

Cells doi: 10.3390/cells15111030

Authors: Douglas M. Ruden

Environmental exposures can influence phenotypes across generations, yet the cellular routes by which somatic stress signals reach the germline remain poorly defined. piRNAs are attractive candidates for transgenerational epigenetic inheritance because they silence transposable elements, guide chromatin regulation, carry a stabilizing 3′ 2′-O-methyl modification, and participate in self-reinforcing amplification pathways, including ping-pong amplification in animals and RNA-dependent RNA polymerase (RdRP)-mediated secondary small-RNA amplification in systems such as C. elegans. This review examines evidence linking piRNAs, macrophage biology, and environmentally induced inheritance. We first summarize small-RNA inheritance in animals, plants, and ciliates, emphasizing C. elegans piRNA-triggered epigenetic memory and plant RNA-directed DNA methylation as parallel small-RNA-based inheritance systems. We then discuss emerging evidence that macrophage polarization states contain distinct piRNA signatures and release extracellular vesicles carrying non-coding RNAs. Finally, we revisit the Drosophila ectopic large bristle outgrowth (ELBO) phenotype as a possible example of macrophage-like hemocytes linking stress, tissue remodeling, and heritable morphological variation. We propose the macrophage-mediated morphological evolution (M3) model as a testable framework connecting environmental stress to transgenerational phenotypes.

Cells doi: 10.3390/cells15111029

Authors: James Chmiel Wiktor Gawełczyk Julia Soczyńska Jerzy Leszek

Alzheimer’s disease is characterized by a chronic, long-term neurodegenerative process and an increasing need for easily accessible biomarkers that enable early diagnosis and disease monitoring. For this reason, tears have attracted growing interest as a potential source of such biomarkers, and prolactin-inducible protein is a candidate tear protein of mechanistic interest whose clinical value remains to be established as a biomarker of Alzheimer’s disease. The literature indicates that prolactin-inducible protein is physiologically present in the lacrimal apparatus. Proteomic studies in patients with Alzheimer’s disease have repeatedly demonstrated decreased levels of prolactin-inducible protein in tears, typically accompanied by reduced concentrations of other proteins associated with normal lacrimal gland function. Although the evidence remains inconclusive, these findings suggest that alterations in prolactin-inducible protein levels may reflect lacrimal gland dysfunction related to neurodegenerative processes, autonomic dysregulation, and inflammation. Nevertheless, the lack of specificity of prolactin-inducible protein for Alzheimer’s disease, as well as the influence of various factors on its concentration, limit its value as a standalone biomarker. The most plausible approach is the incorporation of prolactin-inducible protein into multimarker panels, which could enable improved patient stratification and assessment of lacrimal gland dysfunction in Alzheimer’s disease.

Cells doi: 10.3390/cells15111028

Authors: Avizit Das Tatsuya Yoshizawa Daisuke Yamada Tomonori Tsuyama Yoshifumi Sato Tomoya Mizumoto Takeshi Yoneshiro Shingo Kajimura Kazuya Yamagata

Adipose tissue (AT) browning is an inducible cellular phenomenon that promotes lipid oxidation to increase energy expenditure, reducing adiposity. Various transcription regulators involved in the AT browning process have been reported, but their complex molecular mechanisms remain poorly understood. Here, we explore the effects of SIRT7, one of seven mammalian sirtuins, on AT browning and elucidate the underlying mechanisms. SIRT7 deficiency increased the expression of browning genes in beige adipocytes differentiated from subcutaneous white AT (scWAT) stromal vascular fraction (SVF) cells isolated from adipocyte-specific Sirt7 knockout (Sirt7 AdKO) mice. The effect of SIRT7 on beige adipocyte differentiation was confirmed in Sirt7 knockdown (KD) mouse scWAT and human supraclavicular brown AT (scBAT) SVF cell lines. Mechanistically, SIRT7 deacetylated PPARγ2 (peroxisome proliferator-activated receptor γ2) at lysine (K) 382, thereby attenuating interaction with the transcriptional coactivator PRDM16 (PR domain-containing 16). In differentiated beige adipocytes, the acetylation-mimicking mutant PPARγ2K382Q had higher transcriptional activity compared with the deacetylation-mimicking mutant PPARγ2K382R. Furthermore, the interaction between endogenous SIRT7 and PPARγ2 decreased at the onset of beige adipocyte differentiation. Our findings reveal that SIRT7 is an important thermogenic regulator that puts the brake on AT browning by deacetylating PPARγ2.

Cells doi: 10.3390/cells15111027

Authors: Xiang Zhang Ye Dai Yishuo Shi Fang Yuan

Metabolic state strongly shapes social and reproductive behaviors, yet the neural circuits that convert internal energy signals into behavioral responses remain poorly defined. The ventral premammillary nucleus (PMv) of the hypothalamus has been implicated in this process, particularly through leptin receptor-expressing (LepRb) neurons, but its brain-wide circuit organization is still unclear. Here, we used Cre-dependent retrograde (RV) and anterograde (HSV) viral tracing techniques in LepRb-Cre mice to construct a comprehensive, single-cell-resolution input–output map of PMvLepRb neurons. 3D reconstruction showed that these neurons receive dense convergent inputs, mainly from hypothalamic and forebrain regions involved in energy balance, motivation, and limbic processing. In contrast, their outputs extend not only back to several input regions but also prominently to midbrain and pontine autonomic centers, including the periaqueductal gray (PAG) and parabrachial nucleus (PB). Quantitative analysis revealed that forebrain regions were more likely to participate in reciprocal connectivity, whereas brainstem regions were dominated by outgoing projections. This organization suggests that PMvLepRb neurons are positioned to integrate metabolic and motivational signals and relay them to downstream systems controlling instinctive behavioral and autonomic responses. These findings provide a structural basis for understanding how energy state can influence decisions related to social competition and reproduction.

Cells doi: 10.3390/cells15111026

Authors: Vasudevarao Penugurti Rajni Kant Che-Chia Hsu

Mitochondria play essential roles in cellular metabolism and signaling, regulating biosynthetic pathways, calcium homeostasis, redox balance, and cell fate beyond ATP production. Their continual remodeling through fusion, fission, and mitophagy maintains mitochondrial quality control and adapts organelle function to cellular demands. Here, we review how mitochondrial dynamics, fusion, fission, and mitophagy modulate metabolic reprogramming and signaling to drive cancer progression and therapy resistance. Emerging evidence indicates that in cancer, mitochondrial fusion enhances respiratory efficiency and oxidative phosphorylation, whereas fission promotes glycolytic adaptation, rapid biomass accumulation, and stress tolerance. Mitophagy further refines metabolic fitness by eliminating damaged mitochondria and sustaining redox homeostasis. Together, these processes underscore that dysregulation of mitochondrial dynamics is a hallmark of cancer and a key driver of metabolic reprogramming and therapeutic resistance. In this review, we summarize how mitochondrial fusion, fission, and mitophagy govern metabolic circuitry in cancer development and therapy resistance. We highlight their functional impact on tumor progression and discuss emerging therapeutic strategies targeting mitochondrial dynamics and associated machinery. Understanding this dynamic metabolic crosstalk may reveal new vulnerabilities and guide the development of mitochondria-targeted cancer therapies.

Cells doi: 10.3390/cells15111025

Authors: Dina Sofia da Silva Telinhos Markus Maniak

Ether lipids in varying amounts are membrane constituents and storage material in the protist and animal kingdoms, but are largely absent from fungi and plants. Their biosynthesis pathway starts in the peroxisome and involves a set of well-conserved enzymes. Only one step, the reduction of alkyl-dihydroxyacetone-phosphate to alkyl-glycerol-3-phosphate, is mediated by so-called short-chain dehydrogenases/reductases, which are members of huge protein families. Here, using GFP fusions, we identify a peroxisomal enzyme in Dictyostelium, as well as a highly related protein residing in the endoplasmic reticulum. Single-gene knockouts indicate that these enzymes largely compensate for one another, suggesting a flexible redistribution of lipid metabolites between these organelles. The double knockout, however, is severely affected in ether lipid composition and displays a clear growth retardation. The defects can also be reverted by expression of the cognate yeast enzyme, demonstrating conservation of this metabolic step across kingdoms of life.

Cells doi: 10.3390/cells15111024

Authors: Sung Yong Ahn Chris Hyunchul Jo

Mesenchymal stromal cell (MSC) adhesion and retention at sites of cartilage degeneration are critical for improving cartilage repair. This study investigated whether platelet-rich plasma (PRP) enhances the adhesion and short-term retention of bone marrow-derived MSCs (BM-MSCs) and chondrocytes under in vitro and ex vivo conditions. BM-MSCs and chondrocytes were treated with PRP or pretreated with PRP for 10 or 30 min, and cell adhesion to collagen-coated surfaces was evaluated using a cell viability assay. Ex vivo adhesion and short-term retention of BM-MSCs on osteochondral discs with varying lesion severity were assessed by fluorescence imaging analysis. PRP significantly enhanced the adhesion of both BM-MSCs and chondrocytes in a time-dependent manner, with the 30 min PRP pretreatment group showing the greatest effect. BM-MSC attachment in the 30 min PRP pretreatment group was significantly higher than that in the untreated control group after 30 min of incubation (p < 0.001), whereas chondrocyte attachment was also significantly increased following PRP pretreatment. In addition, PRP pretreatment significantly enhanced BM-MSC attachment compared with PRP treatment alone at 20 and 30 min of incubation (both p < 0.001). In ex vivo experiments, adhesion and short-term retention increased significantly with increasing lesion severity from G1 to G3 (p < 0.05 and p < 0.01, respectively). In G2 and G3 lesions, PRP pretreatment for 30 min significantly enhanced BM-MSC adhesion and short-term retention compared with the control group (both p < 0.01). These findings suggest that PRP may improve the early adhesion and retention of MSCs on damaged cartilage and support the potential use of PRP as a biological adjunct for MSC-based cartilage repair strategies.

Cells doi: 10.3390/cells15111022

Authors: Catarina Hilário Sara Raimundo Catarina Dias Joana Saramago Telma Oliveira Rute Pereira Sofia Quental João Parente Freixo Luís Gales Jorge Oliveira Rosália Sá Mário Sousa

Primary ciliary dyskinesia (PCD) is a rare genetic disorder mainly characterized by impaired mucociliary clearance and chronic respiratory symptoms. From a consanguineous family, a male patient, although with respiratory complaints since birth, was diagnosed with PCD only in adulthood. Whole-exome sequencing disclosed a novel homozygous intronic single-nucleotide deletion, NM_001369.3(DNAH5):c.13723+4del, initially classified as of uncertain clinical significance. Digital highspeed videomicroscopy (HSVM) evidenced a null ciliary beating frequency; transmission electron microscopy showed absence of outer dynein arms (class-1); and immunofluorescence (IF) demonstrated markedly absent DNAH5 protein level in the apical cilia region with delocalization to the transition and basal-body regions. Bioinformatic analysis predicted altered splicing at the donor splice site of exon 78, whereas mRNA sequencing revealed two splicing defects: the mainly expressed transcript corresponding to exon 78 skipping and a minor transcript originated from a cryptic splice site in exon 78. The patient was infertile and showed severe oligoteratozoospermia. Sperm IF analysis revealed absence of DNAH5 from the flagellum with accumulation at the neck region. The family study confirmed homozygosity. The present results support a pathogenic role for the c.13723+4del variant and underscore the importance of integrating clinical, ultrastructural, DNA, mRNA and protein analyses to clarify and contribute to PCD diagnosis.

Cells doi: 10.3390/cells15111023

Authors: David Brownell Alexane Thibodeau Guillaume Locatelli Aiden Smith Megan Richer Stéphane Chabaud Stéphane Bolduc

Tissue engineering using the self-assembly approach represents a promising technology. However, age-related reductions in extracellular matrix deposition by stromal cells limit the mechanical robustness of reconstructed tissues what can be critical for midurethral sling reconstruction. Indeed, stress urinary incontinence predominantly affects women over 50 years of age and is commonly treated by implantation of midurethral slings, whose synthetic versions have raised concerns regarding safety and long-term tolerance. In this study, we investigated whether biochemical modulation could enhance collagen deposition and mechanical properties of self-assembled dermal tissues reconstructed from female donors of different ages. Dermal fibroblasts were cultured in the presence of ascorbic acid, and the effects of hormonal supplementation, metabolic and hypoxia-related stimuli, and insulin signaling activation were evaluated using collagen quantification, histological analyses, and mechanical testing. Fibroblasts derived from younger donors deposited significantly more collagen than those from older female donors. Among all tested conditions, insulin like growth factor 1 (IGF 1) markedly increased collagen deposition in a dose-dependent manner, including in fibroblasts from women over 50 years of age, whereas β-estradiol and progesterone had no significant effect on collagen content. Although β-estradiol slightly increased tissue thickness, only IGF-1 supplementation resulted in substantial improvements in perforation strength, stiffness, displacement at break, and toughness. These results demonstrate that IGF-1 is a potent enhancer of extracellular matrix production and mechanical performance in dermal tissues reconstructed by the self-assembly approach, and represents a promising strategy to improve the development of biological midurethral slings.

Cells doi: 10.3390/cells15111021

Authors: Sophia Elisa Wiener Jens Conrad Saman Javid Sophie Gieß Max Jägersberg Harald Krenzlin Naureen Keric Anne Régnier-Vigouroux Carsten Geiß

The rapid expansion of individualized treatment strategies necessitates advanced patient-specific screening platforms recapitulating tumor complexity. Here, we present such a platform preserving heterotypic cellular interactions and three-dimensional architecture which are both critical for predicting therapeutic responses. This cost-effective and versatile platform allows the rapid generation and functional testing of spheroids derived from primary human glioblastoma specimens. To maximize accessibility and ease of integration, we took advantage of commercially available kits to optimize the protocol of spheroid generation we had previously established. This enabled the robust, adaptable, and reproducible formation of homogeneous, multicellular spheroids within a few days. Spheroids generated from one patient specimen were structurally stable and showed a high degree of homogeneity over time. Immunohistochemical and flow cytometric analyses further revealed patient-specific cellular heterogeneity, underscoring the platform’s ability to preserve clinically relevant tumor features. Functionally, we demonstrate the applicability of this system for drug response profiling by assessing invasion dynamics following treatment with clinically relevant compounds. Collectively, our results establish a scalable and adaptable 3D screening platform that enables rapid, patient-specific phenotypic and functional analyses. This approach provides a powerful tool to complement existing clinical workflows and holds promise for improving the prediction of therapeutic responses in glioblastoma.

Cells doi: 10.3390/cells15111020

Authors: Andressa da Silva Cazote Glenda Domingos Mascarenhas Hugo Perazzo Kim Mattos Geraldo Maria Pia Diniz Ribeiro Juliana Arruda de Matos Pedro Emmanuel Alvarenga Americano do Brasil Sandra Wagner Cardoso Beatriz Grinsztejn Valdiléa Gonçalves Veloso Cynthia Machado Cascabulho José Henrique Pilotto Diogo Gama Caetano Milena Neira Guimarães Goulart Nathalia Beatriz Ramos de Sá Dalziza Victalina de Almeida Fernanda Heloise Côrtes Mariza Gonçalves Morgado Carmem Beatriz Wagner Giacoia-Gripp

This study aimed to identify the distinct intrinsic response potential of γδ T cells from COVID-19 patients with different illness severities, to better understand the implication of these cells in COVID-19 disease. Forty-four COVID patients were enrolled at hospitalization and classified as: moderate without oxygen support (MWO2; N = 15), moderate with oxygen support (MO2; N = 15), or severe disease requiring mechanical ventilation (SD; N = 14). γδ T cells were characterized ex vivo, isolated from peripheral blood cells, stimulated in vitro with OKT3 and K562 cells, and evaluated for functional markers by flow cytometry. Ex vivo analysis identified 16.21% of total γδ T cells as Vδ1−Vδ2−. SD patients presented a lower frequency of TRAIL+ and of IL-17-producing Vδ2 cells, as well as lower value of fluorescence intensity values for TNF-α in Vδ2 cells, than MWO2 patients (p < 0.05). In addition, paired analyses showed a lower frequency of IL-17-producing than CD161+ Vδ2 cells in SD patients (p < 0.05). These observations suggest a more restricted response potential of the Vδ2 subset in severe disease, show the impact of general immune dysregulation on these cells, or even suggest some role for IL-17-producing Vδ2 cells in preventing critical symptoms.

Cells doi: 10.3390/cells15111019

Authors: Nora Hosny Houda Cohen John Bauer Jeff Schreifels Rachel Lin Brian R. Thompson Joseph M. Metzger

The landscape of human cardiac biology was transformed by the discovery that adult somatic cells can be reprogrammed into induced pluripotent stem cells, enabling patient-specific disease modeling, drug testing, and regenerative strategies without the prior ethical or biological constraints. Subsequent advances in directed differentiation made the generation of human iPSC-derived cardiac myocytes reliable and scalable. Despite this progress, a central limitation has remained: these cells are developmentally immature, resembling fetal cardiac myocytes in structure, metabolism, and function. This immaturity restricts their utility for modeling adult-onset disease, predicting drug responses, and achieving clinical translation. Maturation is now understood as a multifactorial symphony, requiring coordinated molecular, structural, and environmental inputs rather than single interventions. As a result, the field is shifting toward integrative approaches that incorporate 3D architecture, multicellular systems, and biomimetic environments to better replicate native cardiac tissue. While fully adult-like myocardium remains an ongoing goal, advances in bioengineering and system-level design are narrowing the gap, with success increasingly defined by the generation of functional cardiac tissue rather than isolated cell maturity.

Cells doi: 10.3390/cells15111018

Authors: Svetlana Lebedeva Yan Bravyy Anna Beknazarova Elena A. Smolyarchuk Kerim Mutig

Acute kidney injury (AKI) is a life-threatening event prevalent in hospitalized patients but also not rare among endurance sports athletes. Hypoxia, oxidative stress, and sterile inflammation are the key pathophysiological factors driving kidney damage in AKI. Zinc is an essential trace element required for the intact function of approximately 3000 proteins (~10% of the human proteome), including over 300 enzymes for which zinc serves as a cofactor. Cell biological tasks of zinc signaling include adaptive responses to hypoxia and oxidative stress, as well as anti-inflammatory effects. The underlying molecular pathways involve modulation of hypoxia-inducible factor signaling, suppression of reactive oxygen species (ROS) generation, and inhibition of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), the latter being the major pro-inflammatory transcription factor. As a catalytic cofactor for the “classical” histone deacetylases, zinc is essential for epigenetic control of gene expression, thereby exerting further adaptive effects. Apart from the intracellular zinc signaling, extracellular zinc elicits cytoprotective and anti-inflammatory effects via the G Protein-Coupled Receptor 39 (GPR39). GPR39 activation by zinc binding may exert antioxidant and anti-inflammatory effects mediated by the zinc-finger protein A20 (TNFAIP3) and NF-κB suppression, followed by reduced production of pro-inflammatory cytokines such as tumor necrosis factor (TNF), interleukin-1β (IL-1β), and IL-6. At the same time, GPR39 signaling may stimulates the release of the anti-inflammatory cytokine IL-10, thus shifting the kidney tissue towards an anti-inflammatory milieu, promoting renal recovery. The present review focuses on the role of zinc in AKI to identify potential therapeutic strategies targeting zinc signaling for renoprotection and biomarker-based risk stratification.

Cells doi: 10.3390/cells15111017

Authors: Alessia Filippone Laura Cucinotta Valentina Bova Marika Lanza Giovanna Casili Irene Paterniti Michela Campolo Salvatore Cuzzocrea Emanuela Esposito

In the original publication [...]

Cells doi: 10.3390/cells15111016

Authors: Jianing Fan Meng Tong Yunkun Lu Qianqian Wang Yangyang Xie Kainan Lin Junjie Xu Xiujun Cai Xiao Liang

Intrahepatic cholangiocarcinoma (ICC) is an aggressive liver malignancy with a rising global incidence and limited therapeutic options. Vascular invasion (VI) is a hallmark of advanced disease, correlating with early recurrence and dismal prognosis, yet its tumor microenvironment (TME) drivers remain elusive. We analyzed single-cell RNA sequencing (scRNA-seq) data from 25 ICC samples to systematically characterize the cellular composition and molecular features related to VI. By integrating bulk RNA-seq data, spatial transcriptomics, and multiplex immunofluorescence, we identified a distinct subset of tumor-like cancer-associated fibroblasts (CAFs), termed tCAFs, enriched in VI-positive tumors. Functional enrichment analyses revealed that tCAFs were prominently associated with hypoxia and angiogenesis pathways, findings corroborated by the significant upregulation of tCAF markers (MME and NT5E) in ICC-derived CAFs under hypoxic conditions in vitro. Cell–cell communication analysis and spatial mapping uncovered that tCAFs might promote VI primarily through VEGF signaling interactions with endothelial cells. Integrative bioinformatics and RT-qPCR validation identified three key functional genes in tCAFs: SLC2A1, PTGS2, and PLOD2. In endothelial sprouting assays, pharmacological inhibition of SLC2A1 exerted a pronounced suppressive effect. Consistently, sprouting assays using ICC-derived CAFs with SLC2A1 knockdown confirmed that its downregulation significantly reduced endothelial sprouting capacity. Importantly, administration of the SLC2A1 inhibitor BAY-876 effectively suppressed tumor progression and intrahepatic metastasis in the orthotopic ICC mouse model. Our findings define a VI-associated cellular ecosystem and molecular landscape in ICC, unveiling a novel hypoxia–tCAFs–endothelial cells axis. Furthermore, we identify SLC2A1 as a clinically relevant therapeutic target, offering new insights into tumor VI.

Cells doi: 10.3390/cells15111015

Authors: Wenyi Jiang Eleanor Lynam Juliette Delafosse Graeme M. Birdsey Anna M. Randi Karl Matter Maria S. Balda

Regulation of the endothelial stress response is important for blood vessel homeostasis and angiogenesis, processes disrupted in common vascular diseases and ageing. Here, we discovered that the Y-box factor ZONAB (ZO-1-associated nucleic acid binding protein; YBX3), a gene associated with risk loci for severe vascular disorders, regulates endothelial homeostasis and angiogenesis. By combining cell-based assays with primary endothelial cells and genome-wide expression and methylation measurements, we found that ZONAB depletion results in mitochondrial deregulation, increased reactive oxygen species, and a defective oxidative stress response, which correlates with increased promoter methylation of cell cycle genes. ZONAB depletion triggered cellular senescence via a phosphatidylinositol 3-kinase (PI3K)/Akt-dependent pathway, which was attenuated by PIK3 inhibitors, an antioxidant, or by drugs targeting mitochondrial function or fragmentation. Thus, our results reveal that ZONAB repression in endothelial cells leads to genome-wide changes in gene expression and DNA methylation, regulating endothelial proliferation and inflammation, as well as mitochondrial deregulation to promote cellular senescence. Hence, ZONAB supports endothelial homeostasis and may play a role in vascular health.

Cells doi: 10.3390/cells15111014

Authors: Valentina Naef Michela Giacich Devid Damiani Filippo Maria Santorelli

Hereditary cerebellar ataxias are progressive neurodegenerative disorders for which disease-modifying treatments remain lacking. Although these conditions have traditionally been investigated from a neuron-centered perspective, evidence from several ataxia models indicates that changes in the cerebellar immune microenvironment can arise before overt neuronal loss and may contribute to early circuit dysfunction. This review examines hereditary cerebellar ataxias through the lens of early neuroimmune regulation, with particular attention to the region-specific properties of cerebellar microglia and their roles in synaptic refinement, inflammatory tone modulation and circuit homeostasis. We further discuss zebrafish as a useful experimental system for this question, because they combine in vivo imaging, genetic manipulation, and scalable functional assays in an intact vertebrate model. In this context, flavonoids—and especially naringenin—are not considered as immediate therapeutic candidates, but as mechanistically informative experimental probes to investigate how modulation of neuroimmune signaling affects disease-relevant phenotypes in vivo. By integrating genetic ataxia models with dynamic neuroimmune readouts, functional behavioral assays, and circuit-level analyses, zebrafish-based approaches can help identify early windows during which neuroimmune signaling influences cerebellar resilience and disease progression and can guide subsequent validation in mammalian systems.

Cells doi: 10.3390/cells15111013

Authors: Nobelle I. Sakwe Olga Y. Korolkova Ngoc B. Vuong Alayjha D. Edwards Perrin J. Black Destiny D. Ball Antonisha R. McIntosh Portia L. Thomas Diva S. Whalen Melvin Heather K. Beasley Antentor O. Hinton Josiah Ochieng Amos M. Sakwe

Annexin A6 (AnxA6) is a predominantly intracellular calcium-dependent membrane-binding multifunctional protein that is also detected extracellularly and in small extracellular vesicles (exosomes). We previously demonstrated that lapatinib resistance in triple-negative breast cancer (TNBC) cells is associated with AnxA6 upregulation and accumulation of cholesterol in late endosomes. Here, we investigated the fate of AnxA6 and cholesterol in lapatinib-resistant (LAP-R) cells and whether extracellular AnxA6 influences TNBC cell survival. We demonstrate that reduced expression of AnxA6 in LAP-R cells decreased the secretion of MCP-1/CCL2, CCL8/IL-8, DKK1, TSP-1, and OPN by antibody arrays. The secretion of exosomes was also markedly reduced in AnxA6-depleted LAP-R cells, while AnxA6 upregulation stimulated the release of MCP-1 and exosomes. Compared to the respective controls, exosome-associated AnxA6, Rab7, and cholesterol levels were increased in exosomes isolated from AnxA6-expressing LAP-R cells. Mechanistically, we demonstrated by co-immunoprecipitation, GST pulldown, and proximity ligation assays that AnxA6 interacts with SNAP23, a component of the membrane fusion machinery. Finally, blocking extracellular AnxA6 with neutralizing antibodies reduced the viability of AnxA6-low TNBC cells but had little effect on AnxA6-high cells. These findings suggest that extracellular AnxA6 is critical for the survival of highly proliferative AnxA6-low basal-like breast cancer cells and that AnxA6 influences TNBC progression by facilitating the secretion of pro-inflammatory cytokines and cholesterol-enriched exosomes.

Cells doi: 10.3390/cells15111012

Authors: Mi Eun Kim Jun Sik Lee

Demyelinating diseases are characterized by loss of myelin and impaired neuronal function. Differentiation of oligodendrocyte progenitor cells (OPCs) and neural stem and progenitor cells is regulated by intracellular kinase signaling pathways. PI3K/Akt/mTOR and Wnt/GSK-3β signaling are involved in oligodendrocyte maturation and neurogenesis, and pharmacological modulation of these pathways affects myelin formation and neuronal differentiation. Small-molecule compounds targeting these pathways influence protein synthesis, lipid production, and β-catenin-dependent transcription. Activation of Akt and mTOR is associated with increased myelin-related protein expression, whereas inhibition of mTOR reduces oligodendrocyte differentiation. In contrast, inhibition of GSK-3β affects β-catenin stability and is associated with oligodendrocyte differentiation. These pathways also affect proliferation and differentiation of neural stem and progenitor cells. However, effects observed in experimental demyelination models have not been established as direct evidence of remyelination in patients. In addition, pharmacological agents act on multiple cell populations in the central nervous system (CNS), which complicates interpretation of their effects on specific cell types. This review examines pharmacological targeting of PI3K/Akt/mTOR and Wnt/GSK-3β signaling and describes intracellular mechanisms involved in oligodendrocyte and neuronal differentiation, with consideration of therapeutic application in demyelinating diseases.

Cells doi: 10.3390/cells15111011

Authors: Luke J. Bolstad Mia J. LaRico Thomas S. Zanovich Grant R. Keith Amgad S. Hanna Daniel J. Hellenbrand

Spinal cord injury (SCI) triggers a secondary injury cascade characterized by neuroinflammation, reactive gliosis, and neuronal apoptosis. While many pro-inflammatory cytokines contributing to this cascade reach peak upregulation within 24 h, Interleukin-18 (IL-18) exhibits a delayed upregulation profile, typically peaking 7 days post-injury. This review examines the temporal regulation and cell-specific roles contributing to the rise in IL-18 after SCI. Following primary insult, damage-associated molecular patterns prime and activate the NLRP3 inflammasome, which in turn drives latent IL-18 secretion. Cellularly, microglia function as the primary producers of IL-18 via the TLR4/p38-MAPK pathway, while astrocytes serve as the primary responders through IL-18R/p65-NF-κβ signaling. The microglia-astrocyte cross-talk propagates reactive gliosis, drives neuropathic pain, facilitates neuronal loss, and potentially contributes to the formation of the astrocytic border. Targeted therapeutic interventions such as upstream inhibition of NLRP3 inflammasome assembly or direct IL-18 neutralization successfully mitigate neuroinflammation. By either inhibiting NLRP3 inflammasome activation or directly neutralizing IL-18, these treatments shift the microglial toward a protective state, restrict histological damage, and significantly improve functional recovery.

Cells doi: 10.3390/cells15111006

Authors: Ahmed Abomhya Ronaldo Panganiban Syed Adeel Hassan Brandon B. Phinney Steven McAninch Gregory Yochum Irina V. Pinchuk Terrence Barrett Ellen J. Beswick

Fibrosis in Crohn’s Disease (CD) occurs when there is continuous inflammation and repair mechanisms, which may lead to fibrotic strictures and ultimately intestinal surgical resection. There are limited non-invasive biomarkers for monitoring CD activity, particularly for detecting fibrosis. Thus, we set out to identify biomarkers that could be monitored for the detection and treatment of fibrosis. We used multiplex protein arrays to examine cytokines and pro-fibrotic factors in the serum of CD patients and analyzed their reliability to predict fibrosis. These markers were confirmed in tissues and the role of cytokines in upregulating fibrotic factors was examined. Collagen 1A1, Fibronectin, MMP9, and Timp-1 in patient serum were identified as strong predictors of fibrosis. IL-1β, MCP-1 and TNFα were identified as major regulators of fibrosis factors in human tissues. Overall, we identified four serum markers that are strong indicators of fibrosis and provided some novel insight into the impact of cytokines on fibrotic factors. Taken together, these findings may lead to earlier detection of fibrosis, and improved monitoring and treatment for CD patients.

Cells doi: 10.3390/cells15111010

Authors: Natalie Fischhaber Julian Vogler Ivana Martan Thomas Michler

Air–liquid interface (ALI) cultures recapitulate key features of the airway epithelium by driving basal cell differentiation into ciliated, club, and goblet cells and by generating a functional mucus barrier, thereby representing a highly relevant model of the human respiratory tract. Using a reduced factorial Design of Experiments (DoE) methodology, we simultaneously investigated the effects of seven variables on human coronavirus OC43 (HCoV-OC43) replication in air–liquid interface (ALI)-cultured primary human bronchial epithelial cells (pHBECs) to identify robust conditions that support infection and viral replication. Epithelial differentiation was monitored by measuring transepithelial electrical resistance and determining expression levels of marker genes for basal, goblet, club, and ciliated cells using reverse transcription quantitative PCR (RT-qPCR). HCoV-OC43 replication was monitored by quantifying genomic and subgenomic RNA by RT-qPCR. Viral RNA peaked three days post-infection in cell lysates and four days post-infection in apical washes. Initiation of ALI conditions induced epithelial differentiation, which was complete after 21 days and emerged as the strongest determinant of viral replication. Differentiated pHBEC cultures showed significantly reduced viral RNA compared with undifferentiated cultures, particularly following apical infection. In contrast, basal infection resulted in lower viral RNA levels in undifferentiated cultures than apical infection but was less dependent on epithelial differentiation. However, productive infection following basal exposure was less consistent and more strongly dependent on viral inoculum size. We further demonstrate that repeated mucus washes prior to infection increased HCoV-OC43 replication in mature cultures. In summary, our findings show that epithelial differentiation negatively affects HCoV-OC43 replication and we identify conditions that maximize viral replication in fully differentiated pHBEC cultures.

Cells doi: 10.3390/cells15111009

Authors: Jay J. Oh Xinge Xie Richard D. Dix

Programmed cell death (PCD) pathways of innate immunity serve to protect host cells from invading viruses. Parthanatos is a novel form of PCD triggered by excessive host cell DNA damage that leads to overactivation of poly(ADP-ribose) polymerase-1 (PARP-1) which in turn stimulates poly(ADP-ribose) (PAR) polymer formation. PAR translocates to the cytoplasm, where it induces release of apoptosis-inducing factor (AIF) from mitochondria, that then travels back to the nucleus, where it mediates large-scale DNA fragmentation and cell death. Little information is available regarding parthanatos as a cell death mechanism to dampen herpesvirus replication at the host cell level. A series of studies were therefore performed to clarify a possible role for parthanatos during productive replication of murine cytomegalovirus (MCMV) and herpes simplex virus type 1 (HSV-1) in diverse cell types. These included mouse embryo fibroblasts, mouse lung fibroblasts, mouse microglial (BV-2) cells, and human retinal pigment epithelial (ARPE-19) cells. We report that PAR protein production is surprisingly cell type specific. Moreover, MCMV or HSV-1 infection may suppress parthanatos as observed for other PCD pathways, such as apoptosis, necroptosis, and pyroptosis, in a dose-dependent and cell type-specific manner. We conclude that the operation of parthanatos at the host cell level during herpesvirus replication is more complex than originally thought but offers new targets for possible therapeutic interventions.

Cells doi: 10.3390/cells15111008

Authors: Jingwen Wang Hongyi Wu Tianqi Wang Meng Nie

Therapeutic resistance in cancer arises not only from intrinsic metabolic plasticity within the tumor, but also from the systemic metabolic state of the host organism. This review advances an integrated framework centered on the metabolic network between the host and tumor to examine how host-related factors—particularly aging, nutrition, and psychological stress—remodel systemic metabolism and thereby influence the efficacy of diverse cancer therapies. We highlight a bidirectional metabolic interplay: host physiology establishes a permissive context for tumor metabolic adaptation, whereas anticancer therapies, in turn, perturb host metabolic homeostasis, accelerating aging and compromising neurocognitive health. Ultimately, we propose that overcoming therapeutic resistance will require strategies that simultaneously target tumor metabolic dependencies and reprogram the host metabolic milieu—a systemic approach poised to redefine precision oncology.

Cells doi: 10.3390/cells15111007

Authors: Changli Zhang Gilbert Gu Joshua W. McNulty David Berenfeld Lisbet Haglund Sangwook Tim Yoon Brian Goh Hicham Drissi

Low back pain is closely associated with intervertebral disc (IVD) degeneration, in which inflammation and neovascularization within the annulus fibrosus (AF) contribute to pain generation. Platelet-derived growth factor (PDGF)-BB plays a crucial role in tissue repair and cellular homeostasis, but its role in AF cell biology remains poorly understood. To investigate the effects of PDGF-BB on human AF cells, healthy and degenerated AF cells were treated with PDGF-BB for 3 or 5 days, followed by bulk RNA sequencing. Functional enrichment of differentially expressed genes, transcription factor activity analysis, and protein–protein interaction network analysis was performed. Publicly available single-cell RNA-seq data were used to compare the transcriptomic profiles of native healthy and degenerated AF samples. In addition, TNF-α stimulation was conducted to validate the anti-inflammatory effects of PDGF-BB. Our findings suggest that PDGF-BB induced both common and context-dependent transcriptional responses in healthy and degenerated AF cells. In healthy AF cells, PDGF-BB consistently upregulated genes associated with cell cycle and developmental growth. In degenerated AF cells, PDGF-BB also induced these responses, while additionally it downregulated the genes related to extracellular matrix remodeling and collagen degradation. Meanwhile, PDGF-BB showed common effects in both healthy and degenerated cells by modulating the expression of genes within G protein-coupled receptor (GPCR) networks that are linked to complement, inflammation, and neurotransmitter signaling. In addition, PDGF-BB also suppressed the expression of genes involved in inflammatory-neurogenic signaling, including nerve growth factor (NGF), C-X-C motif chemokine ligand 12 (CXCL12), and apolipoprotein E (APOE). To relate these PDGF-BB induced responses to disc degeneration, we reanalyzed publicly available single-cell RNA-seq datasets from native human AF tissues and found that NGF-positive cells showed increased tumor necrosis factor (TNF)-α signaling activity. When AF cells were stimulated with TNF-α, PDGF-BB treatment significantly inhibited the expression of NGF, endothelin-1 (EDN1), and interleukin 6 (IL6) under both baseline and TNF-α-stimulated conditions. These results suggest that PDGF-BB modulates gene expression associated with inflammatory and neurogenic signaling as well as ECM remodeling in human AF cells, providing a transcriptomic insight into the PDGF-BB’s function in AF biology.

Cells doi: 10.3390/cells15111005

Authors: Cui Zhang Chonkit Lio Nana Li Yang Yu Jinfang Luo

Rheumatoid arthritis (RA) is a systemic immune-related disease characterized by chronic synovial inflammation and progressive joint destruction. However, the molecular mechanisms and diagnostic biomarkers underlying RA remain unclear. In this study, we aimed to identify potential biomarkers for clinical diagnosis of RA and to investigate their association with immune infiltration. By integrating differentially expressed genes analysis (DEGs) and weighted gene co-expression network analysis (WGCNA), we identified PSMB9 as a hub gene associated with RA. Gene set enrichment analysis (GSEA) and immune infiltration analysis revealed a strong association between RA and macrophage infiltration. Single-cell RNA sequencing datasets also suggest that PSMB9 is not only highly expressed in macrophage but is also present in synovial cells. We employed cellular thermal shift assay (CETSA) combined with Western blot to validate the interaction between sinomenine (SIN) and the target protein. CETSA results demonstrated that, compared with the control group, SIN increased the thermal stability of PSMB9, suggesting direct binding between the two. Western blot experiments further confirmed that PSMB9 protein expression was significantly downregulated following SIN treatment. PSMB9 may serve as potential diagnostic biomarker and therapeutic targets for RA. Moreover, our data suggest SIN may exert anti-inflammatory effects through regulation of PSMB9. This study also provides an additional insight into the underlying mechanisms involved in the progression of RA and discover a new molecular target for SIN.

Cells doi: 10.3390/cells15111004

Authors: Koume Nagayama Hiroshi Nango Komugi Tsuruta Hiroko Miyagishi Yasuhiro Kosuge

Neuritogenesis is essential for neuronal development and circuit formation. Although cAMP signaling downstream of Gs-coupled receptors is considered pro-neuritogenic, activation of these Gs-coupled receptors can produce divergent cellular outcomes. We previously showed that prostaglandin E2 (PGE2) induces neurite outgrowth in NSC-34 motor neuron-like cells predominantly through Gs-coupled E-prostanoid receptor 2 (EP2) signaling. The I-prostanoid receptor (IP) is also Gs-coupled, but whether its ligand PGI2 elicits neuritogenesis remains unclear. Here, we compare the neuritogenic and signaling responses to PGE2 and PGI2 in NSC-34 cells. PGE2 and the EP2 agonist butaprost increased the proportion of neurite-bearing cells, whereas PGI2 and the IP agonist beraprost had no effect. PGI2 and PGE2 induced comparable cAMP accumulation and protein kinase A substrate phosphorylation, and elicited peak cAMP response element binding protein (CREB) phosphorylation at 1 h. However, only PGE2 maintained significant CREB phosphorylation at 3–6 h. RNA sequencing at 4 h revealed broadly concordant transcriptional responses, while direct comparison identified Fst as the only gene expressed at higher levels under PGE2 than under PGI2. These findings suggest that the temporal profile of CREB phosphorylation, rather than the magnitude of early cAMP-PKA signaling, may be associated with differences in neuritogenic outcomes of Gs-coupled prostanoid signaling.

Cells doi: 10.3390/cells15111003

Authors: Yunsi Zheng Anqi Luo Kohji Fukunaga Qibing Liu Qingyun Guo

We previously demonstrated that fatty acid-binding protein 3 (FABP3) is significantly upregulated in ischemic neurons, and its inhibition mitigates ischemic brain injury in mice and attenuates mitochondrial damage under rotenone-induced oxidative stress. These findings suggest a potential role for FABP3 in mitochondrial dysfunction in ischemic neurons, although the underlying mechanism remains unclear. In this study, we further investigated the role of FABP3 in mitochondrial injury and apoptosis in ischemic neurons. Our findings indicated that FABP3 deficiency significantly decreased infarct volume following middle cerebral artery occlusion/reperfusion (MCAO/R) in mice, improved cognitive and spontaneous activity deficits, and suppressed BAX activation and mitochondrial translocation, caspase-3 activation, and cytochrome c release. In HT22 cells subjected to oxygen-glucose deprivation/reoxygenation (OGD/R), FABP3 deficiency increased cell viability, reduced apoptosis, and alleviated the loss of mitochondrial membrane potential. Conversely, FABP3 overexpression further exacerbated mitochondrial dysfunction and apoptosis, effects that were partially reversed by the BAX inhibitor BAI1. Furthermore, FABP3 overexpression promoted abnormal mitochondrial lipid accumulation and increased lipid peroxidation. Both the mitochondria-targeted antioxidant MitoQ and the ferroptosis inhibitor Ferrostatin-1 alleviated FABP3 overexpression-induced mitochondrial damage and apoptotic signaling. Collectively, our findings suggest that FABP3 is an important promoter of cerebral ischemia–reperfusion injury. FABP3 may aggravate ischemic neuronal injury by promoting abnormal mitochondrial lipid accumulation and lipid peroxidation, thereby enhancing BAX-dependent mitochondrial apoptotic signaling. Targeting FABP3 may provide a potential therapeutic strategy for neuroprotection in ischemic stroke.

Cells doi: 10.3390/cells15111002

Authors: Cristina Del Seppia Laura Sabatino

Thyroid hormones (THs) critically regulate metabolism and the central nervous system (CNS) functions, acting as key factors in neuronal differentiation, synaptogenesis and myelination. Furthermore, they play a central role in regulation of cognitive process and behavior. Therefore, compromised TH signaling can interfere with normal brain function and promote neurodegenerative progression and dementia. This review explores the role of THs from embryonic development through adulthood, with particular emphasis on their crucial role in neurogenesis. We discuss key components of TH metabolism and signaling, highlighting their neuroprotective functions in maintaining cellular homeostasis. Furthermore, we address how disruptions in TH signaling contribute to cognitive decline observed in dementia with effects that are even more pronounced in Alzheimer’s Disease (AD).

Cells doi: 10.3390/cells15111000

Authors: Katalin Gyurina Ádám Radványi László Sasi-Szabó Enikő Felszeghy Emese Rácz Gábor Méhes Andrea Kádár Csaba Fekete Tamás Röszer

Catecholamines are crucial signaling molecules that initiate thermogenesis in adipocytes through beta-adrenergic receptors (ADRBs). Adipocyte catecholamine resistance is a common feature of pediatric obesity, often impeding weight loss and the maintenance of a healthy body fat percentage. Our aim was to identify possible mechanisms that may be responsible for the development of catecholamine resistance in adipocytes. We demonstrate that Lim homeobox 8 (LHX8), a transcription factor previously known for its role in gametogenesis, is essential for catecholamine-induced thermogenesis in human adipocytes. LHX8 is expressed in developing human adipocytes throughout intrauterine and perinatal life, as well as in adulthood, and its expression levels positively correlate with the expression of key thermogenesis genes. Pediatric obesity diminished adipocyte expression of LHX8. Functionally, ADRB stimulation failed to induce thermogenesis in both mouse and human adipocytes when LHX8 was absent. Conversely, LHX8 overexpression enhanced thermogenesis in murine adipocytes. Mechanistically, LHX8 stimulated adipocyte interleukin-33 (IL-33) synthesis in response to ADRB activation, which subsequently increased thermogenic gene expression in both human and mouse adipocytes. In conclusion, adipocyte LHX8 is indispensable for catecholamine-responsive thermogenesis and represents a promising novel therapeutic target to overcome catecholamine resistance and promote effective weight management.

Cells doi: 10.3390/cells15111001

Authors: Maria Elena Soto Gilberto Vargas-Alarcón Claudia Huesca-Gómez Israel Pérez-Torres José Antonio Arias-Godínez Sergio Enrique Meza-Toledo Regina de la Mora-Cervantes Hugo Rodríguez-Zanella Gabriela Meléndez-Ramírez Linaloe Manzano-Pech Giovanny Fuentevilla-Álvarez Ricardo Gamboa

Marfan syndrome (MS), Loeys–Dietz syndrome (LDS), Beals–Hecht syndrome (BHS), Ehlers–Danlos syndrome (EDS), and individuals with undifferentiated connective tissue disease (UCTD) exhibit phenotypic overlap, suggesting a likelihood of genotypic coexistence. Our objective was to evaluate genetic variants (GVs), encoding 174 genes related to aortopathies, cardiomyopathies, arrhythmias, structural heart disease, and hypercholesterolemia, and their relationship to clinical and cardiovascular damage in these syndromes. This was a prospective study in Mexican patients with MS, LDS, EDS, BHS, and UCTD. One hundred and seventy-four genes related to hereditary diseases were studied using next-generation sequencing targeting coding regions. Of the 136 patients, 25 were identified with the recurrent and coexisting GV of MYBPC3. In the MS group, in addition to the presence of GV in FBN1, eight patients had GV in MYBPC3, six in FBN2, and five in COL3A1 and COL5A1. In the LDS group, in addition to GV in TGFBR1, TGFBR2, and SMAD3, four patients presented with GV in MYBPC3 and two with FBN2. In the BHS group, in addition to FBN2, two patients had GV in MYBPC3 and one with TGFBR2. In the UCTD group, nine patients had GV in MYBPC3 and two in COL5A1 and COL5A2. All syndromes coexisted with GV in genes related to arrhythmias, sarcomeres, and hypercholesterolemia. In EDS, coexistence with several sarcomere proteins was found.

Cells doi: 10.3390/cells15110999

Authors: Bibiana Sgalletta Francesco Agostini Marco Bisaglia

Iron is an essential micronutrient that plays a central role in numerous biological processes. Despite its relatively low abundance in the human body, iron is particularly critical for brain function. Systemic and cerebral iron homeostasis is tightly regulated through coordinated mechanisms involving absorption, transport, storage, and recycling. Within the brain, iron metabolism is further controlled by the blood–brain barrier and specialized neural cell populations, including neurons, astrocytes, oligodendrocytes, and microglia. Iron is indispensable for neurodevelopment, supporting neurogenesis, myelination, and neurotransmitter synthesis. However, both iron deficiency and iron overload have detrimental consequences. Early-life iron deficiency disrupts neural development and leads to long-lasting cognitive, motor, and behavioral impairments, whereas excessive iron accumulation promotes oxidative stress, ferroptosis, and neuroinflammation. These mechanisms have been described to contribute to the pathogenesis of major neurodegenerative disorders, including Alzheimer’s disease, Parkinson’s disease, neurodegeneration with brain iron accumulation, and amyotrophic lateral sclerosis. This review first outlines systemic and brain iron metabolism, highlighting how neural cells regulate homeostasis. Next, it examines iron’s physiological roles, particularly in neurogenesis and neurodevelopment. Finally, it explores iron’s involvement in neurodegenerative diseases, emphasizing neuroinflammation as a primary mechanism of iron toxicity.

Cells doi: 10.3390/cells15110998

Authors: Masayuki Nagasawa

C-reactive protein (CRP) was discovered as a protein that reacts with C-polysaccharides to form precipitates in the serum of patients with pneumococcal infection. Subsequently, it was found to increase in the serum of patients with bacterial infections and rheumatic diseases, and it has since been widely recognized as a nonspecific biomarker of acute inflammation and utilized in clinical medicine. Meanwhile, CRP-like proteins are also present in the hemolymph of horseshoe crabs, and it has become clear that these proteins have long played a crucial role in the humoral innate immune response against foreign microorganisms. In recent years, advances in molecular analysis have revealed the details of the complex biological functions performed by CRP. Furthermore, with the development of highly sensitive CRP measurement methods, its importance as a biomarker is gaining attention not only in acute inflammatory diseases but also in chronic inflammatory diseases such as cardiovascular disease, diabetes, cancer and neurological disorders. New treatment strategies targeting CRP, based on recent findings, are also being explored.

Cells doi: 10.3390/cells15110997

Authors: Dominika Pigoń-Zając Maria Bryczek Agata Leszczuk Adrian Zając

Cancer stem cells (CSCs) are a highly influential population of tumor cells involved in tumor initiation, progression, metastasis, recurrence, and resistance to therapy. Although CSCs have been widely investigated, their behavior cannot be understood solely through intrinsic cellular features, as these cells strongly depend on a specialized supportive microenvironment known as the CSC niche. In this review, we discuss the CSC niche as a dynamic and therapeutically relevant ecosystem that is distinct from, but closely connected with, the broader tumor microenvironment. Particular attention is given to stromal cells, immune cells, endothelial cells, extracellular matrix components, hypoxia, cytokines, chemokines, and metabolic stress as regulators of CSC self-renewal, plasticity, dormancy, immune escape, epithelial–mesenchymal transition, metastatic dissemination, and survival under therapeutic pressure. We further consider how CSC–niche interactions contribute to pre-metastatic niche formation and tumor relapse. Finally, we outline emerging therapeutic strategies aimed at disrupting CSC-supportive signals, including approaches targeting developmental pathways, angiogenesis, hypoxia, extracellular matrix remodeling, immunosuppressive networks, and cytokine-mediated communication. Overall, this review emphasizes that targeting the CSC-supportive microenvironment is essential for limiting metastasis, recurrence, and long-term treatment failure.

Cells doi: 10.3390/cells15110996

Authors: Ximena L. Ruden Campbell Coddington Lynessa Asplund Anjie Dinakin Awoniyi O. Awonuga Douglas M. Ruden Steven J. Korzeniewski Lijun Zhang Elizabeth E. Puscheck Daniel A. Rappolee

Early mammalian embryos are highly sensitive to environmental, metabolic, hormonal, and genomic stress, yet embryo assessment during In Vitro Fertilization (IVF) relies largely on morphology and ploidy for embryo assessment, but these tests incompletely predict miscarriage. We present a transcriptomics based framework to classify and quantify lineage-specific stress in early embryos by benchmarking human preimplantation embryos against dose-, time-, and quality-dependent stress programs defined in Embryonic and placental Trophoblast Stem Cells (ESCs, TSCs) from the implanting blastocyst. Human embryos and stressed ESCs and TSCs are screened using transcriptomic markers from eleven biologically distinct stress Gene Ontology (GO) groups that define functional stress states and enable quantification of pathway presence and upregulation, pathway activity, and downstream outcomes. This framework determines whether the Integrated Stress Response (ISR), once initiated, resolves to enable the Developmentally Associated Stress Response (DASR). High-throughput screening (HTS) titrates stress to define increasingly risky yet biologically equivalent doses for levels of diminished stem cell growth across mechanistically diverse stressors. Then bulk RNA seq derives lineage specific transcriptomic markers putatively respond to common levels of diminished growth and that distinguish weak vs. strong stress and resolved vs. unresolved ISR. These stem cell transcriptomic signatures are applied to bulk RNA seq data from IVF embryos graded for morphology or adhesion, enabling quantitative inference of stress burden, lineage vulnerability, and developmental trajectory.

Cells doi: 10.3390/cells15110995

Authors: Jian-Hong Lin Yu-Po Luo Pei-Ching Ting Min-Pei Ko Kun-Ta Yang

Bone marrow-derived mesenchymal stem cells (BMSCs) maintain skeletal homeostasis by balancing adipogenic and osteogenic differentiation, yet clinically used drugs that bias this fate choice and their mechanisms remain incompletely defined. Here, we investigated whether dextromethorphan (DXM), a widely used antitussive, modulated lineage commitment in rat BMSCs and interrogated candidate upstream signaling modules. Rat BMSCs were induced with adipogenic medium or osteogenic medium in the presence of DXM (30 μM). Adipogenesis and osteogenesis were quantified using Oil Red O and Alizarin Red S staining with elution-based quantification, and lineage markers were measured by RT-qPCR. Intracellular Ca2+ and ROS were analyzed using flow cytometry, and the levels of p-AKT and p-ERK were assessed through Western blotting analysis. Under adipogenic induction, DXM increased lipid droplet accumulation and the mRNA levels of Pparγ and Fabp4. Although DXM elevated Ca2+ and ROS, the chelation of intracellular Ca2+ and pharmacological inhibition of Sig-1R/PLC–IP3R signaling, redox/ROS, NMDA receptors, AKT/ERK, Kv channels, bitter taste receptor-related signaling, and mTOR did not attenuate the DXM-enhanced adipogenesis. DXM reduced p-ERK without increasing p-AKT; U0126 lowered basal adipogenesis but did not block the DXM effect. Under osteogenic induction, DXM reduced matrix mineralization and downregulated Runx2 and Bglap mRNA levels, while Wwtr1 mRNA levels were not significantly changed. DXM also partially reversed the osteogenic induction-associated reduction in Mtor mRNA. Separately, under adipogenic induction, rapamycin attenuated baseline adipogenesis but did not prevent the additional lipid accumulation induced by DXM. Collectively, DXM shifted the osteogenic–adipogenic balance toward adipogenesis through a non-canonical mechanism.

Cells doi: 10.3390/cells15110993

Authors: Sara Soltani Tehrani Samuel Isaac Olson Karishma Kundu Sylvain Ferrandon Matthew F. Kalady

Locally advanced rectal cancer is commonly treated using total neoadjuvant therapy (TNT), which integrates radiotherapy with systemic chemotherapy to improve tumor downstaging, local control, and long-term oncologic outcomes. Despite its central role in treatment, responses to radiotherapy remain highly heterogeneous. While some tumors undergo complete regression, others exhibit intrinsic or acquired treatment resistance, resulting in incomplete tumor control while experiencing treatment-related toxicity. Understanding the biological determinants that govern radiation sensitivity in rectal cancer, therefore, represents a major clinical challenge. Ionizing radiation induces tumor cell death primarily through the generation of reactive oxygen species (ROS) and DNA damage, particularly DNA double-strand breaks. In addition to nuclear DNA injury, radiation-induced oxidative stress can initiate lipid peroxidation within cellular membranes. When lipid peroxide accumulation exceeds the capacity of cellular antioxidant systems, this process can trigger ferroptosis, an iron-dependent form of regulated cell death driven by phospholipid oxidation. Ferroptotic susceptibility is regulated by interconnected metabolic pathways, including cystine transport through system Xc− (SLC7A11/SLC3A2), glutathione synthesis, glutathione peroxidase-4 (GPX4) activity, iron metabolism, and membrane lipid remodeling. Recent evidence further indicates that ferroptosis intersects with antitumor immunity. Ferroptotic tumor cells release oxidized lipid mediators and damage-associated molecular signals that can influence immune activation, while interferon-γ produced by activated CD8+ T cells during immune checkpoint blockade suppresses SLC7A11 expression, limiting cystine uptake and promoting ferroptotic tumor cell death. These findings suggest that ferroptosis represents a mechanistic interface between tumor metabolic vulnerability and immune-mediated cytotoxicity. This interaction is particularly relevant in colorectal cancer biology, where immune checkpoint inhibitors demonstrate clinical benefit primarily in tumors with deficient mismatch repair or microsatellite instability-high (MSI-H) status. The vast majority of rectal cancers are microsatellite stable (MSS) and exhibit limited responsiveness to immunotherapy due to reduced immunogenicity and immune exclusion within the tumor microenvironment. Strategies capable of increasing tumor immunogenicity in this setting are therefore of considerable interest. In this review, we examine the molecular mechanisms linking radiation-induced oxidative stress to ferroptosis and tumor immunity in colorectal cancer, while focusing on the clinical context of radiotherapy in rectal cancer. We discuss how lipid metabolism, iron homeostasis, cysteine-dependent antioxidant systems, and immune signaling pathways converge to regulate ferroptotic vulnerability and radiation response. We further explore the therapeutic potential of integrating radiotherapy, ferroptosis-targeting strategies, and immunotherapy to overcome radioresistance and improve treatment outcomes in colorectal cancer.

Cells doi: 10.3390/cells15110994

Authors: Keith Ireton

Although evolutionarily distant, the bacteria Listeria monocytogenes, Shigella flexneri, and Burkholderia thailandensis each undergo a “cell-to-cell” spreading process that allows these pathogens to disseminate within human tissues. Spread initiates when bacteria polymerize actin filaments that propel them through the host cell cytosol. The pathogens then remodel the plasma membrane into protrusions that are internalized by adjacent cells and resolved into double membranous vacuoles (DMVs) which lyse to liberate bacteria. In this review, we discuss recent publications indicating that L. monocytogenes, S. flexneri, and B. thailandensis each enhance their spread by altering the subcellular localization of human Dynamin 2—a GTPase that regulates endocytosis and other trafficking pathways. Interestingly, Dynamin 2 controls distinct steps in spread of L. monocytogenes, S. flexneri, and B. thailandensis. In the case of L. monocytogenes, the GTPase has the potential to restrict protrusion formation by generating tension at tight junctions. However, L. monocytogenes secretes a protein that relieves this restriction of protrusions, allowing efficient spread. During dissemination of S. flexneri and B. thailandensis, Dynamin 2 is co-opted to resolve protrusions into DMVs. B. thailandensis also mobilizes Dynamin 2 to lyse DMVs. These findings highlight diverse ways in which bacteria control Dynamin 2 to augment spread.

Cells doi: 10.3390/cells15110992

Authors: Tzu-Wen Hong Rosie Sullivan Ryea Arora Adya Lonsane Zekun Lyu Sara Caxaria Tien-Chi Huang Lydia F. Daniels Gatward Thomas Burgoyne Aileen J. F. King Shanta J. Persaud Peter M. Jones

Type 1 diabetes is caused by autoimmune destruction of insulin-secreting β-cells within islets of Langerhans. Transplantation of donor islets can improve glycaemic control, but current clinical islet transplantation protocols are compromised by extensive loss of β-cell functional mass soon after implantation. Co-incubation in vitro or co-transplantation in vivo of mesenchymal stromal cells (MSCs) with isolated islets improves their functional survival, although the underlying mechanisms remain obscure. Here, we show that MSC-derived extracellular vesicles (MSC-EVs) are alone sufficient to recapitulate many of the beneficial effects of MSCs on islet functional survival, offering the possibility of simple cell-free treatments to improve the outcomes of islet transplantation. We used LC- analysis and small RNA sequencing to analyse the protein and microRNA (miRNA) molecular cargos of MSC-EVs. Proteomic analysis identified >100 proteins from the Uniprot Mouse Database, including β-cell G protein-coupled receptor (GPCR) agonists which we have previously shown to enhance β-cell functional survival. MSC-EVs contained ~300 distinct miRNAs and we identified five highly enriched miRNAs that were significantly upregulated in MSC-EV-treated islets, notably miR-21a-5p. MSC-EV treatment also altered the expression of a distinct set of islet mRNAs known to be involved in islet metabolism and function. These observations may enable the further simplification of the islet pretreatment strategy by focusing on defined GMP-grade biologically active molecules rather than whole heterogeneous EV populations.

Cells doi: 10.3390/cells15110991

Authors: Farah Ahmady-Nield Blaine M. H. Carnie Grace E. C. Anderson Emerson Achari Amit Sharma Adrian A. Achuthan George Kannourakis Rodney B. Luwor

Glioblastoma is the most aggressive form of brain tumor resulting in low overall patient survival rates of 12–15 months post diagnosis. Several factors contribute to the complexity of the tumor, including tumor heterogeneity, blood–brain barrier complications, genetic defects, cancer stem cell generation, and immune evasion. These factors can result in the progression of glioblastoma and are controlled by signaling pathways. Some of the signaling pathways involved in glioblastoma progression include ERK, NF-κB, Wnt, and PI3K/AKT/mTOR. Our and others’ previous studies have found that TIM-3 and TGF-β signaling is altered in glioblastoma patients and may contribute to cancer progression. Immune promoting pathways such as STING have also been studied in glioblastoma to enhance anti-tumor immunity; however the interconnecting roles of these pathways are not well described. This review highlights the role of these three key cancer-related pathways in glioblastoma and their mechanistic link. Better understanding these links may result in improved treatment targets or disease progression biomarkers.

Cells doi: 10.3390/cells15110990

Authors: Jasbir Bisht Priyanka Rawat Andrew C. Shin Vijay Hegde

Alzheimer’s disease (AD) is the most prevalent form of dementia and is characterized by progressive cognitive decline due to the loss of neurons. The accumulation of extracellular senile plaques (Aβ) and intracellular tau neurofibrillary tangles (NFTs) is a key pathological feature of AD. Mitochondrial dysfunction is implicated in all key AD pathologies, whether as a cause or a consequence of disease progression. Growing evidence indicates that mitochondrial impairment plays a central role in AD pathogenesis by disrupting cellular homeostasis, promoting oxidative stress, and contributing to progressive neuronal death. Therefore, targeting mitochondria may offer promising insights into the development of disease-modifying therapies. In this review, we summarize current evidence on the role of mitochondrial dysfunction in the pathophysiology of AD and on its therapeutic potential.

Cells doi: 10.3390/cells15110989

Authors: Raul Canal Elizabeth F. Martinez Jamie S. Foster Fernanda Carla Bombaldi de Souza Renata Francielle Bombaldi de Souza Marcelo Marcos Morales Marcos Cesar Pontes Daniel N. da Rocha Lexie S. Holliday Fahong Yu Anderson Tadeu Silva Roberto D. Fanganiello José Ricardo M. Ferreira André A. Pelegrine

This study aimed to evaluate periosteum-derived mesenchymal stem cells (P-MSCs) cultured under simulated microgravity (SMG) conditions. P-MSCs were induced toward osteogenic differentiation and then exposed to SMG for up to 48 h. As a control, P-MSCs were maintained under identical conditions but without SMG exposure. Cell viability, osteogenesis-related analytes, and gene expression were analyzed at 3, 24 and 48 h. Cell viability under SMG was lower after 3 h but was significantly higher after 24 h, with no difference at 48 h. There was a higher expression of pathways associated with inflammation at 3 h, which was attenuated by 24 h and neutralized at 48 h. P-MSCs under SMG demonstrated three characteristics in at least one timepoint, which supports a pro-osteogenic signaling response: (1) higher osteoprotegerin levels; (2) lower DKK1 and TNF levels; (3) upregulation of genes related to osteogenesis. Our data suggest that P-MSCs exhibit enhanced pro-osteogenic regulatory modulation in SMG.

Cells doi: 10.3390/cells15110988

Authors: Lynn Michelle Grodzki Stefanie Schlichting Yue Hu Sabine Helbing Udo Bartsch

Progranulin (PGRN) is a secreted protein composed of 7.5 granulin domains. The protein is implicated in various functions, including cell survival, inflammation, lysosomal homeostasis, tumorigenesis, and aging. Haploinsufficiency and complete loss of PGRN function cause the neurodegenerative disorders frontotemporal lobar degeneration and neuronal ceroid lipofuscinosis type 11, respectively. In the nervous system, administration of exogenous PGRN has been shown to promote the survival of various nerve cell types under different pathological conditions and to stimulate neurite outgrowth in vitro and axonal regeneration in vivo. In the retina, PGRN dysfunction results in photoreceptor and retinal ganglion cell (RGC) loss, whereas PGRN administration promotes photoreceptor cell survival. In the present study, we analyzed whether a sustained intravitreal administration of PGRN promotes the survival of axotomized RGCs and the regrowth of the lesioned axons. To this end, we generated a PGRN-overexpressing clonal neural stem cell line and injected the cells into the vitreous cavity of a mouse optic nerve crush model. The progression of the lesion-induced degeneration of RGCs was studied at different time points after the nerve crush. The regeneration of the injured RGC axons into the distal optic nerve stump was analyzed one month after nerve lesioning. We found that the intravitreally administered PGRN slowed the degeneration of the injured RGCs for up to four months, the latest post-lesion interval analyzed. Furthermore, PGRN stimulated the regeneration of some RGC axons over long distances into the distal optic nerve stumps. Taken together, our results identify PGRN as a novel neurotrophic factor for retinal ganglion cells.

Cells doi: 10.3390/cells15110987

Authors: Prakash Gangadaran Sanjuda Subramaniyan Ramya Lakshmi Rajendran Chae Moon Hong Kumari Swati Saurabh Kumar Jha Shazia Rashid Byeong-Cheol Ahn

Macrophage-derived extracellular vesicles (EVs) have emerged as promising biomimetic platforms for targeted cancer drug delivery due to their biocompatibility, immune-modulatory properties, and tumor-homing capabilities. Among macrophage subtypes, M1-polarized macrophages exhibit potent anti-tumor functions characterized by pro-inflammatory cytokine secretion, improved antigen presentation, and the ability to remodel the tumor microenvironment (TME). Utilizing these properties, M1-polarized macrophage-derived EVs serve as cell-free therapeutic systems capable of delivering bioactive cargo while simultaneously promoting anti-tumor immune responses. However, the clinical application of natural EVs is limited by low yield, heterogeneity, and challenges in large-scale production. Artificial nanovesicles (ANVs) have been developed to address these limitations, offering improved scalability, compositional control, and reproducibility. This review provides an overview of macrophage differentiation and polarization, with a focus on the immunological profile and anti-tumor mechanisms of M1-polarized macrophages. It further discusses current methodologies for EV isolation and ANV generation, along with cargo loading strategies that balance encapsulation efficiency and vesicle stability. In addition, this review also emphasizes their targeting approaches, cellular uptake pathways, and the intracellular trafficking mechanisms that influence delivery efficiency and therapeutic outcomes. Key challenges, including standardization, biological barriers, and functional consistency, are critically evaluated. Emerging strategies that integrate vesicle engineering with personalized medicine underscore the potential of these systems to advance precision oncology.

Cells doi: 10.3390/cells15110986

Authors: Ivana Persico Maria Grazia Doro Laura Frogheri Maria Cristina Sini Giovanni Battista Maestrale Antonella Manca Domenico Mallardo Paolo Antonio Ascierto Giuseppe Palmieri

Messenger RNA (mRNA) vaccines are emerging as promising tools capable of reshaping how cancer interacts with the immune system and responds to immunotherapy. These vaccines not only act as platforms for antigen delivery but can also influence the tumor microenvironment (TME), fostering a shift from immunologically “cold’’ conditions toward “hotter’’ and treatment-responsive states. In melanoma, this capability has been found to enhance the efficacy of the immune checkpoint inhibitors (ICIs), as mRNA-based priming can provide the robust antitumor activation needed for more effective checkpoint blockade. Early clinical studies with personalized or off-the-shelf vaccines showed benefits in patients with high-risk resected melanoma or refractory to PD-1 inhibition. Combining mRNA vaccines with ICIs, along with other immunomodulatory strategies, may be helpful to overcome resistance arising from the TME and achieve more durable clinical benefits. Besides these advances, computational and in silico modeling are providing new insights into how mRNA vaccines modulate the TME, helping to identify factors such as antigen-presenting cell (APC) density, CD8+ T-cell infiltration, and macrophage polarization that may predict treatment success and guide personalized strategies. Together, these developments indicate that combining mRNA vaccination with ICIs, supported by computational tools, may improve clinical outcomes in melanoma and, potentially, in selected tumor types with favorable immunological features, although important biological constraints limit direct extrapolation to less immunogenic malignancies.

Cells doi: 10.3390/cells15110985

Authors: Satoshi Kamijo Hideki Miwa Kazutaka Ikeda

Autism spectrum disorder is a common neurodevelopmental condition, defined by persistent deficits in social interaction and communication, as well as restricted repetitive patterns of behavior, interests, or activities. Autism spectrum disorder is highly heterogeneous, encompassing a broad range of clinical presentations and suggesting it includes multiple etiological subtypes. Although no unified cause has been established, accumulating evidence indicates that genetic susceptibility interacts with environmental and developmental factors to shape diverse phenotypic outcomes. This review summarizes epidemiological findings and discusses major proposed etiological mechanisms, integrating evidence from human studies and animal models. Although animal models are not directly translatable to humans, their findings provide mechanistic insights that bridge epidemiological observations with neurobiological hypotheses.

Cells doi: 10.3390/cells15110984

Authors: Orion N. Schuldt Sydney R. Leitch Lauren K. Jones Porter R. Buckley Brad E. Morrison

The hallmarks of allergic diseases are Type 2 immunity, including IL-4 and IL-13 production, IgE antibody generation, mast cell and basophil activation, histamine release, and eosinophil activation. There are many routes by which such mediators can influence CNS biology, including cytokine entry or signaling via brain barrier receptors; leukocyte trafficking across activated barriers; cytokine signaling via circumventricular organ sites or dural immune compartments; vagus nerve afferent signaling; mast cell degranulation; and histamine neuromodulation. Neuroinflammation is a common hallmark of many neurodegenerative diseases, but whether and to what degree allergic/type 2 immune biology may be involved depends on the specific disease stage and pathology. Here, we assess studies connecting the roles of IL-4/IL-13 signaling, IgE/mast cell activation, eosinophil-attractive chemokines, and histamines in Parkinson’s disease, Alzheimer’s disease, multiple sclerosis, amyotrophic lateral sclerosis, dementia with Lewy bodies, Huntington’s disease, prion disease, and tauopathy/atypical parkinsonism. Mechanisms appear most clear in the case of Parkinson’s disease, where epidemiology suggests an important role in dementia/Alzheimer’s disease, while for other neurodegenerative conditions the evidence is less compelling and may be either mechanistic or modulatory. Confounding issues include sex differences, drug exposures, comorbid conditions, socioeconomic factors, and coexisting inflammatory diseases. Finally, we suggest a strategy based on longitudinal immune phenotyping, CNS biomarkers, and pathway manipulation to assess the relationship between allergic immune signaling and neurodegeneration.

Cells doi: 10.3390/cells15110983

Authors: Cui Wang Yi Liu Guitao Jiang Chuang Li Kai Shi Shufang Chen Huiying Wang Daqian He

Skeletal muscle satellite cells (SMSCs) are essential for embryonic myogenesis and postnatal muscle regeneration; however, their cellular heterogeneity and transcriptional dynamics during avian development remain largely unexplored. Here, we performed single-cell RNA sequencing (scRNA-seq) on 42,886 cells isolated from goose leg muscles across four embryonic stages (E13, E15, E18, and E23), with each stage comprising pooled tissues from four female embryos. Unbiased clustering resolved 22 transcriptionally distinct clusters representing six major cell types—satellite cells, myocytes, fibro-adipogenic progenitors, endothelial cells, immune cells, and Schwann cells—with satellite cells being the most abundant. Satellite cells were further subdivided into three functional states (quiescent, activated, and proliferative/differentiating), which followed a continuous, linear pseudotime trajectory from early to late embryonic stages. This trajectory was marked by a progressive downregulation of stemness-associated regulators (e.g., PAX7) and upregulation of myogenic commitment and differentiation factors (e.g., MYF5, MYOD1, and MYOG), faithfully mirroring chronological development. Cell–cell communication analysis revealed that quiescent satellite cells exhibited the most extensive intercellular signaling networks (e.g., FGFR, Ephrin, collagen, CADM), whereas activated and proliferative/differentiating cells showed progressively diminished communication capacity. Across developmental stages, the contribution intensities of key signaling pathways—including SEMA6, CDH, FGF, LAMININ, MK, MPZ, CADM, FN1, and COLLAGEN—varied significantly among satellite cell states, indicating state-specific responsiveness to microenvironmental cues. Collectively, these findings demonstrate that satellite cells dynamically coordinate extrinsic signal integration with intrinsic differentiation programs to achieve orderly myogenic progression. This study provides a high-resolution single-cell atlas of goose SMSC development, uncovering subpopulation heterogeneity, state-specific molecular signatures, and key signaling pathways, with important implications for avian muscle biology and genetic improvement of poultry.

Cells doi: 10.3390/cells15110982

Authors: Pravin B. Sehgal Huijuan Yuan

Biomolecular condensates in the cytoplasm and nucleus contribute to carcinogenesis through aberrant signaling by assorted transcription factors and fusion oncoproteins. Oral cancer, which is highly prevalent worldwide, frequently occurs in a U-shaped “high-risk” zone (floor of mouth, side of tongue, and anterior fauces) which forms the path of liquid transit through the mouth. We previously reported that environmental stresses of saliva-like hypotonicity and beverage-like temperature changes triggered cycles of disassembly/reassembly of biomolecular condensates of GFP-tagged human myxovirus resistance protein (MxA; alias Mx1) in oral cancer cells. In the present study, we identified some of the constituents of GFP-MxA cytoplasmic condensates in oral cells. These condensates were isolated from interferon (IFN)-λ1-treated GFP-MxA expressing OECM1 human oral cancer cells using magnetic bead-based immunoisolation. Unbiased peptide identification confirmed the presence of MxA/Mx1 peptides; however, the strongest intensity was for the BACH1 transcription factor family. Immunofluorescence analyses confirmed the association of BACH1 and the family member Nrf2 with cytoplasmic human GFP-MxA condensates. Moreover, GFP-BACH1 and GFP-Nrf2 colocalized with cytoplasmic human HA-MxA condensates in transiently transfected OECM1 cells. Western blot assays confirmed the presence of BACH1 and Nrf2 proteins in complexes isolated using anti-MxA pAb. As much as BACH1 and Nrf2 regulate oxidative stress response genes, it was remarkable that immunofluorescence assays revealed the presence of heme oxygenase 1 (HO1)—a downstream redox regulator—in GFP-MxA condensates. However, these condensates were devoid of p62, KEAP1 and Cul3. In terms of aberrant function, in live cells, the Nrf2 transcription factor underwent rapid disassembly and reassembly cycles driven by saliva-like hypotonicity, and was also disassembled by sulforaphane. The data highlight the unexpected intersections in oral cells between MxA condensates and BACH1, Nrf2 and HO1—proteins well known to be involved in pathways regulating cellular responses to environmental and oxidative stresses, antiviral defense, oral epithelial dysplasia, and cancer progression and metastases.

Cells doi: 10.3390/cells15110981

Authors: Na Jiang Shiyi Wang Jiaqiao Zhang Dandan Jia

Leukemia inhibitory factor (LIF), a member of the interleukin-6 (IL-6) cytokine family, is a well-characterized myokine with pleiotropic regulatory effects on skeletal muscle. LIF modulates several fundamental cellular processes, including myoblast proliferation, apoptosis, angiogenesis, and energy metabolism. Exercise upregulates LIF expression in skeletal muscle, thereby promoting satellite cell activation, proliferation, myoblast differentiation, and angiogenesis, facilitating physiological muscle hypertrophy, and suppressing myocyte apoptosis and muscle atrophy. In addition, LIF plays a critical role in modulating the inflammatory and extracellular matrix remodeling following exercise-induced muscle damage, thereby supporting efficient muscle repair and regeneration. This review elaborates on the biological mechanisms by which LIF regulates skeletal muscle atrophy and contributes to the enhancement of skeletal muscle function. It also highlights the biological characteristics of myogenic LIF and discusses future directions for basic and applied research on exercise interventions targeting LIF signaling pathways.

Cells doi: 10.3390/cells15110980

Authors: Yang Yi Md Nasim Uddin Keira Killeen Donald L. Gill Yandong Zhou

Store-operated calcium entry (SOCE), mediated by the endoplasmic reticulum (ER) Ca2+ sensors stromal interaction molecule (STIM) proteins and the plasma membrane (PM) Orai channels, is essential for calcium signaling and a wide range of physiological processes. Precise regulation of SOCE is critical for maintaining tissue homeostasis, whereas its dysregulation contributes to diverse pathological conditions, particularly organ fibrosis. In this review, we outline the molecular basis of SOCE and discuss how its dysregulation is implicated in human disease. We further emphasize the pivotal role of SOCE in driving fibrotic progression across major organ systems. Finally, we summarize current therapeutic strategies targeting SOCE and highlight their potential for the treatment of fibrosis.

Cells doi: 10.3390/cells15110979

Authors: Jingzhe Xiao Yuwei Ding Songbo Li Yi Yan Ziyue Yu Pengyu Fu Chunyan Xu Lijing Gong

Physical inactivity contributes to type 2 diabetes (T2D), but the molecular links between exercise and metabolic improvement remain incompletely understood. We meta-analyzed genome-wide association studies of vigorous physical activity and T2D (combined n ≈ 1.95 million) and integrated eQTL/sQTL maps with single-cell and spatial transcriptomic datasets to connect genetic risk with tissues, cell types, and regulatory programs. Tissue and cell-type enrichment, colocalization, and network analyses were performed. Computational findings were further examined in male 10-week-old C57BL/6J mice with high-fat diet-induced diabetes. After 1 week of acclimatization, mice were randomly assigned to normal chow, high-fat diet, or high-fat diet plus exercise groups (n = 6 per group; high-fat diet with 60% of total energy from fat). The exercise intervention consisted of treadmill running (10 m/min for 50 min per day, 5 days per week, total 16 weeks), followed by metabolic phenotyping, skeletal muscle histology, bulk RNA sequencing, alternative splicing analysis, and RT-qPCR of Mau2 isoforms. Exercise- and T2D-associated variants showed joint enrichment in skeletal muscle and adipose eQTL/sQTL signals. Integrated single-cell analyses prioritized fibro-adipogenic progenitors and endothelial cells, and identified an extracellular matrix- and collagen-related module in fibro-adipogenic progenitors associated with both exercise and T2D. Mau2 emerged as a shared candidate gene with tissue-specific splicing signals. In diabetic mice, exercise improved glucose homeostasis and muscle fiber structure, and reduced Mau2 intron retention in skeletal muscle without changing total Mau2 expression. These findings support a multiscale framework linking exercise-responsive regulation to T2D-related tissue remodeling and splicing plasticity.

Cells doi: 10.3390/cells15110977

Authors: Tomas Novotny Adam Eckhardt Jarmila Knitlova Martina Doubkova Roman Stachon Filip Hrdina Tatyana Kobets Martin Ostadal

Although Dupuytren’s disease (DD) and relapsed Clubfoot (RC) are clinically distinct conditions, both exhibit fibrotic tissue remodeling and contracture. This exploratory study investigated whether DD and RC share molecular features associated with fibroproliferative contracture. Pathological tissues from DD nodules and contracted tissues from RC together with their respective control tissues (n = 6/group), were analyzed using label-free quantitative proteomics. The analysis identified 12 significantly upregulated proteins shared between both pathological conditions relative to their controls (|log2FC| ≥ 1, p ≤ 0.05). These proteins included structural, signaling and tensile stress ECM proteins. Functional enrichment and network analyses revealed partially overlapping dysregulation of pathways associated with ECM organization and degradation, ECM–receptor interaction, matricellular signaling and mechanobiological processes. In DD samples (n = 10), immunohistochemistry confirmed increased expression of fibrosis-associated proteins (α-SMA, TGF-β1, TGFBI, COL III, COL VI, and COL XII) (at least p < 0.01). Despite these similarities, differences in individual protein abundance and collagen crosslinking were observed between tissues. The findings suggest that DD and RC may share aspects of fibrotic ECM-remodeling despite differences in age, localization, and disease origin. These findings provide initial insights into shared ECM-remodeling processes, although their interpretation should consider the relatively small sample size and biological heterogeneity of the analyzed tissues.