Artificial intelligence (AI) is reshaping biomedical research as both a key analytical tool and an efficient copyeditor. Large language models and related algorithms can help investigators sift through the ever-expanding bodies of scientific literature, connect fragmented biological and clinical datasets, and uncover/recognize meaningful patterns buried within complex multidimensional data. Beyond the bench, these tools can improve and refine manuscript architecture and streamline the flow of information by honing the articulation of simple or complex scientific ideas and sharpening the clarity and cadence of scientific information, which ultimately strengthens the writing in publications and grant proposals alike. For researchers working across languages or at early career stages, these tools allow them to express ideas with greater clarity and fluency, ultimately ensuring that intellectual contributions are assessed on the basis of their conceptual merit and experimental validity. However, the growing persuasiveness of AI-generated writing raises an important concern as to whether persuasive language can create an impression of rigor that is not necessarily matched by rigorous science. AI tools can improve the clarity and presentation of scientific writing, but they cannot establish biological relevance, experimental validation and/or the nuanced judgment required to identify confounding variables, and draw meaningful conclusions. Similarly, an eloquently written grant proposal cannot substitute for an innovative idea, testable hypothesis, feasible experimental approach, and meaningful anticipated contribution. It is therefore essential to view AI tools as instruments of improvement not replacement. They can enhance clarity, efficiency, and readability but they should not replace investigators' intellectual ownership, accountability, and critical interpretation. Disclosure of AI use, together with continued investigators' responsibility for originality, accuracy, citations, data integrity, and interpretation, is essential to maintain trust in scientific discoveries. Similarly, editors, peer reviewers, authors, and funding panels bear a renewed obligation to distinguish writing quality from scientific quality and continue to evaluate approach, innovation, experimental rigor, reproducibility, investigator skills, and scientific and/or clinical significance. Used judiciously and with discernment, AI can free researchers to invest more deeply in the irreplaceable human dimensions of scientific discovery.

Dermal fibroblasts synthesize, organize, and reside within the dermal extracellular matrix (ECM), which provides mechanical support to the skin and a microenvironment critical for dermal cell function. During aging, the ECM becomes fragmented, a change associated with clinical manifestations such as thin, fragile skin and altered fibroblast function, including reduced expression of ECM structural genes and increased expression of pro-inflammatory mediators. We investigated whether functional alterations of fibroblasts in aged skin reflect adaptation to the aged dermal ECM microenvironment or intrinsic aging responses. Primary dermal fibroblast cultures were established from young (20-30 years; 6 females, 6 males) and aged (>80 years; 6 females, 6 males) sun-protected skin. No differences in early growth kinetics were observed between groups; however, fibroblasts from aged individuals grew more slowly after approximately 25 population doublings and had a 20% lower final population doubling level. Prior to passage, after brief primary culture (6-8 days), the average telomere length of fibroblasts from aged donors was reduced by approximately 1000 base pairs (∼1 kb). Global gene expression revealed 477 differentially expressed genes (DEGs; padj ≤0.05, absolute log2FC ≥ 0.58), comprising 300 upregulated and 177 downregulated transcripts. Upregulated DEGs and associated protein-protein interaction networks were enriched in cytokine- and chemokine-mediated signaling pathways (p < 10[-5]). Downregulated DEGs were enriched in lipid biosynthesis and fatty acid metabolism. Notably, fibroblasts from young and aged skin expressed similar levels of genes encoding ECM structural proteins, including collagens (42 genes), proteoglycans (29 genes), and glycoproteins (157 genes). These findings indicate that fibroblasts from aged skin retain a pro-inflammatory transcriptional phenotype in primary culture, resembling their phenotype in vivo. In contrast, primary fibroblasts from aged skin express youthful levels of ECM structural genes in primary culture, suggesting that reduced ECM gene expression in vivo reflects dynamic adaptation to the aged dermal microenvironment rather than permanent reduction of synthetic capacity.

Ferroptosis is a non-apoptotic form of programmed cell death driven by iron-dependent lipid peroxidation. Induction of ferroptosis has been shown to overcome chemoresistance in cancer cells. Given the importance of the nuclear factor erythroid 2-related factor 2 (NRF2) pathway in the regulation of cellular oxidative stress, this study was designed to search for NRF2-regulated genes involved in ferroptosis and chemoresistance in non-small cell lung cancer (NSCLC). Bioinformatics analysis and experimental validation indicated cadherin-6 (CDH6) as a candidate gene. The expression and role of CDH6 in NSCLC were explored. CDH6 protein partners were identified and investigated after CDH6 co-immunoprecipitation assays and mass spectrometry. Our data demonstrated that NRF2 binding to the promoter of the CDH6 gene inhibited RUNX2-mediated transcription of CDH6 in NSCLC. Clinically, CDH6 was downregulated in NSCLC tissues and correlated with more advanced tumor-node-metastasis (TNM) stage and shorter overall survival. Knockdown of CDH6 enhanced the proliferation, colony formation, tumorigenesis, and cisplatin (CDDP) resistance in NSCLC cells. Moreover, overexpression of CDH6 induced ferroptosis and increased CDDP sensitivity in CDDP-resistant NSCLC cells. Mechanistically, CDH6 associated with and sequestered CCAAT enhancer binding protein delta (CEBPD) protein in the cytoplasm and impaired the CEBPD-dependent transcription of target genes. Depletion of CEBPD increased CDDP sensitivity of CDH6-depleted NSCLC cells, whereas overexpression of CEBPD restored CDDP resistance in CDH6-overexpressing NSCLC cells. Additionally, CEBPD upregulation was associated with poor prognosis in NSCLC patients. In conclusion, our data underscore the critical role of the NRF2-CDH6-CEBPD axis in modulating CDDP sensitivity in NSCLC and offer a potential therapeutic target for improving chemotherapy.

[This corrects the article DOI: 10.1002/ccs3.70048.].

Atherosclerotic plaque progression is shaped by coordinated inflammatory and remodeling programs involving immune cells and vascular wall cells. Inflammatory macrophage activation and vascular smooth muscle cell (VSMC) phenotypic remodeling are central features of human atherosclerosis, but their transcriptomic relationships during plaque progression remain incompletely characterized. This study integrated single-cell and bulk transcriptomic datasets to examine highly inflammatory macrophage states, VSMC remodeling-related transcriptional programs, and candidate ligand-receptor expression patterns in human atherosclerotic plaques. Human atherosclerotic plaque single-cell RNA sequencing data from GSE260657 and bulk transcriptomic data from GSE28829 were analyzed. After quality control, 7628 cells were retained for single-cell analysis. Major cell types were annotated using canonical markers, followed by reclustering of macrophages and VSMC-related cells. Functional module scoring, differential expression analysis, Gene Ontology biological process enrichment, and Kyoto Encyclopedia of Genes and Genomes pathway analyses were performed to characterize macrophage transcriptional states. Slingshot was applied to infer VSMC pseudotime ordering. CellChat and NicheNet were used to prioritize candidate ligand-receptor expression patterns and ligand-associated VSMC target gene programs. External bulk transcriptomic analysis was performed to examine whether single-cell-derived inflammatory and remodeling signatures were represented at the tissue-transcriptome level during plaque progression. Macrophage reclustering identified a highly inflammatory macrophage state characterized by prominent inflammatory activation, cytokine-response, and stress-response features. Genes upregulated in this population were enriched in pathways related to tumor necrosis factor (TNF) response, nuclear factor kappa B signaling, leukocyte activation, cytokine signaling, lipid and atherosclerosis, toll-like receptor signaling, and inflammasome-associated inflammation. VSMC reclustering revealed contractile VSMCs, PTHLH+ synthetic VSMCs, KRT7+ VSMC-like cells, interferon-responsive VSMCs, pericyte-like mural cells, and osteogenic/modulated VSMCs. Pseudotime analysis showed a broad contractile-to-osteogenic/modulated transcriptional continuum accompanied by increased expression of remodeling-associated genes and selected inflammatory or remodeling-associated receptor genes. CellChat and NicheNet analyses prioritized candidate ligand-receptor and ligand-associated target gene expression patterns involving SPP1-CD44, TNF-TNFRSF1A, IL1B-IL1R1/IL1RAP, MIF-ACKR3, PDGFB-PDGFRB, and FN1-SDC1/ITGB1. In GSE28829, inflammatory macrophage-, osteogenic/modulated VSMC-, candidate ligand-receptor expression-, SPP1-CD44 candidate axis-, and NicheNet-prioritized target program-related signatures were more prominent in advanced plaques and were positively correlated with each other. This integrative transcriptomic analysis identified a highly inflammatory macrophage state and a VSMC remodeling continuum in human atherosclerotic plaques. Candidate ligand-receptor and ligand-associated target gene expression patterns linked inflammatory macrophage activation with osteogenic/modulated VSMC remodeling at the computational level. External bulk data further showed coordinated enrichment of inflammatory and remodeling signatures in advanced plaques. These findings provide a descriptive and hypothesis-generating transcriptomic framework for understanding inflammatory macrophage activation and VSMC remodeling in human atherosclerosis.

Multiorgan fibrosis is a complex, maladaptive response to chronic injury, where the extracellular matrix (ECM) shifts from serving merely as a structural framework to becoming an active, sustained contributor to disease progression. We propose a unifying mechanobiological framework in which chronic injury promotes fibroblast activation and ECM remodeling, whereas the stiffened and remodeled ECM feeds back through integrins, mechanosensitive ion channels, collagen receptors, and Hippo-YAP/TAZ signaling to sustain fibrogenesis. Regulation of this process is governed by bidirectional feedback between cellular signaling and the matrix's physical properties across organs such as the lung, heart, liver, and kidney. This narrative review article presents recent advances in understanding mechanisms of cell-cell and ECM communication, paratensile signaling, mechanical feedback loops, cellular crosstalk and secretomes, quantitative therapeutic thresholds, and translational strategies for tissue remodeling. These developments collectively offer promising insights for the development of novel pharmacological agents targeting mechanotransduction pathways, ECM normalization, and cell-specific sensitization. We further define quantitative thresholds as diagnostic, action, and response thresholds that may support clinical decisions such as referral, treatment initiation, therapeutic monitoring, and clinical-trial enrichment. Finally, we provide specific details regarding the treatment of liver fibrosis before concluding with research agenda.

High mobility group box 1 (HMGB1) is a nuclear protein that functions as a damage-associated molecular pattern molecule upon extracellular release. It has an established role in sterile inflammation, yet its involvement in UV-induced human skin inflammation remains poorly characterized. This study investigated whether UV irradiation and chronic photoaging trigger HMGB1 release in human skin and whether this release drives proinflammatory cytokine expression and photoaging. Both acutely UV-irradiated and chronically photoaged human skin exhibited HMGB1 release, which temporally correlated with the upregulation of photoaging-related critical proinflammatory cytokines, IL-1β, IL-6, IL-8, and TNF-α. In cultured cells, UV irradiation reduced intracellular HMGB1 protein without altering mRNA levels while simultaneously increasing extracellular HMGB1 in supernatants, confirming active secretion rather than passive leakage. HMGB1 knockdown and blockade of TLR4, its principal receptor, significantly attenuated UV-induced proinflammatory cytokine expression in both cell types. In mouse skin, disruption of the HMGB1-TLR4 axis reduced neutrophil infiltration, suppressed cytokine expression, and mitigated dermal damage (photoaging). Collectively, these findings demonstrate that UV irradiation promotes active HMGB1 release from skin cells, amplifying cutaneous inflammation and exacerbating dermal damage via TLR4 signaling. Therapeutic targeting of the HMGB1-TLR4 axis therefore represents a promising strategy for mitigating UV-induced skin inflammation and its downstream pathologies, including photoaging.

Iron dysregulation has emerged as a contributor to osteoarthritis (OA), yet the cell-communication mechanisms connecting iron-related genetic signals to joint degeneration remain insufficiently defined. Here, we established an integrative computational framework combining transcriptomic screening, machine learning, Mendelian randomization, immune and metabolite mediation analysis, single-cell transcriptomics, virtual gene perturbation, molecular docking, molecular dynamics simulation, and experimental validation to identify signaling regulators involved in iron metabolism-associated OA. By intersecting iron metabolism-related genes, OA differentially expressed genes, and eQTL-supported genes, we identified 27 shared candidates. Machine learning-based prioritization and genetic causal inference further highlighted CSF1R as a central regulatory gene. Single-cell analysis localized CSF1R expression predominantly to macrophages, indicating a macrophage-centered role in the osteoarthritic microenvironment. Mediation analysis integrating 731 immune-cell traits and 1400 circulating metabolites identified CD14[+]CD16[+] monocytes as a significant cellular mediator linking CSF1R activity to OA susceptibility, suggesting that CSF1R may promote disease progression mainly through monocyte-macrophage remodeling rather than isolated metabolic alteration. Genetic colocalization further supported a shared regulatory signal between CSF1R expression and OA risk. Virtual single-cell perturbation revealed distinct downstream consequences of CSF1R modulation. Simulated CSF1R depletion enhanced antigen processing, major histocompatibility complex class II presentation, and phagosome-related programs, whereas simulated CSF1R overexpression preferentially activated extracellular matrix organization, integrin signaling, and cartilage development-associated pathways. Structural analyses identified stable interactions between CSF1R and candidate inhibitory compounds, and inflammatory stimulation of macrophages confirmed increased CSF1R protein expression. Collectively, this study identifies CSF1R as a macrophage-associated immunometabolic signaling hub linking iron dysregulation to OA. These findings provide a mechanistic basis for targeting CSF1R-mediated monocyte-macrophage communication and offer a computational strategy for prioritizing therapeutic targets in degenerative joint disease.

There is a known reciprocal inhibitory relationship between adipogenesis and osteogenesis in mesenchymal stem cells (MSCs). Our previous study showed that low-intensity pulsed ultrasound (LIPUS) did not promote osteogenesis of a murine MSC line C3H10T1/2, although LIPUS treatment suppressed adipogenesis. Therefore, we investigated the effect of a combination of an osteoinductive factor such as bone morphogenic protein-2 (BMP-2) and LIPUS treatment on osteogenesis. This combination significantly upregulated the gene expressions of BMP receptors and RUNX2, which is a key early osteogenic transcription factor. Interestingly, cellular communication network factor 1 (CCN1) was significantly increased at the mRNA and protein levels. Additionally, Runx2 expression was increased by a recombinant CCN1 protein and decreased in CRISPR-Cas9-generated Ccn1 knockout (KO) cells treated with the combination of LIPUS treatment and BMP-2. Moreover, Runx2 expression was recovered through the transfection of a Ccn1 expression plasmid into Ccn1 KO cells. These findings indicate that treatment with the combination of LIPUS treatment and BMP-2 promoted CCN1 production, and the CCN1 regulates the Runx2 expression in C3H10T1/2 cells, thus suggesting that CCN1 induced by mechanical stimulation and an osteoinductive factor plays a pivotal role in osteogenesis of MSCs.

Diabetic retinopathy (DR) is the main cause of vision loss, with retinal pigment epithelial (RPE) cell dysfunction as a key contributor. Ferroptosis is implicated in RPE injury under hyperglycemia; however, its upstream regulatory mechanisms remain unclear. High glucose (HG)-induced human RPE ARPE-19 cells and a diabetic mouse model were used. Ferroptosis was assessed through biochemical assays, Western blot, fluorescence probes, and histological staining. Protein interactions and transcriptional regulation were examined using Co-IP, glutathione S-transferase pull-down, ChIP, and luciferase reporter assays. Immunofluorescence staining was used as key indicator expression in retinal tissues. Nuclear receptor 4A1 (NR4A1) expression was upregulated under HG-induced ARPE-19 cells. NR4A1 knockdown alleviated HG-induced iron accumulation, lipid peroxidation, and ferroptosis marker changes. Mechanistically, SMAD-specific E3 ubiquitin protein ligase 2 (SMURF2) interacted with NR4A1 and promoted its ubiquitination and degradation. Forkhead box O6 (FOXO6) was upregulated by HG and repressed SMURF2 transcription. SMURF2 overexpression mitigated HG-induced ARPE-19 ferroptosis and retinal injury in DR mice, an effect reversed by NR4A1 co-overexpression. SMURF2 knockdown reversed the effect of FOXO6 knockdown on HG-induced ferroptosis in ARPE-19 cells and retinal injury in DR mice. The FOXO6-SMURF2-NR4A1 axis critically regulates ferroptosis in RPE cells during DR.

Normal fibroblasts (NFs) are among the earliest stromal cells encountered by bladder cancer cells during invasion into the lamina propria, yet their contribution to tumor progression and intravesical chemotherapy response remains poorly defined. Here, we investigated whether NFs are passive stromal bystanders or active participants in early bladder cancer microenvironmental remodeling. Analysis of an early-stage non-muscle-invasive bladder cancer transcriptomic cohort revealed that fibroblast-associated scores were higher in T1 than Ta tumors, enriched in European Organisation for Research and Treatment of Cancer high-risk disease, positively correlated with epithelial-mesenchymal transition (EMT) scores, and associated with increased expression of resistance-linked genes. Experimentally, fibroblast-conditioned medium reduced bladder cancer cell proliferation while accelerating wound closure, indicating a less proliferative but more motile phenotype. This was accompanied by EMT-like cadherin remodeling, including reduced E-cadherin and increased N-cadherin staining. Reciprocally, tumor-derived signals rapidly induced cancer-associated fibroblast (CAF)-like activation features in NFs within 48 h, including increased α-smooth muscle actin, fibroblast activation protein, and PDGFRβ expression. Functionally, increasing fibroblast-to-tumor cell ratios progressively attenuated mitomycin C (MMC)-induced cytotoxicity, demonstrating that stromal context can modify chemotherapy response in vitro. Together, these findings show that NF-tumor crosstalk is rapid, bidirectional and functionally relevant to MMC response in vitro. Rather than acting only after stable CAF formation, NFs can rapidly acquire CAF-like activation features following tumor-derived stimulation and contribute to a stromal context associated with EMT-like plasticity and reduced MMC-induced cytotoxicity in vitro.

[This corrects the article DOI: 10.1002/ccs3.70077.].

The scaffold protein TKS4 plays a role in the development of several cancers. Alternative splicing of the TKS4 gene generates two isoforms, TKS4L and TKS4b; however, their distinct expression patterns and functional roles have not yet been characterized. We have shown that TKS4 isoforms were differentially expressed across human cell lines and breast cancer (BC) tumor samples. Both TKS4L and TKS4L/TKS4b mRNA ratios were significantly altered in tumors compared with adjacent tissues. We identified six novel binding SRC homology 3-domain-containing partners for TKS4L, none of which interact with TKS4b, suggesting their functional differences. Tyrosine phosphorylation of both isoforms was induced by Src(Y527F) kinase overexpression, enabling binding to the SH2 domains of signaling proteins. Interestingly, TKS4b significantly accumulated in the nucleus, whereas TKS4L was primarily present in the cytosol in MCF-7 cells. TKS4b overexpression enhanced MCF-7 cell migration. Both TKS4 isoforms exhibit oncogenic properties by promoting epithelial-mesenchymal transition in BC cells, highlighting their potential as targets for therapeutic intervention.

Pregnancy-induced hypertension (PIH) severely impairs placental function, yet its microenvironmental alterations and potential interventions remain unclear. Here, we combined single-cell RNA sequencing (scRNA-seq) with an in vivo model to investigate the protective effects of Prevotella loescheii (PL) in PIH. scRNA-seq revealed that PIH reshaped placental cellular composition, reducing trophoblasts and myeloid cells while increasing smooth muscle cells, and disrupted cell-cell communication. Functional analyses identified oxidative stress (OS) and macrophage-driven pyroptosis via the NLRP3 inflammasome as key pathological features. Trajectory analysis further demonstrated the depletion of Hofbauer cells, which was restored by PL. In vivo, PL significantly alleviated PIH phenotypes, including proteinuria and placental structural damage. Mechanistically, PL reduced proinflammatory cytokines, suppressed mitochondrial OS and pyroptosis, and partially restored short-chain fatty acid metabolism. Collectively, these findings indicate that PL ameliorates PIH by modulating the placental immune microenvironment and metabolic homeostasis, providing a potential therapeutic strategy.

Renal ciliopathies encompass a spectrum of genetic disorders arising from structural or functional impairments of primary cilia, specialized organelles critical for mechanosensation and signal transduction within renal epithelial cells. These disorders are characterized by cystogenesis, driven by dysregulated ciliary signaling, leading to uncontrolled epithelial proliferation, aberrant growth, and loss of cellular polarity. The clinical trajectory evolves from initial cyst formation to advanced tubulointerstitial fibrosis and progressive renal failure. This progression is governed by pathogenic variants in genes encoding ciliary proteins. While advancements in genetic testing have established prenatal diagnosis as a pivotal tool for early identification, definitive diagnosis and therapeutic intervention remain challenging. These difficulties stem from several factors: incomplete understanding of the molecular mechanisms underlying cyst formation and fibrosis; limitations in prenatal diagnostic accuracy owing to phenotypic overlap and incomplete penetrance; and the marked genetic heterogeneity and diverse clinical trajectories of renal ciliopathies. Existing studies have primarily focused on unidirectional modulation of individual pathways, whereas the systematic integration of signaling network cascades remains largely unaddressed. This review systematically elucidates the molecular mechanisms and aberrant signaling pathways in renal ciliopathies, links genetic heterogeneity to clinical phenotypes, and lays a theoretical basis for prenatal diagnosis and novel therapies.