PLoS One
·
2025
·Vol. 20
·期 6
·e0326978
·
IF 2.6
Chang Chen,Qiqi Yun,Juanli Ran,Ziyao Zhou,Pengxiang Zhang,Rong Li
Objective
This study investigated the interplay between Hippo-YAP and β-catenin signaling in regulating odontogenic differentiation of human dental pulp stem cells (DPSCs) and explored its potential implications for dentin-pulp regeneration.
Methods
Using lentivirus-mediated YAP overexpression/silencing, β-catenin siRNA knockdown, and pharmacological Wnt inhibition (via WIF-1), we assessed DPSC proliferation, migration, mineralization, and molecular markers (via qRT-PCR, immunofluorescence). In vivo validation employed subcutaneous transplantation of DPSC-seeded scaffolds in immunocompromised mice.
Results
YAP activation enhanced DPSC proliferation (1.44-fold), migration (1.39-fold), invasion (1.54-fold), and differentiation, as evidenced by elevated ALP activity (1.46-fold) and mineralization (1.36-fold). We observed transcriptional upregulation of odontogenic markers (RUNX2, DSPP, DMP1, OCN, ALP) and Wnt pathway components (β-catenin, Cyclin D1, c-Myc). Immunofluorescence revealed coordinated YAP and β-catenin expression patterns during differentiation. β-catenin silencing or Wnt inhibition abolished YAP-mediated functional enhancements and simultaneously suppressed YAP expression, partially confirming bidirectional regulation. In vivo , YAP-overexpressing DPSCs exhibited 1.27- to 1.62-fold induction of dentin-specific markers and β-catenin, whereas YAP silencing impaired these markers expression.
Conclusions
Our findings demonstrate that coordinated YAP and β-catenin signaling drives DPSC odontogenesis, with potential implications for dentin regeneration. Although reciprocal regulation is evident, direct molecular interactions require further validation.
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高影响因子
Cell Regeneration
·
2020
·Vol. 9
·期 1
·1-16
·
IF 0
Guo Anqi,Wang Bingjie,Lyu Cheng,Li Wenjing,Wu Yaozu,Zhu Lu,Bi Ran,Huang Chenyu,Li Jiao Jiao,Du Yanan
Apparent Young’s modulus (AYM), which reflects the fundamental mechanical property of live cells measured by atomic force microscopy and is determined by substrate stiffness regulated cytoskeletal organization, has been investigated as potential indicators of cell fate in specific cell types. However, applying biophysical cues, such as modulating the substrate stiffness, to regulate AYM and thereby reflect and/or control stem cell lineage specificity for downstream applications, remains a primary challenge during in vitro stem cell expansion. Moreover, substrate stiffness could modulate cell heterogeneity in the single-cell stage and contribute to cell fate regulation, yet the indicative link between AYM and cell fate determination during in vitro dynamic cell expansion (from single-cell stage to multi-cell stage) has not been established.Here, we show that the AYM of cells changed dynamically during passaging and proliferation on substrates with different stiffness. Moreover, the same change in substrate stiffness caused different patterns of AYM change in epithelial and mesenchymal cell types. Embryonic stem cells and their derived progenitor cells exhibited distinguishing AYM changes in response to different substrate stiffness that had significant effects on their maintenance of pluripotency and/or lineage-specific characteristics. On substrates that were too rigid or too soft, fluctuations in AYM occurred during cell passaging and proliferation that led to a loss in lineage specificity. On a substrate with ‘optimal’ stiffness (i.e., 3.5 kPa), the AYM was maintained at a constant level that was consistent with the parental cells during passaging and proliferation and led to preservation of lineage specificity. The effects of substrate stiffness on AYM and downstream cell fate were correlated with intracellular cytoskeletal organization and nuclear/cytoplasmic localization of YAP.In summary, this study suggests that optimal substrate stiffness regulated consistent AYM during passaging and proliferation reflects and contributes to hESCs and their derived progenitor cells lineage specificity maintenance, through the underlying mechanistic pathways of stiffness-induced cytoskeletal organization and the downstream YAP signaling. These findings highlighted the potential of AYM as an indicator to select suitable substrate stiffness for stem cell specificity maintenance during in vitro expansion for regenerative applications.
研究领域:
干细胞生物学
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生物物理学
·
再生医学
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PubMed
ACTA PHARMACOLOGICA SINICA
·
2026
·
IF 8.4
Zhao Xiao-jun,Xu Yang,Zhang Cong-hui,Zhao Cong,Liu Li,Wang Xiao-wei,Wang Meng-yan,Gao Zi-xiang,Shao Rong-guang,Qi Li-qiang,Che Yong-sheng,Zhao Wu-li
Mitochondrial DNA (mtDNA) mutations are the most common cause in aberrant mitochondrion-leading cancer, exploration of direct targeting mutated mtDNA still remains incomplete. Secoemestrin C (Sec C) is epitetrathiodioxopiperazine derived from the endophytic fungus, which exhibited a rapid and prominent anti-breast cancer effect in triple-negative breast cancer (TNBC). In this study we investigated the anticancer mechanism of Sec C, especially its effect on TNBC cells. We showed that Sec C potently inhibited the viability of both TNBC (MDA-MB-231, HS578T, BT-549) and non-TNBC (MCF-7, T47D, SK-BR-3) cells in vitro with IC 50 values of 1−2 μM. In MDA-MB-231 cells, treatment with Sec C (2 μM) induced DNA breakage and subsequent apoptosis. Furthermore, treatment with Sec C (2 μM) caused mtDNA damage, mitochondrial ubiquitination and subsequent mitophagy in MDA-MB-231 and MCF-7 cells. RNA-seq analysis revealed that Sec C mitigated YAP level in time and dose-dependent manner either in MDA-MB-231 and MCF-7 cells. By re-analyzing the Sec C-responsive gene network proteins, we identified SLX4 as an oncogene promoting breast cancer development, potentially by stabilizing mtDNA to suppress pathologic mitochondrion mitophagy. Specifically, Sec C initiated MDA-MB-231 cells to yield ROS that induced SLX4 ubiquitination and degradation, leading to mtDNA damage and exacerbated mitophagy and promoted YAP degradation bypassing YAP-driven DNA repair pathways. This study not only demonstrates that Sec C is a rapid and prominent anti-breast cancer drug for TNBC, but also reveals SLX4 as a novel mtDNA stabilizer supporting breast cancer progression, positioning it as both a prognostic biomarker and therapeutic target.
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Frontiers in Cell and Developmental Biology
·
2021
·
IF 6.684
Li Jiawei,Sun Ziying,Lv Zhongyang,Jiang Huiming,Liu Anlong,Wang Maochun,Tan Guihua,Guo Hu,Sun Heng,Wu Rui,Xu Xingquan,Yan Wenjin,Jiang Qing,Ikegawa Shiro,Shi Dongquan
Mesenchymal stem cells (MSCs) are well known for their multi-directional differentiation potential and are widely applied in cartilage and bone disease. Synovial mesenchymal stem cells (SMSCs) exhibit a high proliferation rate, low immunogenicity, and greater chondrogenic differentiation potential. Microtubule (MT) plays a key role in various cellular processes. Perturbation of MT stability and their associated proteins is an underlying cause for diseases. Little is known about the role of MT stabilization in the differentiation and homeostasis of SMSCs. In this study, we demonstrated that MT stabilization via docetaxel treatment had a significant effect on enhancing the chondrogenic differentiation of SMSCs. MT stabilization inhibited the expression of Yes-associated proteins (YAP) and the formation of primary cilia in SMSCs to drive chondrogenesis. This finding suggested that MT stabilization might be a promising therapeutic target of cartilage regeneration.
研究领域:
干细胞生物学
·
骨科
·
再生医学
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高影响因子
JOURNAL OF NANOBIOTECHNOLOGY
·
2024
·Vol. 22
·期 1
·1-16
·
IF 10.6
Qi Ji,Zhang Long,Ren Zhongyu,Yuan Yi,Yu Jiahao,Zhang Yining,Gu Linbo,Wang Xu,Wang Yan,Xu Haoyue,Yu Rutong,Zhou Xiuping
Background The Hippo pathway is a conserved tumour suppressor signalling pathway, and its dysregulation is often associated with abnormal cell growth and tumorigenesis. We previously revealed that the transcriptional coactivator Yes-associated protein (YAP), the key effector of the Hippo pathway, is a molecular target for glioblastoma (GBM), the most common malignant brain tumour. Inhibiting YAP with small interfering RNA (siYAP) or the specific inhibitor verteporfin (VP) can diminish GBM growth to a certain degree. Results In this study, to enhance the anti-GBM effect of siYAP and VP, we designed stepwise-targeting and hypoxia-responsive liposomes (AMVY@NPs), which encapsulate hypoxia-responsive polymetronidazole-coated VP and DOTAP adsorbed siYAP, with angiopep-2 (A2) modification on the surface. AMVY@NPs exhibited excellent blood‒brain barrier crossing, GBM targeting, and hypoxia-responsive and efficient siYAP and VP release properties. By inhibiting the expression and function of YAP, AMVY@NPs synergistically inhibited both the growth and stemness of GBM in vitro. Moreover, AMVY@NPs strongly inhibited the growth of orthotopic U87 xenografts and improved the survival of tumour-bearing mice without adverse effects. Conclusion Specific targeting of YAP with stepwise-targeting and hypoxia-responsive liposome AMVY@NPs carrying siYAP and VP efficiently inhibited GBM progression. This study provides a valuable drug delivery platform and creative insights for molecular targeted treatment of GBM in the future.
研究领域:
肿瘤学
·
药学
·
分子生物学
·
神经科学
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高影响因子
JOURNAL OF CELLULAR AND MOLECULAR MEDICINE
·
2026
·Vol. 30
·期 17
·e71361
·
IF 4.7
Xin Ma,Yiting Huang,Xiao Lv,Xiumei Mo,Junfeng Liu,Fenggen Yan,Siqi Ye,Yu Zhang,Dacan Chen,Jinjing Jia
Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by skin barrier dysfunction and immune balance disorders. Currently, drugs for repairing the skin barrier in AD are limited. The gu-ben-hua-shi formula (AESS) has been shown to regulate Th1/Th2/Th17/Treg imbalance with good clinical effect. However, its effects on regulating autophagy, repairing the skin barrier, and interacting with Yes-associated protein (YAP) remain unknown. To elucidate the therapeutic effects of AESS on AD and to investigate its mechanism of restoring skin barrier function through the Hippo-YAP pathway. We conducted a single-arm observational study to evaluate the clinical efficacy of the AESS in patients with AD. Network pharmacology was used to predict the potential therapeutic targets and mechanisms. AD mouse models were established by house dust mite stimulation in Nishiki-nezumi Cinnamon/Nagoya mice, and the effects on autophagy and the skin barrier were measured using western blotting, immunofluorescence, mRFP-GFP-LC3 tandem fluorescence assay, and transmission electron microscopy. The effects of AESS serum on an AD-like inflammatory cell model were detected in HaCaT cells stimulated with interferon-γ (IFN-γ)/tumour necrosis factor-α (TNF-α). Activity of the Hippo-YAP signalling pathway was also investigated after YAP knockdown. AESS improved skin rash and barrier function, reduced eosinophil counts, and increased Th1/Th2 and Treg/Th17 ratios in patients with AD. AESS increased autophagy, improved skin barrier function, and inhibited Hippo-YAP signalling pathway activity. YAP knockdown reduces these therapeutic effects. Besides regulating immune imbalance, AESS increases keratinocyte autophagy levels, thereby restoring skin barrier function in AD through the Hippo-YAP signalling pathway. Trial Registration: The Chinese clinical trial registration website: ITMCTR202500042
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EXPERIMENTAL NEUROLOGY
·
2026
·
IF 4.8
Enzhuang Pan,Juan Chen,Yue Xin,Yaxin Gao,Jingquan Dong,Yingda Xu,Jingyuan Zhou,Xinyu Zhou,Huizhen Chen,Panpan Zhao
Ischemic stroke (IS) is one of the leading causes of global disability and mortality, yet effective treatments targeting oxidative stress (OS) and neuronal apoptosis remain lacking. This study investigates the neuroprotective role of Yes-associated protein (YAP) in IS and its regulatory mechanisms. Using clinical IS patient serum samples, a mouse middle cerebral artery occlusion (MCAO) model, and an in vitro oxygen-glucose deprivation/reoxygenation (OGD/R) model, we found that YAP protein levels were significantly downregulated post-ischemia, correlating with aggravated OS and neuronal apoptosis. Overexpression of Yap1 via adeno-associated virus (AAV) vectors reduced infarct volume, restored cerebral blood flow, and improved neurological and cognitive functions in MCAO mice. Mechanistically, YAP alleviated oxidative damage by enhancing the activity of antioxidant enzymes ( e.g. , GSH-Px, SOD) and inhibited apoptosis, as validated by TUNEL and Annexin V/PI assays. Intriguingly, under ischemic conditions, YAP phosphorylation was regulated by TAK1 rather than the canonical Hippo signaling pathway. Knockdown of TAK1 synergized with YAP activation to mitigate oxidative damage and neuronal apoptosis. These findings reveal the critical neuroprotective role of YAP in IS and identify the TAK1-YAP interaction as a therapeutic target. This study provides a mechanistic foundation for developing YAP-based interventions for IS.
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Cell Death & Disease
·
2026
·
IF 12.2
Wu Haibin,Liu Shoupei,Chen Sen,Qin Changlu,Yan Wenjiao,Cao Xiangting,Zhou Yongjian,Duan Yuyou
Primary human hepatocyte (PHH)-derived organoids form grape-like clusters with proliferative capacity, hepatocyte functionality, and multipolar polarity, serving as valuable models for liver biology and therapeutics. However, deriving comparable organoids from human embryonic stem cells (hESCs) remains difficult. Here, we established a defined system to differentiate hESC-derived hepatoblast organoids into hepatocyte organoids (heporgs) with two morphologies: spheroid-like (S-heporgs) and grape-like (G-heporgs). S-heporgs predominated but displayed senescence and apoptosis, generating an inflammatory niche that facilitated G-heporg emergence. G-heporgs exhibited mature hepatocyte markers, binucleation, proliferative activity, and multipolar structures with branched bile canaliculi, closely resembling PHH-derived organoids. Transcriptomic and functional analyses identified IGF2-driven PI3K-AKT activation as essential for G-heporg formation, while YAP signaling supported their long-term expansion. IGF2 supplementation combined with YAP agonist treatment enabled stable G-heporg propagation for over 60 days. These expandable G-heporgs demonstrated regenerative competence and faithfully recapitulated hepatocyte polarity and functional bile canalicular networks, as evidenced by ATP7B copper-dependent translocation and drug-induced cholestasis assays. Our findings establish hESC-derived G-heporgs as expandable, functional counterparts to PHH-derived organoids, providing a robust platform for studying hepatocyte polarity, metabolite trafficking, and liver disease modeling.
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JOURNAL OF NANOBIOTECHNOLOGY
·
2026
·
IF 15
Xu Jie,Zhuang Xiuxiu,Wu Zichang,Yan Fuyi,Shi Deen,Ye Fan,Ding Ruihua,Gao Qi,Yao Qimin,Liu Yiying,Dong Xiaoyu,Zhang Fan,Li Binbiao,Xu Xiangwei,Tong Hongfei,Yao Yanzhi,Jiang Wenbing,Xiao Jian,Li Zhenglin,Chen Wei
Ischemic skin flaps are highly susceptible to distal necrosis because of insufficient perfusion, oxidative stress, and dysregulated cell death. Here, we investigated the role of Hippo/YAP signaling in ischemic flap survival and identified its upstream regulatory mechanism. Hippo pathway inactivation, characterized by reduced MST1/2 and LATS1/2 phosphorylation and enhanced YAP nuclear localization, was preferentially associated with viable flap regions. Mechanistically, ischemia-induced HIF-1α upregulated miR-210-3p, which suppressed ephrin-A3 (EFNA3), weakened the EFNA3–YAP interaction, and promoted YAP nuclear translocation. Re-expression of EFNA3 partially reversed miR-210-3p-induced YAP activation and cytoprotection, confirming its functional involvement. Mechanistically, ischemia-induced HIF-1α increased miR-210-3p expression, which suppressed EFNA3 and facilitated YAP nuclear localization. EFNA3 re-expression partially reversed miR-210-3p-induced YAP activation, supporting its functional involvement. MiR-210-3p treatment was further associated with reduced oxidative injury, lipid peroxidation, mitochondrial dysfunction, and ferroptosis-related molecular alterations, together with improved perfusion and flap survival. In vivo, miR-210-3p improved local perfusion and flap survival. We further developed a virus-inspired biomimetic nanoparticle for miR-210-3p delivery, which achieved protective effects comparable to AAV-mediated delivery in vitro and in vivo. These findings identify the miR-210-3p–EFNA3–YAP axis as an important regulator of ischemic tissue adaptation and provide a non-viral miRNA delivery strategy for ischemic flap repair.
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Materials Today Advances
·
2022
·
IF 9.918
Siyong Gao,Yuxin Tang,Wei Sun,Zhixin Liu,Tianyu Zhao,Xiang Li,Tianlu Wang,Guiqing Liao,Tao Xu,Guangsen Zheng,Yujie Liang
Schwann cells (SCs)-based nerve guidance conduits (NGCs) is a promising strategy for repairing long-gap peripheral nerve injury (PNI). But the number of SCs is limited as terminally differentiated cells. Matrix stiffness is able to direct cell fate of stem cells. And nerve-special stiffness (NSS) may contribute to stem cells converting into SCs. However, the potential mechanisms remain uncertain. The present study aimed to elucidate whether the given NSS was able to promote mesenchymal stem cells (MSCs) trans -differentiating into Schwann cell-like cells (SCLCs), facilitating cell-based NGCs repairing PNI. Gelatin methacryloylate (GelMA) hydrogels with different stiffness were manufactured, and their roles in the trans -differentiation of MSCs into SCLCs were investigated in vitro study. The most favorable stiffness (0.9–2.9 kPa) for trans -differentiation and neurotrophic factors expression was confirmed by western blot and immunofluorescence assays. The reason might lie in softer stiffness stretched cell morphology, leading to up-regulation of PIEZO1 and then activating YAP nuclear translocation. As for in vivo study, multi-channel NGCs containing SCLCs were fabricated with extrusion-based bioprinting and then filled a 5 mm gap in sciatic nerve defect of SD rats. It turned out that GelMA-made NGCs with stiffness of 2.9 kPa achieved promising neurogenerative capacity, not only promoting recovering sensory and motor functions, but also improving myelinated nerve fiber regeneration. Our findings demonstrated that 0.9–2.9 kPa might be the desired mechanical strength inducing trans -differentiation into SCLCs via PIEZO1/YAP axis, and the 3D-printed GelMA NGCs combined with SCLCs could be a potential candidate for long-gap peripheral nerve injury.
研究领域:
生物医学工程
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组织工程
·
神经科学
·
干细胞生物学
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高影响因子
ACS Biomaterials Science & Engineering
·
2025
·
IF 5.5
Shiyang Wu,Zhen Chai,Yanshen Yang,Rong Ding,Bei Gao,Chen Chen,Siyu He,Zhuofu Zhang,Wanting Wan,Rui Zou
Two-dimensional (2D) models fail to mimic a three-dimensional (3D) environment in studying mechanosensing of stem cells. Here, we present a 3D culture model to investigate how 3D matrix stiffness influences YAP activation and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) and evaluate the osteogenic potential of hydrogels in vivo. In this study, a 3D culture model with an adjustable matrix stiffness was established. In the in vitro study, first the osteogenic differentiation of hPDLSCs by the expression of OCN, ALP, COL-1, and RUNX-2 was assessed using qRT-PCR, accompanied by ALP staining, and then YAP expression was evaluated by immunofluorescence. In the in vivo study, hPDLSCs, together with gelatin methacrylate (GelMA) hydrogels of different stiffnesses, were implanted into a rat alveolar bone defect model. As matrix stiffness increased, hPDLSCs showed reduced spreading and significantly decreased expression of OCN, ALP, COL-1, RUNX-2, and YAP activation. Specifically, COL-1 expression in the low-stiffness group was 4.3-fold higher than that in the high-stiffness group, and the YAP nuclear/cytoplasmic ratio under low stiffness was 5.5-fold greater than that under high stiffness at day 7. In vivo, the soft-matrix-cell-laden group exhibited more new bone (86.04%) and collagen formation (74.43%) in the defect area than other groups at week 6. Reduced matrix stiffness likely promotes hPDLSC proliferation, spreading, and osteogenic differentiation through YAP activation, and low-stiffness GelMA hydrogels loaded with hPDLSCs significantly enhance alveolar bone regeneration in vivo.
研究领域:
生物医学工程
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干细胞研究
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组织工程
·
力学生物学
·
再生医学
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高影响因子
FASEB JOURNAL
·
2022
·Vol. 36
·期 10
·e22530
·
IF 5.834
Chengyu Yang,Yang Liu,Ziyan Wang,Minmin Lin,Chao Liu
Despite the best treatment, approximately 10% of fractures still face undesirable repair and result in delayed unions or non-unions. Dynamic mechanical stimulation promotes bone formation, when applied at the correct time frame, with optimal loading magnitude, frequency, and repetition. Controlled mechanical loading significantly increases osteogenic cells during the matrix deposition phase of bone repair. In the bone defect, the blood vessel network guides the initial bone formation activities. A unique blood vessel subtype (Type H) exists in bone, which expresses high levels of CD31 and endomucin, and functions to couple angiogenesis and osteogenesis. However, how this form of controlled mechanical loading regulates the Type H vessels and promotes bone formation is still not clear. Sphingosine 1-phosphate (S1P) participates in the bone anabolic process and is a key regulator of the blood vessel. Its receptor, sphingosine 1-phosphate receptor 1 (S1Pr1), is a mechanosensitive protein that regulates vascular integrity. Therefore, we hypothesis that controlled anabolic mechanical loading promotes bone repair by acting on Type H vessels. To study the effect of S1Pr1 on loading induced-bone repair, we utilized a stabilized tibial defect model, which allows for the application of anabolic mechanical loading. Mechanical loading upregulated S1Pr1 within the entire defect, with up to 80% expressed in blood vessels, as observed by deep tissue imaging. Additionally, S1Pr1 antagonism by W146 inhibited the anabolic effects of mechanical loading. We showed that mechanical loading or activating S1Pr1 could induce YAP nuclear translocation, a key regulator in the cell's mechanical response, in endothelial cells (ECs) in vitro. Inhibition of S1Pr1 in endothelial cells by siRNA reduced loading-induced YAP nuclear translocation and expressions of angiogenic genes. In vivo, YAP nuclear translocation in Type H vessels was up-regulated after mechanical loading but was inhibited by antagonizing S1Pr1. S1Pr1 agonist, FTY720, increased bone volume and Type H vessel volume, similar to that of mechanical stimulation. In conclusion, controlled anabolic mechanical loading enhanced bone formation mainly through Type H vessels in a S1Pr1-dependent manner.
研究领域:
骨科
·
再生医学
·
生物力学
·
分子生物学
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