Calcium/calmodulin-dependent serine protein kinase (CASK), a member of the membrane-associated guanylate kinase (MAGUK) family, functions as a multifunctional scaffold protein by engaging diverse binding partners. Although CASK has been implicated in tumorigenesis, its molecular role inthe pathogenesis of non-small cell lung cancer (NSCLC) remainspoorly characterized. CASK expression was assessed by immunohistochemistry in lung adenocarcinoma tissues and matched non-tumor samples. Bioinformatics analyses were performed using Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) datasets. Functional studies were conducted in epidermal growth factor receptor (EGFR) wild-type H1299 cells and EGFR exon 19-deleted PC9 cells following CASK silencing. RNA-Seq was used to identify differentially expressed genes, and assays of cell growth, cell cycle progression, EGFR trafficking, and downstream signaling were performed. CASK expression was significantly elevated in early-stage NSCLC, and higher levels correlated with reduced overall and first progression survival. Silencing CASK inhibited cell growth by blocking the G1-S transition and inducing p21waf1/Cip1, a key mediator of cell cycle arrest, at both transcript and protein levels. Growth suppression was rescued by treatment with a p21 inhibitor UC2288, confirming its central role. Elevated p21 in CASK-deficient cells was reversed by EGFR, AKT, and ERK inhibitors, suggesting that CASK is involved in downregulating p21 via suppressingEGFR/ERK-dependent transcription and EGFR/AKT-dependent protein stabilization. Although p53 is constitutively involved in upregulation of p21 gene expression, the findings from the p53 inhibitor and p21 promoter activity assay excluded the role of p53 in CASK silencing-induced p21 upregulation. Furthermore, CASK modulated the autocrine EGFR loop. CASK silencing enhanced EGFR expression through genetranscription and post-transcriptional proteinstabilization and promoted EGFR activation by upregulating TGF-α in p53-, ERK-, and AKT-dependent manners. Notably, CASK loss did not affect EGFR trafficking to early endosomes but delayed its transition to late endosomes in an ERK-dependent fashion, thereby reducing receptor degradation. We identified CASK as a previously unrecognized driver of NSCLC growth and a potential prognostic biomarker. By regulating EGFR trafficking to late endosomes and attenuating AKT and ERK signaling, CASK suppresses p21 expression and promotes NSCLC cell proliferation, revealing a novel proliferation regulator in NSCLC.

Articular cartilage, as a mechanosensitive tissue, supports and distributes various mechanical forces-including compression, shear, hydrostatic pressure, and tensile strain-during joint loading and motion. These external forces deform not only the chondrocytes but also their pericellular matrix and the surrounding extracellular matrix (ECM). Those mechanical cues are detected by mechanosensors on the plasma membrane (e.g., integrins) and transmitted through the cytoskeleton, ultimately being converted into biochemical signals. These signals activate key mechanoresponsive intracellular pathways-including TGF-β-induced SMAD, Rho-GTPase, MAPKs (ERK, JNK, p38), PI3K/AKT/mTOR, MRTF-SRF, and YAP/TAZ-that regulate chondrogenic differentiation and cartilage-specific matrix synthesis. This field of study is known as mechanobiology. Over the past decades, it has gained increasing recognition, particularly with the emergence of tissue-engineering constructs as a novel strategy for cartilage repair. However, progress in chondrogenic mechanobiology has primarily centred on intrinsic substrate- or matrix-derived cues, while overlooking the role of extrinsic mechanical forces. This review therefore provides an updated perspective on chondrogenic mechanobiology, with a particular focus on the cellular responses to external mechanical stimuli. It also emphasizes the therapeutic potential of incorporating mechanical stimulation into tissue-engineering strategies for cartilage repair, an emerging filed referred to as Regenerative Rehabilitation (RR). Since this concept has so far been investigated mainly in vitro, we highlight only those studies and refer to it as In vitro Regenerative Rehabilitation. Moreover, this review also addresses post-traumatic osteoarthritis (PTOA), a common joint disorder that frequently results from traumatic cartilage damage. It explores the mechanobiological mechanisms underlying OA and discusses in vitro regenerative rehabilitation studies, highlighting how external forces could serve as an alternative to conventional biochemical treatments for preventing OA progression.

Neurogenic heterotopic ossifications (NHOs) are heterotopic bones that develop in periarticular muscles after traumatic brain (TBI) and spinal cord injuries (SCI). The mechanisms leading to NHO are incompletely understood and the only effective treatment to-date remains surgical resection. We previously established that several inflammatory pathways drive NHO pathogenesis in injured muscles in a mouse model of NHO and in humans. We also demonstrated a functional association between gram-negative bacterial infections and NHO development via lipopolysaccharide (LPS), a pathogen associated molecular pattern (PAMP), which exacerbated NHO in a Toll-like receptor-4 (TLR4)-dependent manner in mice. Using our mouse model of NHO induced by SCI and muscle injury in mice, we tested the effect of a large array of purified PAMPs post-surgery to mimic fungal, viral and bacterial infections and measured NHO bone volumes by micro-computerized tomography (microCT). The effect of PAMPs was also tested in vitro on human muscle progenitors and monocyte/macrophage populations. Muscle progenitors and monocyte/macrophage populations from humans and mice express numerous pattern recognition receptors. In mice, numerous PAMPs produced by bacteria, viruses and fungi exacerbated NHO formation, and the majority of these PAMPs indirectly stimulated fibro-adipogenic progenitor (FAP) calcium mineralization in vitro via macrophages. Likewise, in humans, some PAMPs, particularly those binding to TLR2, directly and indirectly increased the calcium mineralization and osteogenic differentiation of human FAPs isolated from muscles surrounding human NHO. Finally, we established that the indirect stimulation of human FAP mineralization was mediated by inflammatory cytokines IL-1 and oncostatin M secreted by monocytes in response to PAMPs. Overall, our findings suggest that numerous types of infection have the potential to exacerbate NHO development and further highlight the role of oncostatin M and IL-1 signaling pathways in NHO pathophysiology.

BackgroundTemozolomide (TMZ) resistance in glioblastoma (GBM) remains a critical barrier to treatment success, driven by O6-methylguanine-DNA methyltransferase (MGMT) overexpression, glioma stem cell (GSC) persistence, and redox adaptation.MethodsWe developed cp8, a first-in-class abiraterone-based histone deacetylase (HDAC) inhibitor, to simultaneously target these resistance mechanisms. The orthotopic mouse models were used to evaluate the efficacy of cp8 compared to SAHA (vorinostat). The mouse survival period was recorded, and the tumor growth was monitored using the IVIS imaging system.ResultsCp8 demonstrated approximately tenfold greater potency than SAHA, with IC₅₀ values ≤ 3 µM against TMZ-resistant GBM cells (compared with ≥ 30 µM for SAHA). Transcriptomic analysis revealed a unique ability of cp8 to upregulate filaggrin (FLG), a structural protein whose expression correlated with improved patient survival in TCGA and CGGA datasets (p = 0.001). Functional studies showed that FLG knockdown increased GSC-associated markers (Oct4, 2.1-fold; SOX2, 1.8-fold) and enhanced TMZ resistance, whereas cp8 treatment reduced MGMT protein expression by 68% and significantly decreased glioma sphere size by 54% (p < 0.01). In orthotopic models, cp8 extended median survival to 59 days compared with 34 days for controls (p < 0.001) and 49 days for SAHA (p < 0.01), while reducing tumor volume by 72% (p < 0.001) without systemic toxicity. Mechanistically, dual inhibition of HDAC6 and CYP17A1 by cp8 disrupted redox homeostasis and stemness-associated pathways, leading to altered ROS metabolism, reduced MGMT expression, and attenuation of GSC-driven tumor growth while restoring FLG-mediated tumor suppression.ConclusionThis study establishes FLG as a novel therapeutic target in GBM and validates the suppressive efficacy of cp8 on the characteristics of TMZ resistance, highlighting the translational potential as a multitargeted therapy against TMZ-resistant GBM.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12929-026-01241-2.

Candida auris has emerged over the past fifteen years as a multidrug-resistant human fungal pathogen responsible for hospital-acquired infections associated with high mortality rates. Clinical isolates frequently exhibit resistance to one or more first-line antifungal drugs, and management is further complicated by persistent colonization, biofilm formation, and historical misidentification in diagnostic laboratories. Rapid species-level identification and accurate antifungal susceptibility testing are critical for effective patient care and infection control. In this article, we summarize the clinical spectrum of C. auris infections, highlight key pathogenic traits and clade-specific biological differences, and discuss emerging diagnostic, therapeutic, and preventative strategies—including novel antifungal agents, host-directed therapies, and vaccine development—that may improve detection and expand treatment options for C. auris infections.

BackgroundAdvanced metastasis produces cachexia, a complex skeletal muscle wasting syndrome that accounts for one-third of patient deaths. There is currently no approved drug therapy for cancer cachexia. Cancer-associated fibroblasts (CAF) within tumors have been hypothesized to contribute to cachexia, but the detailed mechanism(s) are unknown.MethodsMyotubes were treated with conditioned media (CM) from CAF or CAF activated by cancer cells. Upregulated chemokines in the cancer-activated CAF CM were identified by cytokine array. The effects of chemokine neutralization were investigated using in vitro myotube cultures and in vivo mouse models. The mechanism of action was characterized by in vivo RNA Seq and validated in human muscle cells. Immunostaining delineated the chemokine expression pattern in a patient tumor type highly associated with cachexia.ResultsCancer-activated CAF induced myotube atrophy. CXCL5 was as the major chemokine highly upregulated in the cancer-activated CAF. CXCL5 treatment alone induced myotube atrophy and inhibited myogenic ERK1/2 signaling, similar to cancer-activated CAF treatment. CXCL5 neutralization inhibited cachexia in mice co-injected with HCT 116 colon cancer cells and CAF. RNA Seq showed that CXCL5 neutralization upregulated hypertrophy-related PI3K-AKT-MyoG signaling and remodeled the muscle ECM. CXCL5 neutralization ameliorated muscle wasting induced by CXCL5 and IL-6 co-treatment, and also prevented atrophy in cancer-activated CAF CM-treated human myotubes. CAF were the major detectable source of CXCL5 in a patient tumor highly associated with cachexia.ConclusionCAF contribute to cachexia via cancer cell crosstalk that upregulates CXCL5 secretion. CXCL5 neutralization offers a novel therapeutic approach to maintain muscle mass in cancer patients.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12929-025-01192-0.

Early-onset pre-eclampsia (ePE) is a severe pregnancy complication characterized by dysregulated trophoblast functions and impaired placentation during early pregnancy, leading to substantial maternal and fetal morbidity. While circumstantial evidence indicates defective secretion from endometrial glands impairs placental development, direct evidence linking maternal glandular dysfunction to ePE pathogenesis remains elusive. We established endometrial glandular organoids from women with ePE and healthy pregnancies, analyzing their secretomes by iTRAQ-based proteomics, RNAseq, and spatial transcriptomics. Functional effects of organoid secretomes on trophoblasts were examined in vitro. An endometrial-specific apolipoprotein D (APOD) knock-in mouse model was studied in vivo. APOD levels in first-trimester serum samples from women who later developed ePE were compared to healthy pregnancies. Secretomes from ePE derived endometrial organoids impeded spiral artery remodeling. Multiomic analyses revealed increased APOD production in both ePE organoids and decidual tissues. APOD overexpression disrupted trophoblast functions and endothelial vascular remodeling in vitro, and recapitulated ePE phenotypes in an APOD knock-in mouse model through PI3K/Akt-mediated placental ferroptosis and potential ER stress induction. Ferroptosis inhibition with Fer-1 rescued placental defects and PE symptoms in APOD knock-in mice. Elevated APOD levels in first-trimester serum samples from women who later developed ePE suggest its potential as an early biomarker. This study provides the first direct evidence linking dysregulated endometrial gland function to defective placentation and ePE. APOD was identified as a crucial endometrial gland-secreted factor contributing to ePE, suggesting its potential as an early biomarker and therapeutic target.

BackgroundIn gene therapy via genome editing, it is essential to precisely repair disease-associated gene sequences without introducing random mutations. However, achieving highly accurate genome editing remains challenging owing to the low efficiency of homology-directed repair (HDR)-mediated gene repair, which relies on template DNA. Therefore, if Cas9 mutants capable of enhancing HDR can be identified, they could enable more precise gene therapies.MethodIn this research project, we developed a screening system that uses the acquisition of diphtheria toxin resistance as an indicator of HDR efficiency in human cells and EGFP disruption as an indicator of off-target effect.ResultsBy screening a library of SpCas9 variants with random mutations introduced into its nuclease domain, we identified a novel SpCas9 mutant with higher HDR efficiency than wild-type Cas9.ConclusionWe explored the possibility of obtaining Cas9 mutants with high HDR efficiency via this screening system.Graphical Supplementary InformationThe online version contains supplementary material available at 10.1186/s12929-025-01197-9.