Spatial transcriptomic modeling of vascular remodeling in aortic aneurysm using integrated single-cell RNA sequencing analysis.

Antibiotic-mediated prophage activation: Mechanisms, consequences, and therapeutic implications.

Lactate dehydrogenase inhibitors: A promising candidate against aging and fibrosis.

Cadmium induces ferroptosis in B cells via ATP6V0A1-upregulated lysosomal ferritinophagy: insights from murine transcriptomics and human cellular models.

Inhibiting VEGFC - mediated hepatocyte - macrophage regulatory axis contributes to protective effects of naringin against high - fat diet - induced hepatic fibrosis.

Higenamine alleviates abdominal aortic aneurysm by regulating oxidative stress and inflammation against VSMC apoptosis.

This study investigates the roles of hypoxia-inducible factor (HIF-1α), SMAD2, and vascular endothelial growth factor (VEGF) in renal repair under hypoxic conditions, focusing on their impact on macrophage phenotype transformation. Bioinformatics analysis identified SMAD2 as a key gene in renal injury, correlating with HIF-1α and VEGF levels. In a cohort of 60 pediatric patients, non-AKI individuals exhibited higher VEGF and SMAD2 levels but lower HIF-1α compared to AKI patients. Using a chronic hypoxia rat model, bevacizumab treatment exacerbated renal damage, as indicated by elevated serum creatinine (79.4 ± 61.7 μmol/L), increased inflammatory markers, and heightened HIF-1α expression. Bevacizumab’s inhibition of VEGF also impaired macrophage phenotype modulation. In vitro experiments revealed that hypoxia alone had minimal direct effects on macrophage polarization but enhanced IL-4-induced M2 polarization, further amplified by SMAD2 and VEGF overexpression. These findings underscore the distinct yet interconnected roles of HIF-1α, SMAD2, and VEGF in shaping the hypoxic renal microenvironment and influencing macrophage polarization. The study highlights VEGF’s critical role in renal repair and its interaction with hypoxia and inflammatory pathways, which may modulate macrophage polarization and ultimately impact renal outcomes. These insights suggest a promising therapeutic strategy for kidney diseases.

The role of the lung's microcirculation and capillary endothelial cells in normal physiology and the pathobiology of pulmonary diseases is obviously vital. The recent discovery of molecularly distinct aerocytes and general capillary (gCaps) endothelial cells by single-cell transcriptomics (scRNAseq) advanced the field in understanding microcirculatory milieu and cellular communications. However, increasing evidence from different groups indicated the possibility of a more heterogeneous nature of lung capillaries. Therefore, we investigated enriched lung endothelial cells by scRNAseq and identified five novel populations of gCaps with distinct molecular signatures and roles. Our analysis suggests that two major populations of gCaps that express Scn7a(Na+) and Clic4(Cl-) ion transporters form the arterial-to-vein phenotypic transition. We also discovered and named mitotically-active "root" cells (Flot1+ ) on the interface between arterial, Scn7a+ , and Clic4+ endothelium, responsible for the regeneration and repair of the adjacent endothelial populations. Furthermore, the transition of gCaps to a vein requires a venous-capillary endothelium expressing Lingo2. Finally, gCaps disconnected from the zonation represent a high level of Fabp4, other metabolically active genes, and tip-cell markers showing angiogenesis-regulating capacity. The hypoxia-induced models demonstrated that "root" cells exhibit a marked expansion in hypoxia, supporting their role in vascular regeneration and neocapillarization. We also showed a developmental time-course analysis demonstrating an evolution of progenitor (FoxM1+ ) cells, which are progressively replaced by "root" cells during lung maturation, revealing a switch in vascular homeostasis. The discovery of these populations will translate into a better understanding of the involvement of capillary phenotypes and their communications in lung disease pathogenesis.

MITF, TFEB, and TFE3 drive distinct adaptive gene expression programs and immune infiltration in melanoma.

AimVascular smooth muscle cells (VSMCs) are characterized by a considerable plasticity. Their phenotypic switch (from contractile to synthetic) plays a crucial role in the atherosclerotic process, explaining that numerous studies focus on this phenotypic transition. Thus, it is essential to use VSMCs that have been finely phenotyped for experimental purposes. The use of MOVAS cell line is suitable because, unlike primary cells, it is believed that these cells retain their phenotype and avoid cell senescence in culture. This study aimed to assess the phenotype of MOVAS cells over culture passages to ensure that they retained a contractile phenotype, before using them for further investigations.MethodsThe phenotype of MOVAS cells at different culture passages (P3, P5 and P8) was analysed morphologically and by studying the expression of genes that indicate a contractile (Acta2, Myocd and Cnn1) and synthetic (Klf4 and Lgals3) VSMC phenotype by RT-qPCR. Cell stiffness was analysed by atomic force microscopy and cell adhesion and migration.ResultsOur results showed that MOVAS cells rapidly changed morphologically and that the gene expression of contractile markers was significantly reduced in favor of markers specific to the synthetic phenotype. These changes were associated with a reduction in cell stiffness and a significant increase in adhesion and migration properties.ConclusionMOVAS cells undergo a transition from contractile to synthetic phenotype with increasing number of passages in vitro, which means that these cells should be used with caution, at a low number of passages, while being regularly characterized.

Vascular smooth muscle cells (VSMCs) are characterized by a considerable plasticity. Their phenotypic switch (from contractile to synthetic) plays a crucial role in the atherosclerotic process, explaining that numerous studies focus on this phenotypic transition. Thus, it is essential to use VSMCs that have been finely phenotyped for experimental purposes. The use of MOVAS cell line is suitable because, unlike primary cells, it is believed that these cells retain their phenotype and avoid cell senescence in culture. This study aimed to assess the phenotype of MOVAS cells over culture passages to ensure that they retained a contractile phenotype, before using them for further investigations. The phenotype of MOVAS cells at different culture passages (P3, P5 and P8) was analysed morphologically and by studying the expression of genes that indicate a contractile (Acta2, Myocd and Cnn1) and synthetic (Klf4 and Lgals3) VSMC phenotype by RT-qPCR. Cell stiffness was analysed by atomic force microscopy and cell adhesion and migration. Our results showed that MOVAS cells rapidly changed morphologically and that the gene expression of contractile markers was significantly reduced in favor of markers specific to the synthetic phenotype. These changes were associated with a reduction in cell stiffness and a significant increase in adhesion and migration properties. MOVAS cells undergo a transition from contractile to synthetic phenotype with increasing number of passages in vitro, which means that these cells should be used with caution, at a low number of passages, while being regularly characterized.

Myocardial infarction (MI) is a cardiovascular disease characterized by the irreversible necrosis of myocardial cells and high morbidity and mortality rates. The vascular contributions and diversity of endothelial cells in this fatal disease are poorly characterized. Here, we integrated multiple mouse cardiac single-cell transcriptomic data from six time points after MI and characterized the transcriptome reprogramming of endothelial cell subsets. Endothelial cell subsets differentiation trajectory analysis was conducted using scVelo, CellRank, and Dynamo. In silico simulation of Sox17 depletion perturbed the phenotypic transition from tip-like cells to capillary endothelial cells. Endothelial cell and immune cell interaction analyses identified a Mif-Cd44 interaction between tip-like cells and monocytes in the acute phase. Invivo pharmacological inhibition of Mif exacerbated cardiac dysfunction in the mouse MI model.

PurposeThe coronal plane alignment of the knee (CPAK) classification helps understand knee alignment variability, guiding personalised total knee arthroplasty (TKA) strategies. However, evidence regarding the impact of postoperative CPAK classification changes on patient‐reported functional outcomes after restricted kinematic alignment (rKA) TKA remains limited. This study aimed to investigate whether CPAK classification changes after TKA with rKA influence functional outcomes, measured by the International Knee Society score (IKS).MethodsA retrospective cohort study included 464 patients who underwent primary TKA with a posterior stabilised implant (KNEO®) between January 2020 and May 2024. The inclusion criteria were primary or secondary osteoarthritis with complete radiographic and clinical follow‐up at 2 years; patients with incomplete data or intraoperative complications requiring implant change were excluded. Pre‐ and postoperative CPAK classifications were compared, and functional outcomes were assessed using IKS knee and function scores at 2 years of follow‐up. Radiographic assessment was performed on standing long‐leg radiographs by a single experienced observer.ResultsA significant redistribution of CPAK classifications was observed postoperatively (p < 0.001), with 22.2% of CPAK I changing to CPAK II and 19.1% changing to CPAK V. In total, 33.3% of all knees retained their initial CPAK classification. No significant differences were observed in the postoperative IKS Knee (85.2 ± 12.3 vs. 83.9 ± 11.8; p = 0.62) or IKS Function scores (78.5 ± 13.1 vs. 76.9 ± 12.7; p = 0.54). Given the small sample sizes within certain CPAK subtype transitions, subgroup analyses were not feasible.ConclusionsChanges in CPAK classification following rKA‐TKA were common but did not significantly influence functional outcomes at 2 years. These findings suggest that CPAK phenotype transition alone may not be a reliable predictor of clinical success, although larger studies are needed to explore subtype‐specific effects.Level of EvidenceLevel IV.

RB1 was the first identified tumour suppressor gene, named for its crucial role in opposing retinoblastoma oncogenesis. The RB1 gene encodes the retinoblastoma protein pRB, which is a well-known negative regulator of the cell cycle. However, pRB also contributes to cell differentiation by restricting reprogramming and stem cell properties. Accordingly, RB1 inactivation in tumours can induce phenotypic modifications, contributing to tumour progression. Indeed, RB1 pathogenic alterations, either point mutations or deletions, leading to pRB loss of function are observed in 5% of all human cancers. Mutations are much more prevalent in some histologic subgroups, including retinoblastoma, spindle cell lipoma, neuroendocrine prostate cancer and small cell lung carcinoma. In such entities, molecular investigation of tumour samples and mechanistic studies strongly suggest that early RB1 inactivation contributes not only to dysregulation of cell cycle control, but also to the tumour cell phenotype. Among skin carcinomas, RB1 inactivation is the hallmark of primary cutaneous neuroendocrine carcinoma commonly known as Merkel cell carcinoma (MCC), but it has also been described in other tumours including a subset of squamous cell carcinomas, sebaceous carcinomas and the recently described Wnt/beta-catenin-activated non-pilomatrical carcinomas. In this context, we provide a brief overview of the contribution of RB1 inactivation to oncogenesis and tumour cell phenotypes in general and summarise current knowledge regarding RB1-deficient cutaneous carcinomas, highlighting the potential uses of RB1 pathway characterisation for diagnosis, prognosis and therapeutic purposes.

Gallbladder cancer (GBC) is an aggressive malignant tumor that seriously threatens the survival of GBC patients. Laminin-γ2 (LAMC2), an important component of laminins, has been reported to facilitate cancer development and chemoresistance in several cancers. However, the biological effect of LAMC2 on GBC is still unclear. LAMC2 expression was assessed in GBC specimens through proteomics and RNA-seq data. To evaluate the biological effects of LAMC2, we performed colony formation, sphere formation, wound healing, and transwell and invasion assays, along with an in vivo metastasis model. Additionally, we also explored the role of LAMC2 in GBC organoid. Finally, Western blot and RT-qPCR were employed to identify the molecular mechanism underlying LAMC2-mediated regulation of aggressive behavior in GBC. Here, we revealed that LAMC2 was overexpressed in GBC tissues and was positively correlated with poor patient outcomes. Furthermore, the genetic silencing of LAMC2 significantly inhibited GBC cell colony formation, sphere formation, and organoid growth. Additionally, LAMC2 deficiency impaired GBC cell metastasis in vitro and in vivo, accompanied by a reversal of the epithelial-mesenchymal transition (EMT) phenotype. Mechanistically, LAMC2 expression was induced by the canonical TGF-β/SMAD2 signaling pathway, and in turn, LAMC2 feedback enhanced the activity of TGF-β signaling by binding to TGF-β receptor II (TGF-βRII). Our findings suggest that LAMC2 promotes GBC progression and metastasis, potentially by regulating the TGF-β signaling pathway. Therefore, LAMC2 could serve as a potential prognostic biomarker and therapeutic target for GBC.

Diabetic wound healing is critically impaired by dysregulated macrophage polarization, compromised endothelial angiogenic function, and diminished fibroblast proliferation/migration under persistent hyperglycemia. Current therapies, predominantly focused on single-cell targeting, lack coordinated modulation across these key cellular components. We developed a novel triple-targeting core-shell nanoparticle (miR-RPC) leveraging the shared integrin αvβ3 receptor on macrophages, endothelial cells, and fibroblasts to address this limitation. miR-RPC features an RGD/phosphatidylserine (PS)-modified lipid shell encapsulating a chitosan/​​miR-146a-5p​​ core. This miRNA was selected as a model RNA because of its widely recognized beneficial role in three key cell types in wound healing. The RGD peptide enables specific αvβ3-mediated triple-targeting. The anionic lipid PS facilitates core-shell assembly via electrostatic interaction with the cationic chitosan/RNA core and mimics apoptotic signals to enhance macrophage phagocytosis and phenotypic transition. miR-RPC effectively reprogrammed macrophages towards the M2 phenotype, restored endothelial angiogenic capacity under high glucose, and stimulated fibroblast proliferation, migration, and collagen secretion. Incorporated into a gelatin methacrylate (GelMA)/oxidized hyaluronic acid (OHA) double cross-linked hydrogel (GelO), miR-RPC@GelO significantly accelerated diabetic wound healing in rat models, demonstrating reduced inflammation, increased vascular density, and enhanced collagen deposition. This innovative triple-targeting system achieves coordinated diabetic wound repair through synergistic “immunomodulation-angiogenesis-collagen deposition” mechanisms, offering a promising therapeutic approach. Furthermore, the successful preparation of miR-RPC expands the application of anionic lipids in RNA delivery systems and highlights its potential as a versatile gene delivery vector.Graphical Supplementary InformationThe online version contains supplementary material available at 10.1186/s12951-025-03786-0.

Alzheimer's disease is a multifaceted neurodegenerative condition marked by the build-up of amyloid plaques and neurofibrillary tangles that lead to progressive cognitive impairment. Neuroinflammation, especially the activation of microglia, plays a pivotal part in driving this pathology. Microglia are the brain's resident immune cells and can adopt a spectrum of activation states that support either neuroprotection or neurodegeneration. Evidence shows that their phenotypes are highly dynamic and shaped by environmental influences and pathological signals. During the early phases of the disease, microglia tend to assume anti-inflammatory roles that facilitate plaque clearance and promote tissue recovery. Prolonged or dysregulated activation, however, shifts them toward a pro-inflammatory state that amplifies neuronal damage. Several molecular pathways including JAK STAT, PI3K AKT, and MAPK are central to regulating these processes and have emerged as promising therapeutic targets. This review summarizes current insights into microglial phenotypic transitions, the signaling mechanisms governing their activation, and the therapeutic potential of modulating neuroinflammation. Enhancing the neuroprotective capacity of microglia, suppressing chronic inflammatory responses, and targeting key receptors such as TREM2 and P2 × 7 represent potential strategies. A deeper understanding of microglial interactions with other glial cells and the molecular drivers of their activation may provide new avenues for slowing or halting the progression of Alzheimer's disease and related neurodegenerative disorders.

Evogliptin, an anti-diabetic drug had positive impact on various cardiovascular events including inflammation and vascular calcification (VC), an active process driven by vascular smooth muscle cell (VSMC) phenotypic transition. Sphingolipids such as ceramide (CER) mediates inflammation and VC in the vascular tissue. We investigated whether evogliptin ameliorate phenotypic transition and pyroptosis in VSMCs as underlying cause of VC. In cultured VSMCs, isolated from the aorta of (C57/BL6) mouse, we observed more severe calcification with prior treatment of CER in Pi-treated VSMCs as detected by Alizarin Red Staining. Prior CER- stimulation led to a marked upregulation of osteogenic markers such as RUNX2, OPN, BMP2 and decreased contractile markers SM22-α and α- SMA in Pi-treated VSMCs as compared to control cells. In addition, increased expression of pyroptotic markers such as NLRP3, GSDM-D, IL-1β, IL-18, and LDH release was observed with prior treatment of CER in Pi-treated VSMCs as compared to control cells. Furthermore, MCC950 (NLRP3 inhibitor), disulfiram (GSDM-D inhibitor) and evogliptin significantly downregulated osteogenic and pyroptotic markers including LDH release in both Pi‐induced only and CER + Pi-treated VSMCs. Moreover, GW4869 (SMase inhibitor) and evogliptin significantly reduced SMase activity in sphingomyelin (SM)-induced VSMCs as compared to both Pi and SM only-treated groups. Also, the cleavage efficiency of GSDM-D was high in Pi and CER + Pi groups which was reduced with prior treatment of evogliptin. Hence, our data demonstrate that evogliptin alleviates VC by blocking phenotypic transition and associated pyroptosis via modulation of NLRP3/GSDM-D mediated pathway in CER-induced VSMCs.

Retraction Note: Modulation of microglial phenotypes by dexmedetomidine through TREM2 reduces neuroinflammation in heatstroke

Hypoxia and immune-suppressive microenvironments in glioblastoma drive transcriptional plasticity and phenotypic transition. Understanding these processes is crucial for overcoming therapy resistance and tumor relapse. This study investigates the expression pattern of sterile alpha motif domain-containing 9 (SAMD9) under these conditions, evaluates its prognostic value and spatial distribution, and explores its therapeutic implications. We analyzed SAMD9 expression and its prognostic value across three independent IDH-wildtype glioblastoma cohorts and validated findings via immunohistochemistry in human glioma tissues. We mapped its spatial distribution using the IvyGAP database and leveraged integrated single-cell and spatial transcriptomic data to delineate local cellular interactions. Drug sensitivities were predicted using the oncoPredict package, and molecular docking was performed with Autodock for drug screening. Key findings regarding SAMD9 expression and its inducers were experimentally validated. SAMD9 was prominently expressed in an immune-suppressive subset of glioma tumor cells and was strongly associated with an interferon (IFN) signature. Elevated levels of SAMD9 correlated with reduced efficacy of anti-PD-1 treatment. Spatial mapping revealed that SAMD9 was predominantly distributed in regions of microvessel proliferation and peri-necrotic niches, where SAMD9-positive tumor cells actively interacted with vascular cells and tumor-associated macrophages (TAMs). Hypoxia and TAM co-culture significantly upregulated SAMD9, suggesting a mechanism for Bevacizumab resistance. SAMD9-high tumors exhibited TAM-dominated immune infiltration, confirmed by immune signatures profiling and histological staining. Drug prediction and molecular docking identified the multi-kinase inhibitor Dasatinib as a promising therapeutic agent for SAMD9-high IDH-wildtype glioblastoma. SAMD9 emerges as an adaptive response gene to environmental changes, exhibiting a significant immunomodulatory function, which highlights its promise as a therapeutic target for IDH-wildtype glioblastoma.