Jul 2026· Life Science· Vol 402, pp.
124579
· 0 citations· 51 references
Medicine
TL;DR
A non-canonical role for SLFN11 safeguarding MSCs from senescence via the DDX1/mTOR/eIF4E axis is delineated and SLFN11-overexpressing MSCs constitute a promising and safe cell therapy source, characterized by preserved pluripotency and augmented therapeutic efficacy.
Abstract
Aims
Mesenchymal stem/stromal cells (MSCs) are indispensable for cellular therapy; however, long-term in vitro expansion triggers replicative and stress-induced senescence, which undermines their stemness and therapeutic potential. This study aimed to investigate the protective effect and underlying mechanism of the Schlafen (SLFN) 11 gene against human MSC senescence in vitro and to evaluate the efficacy and safety of SLFN11-overexpressing MSCs.
Materials And Methods
SLFN11 was overexpressed in human MSCs via lentiviral transduction. Protective effects were comprehensively assessed in replicative and stress-induced senescence models by evaluating cell cycle arrest, proliferative and clonogenic capacities, DNA damage, pluripotency, and β-galactosidase activity. Molecular interactions were elucidated through co-immunoprecipitation (Co-IP) and LC-MS/MS, whereas therapeutic efficacy was determined using a DSS-induced ulcerative colitis mouse model.
KEY
Findings
SLFN11 overexpression significantly attenuated senescence by reducing DNA damage accumulation, reversing cell cycle arrest, and enhancing proliferation, clonogenicity, and stemness properties. Mechanistically, SLFN11 bound the RNA helicase DDX1 to attenuate mTOR/eIF4E signaling, thereby resisting senescence. Compared to control MSCs, SLFN11-overexpressing MSCs exhibited superior protection against ulcerative colitis, as evidenced by reduced colonic lesions, maintenance of mucosal integrity, and inhibition of proinflammatory factor secretion. SLFN11-overexpressing MSCs also showed no detectable tumorigenicity or chromosomal aberrations.
Significance
These findings delineate a non-canonical role for SLFN11 safeguarding MSCs from senescence via the DDX1/mTOR/eIF4E axis. SLFN11-overexpressing MSCs therefore constitute a promising and safe cell therapy source, characterized by preserved pluripotency and augmented therapeutic efficacy.
Crocin I has a protective effect against D-galactose-induced aging in rat models; however, its specific mechanism on delaying cellular senescence remains unclear. The aim of the present study was to investigate the interventional effect and related molecular mechanism of Crocin Ⅰ on replicative senescence and premature senescence of human embryonic lung diploid fibroblasts (WI-38) cells. Replicative senescence model was established through long-term culture and passaging to >55 PD. The premature senescence model was established using WI-38 cells treated with 2,2'-azobis-2-methy-propanimidamide dihydrochloride. The cell viability was detected by using a cell counting kit-8 assay, and the effects of Crocin Ⅰ on senescent phenotype of WI-38 cells were detected by senescence-associated β-galactosidase (SA-β-Gal) staining. In addition, the intracellular reactive oxygen species (ROS) level was detected by flow cytometry, and gene expression levels were detected by quantitative PCR. RNA sequencing analysis was performed to explore the alteration genes in senescent cells and the Crocin Ⅰ intervention group. The proteins PKM2, SIRT1, p53 and p21 in WI-38 cells were detected by western blotting, whereas cellular lactate level was detected by L-lactic acid content detection kit and pyruvate kinase (PK) enzyme activity was measured using PK activity assay kit. Crocin Ⅰ promoted cell proliferation within the range of 10-40 µM. In addition, Crocin Ⅰ decreased the SA-β-Gal staining positive rate on both replicative and premature senescent WI-38 cells. Furthermore, the intracellular ROS level was decreased by Crocin Ⅰ intervention. The differentially expressed genes were mainly enriched in phosphorylation, oxidation-reduction process, glucose metabolic process, cellular senescence pathway, glycolysis, hippo signaling pathway and pyruvate metabolism. RNA sequencing analysis and molecular docking revealed that PKM2 was one of the potential targets in Crocin Ⅰ against senescence activity. Furthermore, Crocin Ⅰ could enhance the expression of SIRT1 following with decreasing the level of PKM2, p21 and p53. Meanwhile, Crocin Ⅰ significantly decreased the cellular lactate level in WI-38. In conclusion, Crocin Ⅰ alleviated the replicative and premature senescence of WI-38 cells partially through regulating SIRT1 and PKM2 expression. Furthermore, this function may involve in glycolysis pathway.
Yicheng Fu, Zhiwei Ding, Yunchuang Chang et al.· Experimental and Therapeutic...· 0 citations
Human mesenchymal stromal cells (hMSCs) remain the most clinically advanced adult stem cell source; however, their therapeutic potential is limited by rapid replicative senescence during ex vivo expansion. Replicative senescence in hMSCs is characterized by cell cycle arrest, acquisition of senescence-associated β-galactosidase (SA-β-Gal) activity, and secretion of the senescence-associated secretory phenotype (SASP) factors. We investigated whether conditioned medium derived from human extended pluripotent stem cells (hEPSCs), which possess both embryonic and extra-embryonic developmental potential beyond that of conventional embryonic stem cells (hESCs), could attenuate replicative senescence in human Wharton’s Jelly-derived MSCs (WJMSCs). Using sequential ultrafiltration (10 kDa and 3 kDa) followed by liquid chromatography-tandem mass spectrometry, we identified several proteins from hEPSC-conditioned medium. We then tested the combination of S100A9/GAPDH/S100A8 proteins for their effects on doxorubicin (DOXO)-induced and replicative senescence. hEPSC-conditioned medium markedly attenuated replicative senescence in hMSCs. Notably, the combination of S100A9/GAPDH/S100A8 proteins not only mitigated doxorubicin-induced senescence but also counteracted replicative senescence, as evidenced by a significant reduction in SA-β-Gal-positive cells and downregulated mRNA expression of senescence-associated genes, including p16, p21, and the SASP factor IL-6. Furthermore, EdU incorporation assays revealed significantly enhanced proliferative capacity following treatment. Collectively, our findings establish a defined protein combination (S100A9/GAPDH/S100A8) that counteracts both replicative and stress-induced senescence, offering a novel, cell-free strategy to enhance the clinical utility of WJMSCs.
Qingcai Feng, Qianwen Pang, Hongyu Lu et al.· Stem cell research & therape...· 0 citations
Background: Drug repurposing represents an accelerated and cost-effective approach to discovering novel oncologic therapeutics. Here, we investigated the anticancer potential and underlying mechanisms of marbofloxacin (MBF), a veterinary fluoroquinolone (FQ), against breast cancer (BC) cells. Methods: The cellular impacts of MBF on cell viability, anchorage-dependent growth, tumorigenicity, migration, apoptosis, proliferation, senescence, and mitochondrial function were thoroughly characterized. To further elucidate its mechanistic activity, real-time qRT-PCR, untargeted LC-MS/MS-based metabolomics, network pharmacology, and molecular docking analysis were integrated. Results: MBF suppressed BC cell growth by inhibiting cellular proliferation and migration, disrupting mitochondrial membrane potential, and inducing ROS-mediated apoptosis and irreversible cellular senescence. These phenotypic impacts were accompanied by upregulation of tumor suppressors such as CDKN1A and PUMA and downregulation of oncogenes including MKI67, BIRC5, and BCL-2. Metabolomic analysis revealed broad suppression of biosynthesis-related metabolic pathways, characterized by the depletion of critical polyamines and nucleotide pathways. Network pharmacology and molecular docking analyses identified EGFR and HSP90AA1 as putative hub proteins potentially associated with the observed anticancer phenotype. Conclusions: These results provide initial evidence that MBF induces metabolic and molecular rewiring in BC, highlighting its promise as a repositionable therapeutic candidate.
M. Yavuz, F. R. P. Dewi, Ilknur Keskin et al.· Pharmaceuticals· 0 citations
OBJECTIVE
This study aims to elucidate the molecular mechanisms by which Polyphyllin I (PPI), a potent steroidal saponin, attenuates non-small cell lung cancer (NSCLC) progression via mechanistic reprogramming of an autophagy-dependent immunogenic response.
METHODS
Integrated in vitro (A549, H460) and in vivo (LLC xenograft) models were deployed to evaluate PPI's efficacy on autophagic flux and the tumour immune microenvironment. The regulatory role of autophagy in macrophage-mediated antigen presentation was scrutinised via ATG3-mediated genetic silencing or overexpression in tumour-macrophage co-culture systems. Concurrently, the capacity of PPI to sensitise NSCLC cells to cisplatin (DDP) and counteract chemoresistance was evaluated.
RESULTS
PPI activated the AMPK/p53/mTOR signalling axis, robustly inducing core autophagic markers (LC3-II and Beclin-1) in a dose-dependent manner. Mechanistically, PPI-induced autophagic flux served as a prerequisite for antitumoural M1 macrophage polarisation, characterised by significant upregulation of iNOS and MHC-II in co-cultured THP-1 cells. Genetic knockdown of ATG3 effectively abrogated these immunostimulatory profiles, whereas ATG3 overexpression potentiated PPI-driven antigen presentation. Furthermore, PPI administration markedly delayed the onset of DDP resistance sustained by functional autophagic flux. In vivo, PPI significantly suppressed tumour burden, accompanied by enhanced CD8+ T-cell infiltration and elevated cytotoxic effector levels (IFN-γ and Granzyme B).
CONCLUSION
Our findings establish PPI as a dual autophagic-immune modulator that re-engineers the immunosuppressive microenvironment. By coupling intracellular autophagic stress with macrophage-mediated antigen presentation, PPI reinstates antitumour immunity and abrogates chemoresistance, offering a compelling therapeutic framework for managing recalcitrant NSCLC.
Zongxu Liu, Yanping Li, Shumin Li et al.· Clinical and Experimental Ph...· 0 citations
Colorectal cancer (CRC) remains a major cause of cancer-related mortality due to therapeutic resistance. Because colorectal cancer stem-like cells (CRCSCs) play a central role in tumor initiation and progression, therapeutic strategies addressing CSC-enriched populations are urgently needed. In this study, we investigated the anticancer effects of emodin, a natural anthraquinone, in CSC-enriched tumorsphere models. Emodin significantly suppressed the viability and self-renewal capacity of HCT116- and SW480-derived CSCs. It induced G0/G1 cell cycle arrest and markedly downregulated stemness-associated markers (CD44, CD133, ALDH1A1, SOX2, NANOG, and OCT4). Importantly, emodin-induced cell death was characterized by mitochondrial dysfunction, increased mitochondrial reactive oxygen species, loss of membrane potential, and nuclear translocation of apoptosis-inducing factor (AIF). This cytotoxicity was not rescued by the pan-caspase inhibitor Z-VAD-FMK, confirming caspase-independent apoptosis. Furthermore, network pharmacology and experimental validation identified GSK3β as a key target. Emodin reduced Wnt/β-catenin signaling by decreasing β-catenin stabilization and nuclear accumulation. Crucially, a rescue experiment utilizing LiCl confirmed that emodin’s suppressive effects are mechanistically dependent on the GSK3β/Wnt/β-catenin axis. Collectively, emodin suppresses CRCSC characteristics in vitro by downregulating Wnt/β-catenin signaling and inducing AIF-associated caspase-independent apoptosis, highlighting its therapeutic potential against CRC.
M. Ahmadi, Hong Lae Kim, H. Jung· Biomolecules & Therapeutics· 0 citations
Mesenchymal stem cells (MSCs) are widely investigated for regenerative medicine, tissue repair, immunomodulation, and selected cancer-related applications because of their multilineage differentiation potential, paracrine activity, immunoregulatory properties, and capacity to home to sites of injury. However, the therapeutic potential of MSCs depends not only on their tissue source or surface-marker expression but also on their functional state. MSC senescence is not a single phenotypic alteration; rather, it reflects the combined effects of persistent DNA damage response activation, disruption of mitochondrial and metabolic homeostasis, remodeling of epigenetic and secretory networks, and loss of proteostasis. Collectively, these changes drive MSCs toward reduced proliferative capacity, dysregulated paracrine signaling, impaired immunomodulatory activity, and diminished tissue-repair potential, thereby compromising cell-product potency and batch-to-batch consistency. This review summarizes the molecular mechanisms underlying MSC senescence, its source-specific functional consequences, and emerging rejuvenation strategies, with an emphasis on MSC quality control, potency assessment, and clinical translation.
S. Zhou, Yu Xie, Yanjie Guo et al.· Frontiers in Aging· 0 citations
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