Stem Cells Translational Medicine最新文献

Juvenile systemic sclerosis (jSSc) is a rare, chronic, autoimmune disease in children/adolescents and is associated with significant morbidity, skin thickening/hardening (scleroderma), organ toxicity and sub-optimal therapeutic options. In this report, autologous stem cell transplantation is associated with clinical improvement in a 17-year-old with refractory jSSc.

Background/aim: Adipose tissue-derived mesenchymal stromal cells (ASC) are used in advanced therapy medicinal products due to their regenerative and immunomodulatory properties. Increasing the number of dosages derived from each donor product is essential to reduce variability and improve scalability of cell therapy. However, extended in vitro expansion may induce cellular senescence, potentially compromising therapeutic efficacy. This study aimed to assess the remaining proliferative potential of a cryopreserved ASC product and identify robust transcriptomic -biomarkers of senescence.

Methods: ASC from five donors were cultured until replicative senescence or passage 10. Morphology, growth kinetics, and confluence were monitored. Bulk RNA sequencing was performed on samples from passage 1, 3, 6, and final passage. Principal component analysis, differential expression, gene set variation analysis, and variance partitioning were used to characterize transcriptional changes and identify biomarkers.

Results: ASC maintained stable proliferation and morphology for at least three passages post-thaw. Major transcriptional shifts occurred between passage 3 and later passages. Senescence-associated gene enrichment increased progressively, with donor-specific variation evident at intermediate passages. Forty biomarkers (20 upregulated, 20 downregulated) were identified with expression changes primarily attributable to passage rather than donor identity.

Conclusion: Cryopreserved ASC retain substantial proliferative capacity post-thaw. Senescence develops gradually and is detectable through consistent transcriptomic changes. These findings relate to proliferative and senescence-associated molecular changes and do not directly assess therapeutic efficacy. The identified biomarkers provide a foundation for developing senescence-focused quality control assays to support safe and effective ASC-based therapies.Significance statementThis study identifies promising biomarker candidates for developing a quality control assay to detect cellular senescence and evaluates the remaining replicative potential of a GMP-approved investigational medicinal product. These findings contribute to improving the safety and consistency of cell-based therapies by enabling detection of senescence-related changes.

Chondrocytes play a pivotal role in cartilage tissue engineering. ADAMTS4 gene encodes aggrecanase-1, which is known to affect chondrocyte biology by regulating aggrecan degradation. However, the molecular mechanism by which ADAMTS4 regulates chondrocyte phenotype remains unclear. To comprehensively investigate Adamts4-regulated genes in primary rat costal chondrocytes, we conducted siRNA-mediated Adamts4 knockdown alongside RNA sequencing (RNA-seq), co-immunoprecipitation coupled with mass spectrometry (CO-IP/MS), and enhanced RNA immunoprecipitation sequencing (iRIP-seq). Our results demonstrated that Adamts4 knockdown did not affect chondrocyte apoptosis. However, Adamts4 silencing markedly changed the expression levels of numerous genes linked to cell differentiation and cell cycle progression. CO-IP/MS experiments showed that Adamts4 extensively interacted with RNA-binding proteins (RBPs) in rat chondrocytes. iRIP-seq data suggested that Adamts4 bound to a large number of transcripts, especially those with AU-rich motifs at coding regions. Most interestingly, we found three genes Hmox1, Acan, and Col2a1, which were deregulated upon Adamts4 silencing and enriched in the Adamts4 RIP samples. Altogether, these results indicate that Adamts4 knockdown remarkably modulates the transcriptomic profile of rat chondrocytes. Interactions with RBPs or target mRNAs might contribute to Adamts4-mediated alterations in gene expression. These findings warrant further investigation of the crucial target genes of ADAMTS4 in the regulation of human chondrocyte behaviors.

Calvarial sutures, the major growth centers for skull morphogenesis, are currently regarded as "niches" for calvarial stem cells. Our previous study has identified a skeletal stem/progenitor cell population resident within the suture mesenchyme. Moreover, we have shown that decrease and/or imbalance of their representation in the "niche" impact the fate of a non-fusing suture to fusing-suture and vice versa. Herein, taking advantage of an our established ex vivo calvarial suture explant model we investigated the impact triggered by FGF2, a pro-osteogenic and pro-angiogenic factor, on our skeletal stem/progenitor cell population resident in the suture mesenchyme. Multi-omics data integration combined with cell biology identifies dynamic changes in the representation of skeletal stem/progenitor cell population thus, unveiling within them functionally distinct populations with angiogenesis-competent properties. Findings altogether indicate that FGF2 stimuli may alter the suture "niche" homeostasis and that coordinate an osteogenesis-angiogenesis coupling within skeletal stem/progenitor cell sub-populations.

Background: Allergic airway inflammation is one of the chronic inflammatory diseases and is generally dominated by T helper 2 cells (Th2). Dendritic cells (DCs) are essential to mounting the Th2-mediated airway inflammation by presenting inhaled antigens to prime CD4+ T cells. Small extracellular vesicles (sEV) derived from mesenchymal stem cells (MSCs) exhibited great interest in intractable diseases. However, the effects and mechanisms of MSC-sEV on DCs in airway inflammation is still unclear.

Methods: We isolated MSC-sEV using anion-exchange chromatography. Mouse bone marrow-derived DCs (BMDCs) and human monocyte-derived DCs (moDCs) were used to study the effects of MSC-sEV on dendritic cell surface molecules and their cytokine release. Mice were sensitized with house dust mites (HDM) to induce airway inflammation, and treated with MSC-sEV, the effects of sEV on murine DCs were identified. Extracellular flux analysis techniques were used to study the effects of MSC-sEV on the metabolic state of dendritic cells. RNA sequencing to study altered gene expression in BMDCs after MSC-sEV treatment.

Results: MSC-sEV mitigated the accumulation of Th2-associated cDC2s and moDCs in mouse lung in response to HDM. In vitro, MSC-sEV treatment significantly attenuated the activation of BMDCs. Furthermore, we identified that DCs were able to take MSC-sEV in vitro and in vivo. Mechanistically, MSC-sEV exerted regulatory effects on the metabolic pathways of murine DCs, specifically enhancing the reliance on oxidative phosphorylation of BMDCs. Importantly, MSC-sEV displayed similar effects on human moDCs.

Conclusions: MSC-sEV are able to alter the metabolic state of DCs, favoring DCs to maintain OXPHOS (oxidative phosphorylation) rather than glycolysis, thereby reducing DCs-initiated inflammatory responses and attenuating Th2 lung inflammation, suggesting MSC-sEV can be a potential clinical therapy for airway inflammation.

Extracellular vesicles (EVs) derived from mesenchymal stem cells have therapeutic potential for optic nerve injury. However, further investigations are needed to increase their efficacy. In this study, we tried to enhance targeting and recovery function of EVs using conjugation with integrin αVβ3 antagonist-c(RGDyk) peptide. The molecular mechanism of neuronal repair was investigated as a potential treatment for optic nerve injury. EVs were shown to restore effectively the abnormal regulations of neuronal markers in optic nerve injury models. Notably, the functionally optimized c(RGDyK)_EVs exhibited superior targeting capabilities and modulated neuroregeneration in cases of hypoxic damage and inflammation. Single-cell RNA-seq analysis of R28 cells revealed significant regulation of transcription of genes involved in retinal ganglion cell (RGC) regeneration, neuronal growth, and inflammation by c(RGDyK)_EVs via the Yap-Taz signaling pathway. This study highlighted the enhanced therapeutic potential of c(RGDyK)_EVs over naïve EVs in the context of optic nerve disease. The findings suggested that c(RGDyK)_EVs hold promise as an alternative therapy for optic nerve injury.

Severe ocular damage, such as corneal alkali-injury, can compromise the limbal stem cell (LSC) niches, leading to LSC deficiency (LSCD) and resulting in significant vision impairment. Current insights into the wound healing process associated with simple limbal epithelial transplantation (SLET), a therapeutic intervention for LSCD, are derived solely from clinical studies, which impose methodological limitations and hinder a complete understanding of the cellular dynamics underlying SLET. For the first time, we adapted the clinical SLET procedure to a mouse model of LSCD, enabling a robust and detailed assessment of short- and long-term morphological, immunomodulatory and cell-fate outcomes in corneal regeneration. We demonstrated that simple limbal-corneal epithelial transplantation (SLCET) treatment effectively reduces corneal opacification, vascularization, and inflammation induced by alkali-injury, with persistent benefits. We used transgenic GFP-expressing mice to isolate GFP-positive limbal-corneal cells and trace their fate in the context of SLCET. Our results revealed that both endogenous and transplanted cells contribute to replenishing the depleted limbal-corneal cell population. Furthermore, we showed that the engrafted-cells retained a long-term proliferative capacity and stemness phenotype, underscoring their role in sustained corneal regeneration. Overall, our study provides a temporal and mechanistic overview of SLCET outcomes, highlighting the engrafted-cells as key players in corneal regeneration. This framework also holds a strong translational potential for enhancing SLET's efficacy and uncovering novel LSC modulators.

Carcinogenesis and acquisition of multidrug resistance within established cancers are both multistep evolutionary processes in which stem cells play a role. This perspective will briefly review two corresponding theoretical constructs under development. Efficiency of carcinogenesis (EOC) considers multistep carcinogenesis and predicts the effect of differing dynamics on the efficiency of generating a transformed founder cell. EOC has been applied to evaluation of the role of genetic instability in carcinogenesis. Dynamic precision medicine (DPM) is a method for providing personalized treatment sequences for cancer while explicitly considering intracancer subclonal heterogeneity and evolutionary dynamics (growth and evolutionary rates). It adapts therapy frequently and proactively by anticipating the kinetics of multidrug resistance prior to its detection, and prioritizing its prevention. Simulations suggest potential to substantially increase survival and cure rates across a broad range of clinical presentations. Both of these problems implicate very small subclones within stem cell and/or differentiated compartments, and evolution may occur over months to years. We describe novel experimental technologies for quantifying longitudinal dynamics of very large numbers of cells for prolonged periods, allowing detection and tracking of rare events and their evolution over time. We further highlight two potential applications. In Fanconi anemia, optimal treatment sequences for minimizing bone marrow failure while not increasing the risk of leukemia may be designed using EOC and DPM and tested in laboratory models. In refractory acute myeloid leukemia, high throughput molecular characterization and drug sensitivity screening of subclones is showing clinical promise, and may be further optimized with DPM.

The clinical use of bone graft materials for alveolar ridge preservation following tooth extraction has become a standard procedure to facilitate subsequent implant restoration and prosthetic rehabilitation. However, the therapeutic efficacy of these materials is substantially limited by their bio-inertness, lack of cellular activity, and unpredictable resorption rates. The development of bioactive osteogenic materials capable of host integration and active promotion of bone regeneration would represent a significant advancement over current clinical protocols. To address this challenge, our research has focused on developing bioactive biomaterials using human dental pulp stem cells (hDPSCs). This study proposes a novel strategy for alveolar ridge repair utilizing lyophilized hDPSC microspheres preconditioned through a three-dimensional (3D) dynamic osteogenic induction system. We cultured hDPSCs into 3D microspheres and subjected them to osteogenic induction within our established dynamic culture system, followed by lyophilization to prepare "off-the-shelf" osteogenic tissues. The resulting lyophilized microsphere constructs were implanted into fresh extraction sockets of SD rats and New Zealand white rabbits and evaluated over 4 weeks. Comparative analysis demonstrated that the lyophilized microsphere group exhibited significantly enhanced alveolar bone preservation, superior new bone formation, and improved bone microarchitecture compared to both control groups and traditional artificial bone powder groups. This preclinical study validates the potential of lyophilized hDPSC microspheres as an efficient and clinically promising bioactive material for post-extraction alveolar ridge reconstruction.

The conserved, pluripotency-associated miR-302/367 cluster coordinates cell fate and aging via epigenetic, cell cycle, and signaling regulation. Highly expressed in pluripotent stem cells and silenced during differentiation, it promotes efficient somatic cell reprogramming by suppressing senescence mediators (eg, p16INK4a, p21) and replacing oncogenes such as c-Myc to minimize tumorigenic risks. Beyond pluripotency, the miR-302/367 cluster reduces oxidative stress, mitochondrial dysfunction, and fibrosis, indicating therapeutic potential in age-associated conditions such as neurodegenerative, ocular, and fibrotic diseases. This review summarizes the dual ability of miR-302/367 cluster in promoting cell state transitions and transiently resetting cellular aging to enable healthspan extension. We critically discuss the pivotal role of miR-302/367 cluster in pluripotency and reprogramming while countering aging hallmarks. Finally, we explore how combining single-miRNA therapeutics with clinically viable delivery systems (lipid nanoparticles and extracellular vesicles) can link cellular reprogramming with targeted rejuvenation therapies.