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BACKGROUND: Treatment benefits of paclitaxel (Px)-based chemotherapy are often offset by dose-limiting side effects. The sigma-1 receptor (Sig1R) is implicated in chemotherapy (ChT)-induced neuropathy but its role in the metastatic potential of breast cancer (BCa) has not been properly explored. METHODS: This work investigated the predictive and prognostic value of Sig1R gene (S1R) expression for pathologic complete response (pCR) and distant relapse-free survival (DRFS) following neoadjuvant ChT (nChT). We further examined the anticancer efficacy of the Sig1R antagonists IPAG and BD1047 in combination with Px in triple-negative breast cancer (TNBC) cell lines. RESULTS: We report that S1R positively associated with pCR in two patient cohorts. Upregulated gene clusters in high-S1R samples of the pCR group are associated with ontology terms related to cell division, DNA replication and microtubule dynamics. High S1R expression was also associated with poor DRFS in TNBC patients. Sig1R knockdown (Sig1R-KD) in MDA-MB-231 and HCC1806 cell lines reduced clonogenic proliferation while treatment with Sig1R antagonists IPAG or BD1047 decreased cell motility. Sig1R-KD decreased Px-induced apoptosis; the synergistic effects of Px in combination with IPAG or BD1047 were evaluated by the Chou-Talalay method. Cytotoxic and antimotility effects of Px were enhanced when combined with Sig1R antagonists. CONCLUSION: Taken together, our results indicate that S1R is a potential biomarker for the response to nChT and that targeting Sig1R could enhance Px efficacy while deterring key metastatic mechanisms.

IntroductionAdvanced glycation end products are a diverse, complex group of chemicals that are mostly produced by the Maillard reaction. The Maillard reaction happens when reducing sugar interacts non-enzymatically with amino acids found in proteins, lipids, or DNA. AGEs can be produced not only endogenously but also exogenously, as a result of certain cooking conditions or food processing. AGEs can interact with their receptor RAGE (receptor for AGE), expressed in various tissues including lung, inducing inflammatory effects and oxidative stress conditions that could damage target organs. Recent studies demonstrated that AGEs could be implicated in pathophysiology of chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis (IPF).ObjectiveConsidering that human lung macrophages (HLMs) are the most abundant immune cells in the lung, the aim of this project was to investigate the effects of AGEs on HLM activation and RAGE expression in these cells.ResultsAGEs induce, in a concentration-dependent manner, the release of cytokines (IL-6, IL-1β, TNF-α) and chemokines (CXCL1, CXCL2, CXCL8) from HLMs but not their de novo synthesis, except for IL-6. AGEs affect cellular kinetic proprieties and phagocytosis. HLMs constitutively express both sRAGE (soluble form of the receptor) and flRAGE (membrane-bound form of the receptor) mRNAs, and AGEs interaction slightly upregulate sRAGE expression. RAGE protein is constitutively expressed by about 60% of HLMs. Pre-incubation of HLMs with anti-RAGE antibody inhibits the release of cytokines and chemokines induced by AGEs. Finally, HLMs store and spontaneously release AGEs.ConclusionAGE accumulation promotes a pro-inflammatory phenotype in HLMs, potentially contributing to lung injury and respiratory dysfunction in AGEs-related diseases through the release of pro-inflammatory mediators. These findings highlight HLMs as a potential target in AGE-mediated pulmonary inflammation and suggest novel therapeutic strategies targeting the AGE-HLM axis. Graphical Abstract

ObjectiveRegulatory T cells (Tregs) play a pivotal role in tumor immune evasion, and strategies to overcome their immunosuppressive activity are urgently needed. This study investigates the immunomodulatory effects of microwave ablation (MWA) on TNFRSF4+ Tregs, focusing on the OX40L/TNFRSF4 signaling axis as a potential therapeutic target.MethodsTNFRSF4+ Tregs were isolated from C57BL/6 mice and subjected to MWA-mimetic thermal stress. In vitro functional assays and in vivo LLC xenograft models were employed, with OX40 agonist intervention. Molecular mechanisms were analyzed via RT-qPCR, Western blot, and immunohistochemistry. The balance of tumor-infiltrating immune cells was quantified by multi-color flow cytometry.ResultsMWA induced three key effects: (1) Phenotypic shift: decreased CTLA-4+ (P<0.0001) Treg subsets, but increased OX40L+ (P<0.01) in LLC cells; (2) Functional impairment: reduced Treg-mediated support for LLC proliferation, migration, and invasion; (3) Enhanced CD8+ T cell cytotoxicity. In vivo, MWA reshaped the tumor microenvironment by significantly increasing the intratumoral CD8+/Treg ratio (P<0.001), indicating a shift toward an anti-tumor inflammatory state. Mechanistically, MWA suppressed NF-κB/IκBα/TRAF6 signaling, and these effects were amplified by an OX40 agonist, suggesting the pathway is potentially OX40L-dependent.ConclusionThis study demonstrates that MWA disrupts Treg immunosuppression, likely by activating OX40L/TNFRSF4 signaling, and favorably alters the balance of effector to suppressor cells, providing a novel rationale for combining thermal ablation with OX40-targeted immunotherapies in cancer treatment.

ABSTRACTMYO1F, a long-tailed myosin of class I, is selectively expressed in immune cells and upregulated in microglia associated with neurodegenerative pathogenesis. Myosin motor functions are regulated by adaptor proteins that mediate cargo attachment and motor recruitment. To define the MYO1F interactome, we used in situ proximity labelling and proteomics in human myeloid cells. We identified a distinct SH3-domain-dependent adaptor module comprising CD2AP, ASAP1, SH3BP2 and SH3KBP1 (herein termed the CASS group of proteins). Interestingly, CD2AP is an Alzheimer's disease (AD) risk gene upregulated in the microglia of individuals with AD, which are implicated in phagocytic responses to amyloid-β. Structural modelling and mutagenesis confirmed multivalent proline-rich motif interactions between the CASS group of proteins and the MYO1F SH3 domain. Additional binding partners associate with the MYO1F pleckstrin homology (PH) domain. Immunofluorescence revealed colocalisation of MYO1F and the CASS group of proteins at actin-rich podosomes and phagocytic cups in macrophages and microglia. Functional assays demonstrated that MYO1F recruitment to the phagocytic cup requires motor activity and intact PH and SH3 domains. We provide the first MYO1F interactome identifying adaptor proteins for MYO1F in podosomes and during phagocytosis, offering new insights into its function in disease-associated microglia during neurodegeneration. Highlighted Article: The MYO1F interactome identifies ASAP1, CD2AP and SH3BP2 as novel adaptor proteins in podosomes and during phagocytosis, offering new insights into MYO1F function in neuroinflammatory microglia.

Long non-coding RNAs (lncRNAs) and RNA N⁶-methyladenosine (m6A) have been linked to leukemia drug resistance. However, whether and how lncRNAs and m6A coordinately regulate resistance remain elusive. Here, we show that many differentially expressed lncRNAs enrich m6A, and more lncRNAs tend to have higher m6A content in CML cells resistant to tyrosine kinase inhibitors (TKIs). We demonstrate the broad clinical relevance of our findings, showing that upregulation of top-ranked lncRNAs (e.g., SENCR, PROX1-AS1, LINC00892) in TKI-resistant cell lines occurs in CML patients at the diagnostic stage, blast crisis phase, or not responding to TKIs compared to the chronic phase or TKI responders, respectively. Higher lncRNAs predict drug resistance and shorter survival duration. The knockdown of SENCR, PROX1-AS1, or LINC00892 restores TKI sensitivity. Mechanistically, upregulation of PROX1-AS1, SENCR, and LINC00892 results from FTO-dependent m6A hypomethylation that stabilizes lncRNA transcripts and empowers resistant cell growth through overexpression of PI3K signaling mediators (e.g., ITGA2, F2R, COL6A1). Treatment with PI3K inhibitor alpelisib eradicates resistant cells in vitro and in vivo, with prolonged survival of leukemic mice through downregulation of F2R, ITGA2, and COL6A1. Thus, the lncRNA-m6A-PI3K cascade represents a new non-genetic predictor for drug resistance and poorer prognosis in cancer, and a pan-cancer mechanism underlying TKI resistance.

BackgroundCell therapy can be utilized to induce operational tolerance following solid organ transplantation. In thi study, donor-specific immunomodulatory cells (DSIMC) are generated by co-culturing recipient peripheral blood mononuclear cells (PBMC) with irradiated donor PBMC in the presence of belatacept, a CTLA-4-IgG1 fusion protein. DSIMC promote a regulatory response to donor cells. Reinfusion of these cells into the recipient may induce donor-specific tolerance, enabling weaning or complete cessation of immunosuppression (IS).AimThis study aims to determine optimal culture conditions for DSIMC production.MethodsDSIMC were generated by culturing PBMCs from healthy volunteers with irradiated allogeneic PBMC and belatacept. We evaluated the choice of medium, plasma supplementation, costimulation blocker concentration, and red blood cell (RBC) lysis, using automated cell counts, cytokine assays, PCR, and flow cytometry.ResultsIncreasing belatacept concentration (0-80 μg per million cells) resulted in a significant reduction in PD-1 expression in regulatory T cells. RBC lysis reduced inflammatory cytokine production and improved DSIMC generation, as indicated by increased IL-10 and decreased IFN-γ production. Between culture days 9-14, the total cell yield and IFN-γ-producing cell numbers declined, while IL-10-producing cell numbers increased.ConclusionTreating responder and irradiated stimulator PBMC with RBC lysis buffer before co-culture in TexMACS medium supplemented with 1% autologous plasma and 40 μg belatacept per million cells for 14 days produces DSIMC which meet established quality criteria. The protocol is currently currently under evaluation in a clinical study.

Non-small cell lung cancer (NSCLC) is a leading cause of cancer-related mortality, with resistance to PD-1 immune checkpoint inhibitors presenting a significant clinical challenge. Although the protein tyrosine phosphatase SHP2 has been implicated in immune evasion, its precise role in NSCLC and contribution to anti-PD-1 resistance remain poorly understood. To address this, we conducted a CRISPR-based screen which identified SHP2 as a pivotal factor promoting tumor escape from CD8 + T cell-mediated killing. SHP2 expression in NSCLC tissues was analyzed through immunohistochemistry (IHC), qRT-PCR, and Western blotting. Functional assays, including CCK-8 and colony formation, were employed to assess SHP2’s role in tumor proliferation under IFN-γ stimulation. Co-culture experiments with CD8 + T cells evaluated the modulation of immune responses. Mechanistic investigations focusing on IFN-γ/STAT1/IRF1 signaling and CCL5 secretion were analyzed using bulk RNA sequencing, Western blotting, qRT-PCR, ELISA, and proximity ligation assays. We found that SHP2 overexpression correlated with advanced disease and poor prognosis. Mechanistically, SHP2 suppressed IFN-γ/STAT1/IRF1 signaling, reducing CCL5 secretion and impairing CD8 + T cell cytotoxicity. SHP2 knockdown restored immune responses and sensitized tumors to anti-PD-1 therapy. Additionally, pharmacological inhibition of SHP2 with JAB-3312 reversed this immunosuppressive phenotype in NSCLC cell lines and patient-derived organoids (PDOs). Furthermore, in a syngeneic mouse model, JAB-3312 acted synergistically with anti-PD-1 antibodies to suppress tumor growth, an effect driven by a potent T-cell-intrinsic mechanism. These findings establish SHP2 as a key mediator of immune evasion and PD-1 resistance in NSCLC, and targeting SHP2 offers a promising therapeutic strategy to overcome immune resistance and improve responses to checkpoint blockade therapy.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12964-025-02498-0.

ABSTRACTZinc finger E‐box binding protein 2 (ZEB2) is a key factor in the differentiation of naïve CD8+ T cells into effector and memory T cells. However, the precise regulatory role of ZEB2 in cytotoxic CD8+ T cells remains unknown. Our recent DNA methylation analysis of cytomegalovirus (CMV)‐specific human CD8+ T cells revealed two differentially methylated regions (DMRs) within the ZEB2 locus. In the present study, we show that these ZEB2 DMRs undergo pronounced demethylation during T cell differentiation. In particular, terminally differentiated CD8+ T cells and cytotoxic CD4+ T cells show an almost complete demethylation. Demethylation of the ZEB2 DMRs correlates strongly with ZEB2 expression in all T cell subsets. Furthermore, DNA methylation patterns remain stable during long‐term in vitro culture. ZEB2 knockout in CD8+ effector T cells results in altered gene expression profiles, affecting genes related to cell–cell adhesion and impairing the cytotoxic capacity in CMV‐specific killing assays. Our data show that ZEB2 expression contributes to the differentiation of naïve CD8+ T cells into effector and memory T cells and regulates the functional properties of virus‐specific cytotoxic CD8+ T cells. Differentially methylated regions (DMRs) within the ZEB2 locus progressively demethylate during CD4+ and CD8+ T cell differentiation into cytotoxic CD4+ and CD8+ TEMRA subsets, respectively. This epigenetic remodeling tightly correlates with elevated ZEB2 expression, and ZEB2 knockout (KO) impairs the effector and cytotoxic function of CD4+ and CD8+ T cells.

α/β-hydrolase domain-containing protein 11 (ABHD11) is a mitochondrial hydrolase that maintains the catalytic function of α-ketoglutarate dehydrogenase (α-KGDH), and its expression in CD4 + T-cells has been linked to remission status in rheumatoid arthritis (RA). However, the importance of ABHD11 in regulating T-cell metabolism and function is yet to be explored. Here, we show that pharmacological inhibition of ABHD11 dampens cytokine production by human and mouse T-cells. Mechanistically, the anti-inflammatory effects of ABHD11 inhibition are attributed to increased 24,25-epoxycholesterol (24,25-EC) biosynthesis and subsequent liver X receptor (LXR) activation, which arise from a compromised TCA cycle. The impaired cytokine profile established by ABHD11 inhibition is extended to two patient cohorts of autoimmunity. Importantly, using murine models of accelerated type 1 diabetes (T1D), we show that targeting ABHD11 suppresses cytokine production in antigen-specific T-cells and delays the onset of diabetes in vivo in female mice. Collectively, our work provides pre-clinical evidence that ABHD11 is an encouraging drug target in T-cell-mediated inflammation. α/β-hydrolase domain-containing protein 11 (ABHD11) is a mitochondrial hydrolase, and its expression in CD4 + T-cells has been linked to remission status in rheumatoid arthritis. Here the authors report that pharmacological inhibition of ABHD11 modulates T-cell effector function via increased 24,25-epoxycholesterol biosynthesis and subsequent liver X receptor activation.

Psoriasis vulgaris (PV) is a common, T cell mediated dermatosis with substantial systemic footprint. While αβ T cells are well established drivers of PV, the role of γδ T cells, including their abundance, clonal architecture and transcriptional programs in PV remain incompletely understood. To address this, we performed an integrated analysis of circulating and cutaneous γδ cells from 65 patients with PV and 35 healthy controls using TCR repertoire sequencing, bulk transcriptomics, and flow cytometry. In PV, disease severity and age drove contraction of peripheral γδ T cell repertoires, marked by loss of rare clonotypes and hyperexpansion patterns. Subset composition, segment usage, and CDR3 length of both skin and blood clonotypes were further modulated by age, disease severity, and sex, highlighting nuanced repertoire remodeling. TCRγ clonotypes showed partial overlap between blood and skin, whereas TCRδ clonotypes remained private and tissue-specific, with no PV-specific clonotypes identified. Transcriptomic profiling indicated that circulating γδ T cells adopt an activated, cytotoxic, tissue-homing phenotype, consistent with enhanced potential to migrate into and act within lesional skin, especially in a subset of patients. Collectively, these findings demonstrate that PV drives dynamic, clinically modulated remodeling of γδ T cells across compartments, positioning them as dynamic elements of the psoriatic immune landscape and potential targets for future functional and therapeutic investigation.

Lipid-driven inflammation contributes to the development of atherosclerosis, and regulatory T cells (Tregs) have been proposed to influence macrophage responses to lipid stress. While adoptive Treg transfer has been shown to be safe in clinical studies, the mechanisms by which Tregs modulate macrophage lipid handling remain incompletely understood. In this study, we investigated the effects of ex vivo–expanded human Tregs on primary monocyte-derived M2-like macrophages exposed to oxidized low-density lipoprotein (oxLDL) in an in vitro coculture system. We assessed macrophage phenotype, gene expression, and cholesterol accumulation using flow cytometry, RNA sequencing, and western blotting. Our data show that coculture with Tregs attenuated oxLDL-induced pro-inflammatory responses and reduced intracellular lipid accumulation in macrophages. Mechanistically, we found evidence that Tregs transfer cyclic AMP (cAMP) into macrophages, which enhanced the ABCA1-mediated cholesterol efflux pathway and increased expression of paraoxonase-1 (PON1). These findings provide mechanistic insight into how Tregs modulate macrophage responses to oxLDL under controlled in vitro conditions. They highlight potential pathways through which Tregs may regulate macrophage lipid metabolism and inflammatory activity. Further in vivo studies will be essential to determine the physiological significance and therapeutic potential of these mechanisms.

Pancreatic islet transplantation offers great promise for the treatment of type 1 diabetes, yet the functional decline of islets after isolation remains a major obstacle. Increasing evidence highlights the endoplasmic reticulum (ER) as a critical regulator of islet cell survival under stress. We explored how ex vivo culture conditions affect encapsulated islet resilience under ER-stress. Two conditions were assessed: (i) incorporation of decellularized porcine pancreatic extracellular matrix (ECM) into alginate microcapsules, and (ii) free-fall dynamic pre-conditioning culture. Human islets were encapsulated in alginate with or without ECM, cultured under static or dynamic conditions, and exposed to acute ER-stress followed or not by a recovery period. Dynamic culture preserved viability and enhanced glucose responsiveness. ECM-containing capsules showed reduced inflammatory marker expression, while encapsulation in alginate-only capsules led to more pronounced changes associated with ECM remodeling. Under ER-stress, the dynamic culture, especially combined with ECM, maintained cell function and reduced cell death. Gene profiles indicated improved stress adaptation and ECM remodeling. These results highlight ECM enrichment and dynamic culture as good strategies to maintain islet survival and functionality. Graphical AbstractEncapsulated human pancreatic islets were cultured in a free-fall dynamic system that simulates a low-shear environment, enhancing nutrient and oxygen exchange. Upon exposure to endoplasmic reticulum (ER)-stress, dynamic culture promoted a protective and adaptive cellular response, either in alginate-only or in extracellular matrix (ECM)-containing microcapsules. Dynamic culture enhanced insulin secretion, indicating improved β-cell glucose responsiveness. It reduced the expression of inflammatory chemokines, particularly CXCL1, suggesting lower immunogenic signaling. Contributed to reduced cell death, reflecting enhanced islet viability and survival. Additionally, matrix remodeling activity was elevated, as shown by increased MMP2 and MMP9 expression, indicating an adaptive extracellular matrix turnover. Created in BioRender. Silva, I. (2025) https://BioRender.com/nf8b3yv