Diabetic retinopathy (DR), a common microvascular complication of diabetes mellitus, has been associated with ferroptosis-related pathological mechanisms. Evidence indicates that ferroptosis may contribute to retinal injury and disease progression. The present analysis aimed to characterize ferroptosis-associated cell subsets and their roles in the progression of DR. Single-cell RNA sequencing data derived from diabetic rat models of DR were obtained from the Gene Expression Omnibus database. A total of 464 ferroptosis-related genes were used to calculate ferroptosis scores. Data processing and analysis were performed using the Seurat R package, with differentially expressed genes identified via the FindAllMarkers function. Functional enrichment analyses were conducted using the clusterProfiler package. Protein-protein interaction networks were constructed using the STRING database, and intercellular communication was predicted using CellChat. Key findings were validated through reverse transcription quantitative polymerase chain reaction (RT-qPCR). Cell clustering identified nine principal retinal cell types. Compared with normal control groups, the DR group demonstrated altered proportions of Müller cells, vascular endothelial cells, microglia, and cone photoreceptor cells. Ferroptosis-score-related differences were most evident in Müller cells and rod cells. In the revised WT-baseline sensitivity analysis, the high-ratio ferroptosis-score cell distribution remained significantly altered in Rod and Müller cells after multiple-testing correction. This study identified key ferroptosis-associated retinal cell populations, with an emphasis on the functional roles of ferroptosis-associated Müller cell subsets in the context of DR. These findings provide further insight into the cellular mechanisms underlying DR and highlight potential molecular targets for therapeutic intervention.
To investigate the underlying mechanism of Solanum lyratum Thunb. (SLT) against psoriasis. A mouse model of psoriasis was established by topical application of imiquimod (IMQ) cream on the nape and dorsal skin. Mice were randomly allocated into the blank control group, model group, SLT treatment groups (low-, medium- and high-dose), and positive control group, with 6 mice in each group. The administration lasted for 7 consecutive days. The skin lesions and pruritic behaviors of mice were observed. Hematoxylin-eosin (H&E) staining was performed to assess the pathological changes of lesional skin and spleen tissues. Toluidine blue staining was used to detect the alterations of mast cells. Immunofluorescence staining was applied to evaluate the changes of T helper 17 (Th17) cells and neutrophils in lesional skin and spleen tissues. Untargeted metabolomics profiling via ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was conducted to identify differential metabolites in mouse serum, screen potential biomarkers, and analyze the involved metabolic pathways combined with the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Enzyme-linked immunosorbent assay (ELISA) was used to determine the content changes of interleukin (IL)-17A, IL-23, tumor necrosis factor-α (TNF-α), IL-1β, IL-4, interferon-γ (IFN-γ), vascular endothelial growth factor (VEGF), histamine (HIS), and 5-hydroxytryptamine (5-HT) in lesional skin tissues. SLT alleviated IMQ-induced psoriasis-like skin lesions and spleen edema in mice. SLT ameliorated epidermal hyperplasia and mast cell infiltration in psoriatic lesional skin, and improved inflammatory cell infiltration in both lesional skin and spleen tissues of psoriatic mice. Meanwhile, SLT regulated the levels of Th17 cells and neutrophils in the above two tissues. A total of 30 differential metabolites were screened out via serum metabolomics analysis, which were mainly enriched in signaling pathways including glycerophospholipid metabolism, fatty acid biosynthesis, biosynthesis of unsaturated fatty acids, dysregulated fatty acid metabolism, and Fcγ receptor-mediated phagocytosis. In addition, SLT significantly downregulated the contents of IL-17A, IL-23, IL-1β, HIS and 5-HT in psoriatic lesional skin. SLT exerts a dual anti-psoriatic effect of anti-inflammation and anti-pruritus by inhibiting immune cells such as Th17 cells and mast cells, thereby downregulating inflammatory factors associated with the Th17/IL-23 axis and pruritogens released by activated mast cells. Furthermore, untargeted metabolomics analysis revealed that the anti-psoriatic mechanism of SLT may be closely related to lipid metabolism disorder, a hallmark pathological feature of psoriasis.
The biophysical properties of cells determine cellular physiology. Leveraging these properties for biomedical applications demands the ability to measure multiple parameters simultaneously across millions of cells and diverse cell types. However, current technologies are limited by throughput and low dimensionality. Here we introduce spectral biophysical cytometry (SBC), a high-throughput platform that integrates environment-sensitive nanosensors with spectral flow cytometry to resolve multiparametric biophysical properties of immune cells at single-cell resolution. By using fluorescent nanosensors that report membrane order, mitochondrial potential and membrane potential, SBC enables simultaneous quantification of key cellular physical states across diverse immune cell populations. When applied to peripheral blood mononuclear cells, SBC reveals cell-type-specific biophysical heterogeneity and identifies distinct remodelling signatures associated with atherosclerosis. In particular, T-cell subsets exhibit substantial alterations in membrane order and mitochondrial depolarization, reflecting coordinated changes in lipid composition and metabolic pathways. Integration with lipidomics and transcriptomics demonstrates that the nanosensors can detect biophysical shifts that correlate with dysregulated lipid metabolism and mitochondrial function, providing mechanistic insight into immune dysfunction in disease. Importantly, SBC achieves rapid, label-efficient profiling using commercially available instrumentation, enabling scalable biomarker discovery directly from blood samples and establishing a powerful strategy for linking biophysical phenotypes to immune cell function.
Natural killer (NK) cells undergo stepwise differentiation from multipotent progenitors within secondary lymphoid tissues. Despite the central importance of the tissue microenvironment in their development, little is known about cell-cell interactions that regulate human NK cell trafficking and maturation. Here, we identify the chemokine receptor CXCR4 and its ligand CXCL12 as regulators of stromal-NK cell interactions required for NK cell maturation. We demonstrate that CXCR4 is expressed throughout human NK cell development in peripheral blood and tonsil, and CXCL12 is enriched in stromal niches containing developing NK cells. Pharmacologic blockade or genetic disruption of CXCR4 resulted in diminished adhesion to integrin ligands, and high-resolution imaging demonstrated crosstalk between CXCR4 and integrins, providing a mechanistic basis for chemokine-dependent modulation of adhesion. Further, CXCR4 blockade resulted in altered contact-dependent motility on stromal cells and integrin ligands, with decreased stable stromal engagement and increased cell speed. Consistent with a requirement for these interactions, treatment with the CXCR4 antagonist plerixafor (AMD3100) impaired NK cell generation from CD34+ precursors. Analysis of NK cells from WHIM syndrome patients with CXCR4 gain-of-function mutations treated with plerixafor revealed similar defects in migration and adhesion, supporting the in-vivo relevance of CXCR4-dependent regulation of NK cell adhesion and motility.
Chronic oxidative stress has long been implicated in renal pathologies, but whether sustained oxidative damage primarily promotes chronic kidney disease (CKD) or tumorigenesis remains unclear. To address this question, we investigated the long-term effects of oxidative stress on human embryonic kidney (HEK293T) cells chronically exposed to a low dose of hydrogen peroxide (H2O2, 50 μM H2O2) for 9 months, generating two adapted lines, 50R30 and 50R45. These cells exhibited enhanced survival and tolerance to acute high-dose H2O2 challenge, indicating an oxidative stress-resistant phenotype. Despite this adaptation, both cell lines showed markedly reduced proliferation and migration, reflecting loss of cellular vitality and function typical of renal degeneration. Transcriptomic and protein analyses revealed upregulation of genes and proteins involved in cell-cycle arrest (p53 and p21), senescence, and the NF-κB/IL-6-driven senescence-associated secretory phenotype (SASP), oxidative stress responses, together with elevated heat shock factor 1 (HSF1) expression indicative of biomolecular damage and impaired adaptive capacity. Collectively, these findings suggest that chronic oxidative stress drives cellular aging and dysfunction rather than malignant transformation, leading to degenerative changes resembling CKD pathology. Moreover, prolonged oxidative stress alone appears insufficient to induce carcinogenic transformation; additional genetic or epigenetic alterations, together with specific cellular machinery, are likely required to drive kidney malignancy. This study therefore provides mechanistic insight into how sustained oxidative stress promotes renal cell senescence and contributes to CKD progression.
Immune dysfunction, spanning pathogenic autoimmunity and impaired host defense, represents a convergent mechanism across neurological autoimmune and inflammatory diseases and opportunistic infections. Despite advances in immunomodulatory and anti-infective therapies, many patients remain treatment-refractory, reflecting limitations of conventional agents. Adoptive T-cell therapies introduce dynamic "living drugs" capable of in vivo expansion, adaptation, and persistence. These promising characteristics have led to a rapid proliferation of preliminary reports and clinical trials in inflammatory and infectious diseases of the nervous system, placing neurologists at the forefront of this evolving therapeutic landscape. In this Update, we advance a disease-centred conceptual framework designed to reposition T-cell-based therapies within neurological practice. Rather than adopting a technology-driven perspective, we organize disorders according to major patterns of immune dysfunction. Immune deficiency predisposing to opportunistic infection and immune dysregulation driving autoimmunity constitute the principal axes of neurological immune pathology. Within the autoimmune spectrum, distinct immunopathological archetypes (autoantibody-mediated, mixed B- and T-cell-driven, and disorders at the interface of inflammation and neurodegeneration) provide a pragmatic structure for therapeutic reasoning. Building on this classification, we delineate how adoptive T-cells (chimeric antigen receptor T-cells, virus-specific T-cells, and regulatory T-cells) may be differentially aligned with underlying disease biology, linking mechanistic insight to clinical strategy.
Ex vivo expansion of human hematopoietic stem cells (HSCs) holds promise for overcoming their limited availability, a major barrier to broader clinical application. Although recent advances in culture systems can increase HSC numbers, these conditions frequently impair self-renewal and induce myeloid bias, and the underlying molecular mechanisms remain poorly understood. Here, we performed single-cell multiome sequencing (scMultiome-seq) on human umbilical cord blood-derived CD34⁺ hematopoietic stem and progenitor cells to co-profile transcriptional and epigenetic adaptations within the same cells during ex vivo culture. Our analyses revealed reduced transcriptional and epigenetic HSC signatures, accompanied by markedly increased activity of myeloid-associated transcription factor motifs, providing molecular insight into the functional decline and myeloid bias of cultured HSCs. We further observed substantial functional heterogeneity among phenotypically defined HSCs following culture. To address these limitations, we established a niche-mimetic culture system that integrates intrinsic and extrinsic bone marrow regulatory cues, including pharmacologic inhibition of the m6A reader YTHDF2 using the small molecule Y13-27, a three-dimensional microenvironment, and N-cadherin-mediated adhesion. This condition (3D-NcadP-Y) robustly preserved long-term repopulating capacity. When combined with the self-renewal agonist UM729, the resulting platform (3D-NcadP-Y-UM) uniquely enabled the expansion of serially transplantable long-term HSCs with balanced multilineage potential. scMultiome-seq and cellular analyses demonstrated that this condition preserves transcriptional and epigenetic long-term HSC signatures, maintains multilineage-associated transcription factor motifs, and limits excessive cell-cycle activation. Together, these findings elucidate molecular mechanisms underlying culture-induced HSC dysfunction and establish a niche-mimetic strategy for expanding functional human long-term HSCs while preserving key features of stemness.
Zinc (Zn) is essential for immune cell function, while mesenchymal stem cells (MSCs) exert immunomodulatory effects primarily through the secretion of soluble factors. Considering the ability of MSCs and Zn to modulate the immune and inflammatory systems, this study investigated, in vitro, the effects of Zn supplementation on MSC responses to inflammatory stimuli and the subsequent modulation of macrophages and lymphocytes. Using the C3H10T1/2 line as a MSC model, we determined that 1 µM ZnSO4 enhanced MSC metabolic activity without affecting viability or cell-cycle distribution, whereas higher concentrations reduced cell viability. Under lipopolysaccharide (LPS) stimulation, Zn inhibited NFκB phosphorylation and increased AMPK phosphorylation, indicating anti-inflammatory and adaptive metabolic responses. Similarly, under TNF-α stimulation, Zn also reduced NFκB phosphorylation. Zn supplementation altered MSC secretory profiles, reducing IL-6, IL-10, and nitric oxide (NO) production while increasing TGF-β and prostaglandin E2 (PGE2) levels, indicating that Zn modifies MSC-derived soluble factor production under inflammatory conditions. Conditioned media from Zn-treated MSCs attenuated IL-6 and IL-12 production in macrophages, indicating a reduced pro-inflammatory cytokine response, whereas lymphocyte responses were unaffected. Importantly, Zn modulation of cytokine production was observed under LPS stimulation but not under TNF-α exposure, suggesting that Zn preferentially interferes with signaling pathways triggered by microbial stimuli. Overall, this study provides mechanistic insight into how Zn affects the secretory profile and inflammatory signaling pathways of C3H10T1/2 cells. These findings support further studies in primary MSCs to determine whether Zn supplementation may represent a useful strategy for modulating MSC-mediated immune regulation in therapeutic settings.
Immune checkpoint inhibitor (ICI) therapy has become standard of care for late stage non-small cell lung cancer (NSCLC), producing durable responses in a subset of patients. However, inflammatory side eKects termed immune-related adverse events (irAEs) occur in up to 40% of ICI-treated NSCLC patients. Current approaches to alleviate irAEs include treatment with immune-suppressing corticosteroids. However, these treatments may undermine the eKicacy of ICIs by suppressing both the irAE and the anti-tumour immune response. To identify more specific therapeutic targets, a better understanding of the complex immunopathology underlying the development of irAEs in NSCLC is required. In this study, pre-treatment blood samples were prospectively collected from 72 NSCLC patients, including 23 who subsequently developed irAEs. Of these 72 samples, PBMCs from 59 were characterised using high-parameter mass cytometry. Plasma from 30 samples was analysed using the SomaScan platform that provides in depth characterisation of over 10,000 proteins, and the plasma metabolome of 30 samples was explored using liquid chromatograph-mass spectrometry (LC-MS). A unique peripheral immunophenotype was observed in patients who subsequently developed irAEs, characterised by decreased memory B cell abundance, heightened Th2 immunity, and an increase in plasma cytokines. Investigation into baseline metabolites revealed dysregulation of fatty acid metabolism associated with development of irAEs. Analysis of additional paired PBMC (n = 17) and plasma (n = 12) samples collected early on treatment allowed exploration of the immunological, proteomic, and metabolic changes associated with irAE development. ICItreatment of patients who developed irAEs induced a significant increase in the abundance of CD8 memory cells and plasma histones. This points to the induction of a strong and potentially pathogenic immune response early following ICI treatment in patients who subsequently develop overt toxicity. Overall, through application of a high-parameter multiomic approach, we have identified key cellular, proteomic and metabolomic features that predispose patients to developing immunotherapy toxicity. These findings provide insight into the complex biology underlying the development of ICI-related adverse events and inform potential treatment strategies.
Zika virus (ZIKV) is primarily transmitted through mosquito bites, and the skin acts as the initial site of viral entry into the host. Consequently, resident skin cells are among the first targets of infection. The epidermis, mainly composed of keratinocytes, can mount an antiviral response against arboviruses through the production of interferons, interferon-stimulated genes, cytokines, and antimicrobial peptides (AMPs), including the Trappin-2/Elafin (Tr2/E) peptide. However, the antiviral activity of Tr2/E during ZIKV infection remains poorly understood, therefore, this study aimed to investigate the antiviral activity of Tr2/E in human keratinocytes during ZIKV infection. In this study, we evaluated the permissiveness of the human keratinocyte cell line HaCaT to infection with a Mexican isolate of ZIKV and observed that these cells support productive viral infection. We then assessed whether ZIKV infection induces endogenous expression of Tr2/E. Tr2/E transcripts were detected in infected cells and showed increased expression over time post-infection, which correlated with the presence of its corresponding protein. Furthermore, we evaluated the antiviral potential of this peptide through exogenous treatment of infected keratinocytes. A significant reduction in ZIKV infection following Tr2/E treatment was observed. Collectively, these findings provide additional insight into the involvement of AMPs in the antiviral response to ZIKV infection and highlight Tr2/E as a potential antiviral factor.
Datopotamab deruxtecan (Dato-DXd) is a novel antibody-drug conjugate currently under investigation in clinical trials of various solid tumors. In this manuscript, an international steering committee formed by medical oncologists, ophthalmologists, and pulmonologists reached consensus to provide multidisciplinary healthcare professionals (HCPs) with practical recommendations for Dato-DXd adverse event (AE) prevention, monitoring, and management in patients with advanced/metastatic non-small cell lung cancer. Experts' recommendations focused on the following AEs: oral mucositis/stomatitis, interstitial lung disease/pneumonitis, ocular surface events, nausea, and vomiting. Based on available guidelines, clinical trials data, and real-world experience, 55 statements were developed and, as per Delphi methodology, all reached consensus with more than 75% of experts' agreement. The key recommendations include preventive measures, such as proactive communication between patients and caretakers, regular clinical examinations, collaboration with multidisciplinary HCPs; and monitoring measures, such as treatment dosage and care planning decisions based on the level of severity of each AE. Experts pointed out real-world challenges and local differences on AEs management and gave insight into implementation of regional strategies with a multidisciplinary collaboration approach. Dato-DXd AEs awareness, understanding, and proper management via a well-established multidisciplinary collaboration is crucial to guarantee patients' quality of life. This international consensus aims to provide comprehensive recommendations for Dato-DXd AEs management, considering the regional and institutional differences present in the healthcare system, such as access to examinations and treatment possibilities. International expert consensus on the prevention and management of side effects from Dato-DXd treatment in non-small cell lung cancer Datopotamab deruxtecan (Dato-DXd) is a new drug being tested for the treatment of different cancer types. In this publication, an international group of oncologists, eye specialists and lung specialists provides practical recommendations to help healthcare teams prevent, monitor and manage side effects caused by treatment with Dato-DXd in patients with metastatic non-small cell lung cancer. Based on clinical data, existing guidelines, and real-world experience, these experts agreed on 55 recommendations for the management of mouth sores, lung inflammation, eye-related problems, nausea and vomiting. The experts highlight the importance of preventing side effects through clear communication with patients, regular check-ups, and close collaboration among different medical specialties. They give specific recommendations on how to monitor patients and adjust treatment based on the severity of each side effect. The experts discuss real-world challenges in managing side effects, which may vary across regions due to differences in healthcare systems, access to tests, and available treatments, and they suggest ways to adapt these strategies locally based on active collaboration between different specialists. Overall, awareness, a good understanding of how to manage these side effects, early recognition and coordinated multidisciplinary care are essential to ensure patients’ quality of life.
CCR2+ monocytes are recruited to sites of acute myocardial injury, where they play a critical role in clearing necrotic debris and replenishing the depleted resident macrophage population. Although this response is necessary for early tissue repair, prolonged activation of inflammatory pathways and persistent recruitment of CCR2+ monocytes have been associated with accelerated ventricular remodeling and adverse outcomes. Inhibition of CCR2 has shown promise in preclinical models of myocardial injury and represents a potential therapeutic target. Cardiosphere-derived cell extracellular vesicles (CDC-EVs) have demonstrated cardioprotective effects partly through modulation of the immune response. We investigated whether CDC-EVs regulate inflammatory monocyte trafficking through effects on CCR2 signaling. We found that CDC-EVs reduce the surface availability of CCR2 on human monocytes through an miR-146a-dependent mechanism, resulting in decreased monocyte migration toward CCL2. These findings identify a previously unrecognized mechanism by which CDC-EVs modulate CCR2-dependent monocyte trafficking and provide new insight into how EVs regulate innate immune responses after myocardial injury.
Hypoxic stress triggers transcriptional signaling mainly through hypoxia-inducible transcription factors (HIFs), which bind hypoxia response elements (HREs) in gene regulatory regions. However, only a small proportion (~1%) of known HREs are occupied by HIFs during hypoxia, suggesting the involvement of additional hypoxia-responsive factors. To address this gap, we utilized MNase-defined cistrome Occupancy Analysis sequencing (MOA-seq), with the term cistrome referring to all genomic regions where transcription factors and other trans-acting regulators are bound to cis-acting elements across the genome for a particular cell type or treatment. This MNase-based assay enables genome-wide, high-resolution (<30 bp) identification of transcription factor (TF) occupancy footprints embedded within larger regions, most of which were previously annotated as open or accessible chromatin. Applying this in situ cistrome mapping to fixed nuclei from endothelial cells under normoxia or hypoxia (1, 3, or 24 hr) revealed thousands of hypoxia-responsive genomic sites with dynamic TF footprints. The affected genes were enriched in canonical hypoxia-induced pathways, such as angiogenesis. Motif analysis identified over 100 candidate TFs potentially mediating these multifaceted genomic responses. By grouping hypoxia-modified occupancy signals across the hypoxia exposure times, we clustered differentially occupied MOA sites into defined 10 distinct TF kinetic clusters, half of which were associated with HIF1A. HIF1A-proximal binding sites suggested co-activators, while non-HIF1A clusters pointed to additional TFs that may have HIF1A-independent roles. This analysis provides insight into how multiple TF networks coordinate hypoxia responses and highlights the power of cistrome profiling to deepen our understanding of the complex genomic response to low oxygen conditions.
B-cell acute lymphoblastic leukemia (B-ALL) is the most common malignancy occurring in children and a leading cause of cancer-related mortality, thus there is an urgent need for development of novel therapeutic strategies for high risk B-ALL patients. The significance of IKZF1 gene alterations in B-ALL cases is controversial, as some studies have shown those to be associated with poor prognosis, while others have reported that deletion of exons 4-7 of the IKZF1 gene, which results in generation of IKAROS isoform 6, is related favorable prognosis. For the present study, clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 was employed for knockout of the IKZF1 gene and various IKAROS knockout lines were established, which allowed for investigation of precise functions of the gene by comparing four lines with different IKAROS isoform 1 and 6 statuses. The results clearly showed induction of significant differentiation and cell cycle progression in IKAROS isoform 6 knockout clones. Furthermore, knockout of IKAROS isoform 6 resulted in significant upregulation of the expression of IKAROS isoform 1 as well as signal transducer and activator of transcription 5 (STAT5) activity, which are considered to be related to the observed features. Further investigations are warranted to provide greater mechanistic insight regarding IKAROS-mediating pathways, which will lead to development of novel therapy for human B-ALL.
Periodontitis progresses more rapidly and severely in individuals with type 2 diabetes mellitus (T2DM), yet whether the integrated stress response (ISR) contributes to the progression of diabetic periodontitis has not been elucidated. This study aims to clarify the impact and mechanisms of growth differentiation factor 15 (GDF15) in diabetic periodontitis. We performed single-cell RNA sequencing on periodontal tissues and integrated the datasets with bidirectional Mendelian randomization to identify ISR-related alterations associated with impaired periodontal regeneration under diabetic conditions. Functional analyses were conducted in periodontal ligament stem cells (PDLSCs) subjected to diabetic-inflammatory conditions. Genetic deletion, antibody-based neutralization of GDF15, and ISR inhibition were used to evaluate therapeutic rescue effects in diabetic mice. We identify GDF15 as an important effector of ISR signaling in PDLSCs. T2DM induces sustained PERK-eIF2α activation accompanied by GDF15 induction, leading to translational repression, loss of proteostasis, and impaired osteogenic differentiation. Mechanistically, GDF15 silencing preserved BiP-PERK complex stability and attenuated downstream PERK-eIF2α activation under diabetic-inflammatory stress. Genetic ablation or pharmacologic neutralization of GDF15, as well as ISR inhibition, partially restores PDLSC proteostasis and bone-forming capacity, preserving periodontal tissue regenerative capacity in diabetic mice. These findings suggest that GDF15 contributes to ISR-mediated PDLSC dysfunction and impaired periodontal regeneration under diabetic conditions. Targeting pathological GDF15-ISR signaling may provide mechanistic insight for future strategies to improve periodontal regenerative outcomes in T2DM.
To explore circadian involvement in periodontitis and provide insight into mechanisms. Clinical samples from patients with periodontitis and a ligature-induced periodontitis mouse model were used in the present research. The rhythmic pattern of cortisol, the core circadian rhythm proteins BMAL1 and CLOCK, the expression of senescence markers (p16, p21, and p53), and the percentage and length of primary cilia were detected in hPDLSCs (human periodontal ligament stem cells) exposed to cortisol as an in vitro model of cortisol-associated circadian disruption. BMAL1 expression was manipulated by siRNA transfection to knock down BMAL1 expression, and SR8278 treatment was used to modulate BMAL1-related circadian regulation. Moreover, the effects of SR8278 on the progression of periodontitis were evaluated by micro-CT, mouse behavior tests, the number of primary cilia, and senescence assays. Patients with periodontitis and ligature-induced periodontitis mice exhibited elevated cortisol levels, reduced BMAL1 expression, increased senescence-associated markers, and decreased primary cilia abundance. Elevated cortisol was associated with circadian disruption, accompanied by reduced BMAL1 expression and impaired ciliary homeostasis. BMAL1 regulated ARL13B expression, a key regulator of ciliogenesis. BMAL1 knockdown impaired Hedgehog signaling and induced hPDLSC senescence. SR8278 partially recovered BMAL1-related circadian alterations and primary cilia abundance while attenuating cellular senescence in both periodontitis model mice and cortisol-treated hPDLSCs. Collectively, these findings support the involvement of the cortisol-BMAL1-ARL13B pathway in periodontitis-associated primary cilium dysfunction and hPDLSC senescence. Modulation of BMAL1 partially restored primary cilium homeostasis and attenuated periodontal tissue damage, supporting its potential as a therapeutic target for periodontitis.
COVID-19 continues to present ongoing global health challenges driven by diverse immune responses and heterogeneous clinical outcomes. The ACCORD trial evaluated 3 investigational treatments-bemcentinib, tozorakimab, and zilucoplan-in patients hospitalized with COVID-19, each of which has demonstrated clinical efficacy. To better understand their molecular mechanisms, we conducted a mechanistic follow-up study, integrating transcriptomic and clinical data from 65 patients and applying cellular deconvolution, differential expression, coexpression, and pathway enrichment analyses to uncover treatment-specific immune responses. Each therapy induced transcriptional shifts and modulated distinct immune pathways implicated in severe disease. Bemcentinib primarily modulated myeloid cell populations and inflammatory signalling; zilucoplan enhanced B-cell signalling and lymphocyte-associated pathways; and tozorakimab exerted broad immune and cellular responses across immune cell types. Co-expression analysis revealed gene networks associated with clinical improvement, each driven by distinct treatment-specific hub genes, indicating diverse regulatory mechanisms across treatments. Improved outcomes correlated with gene expression shifts in 4 key immunological pathways: B-cell signalling, antiviral defense, innate inflammation, and platelet/coagulation activity. In contrast, nonresponders had persistent dysregulation of 1 or more of these gene signatures. Our findings define molecular signatures of treatment response and failure in COVID-19, providing mechanistic insight into how distinct therapies modulate the immune system. These insights support the need for adaptive precision medicine approaches tailored to individual, evolving immune trajectories. Moreover, the immunological mechanisms targeted by these repurposed immunomodulatory therapies may inform treatment strategies across a broader spectrum of immune-mediated diseases beyond COVID-19.
There is renewed interest in resection of Stage III (N2+) NSCLC given impressive outcomes with neoadjuvant/perioperative chemoimmunotherapy (ChIO). We report surgical outcomes from an exclusively N2+ NSCLC clinical trial following chemotherapy + durvalumab. This was a single arm phase II trial enrolled at 9 US hospitals. Eligible patients had resectable stage III NSCLC, pathologically proven N2+. Patients received 4 cycles of platinum doublet + durvalumab followed by lobectomy or greater, and adjuvant durvalumab for 1 year. Surgical approach, margins, extent of lymphadenectomy, complications, and treatment timeliness were analyzed. From 2021-2023, 37 patients were enrolled; 30 patients underwent resection (81%). Surgical outcomes are notable for R0 resection in 28/30 patients (93.3%), pneumonectomy rate of 6.7% (2/30), median stay 3.5 days, no mortality at 30 and 90 days. Minimally invasive surgery was possible in 19/30 (63.3%: 2 VATS, 17 robotic), with 2 conversions to thoracotomy (total thoracotomy rate: 11/30: 36.7%). Increased surgical difficulty was reported in 14/30 (46.7%). Median interval from neoadjuvant therapy to surgery was 46 days, and from surgery to adjuvant therapy was 35 days. All 23 patients recommended for adjuvant therapy received it. This trial provides insight into surgical outcomes for prospectively documented N2+ NSCLC treated with neoadjuvant ChIO. Surgery in this challenging scenario was accomplished with no mortality, high rates of minimally invasive surgery, R0 resection, and lobectomy, with prompt return to oncologic therapy. Following ChIO, resection of N2+ NSCLC can be achieved with excellent outcomes and warrants stronger consideration within multidisciplinary NSCLC care.
Nutrient limitation is a characteristic feature of poorly perfused tumors. In contrast to well-perfused tissues, nutrient deficits in tumors impose metabolic constraints on cancer cells. The metabolic constraints created by the tumor microenvironment can lead to vulnerabilities in cancers. Identifying the metabolic constraints of the tumor microenvironment and the vulnerabilities that arise in cancers can provide new insight into tumor biology and identify promising anti-neoplastic targets. To identify how the microenvironment constrains the metabolism of pancreatic tumors, we challenged pancreatic cancer cells with microenvironmental nutrient levels and analyzed changes in cellular metabolism. We found that arginine limitation in pancreatic tumors perturbs saturated and monounsaturated fatty acid synthesis by suppressing the lipogenic transcription factor SREBP1, in part via activation of the amino acid sensor GCN2. Synthesis of these fatty acids is critical for maintaining a balance of saturated, monounsaturated, and polyunsaturated fatty acids (PUFAs) in cellular membranes. Because of microenvironmental constraints on fatty acid synthesis, pancreatic cancer cells and tumors are unable to maintain lipid homeostasis when exposed to PUFAs, leading to cell death by ferroptosis. In sum, arginine restriction in the tumor microenvironment constrains lipid metabolism in pancreatic cancers, which renders these tumors vulnerable to polyunsaturated-enriched fats.
In mammals, glutathione peroxidase 7 (GPx-7) is a member of the GPx family that exhibits peroxidase activity. Its immunological functions, especially in host defense against bacterial infection, remain unexplored in lower vertebrates. In this study, we identified a GPx-7 homolog from Paralichthys olivaceus (PoGPx-7) and investigated its roles during Vibrio alginolyticus infection. PoGPx-7 possesses a conserved GSH-Px domain and carries positive net charges. PoGPx-7 was constitutively expressed in various tissues, with significant upregulation upon bacterial challenge. Recombinant PoGPx-7 (rPoGPx-7) exhibited GPx activity and bound to V. alginolyticus via interaction with lipopolysaccharide and peptidoglycan. In addition, rPoGPx-7 could directly kill bacteria by disrupting membrane integrity, leading to severe structural damage and content leakage. The bactericidal activity was modulated by protein concentration, pH, temperature, and Zn2+. Furthermore, rPoGPx-7 bound to peripheral blood leukocytes (PBLs), reduced bacterial attachment and LDH release, and protected PBLs from cell death. It also significantly enhanced phagocytosis, respiratory burst, and acid phosphatase activity of PBLs. In vivo administration showed that rPoGPx-7 reduced bacterial loads in the tissues, and improved fish survival, whereas knockdown of PoGPx-7 increased susceptibility to infection. These findings provide the first evidence that teleosts GPx-7 functions as a dual-effector molecule with direct bactericidal activity and immunomodulatory capacity, providing an immunological insight of GPx family members on resistance bacterial infection.