Cellular and Molecular Life Sciences

Yokoyama, Shigetoshi; Kido, Taketomo; Yoneda, Mitsuhiro; Springer, Danielle A.; Awasthi, Parirokh P.; Chari, Raj; Patterson, Andrew D.; Kimura, Shioko

doi: 10.1007/s00018-025-06045-5pmid: 41469838

Skeletal muscle has an innate ability to restore damaged muscle fibers by contributing specific progenitor cells, called muscle satellite cells. Here we show that secretoglobin (SCGB) 3A1, a tumor suppressor gene in various malignancies including rhabdomyosarcoma, is induced just after muscle injury and contributes to damaged muscle fiber regeneration. Lineage tracing of SCGB3A1 in mice show that SCGB3A1-positive cells highly express myosin heavy chain (MyHC)-IIX in damaged fiber area. Scgb3a1-null and Pax7CreERT2;Scgb3a1f/f conditional-null mice exhibit defective IIX and IIB fiber regeneration, with a concomitant reduction in the expression of Notch3, a gene important for the maintenance of satellite cell self-renewal pools. Aged Scgb3a1-null mice show reduced size of muscle fibers and mass, resulting in compromised muscle performance as compared to the age-matched wild-type mice. This study reveals that SCGB3A1 is an unexpected novel molecule expressed in muscle satellite cells that contributes to fiber type specific muscle regeneration.

Wang, Yingzi; Huang, Haozhong; Liu, Zihao; Gao, Zhenyu; Liao, Bin; Long, Yang; Yu, Fengxu; Nie, Yongmei

doi: 10.1007/s00018-025-06026-8pmid: 41452468

BackgroundAtherosclerosis is a chronic vascular inflammatory disease caused by multiple factors. Anti-inflammatory treatment is an effective approach to treat atherosclerosis. Talin1 is a cell membrane-associated cytoskeletal protein that is widely expressed in mammals and plays essential roles in angiogenesis and endothelial cell barrier function. However, the role of Talin1 in atherosclerosis and the related mechanisms remains unclear.MethodsApoE-KO mice were subjected to partial carotid artery ligation to establish an atherosclerosis model, and the expression of Talin1 in atherosclerotic plaques was verified in vivo. Human umbilical vein endothelial cells (HUVECs) and aortic endothelial cells (HAECs) were treated with tumour necrosis factor α (TNF-α) (10 ng/mL) and subjected to low oscillatory shear stress (OSS) (approximately ± 4 dyn/cm2) to establish cellular inflammation models. A lentivirus was used to regulate Talin1 expression in HUVECs and HAECs.ResultsTalin1 levels were increased in the serum of subjects with coronary heart disease (CHD) compared with those without CHD. We also found that Talin1 levels were increased in the serum of ApoE-KO mice in the operation group compared with the sham operation group. In addition, Talin1 expression was increased in endothelial cells in atherosclerotic plaques. In addition, neither TNF-α nor OSS promoted inflammation in endothelial cells with Talin1 knockdown. Moreover, we found that TNF-α and OSS could activate Piezo1 to mediate Ca²⁺ influx and subsequently activate Talin1 to regulate YAP and promote inflammation.ConclusionsThe results of this study suggest that Talin1 plays a vital role in endothelial inflammation and may be a novel anti-inflammatory therapeutic target for atherosclerosis.

Aiello, Jacopo; Zamarato, Roberta; Moscheni, Claudia; Perrotta, Cristiana; Clerici, Mario; Trabattoni, Daria; Biasin, Mara; Limanaqi, Fiona

doi: 10.1007/s00018-025-05985-2pmid: 41460324

Since the first report in 1972, several studies have documented an association between Parkinson’s disease (PD) and melanoma. Up to 20-fold increased risk of melanoma was reported in PD patients, while a personal/family history of melanoma was linked to a 1.85-fold PD risk. Neurons and melanocytes, which both derive from the neuroectodermal crest, share biological pathways that may be dysregulated in both diseases. In particular, accumulation of the alpha-synuclein (α-syn, SNCA) protein, a pathological hallmark of PD, is also observed in melanoma. Indeed, dysregulated α-syn proteostasis is known to disrupt several biological pathways which can co-incidentally, albeit paradoxically contribute to both neurodegeneration and hyper-proliferative cell growth. These include abnormalities in dopamine (DA), melanin, and iron metabolism, oxidative stress, DNA damage/repair response, inflammation, as well as alterations in mitochondrial function, and cell-clearing machinery. Although α-syn depletion was shown to attenuate melanoma cell proliferation and neurodegeneration, it remains unclear whether α-syn accumulation is a mere culprit of disease, if it represents a common outcome from shared upstream mechanisms, or, finally, a compensatory response to cellular stress. In an effort to elucidate how α-syn bridges melanomagenesis and the neurodegenerative events of PD, this review discusses specific cellular and molecular pathways related to α-syn proteostasis, including environmental factors implicated in melanocytic transformation, such as UV radiation. Addressing open questions and establishing novel experimental models remain essential for developing effective therapeutic approaches to target melanoma and PD without overlooking their comorbidity.

Qian, Zhenyuan; Cai, Xufan; Wu, Fang; Ye, Zaiyuan; Wu, Jianzhang

doi: 10.1007/s00018-025-05955-8pmid: 41288742

BackgroundHistone lactylation represents a novel epigenetic modification that can modulate gene expression, further boosting tumor proliferation, metastasis, and immune suppression. However, whether histone lactylation affects immune evasion in gastric cancer (GC) remains to be elucidated.MethodsWB and IHC were employed to assess the levels of H3K18la in tumor tissues. CCK-8, colony formation, T cell killing, ELISA, cytotoxicity experiments, and T cell chemotaxis experiments were undertaken to determine the biological functions of histone lactylation and METTL3. RNA-seq, WB, and CHIP experiments were utilized to validate the regulatory role of H3K18 lactylation in METTL3 transcriptional activity. The RIP assay was undertaken to verify the post-transcriptional modification of METTL3 protein on CCT2. The IF and flow cytometry were completed to analyze the regulatory mechanism of CCT2 on CD8+T cells’ activity. IHC and IF were performed to detect protein expression in the homograft mouse model.ResultsHistone H3K18 lactylation was elevated in GC, promoting tumor proliferation, inhibiting cytotoxicity, and chemotaxis of CD8⁺T cells. Mechanistically, H3K18 lactylation boosted tumor immune suppression and in vivo tumorigenesis by upregulating METTL3 expression, while METTL3 enhanced CCT2 translation through m6A modification. CCT2 weakened CD8⁺T cell activity by inhibiting Ca²⁺ influx. METTL3 knockdown inhibited the immune escape of GC cells, while overexpression of CCT2 reversed the anti-tumor effect of METTL3 knockdown on CD8⁺T cells.ConclusionHistone lactylation drives upregulation of METTL3 and mediates m6A modification and overexpression of CCT2. CCT2 dampens the activity of CD8+T cells by repressing Ca2+influx, thereby mediating the malignant progression of GC.Graphical abstract[graphic not available: see fulltext]

Liao, Yuxiang; Liu, Bo; Zhang, Zhiping; Zhang, Qian; Xiang, Mingyong; Jin, Chen

doi: 10.1007/s00018-025-06002-2pmid: 41359154

BackgroundGlioma is the most frequently diagnosed brain tumor in adults worldwide which is associated with unfavorable prognosis and survival time. However, the understanding of glioma progression remains limited.MethodsThe cell proliferation in glioma cells were monitored by EdU incorporation and CCK-8 assays. Glioma cell invasion and migration were assessed by Transwell assay. In vivo tumorigenesis were detected by xenograft study with bioluminescence imaging. qRT-PCR, RNA FISH, IHC or western blot were used to detect circEPHB4, MRPS16, YBX1, RBBP6 and other molecules expression. The associations between YBX1 and MRPS16 mRNA, as well as between circEPHB4 and YBX1, were detected by RNA immunoprecipitation (RIP) and RNA pull-down assays. In addition, the ubiquitination of YBX1 and RBBP6-YBX1 interaction were assessed by co-immunoprecipitation (co-IP).ResultsKnockdown of circEPHB4 or MRPS16 inhibited glioma progression in vitro and in vivo. circEPHB4 promoted glioma cell proliferation, migration, and invasion via increasing MRPS16 expression in vitro. At the post-transcriptional level, circEPHB4 enhanced MRPS16 mRNA stability through YBX1-mediated m5C modification in vitro. At the post-translational level, RBBP6 catalyzed the ubiquitination of YBX1, and circEPHB4 competed with RBBP6 to bind YBX1 to inhibit the ubiquitin-proteasomal degradation of YBX1 in vitro. circEPHB4 interacted with YBX1 to promote glioma cell growth via inducing MRPS16 in vitro and in vivo.ConclusioncircEPHB4 bound to YBX1 to inhibit RBBP6-mediated degradation and increase its expression, thus enhancing MRPS16 mRNA stability via m5C modification, and ultimately promoting glioma progression.

Wang, Sijia; Peng, Xueting; Ran, Yutong; Feng, Rongfang; Xia, Yumin; Liu, Yale

doi: 10.1007/s00018-025-05993-2pmid: 41307670

Bullous pemphigoid (BP) is a chronic autoimmune blistering disorder characterized by dermal-epidermal separation and immune cell infiltration. The mechanisms underlying epithelial barrier dysfunction in BP remain incompletely understood. Here, we identify transforming growth factor alpha (TGF-α) as a key regulator of BP pathogenesis through disruption of cellular junctions and induction of pro-inflammatory responses. Elevated TGF-α levels in BP lesions correlated with disease severity. Mechanistically, TGF-α downregulated BP180 expression, upregulated matrix metalloproteinases (MMPs), and enhanced chemokine secretion in keratinocytes via epidermal growth factor receptor (EGFR) activation and downstream phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) signaling. Pharmacological inhibition of EGFR and PI3K/Akt pathways mitigated these effects, highlighting their critical roles in disease progression. Additionally, tumor necrosis factor-like weak inducer of apoptosis (TWEAK) signaling was found to amplify TGF-α secretion, further exacerbating inflammatory responses. These findings suggest that TGF-α drives BP pathogenesis by compromising cell adhesion and promoting tissue inflammation. Targeting TGF-α or its downstream effectors may offer novel therapeutic strategies for BP treatment.

Kang, Jeehoon; Seo, Hyun Ju; Son, HyunJu; Kang, Minjun; Lee, Jaewon; Lee, Eun Ju; Cho, Hyun-Jai; Kim, Hyo-Soo

doi: 10.1007/s00018-025-05930-3pmid: 41441877

Human peripheral blood stem cells (PBSC) that have been most frequently used for repair or regeneration in ischemic cardiovascular disease (CVD) showed limitations in their efficacy. We previously reported that angiopoietin-1 (Ang1) is the cell-priming agent to enhance the vasculogenic potential of PBSC. The limitation was the difficulty to produce Ang1 protein with high efficiency. In this study, we engineered Ang1 structure and made FVA3-Ang1 by adding VASP and COMP sequence for stable tetramer formation as well as FLAG sequence for purification in large scale production and a signal peptide derived from influenza A virus (IAV) for better protein expression. FVA3-Ang1 showed stronger effect on endothelial cells than naïve Ang1 or COMP-Ang1 in terms of gene expression of Ang1, Ang2, VEGFA, FGF2, and KDR, as well as phosphorylation of Tie2, ERK, and Akt. Then we primed PBSC with FVA3-Ang1 and examined the transcriptome analysis. Priming for 1 h did not change whole gene expression profiles of PBSC, whereas priming for 24 h did change the pattern from myeloid toward endotheloid lineage. In mouse models of hind-limb ischemia and myocardial infarction, FVA3-Ang1-primed PBSCs showed superior engraftment and tissue regeneration compared to non-primed cells. A clinical trial is underway to assess efficacy and safety of FVA3-Ang1-primed PBSCs when infused via the culprit coronary artery following emergent stent implantation.

Kalina, Daniel; Potaczek, Daniel P.; Zeng-Brouwers, Jinyang; Boehm, Mario; Nolte, Marc W.; Bielohuby, Maximilian; Schermuly, Ralph T.; Schaefer, Liliana; Wygrecka, Malgorzata

doi: 10.1007/s00018-025-05988-zpmid: 41318746

Kidney fibrosis is a common manifestation of chronic kidney diseases, with parenchymal tissue scarring serving as a histologic predictor of functional deterioration. Considering the relationships between contact-phase system activation and renal fibrosis as well as potential direct profibrotic activities of factor XII (FXII), we hypothesized that FXII inhibition with an anti-FXII/activated FXII (FXIIa) antibody (3F7) demonstrates therapeutic efficacy in a mouse model of unilateral ureteral obstruction (UUO). Treatment of UUO mice with 3F7 attenuated kidney fibrosis, as evidenced by preserved tissue structure, decreased deposition of collagen, and diminished apoptosis, but increased proliferation of tubular epithelial cells. No effect was observed with the administration of C1 esterase inhibitor, which serves as a primary plasma inhibitor of FXIIa and kallikrein. Transcriptome analysis revealed that 3F7 therapy predominantly affects stress-activated protein kinase signaling cascades and signal transduction in response to DNA damage. Exposure of renal epithelial cells to FXII or FXIIa triggered p21 expression in an Akt- and ERK1/2-dependent manner. Accordingly, treatment of UUO mice with 3F7 reduced numbers of p21+ renal tubular epithelial cells. Our work provides proof-of-concept data supporting efficacy of 3F7 in the UUO model and unravels molecular mechanisms underlying the protective role of FXII inhibition in chronic kidney injury.

Zhang, Li; Yang, Wenhui; Zhao, Zhi; Wang, Yingsong; Wang, Haonan; Zhao, Ninghui; Zhou, Jin; Song, Zhibo; Zhang, Zhaoquan; Li, Tao; Zhe, Shangjia; Qian, Genghao; Liu, Yingliang; Xie, Jingming

doi: 10.1007/s00018-025-06004-0pmid: 41389086

This paper investigated the role of cilia gene defects in bone marrow stem cells (BMSCs) in idiopathic scoliosis (IS). IS, a spinal deformity commonly occurring in adolescence, impacts patients’ quality of life. Cilia, axial filaments covered by the cell membrane and composed of microtubules on the eukaryotic cell surface, include motile cilia and primary cilia. Primary cilia on the surface of most cells mediate intracellular signaling pathways. Cilia dysfunction correlates with skeletal diseases. Defects in cilia genes disrupt osteogenic signaling pathways in BMSCs, promoting IS progression. By sample analysis, lentiviral transfection of BMSCs, and in vivo experiments in zebrafish, we explored the impact of cilia gene defects on BMSCs’ osteogenic signaling pathways and IS, revealing that defects impaired ciliogenesis, cellular signaling, and mechanical sensing, and affected vertebral alignment and skeletal patterning. These findings highlight the role of cilia gene defects in IS pathology, providing potential targets for clinical treatment.

Ueyama, Takehiko; Yamaguchi, Kyoko; Aoyama, Yakumo; Aoshima, Kota; Onizuka, Michiho; Tamagawa, Taichi; Kitayama, Shota; Ueyama, Junichi; Okamoto, Kiyoki; Mohri, Hiroaki; Shimazawa, Masamitsu

doi: 10.1007/s00018-025-05876-6pmid: 41339603

NADPH oxidase 3 (Nox3), expressed in the endolymphatic duct and sac within the vestibule of the inner ear, is essential for otoconia formation. Mice with functionally deficient Nox3 exhibit a “head-tilt” phenotype. Recently, we reported that Nox3 expression in the cochlea is induced by aging, cisplatin treatment, and intense noise exposure, contributing to the primary source of reactive oxygen species (ROS) and causing acquired sensorineural hearing loss. However, its expression and function outside the inner ear remain poorly characterized. To explore novel Nox3 functions, we used Nox3-Cre;tdTomato mice, in which Cre recombinase (Cre) is knocked into the ATG site of Nox3, enabling visualization of Nox3 expression via tdTomato fluorescence. We identified Nox3 expression in retinal ganglion cells (RGCs) and GABAergic amacrine cells (ACs). The tdTomato-positive cells increased by 2 months of age and then plateaued in Nox3-Cre+/−;tdTomato+/+ (heterozygous knock-in [KI], hereafter HT Nox3-knockout [KO]) mice, while in Nox3-Cre+/+;tdTomato+/+ (homozygous KI, hereafter Nox3-KO) mice, the increase occurred by 12 months, suggesting that Nox3-derived ROS are toxic to RGCs and ACs. Additionally, Nox3-KO mice showed reduced a-, b-, and scotopic threshold response (STR)-waves in electroretinogram (ERG) compared to WT mice. Treatment with cisplatin, a well-known toxic agent for the retina and optic nerve, reduced tdTomato-positive cells in the retinae of 2-month-old HT Nox3-KO mice but not in Nox3-KO mice, compared to age-matched controls. Nox3-KO retinae developed normally. Thus, Nox3 is expressed in RGCs and ACs to regulate retinal function; however, excessive Nox3-derived ROS cause toxicity under pathological conditions, including aging.