技术资料

This study sheds light on the pivotal role of the oncoprotein DEK in B-cell lymphoma. We reveal DEK expression correlates with increased tumor proliferation and inferior overall survival in cases diagnosed with low-grade B-cell lymphoma (LGBCL). We also found significant correlation between DEK expression and copy number alterations in LGBCL tumors,highlighting a novel mechanism of LGBCL pathogenesis that warrants additional exploration. To interrogate the mechanistic role of DEK in B-cell lymphoma,we generated a DEK knockout cell line model,which demonstrated DEK depletion caused reduced proliferation and altered expression of key cell cycle and apoptosis-related proteins,including Bcl-2,Bcl-xL,and p53. Notably,DEK depleted cells showed increased sensitivity to apoptosis-inducing agents,including venetoclax and staurosporine,which underscores the therapeutic potential of targeting DEK in B-cell lymphomas. Overall,our study contributes to a better understanding of DEK’s role as an oncoprotein in B-cell lymphomas,highlighting its potential as both a promising therapeutic target and a novel biomarker for aggressive LGBCL. Further research elucidating the molecular mechanisms underlying DEK-mediated tumorigenesis could pave the way for improved treatment strategies and better clinical outcomes for patients with B-cell lymphoma. View Publication

SummaryMonocytes are critical to innate immunity,participating in chemotaxis during tissue injury,infection,and inflammatory conditions. However,the migration dynamics of human monocytes under different guidance cues are not well characterized. Here,we developed a microfluidic device to profile the migration characteristics of human monocytes under chemotactic and barotactic guidance cues while also assessing the effects of age and cytokine stimulation. Human monocytes preferentially migrated toward the CCL2 gradient through confined microchannels,regardless of donor age and migration pathway. Stimulation with interferon (IFN)-γ,but not granulocyte-macrophage colony-stimulating factor (GM-CSF),disrupted monocyte navigation through complex paths and decreased monocyte CCL2 chemotaxis,velocity,and CCR2 expression. Additionally,monocytes exhibited a bias toward low-hydraulic-resistance pathways in asymmetric environments,which remained consistent across donor ages,cytokine stimulation,and chemoattractants. This microfluidic system provides insights into the unique migratory behaviors of human monocytes and is a valuable tool for studying peripheral immune cell migration in health and disease. Graphical abstract Highlights•The MAP chip profiles migration of human monocytes under various chemotactic and barotactic cues•Monocytes preferentially migrate toward CCL2 gradients,regardless of migration pathway and donor age•IFN-γ reduces human monocyte chemotaxis,velocity,and CCR2 expression•Human monocytes show biased migration toward low-hydraulic-resistance pathways MotivationCell migration is fundamental to the biological processes that drive health and disease. While in vivo models provide invaluable insights into cell migration within complex biological environments,precise control over the microenvironment and single-cell tracking is essential to deepen our understanding of the fundamental characteristics of cell migration. We present a high-throughput microfluidic platform,termed the migration analysis of peripheral immune cells (MAP) chip,that features four distinct sets of microchannels designed to assess the effects of both chemotactic and barotactic stimuli on cell migration at a single-cell level. By profiling human monocyte migration using the MAP chip,we demonstrated the utility of this device in characterizing migration of human monocytes under diverse conditions. Hall et al. introduce the MAP chip,a microfluidic platform for profiling human monocytes under chemotactic and barotactic guidance cues. It reveals biased migration toward low-hydraulic-resistance pathways,disrupted migration upon cytokine stimulation,and consistent chemotaxis and barotaxis across donor ages—enhancing our understanding of human monocyte migration characteristics. View Publication

The integrity of the blood–labyrinth barrier (BLB) is essential for inner ear homeostasis,regulating the ionic composition of endolymph and perilymph and preventing harmful substance entry. Endothelial hyperpermeability,central in inflammatory and immune responses,is managed through complex intercellular communication and molecular signaling pathways. Recent studies link BLB permeability dysregulation to auditory pathologies like acoustic trauma,autoimmune inner ear diseases,and presbycusis. Polymorphonuclear granulocytes (PMNs),or neutrophils,significantly modulate vascular permeability,impacting endothelial barrier properties. Neutrophil extracellular traps (NETs) are involved in diseases with autoimmune and autoinflammatory bases. The present study evaluated the impact of NETs on a BLB cellular model using a Transwell® setup. Our findings revealed a concentration-dependent impact of NETs on human inner ear-derived endothelial cells. In particular,endothelial permeability markers increased,as indicated by reduced transepithelial electrical resistance,enhanced dextran permeability,and downregulated junctional gene expression (ZO1,OCL,and CDH5). Changes in cytoskeletal architecture were also observed. These preliminary results pave the way for further research into the potential involvement of NETs in BLB impairment and implications for auditory disorders. View Publication

Hidradenitis suppurativa (HS) is a chronic inflammatory skin condition characterized by painful nodules,abscesses,and scarring,predominantly affecting intertriginous regions and it is often underdiagnosed. This study aimed to utilize single cell RNA and cell-surface protein sequencing (CITE-Seq) to delineate the immune composition of circulating cells in Hidradenitis suppurativa (HS) peripheral blood compared to healthy controls. CITE-Seq was used to analyze the gene and protein expression profiles of peripheral blood mononuclear cells (PBMCs) from 9 HS and 29 healthy controls. The study identified significant differences cell composition between HS patients and healthy controls,including increased proportions of CD14+ and CD16+ monocytes,cDC2,plasmablasts,and proliferating CD4+ T cells in HS patients. Differential expression analysis revealed upregulation of inflammatory markers such as TNF,IL1B,and NF-κB in monocytes,as well as chemokines and cell adhesion molecules involved in immune cell recruitment and tissue infiltration. Pathway enrichment analysis highlighted the involvement of IL-17,IL-26 and TNF signaling pathways in HS pathogenesis. Machine learning identified key markers for diagnostics and therapeutic development. The findings also support the potential for machine learning models to aid in the diagnosis of HS based on immune cell markers. These insights may inform future therapeutic strategies targeting specific immune pathways in HS. View Publication

BackgroundMacrophage-based cell therapies have shown modest success in clinical trials,which can be attributed to their phenotypic plasticity,where transplanted macrophages get reprogrammed towards a pro-tumor phenotype. In most tumor types,including melanoma,the balance between antitumor M1-like and tumor-promoting M2-like macrophages is critical in defining the local immune response with a higher M1/M2 ratio favoring antitumor immunity. Therefore,designing novel strategies to increase the M1/M2 ratio in the TME has high clinical significance and benefits macrophage-based cell therapies.MethodsIn this study,we reprogrammed antitumor and proinflammatory macrophages ex-vivo with HDAC6 inhibitors (HDAC6i). We administered the reprogrammed macrophages intratumorally as an adoptive cell therapy (ACT) in the syngeneic SM1 murine melanoma model and patient-derived xenograft bearing NSG-SGM3 humanized mouse models. We phenotyped the tumor-infiltrated immune cells by flow cytometry and histological analysis of tumor sections for macrophage markers. We performed bulk RNA-seq profiling of murine bone marrow-derived macrophages treated with vehicle or HDAC6i and single-cell RNA-seq profiling of SM1 tumor-infiltrated immune cells to determine the effect of intratumor macrophage ACT on the tumor microenvironment (TME). We further analyzed the single-cell data to identify key cell-cell interactions and trajectory analysis to determine the fate of tumor-associated macrophages post-ACT.ResultsMacrophage ACT resulted in diminished tumor growth in both mouse models. We also demonstrated that HDAC6 inhibition in macrophages suppressed the polarization toward tumor-promoting phenotype by attenuating STAT3-mediated M2 reprogramming. Two weeks post-transplantation,ACT macrophages were viable,and inhibition of HDAC6 rendered intratumor transplanted M1 macrophages resistant to repolarization towards protumor M2 phenotype in-vivo. Further characterization of tumors by flow cytometry,single-cell transcriptomics,and single-cell secretome analyses revealed a significant enrichment of antitumor M1-like macrophages,resulting in increased M1/M2 ratio and infiltration of CD8 effector T-cells. Computational analysis of single-cell RNA-seq data for cell-cell interactions and trajectory analyses indicated activation of monocytes and T-cells in the TME.ConclusionsIn summary,for the first time,we demonstrated the potential of reprogramming macrophages ex-vivo with HDAC6 inhibitors as a viable macrophage cell therapy to treat solid tumors.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13046-024-03182-w. View Publication

BackgroundAt present,hepatic ischemia-reperfusion injury (IRI) is an important complication of partial hepatectomy and liver transplantation,and it is an important cause of poor prognosis. Spleen tyrosine kinase(SYK) plays an important role in a variety of signaling pathways in the liver,but its role in hepatic IRI is still unclear. This study aims to investigate the role and mechanism of SYK in hepatic IRI and tumor recurrence.MethodsWe first observed the activation of SYK in the liver of mice in response to hepatic IRI. Subsequently,Pharmacological inhibitions of SYK were used to evaluated the effect of SYK on neutrophil recruitment and NETosis,and further explored the effect of SYK on IRI and tumor recurrence.ResultsOur study shows that SYK is activated in response to hepatic IRI and aggravates liver injury. On the one hand,neutrophils SYK during the early stage of liver reperfusion increases neutrophil extracellular traps (NETs) production by promoting Pyruvate kinase M2(PKM2) nuclear translocation leading to upregulation of phosphorylated STAT3,thereby exacerbating liver inflammation and tumor recurrence. On the other hand,macrophages SYK can promote the recruitment of neutrophils and increase the activation of NLRP3 inflammasome and IL1β,which further promotes the formation of NETs.ConclusionsOur study demonstrates that neutrophil and macrophage SYK synergistically promote hepatic IRI and tumor recurrence,and SYK may be a potential target to improve postoperative hepatic IRI and tumor recurrence.Supplementary InformationThe online version contains supplementary material available at 10.1186/s10020-024-00907-7. View Publication

SummaryAntibodies represent a primary mediator of protection against respiratory viruses. Serum neutralizing antibodies (NAbs) are often considered a primary correlate of protection. However,detailed antibody profiles including characterization of antibody functions in different anatomic compartments are poorly understood. Here we show that antibody correlates of protection against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) challenge are different in systemic versus mucosal compartments in rhesus macaques. In serum,NAbs were the strongest correlate of protection and linked to spike-specific binding antibodies and other extra-NAb functions that create a larger protective network. In bronchiolar lavage (BAL),antibody-dependent cellular phagocytosis (ADCP) proved the strongest correlate of protection rather than NAbs. Within BAL,ADCP was linked to mucosal spike-specific immunoglobulin (Ig)G,IgA/secretory IgA,and Fcγ-receptor binding antibodies. Our results support a model in which antibodies with different functions mediate protection at different anatomic sites. Graphical abstract Highlights•Correlates of protection to SARS-CoV-2 Omicron are highly compartment specific•Antibody effector functions are primary correlates of protection at infection site•Mucosal boosting enhances IgA and functionally levered IgG in lower respiratory tract Health sciences; Biological sciences View Publication

Pain and inflammation contribute immeasurably to reduced quality of life,yet modern analgesic and anti-inflammatory therapeutics can cause dependence and side effects. Here,we screened 1444 plant extracts,prepared primarily from native species in California and the United States Virgin Islands,against two voltage-gated K+ channels - T-cell expressed Kv1.3 and nociceptive-neuron expressed Kv7.2/7.3. A subset of extracts both inhibits Kv1.3 and activates Kv7.2/7.3 at hyperpolarized potentials,effects predicted to be anti-inflammatory and analgesic,respectively. Among the top dual hits are witch hazel and fireweed; polymodal modulation of multiple K+ channel types by hydrolysable tannins contributes to their dual anti-inflammatory,analgesic actions. In silico docking and mutagenesis data suggest pore-proximal extracellular linker sequence divergence underlies opposite effects of hydrolysable tannins on different Kv1 isoforms. The findings provide molecular insights into the enduring,widespread medicinal use of witch hazel and fireweed and demonstrate a screening strategy for discovering dual anti-inflammatory,analgesic small molecules. A dual potassium channel functional screen of 1444 plant extracts uncovers unexpected molecular mechanisms underlying the traditional use of witch hazel and fireweed as analgesic,anti-inflammatory medicines. View Publication

A shift of arginine metabolism from polyamine synthesis to nitric oxide synthesis induces reprogramming of macrophages from pro-tumor M2 to anti-tumor M1 types. HER2+ breast tumors have abundant immune-suppressive cells,including M2-type tumor-associated macrophages (TAMs). Although TAMs consist of the immune-stimulatory M1 type and immune-suppressive M2 type,the M1/M2-TAM ratio is reduced in immune-suppressive tumors,contributing to their immunotherapy refractoriness. M1- versus M2-TAM formation depends on differential arginine metabolism,where M1-TAMs convert arginine to nitric oxide (NO) and M2-TAMs convert arginine to polyamines (PAs). We hypothesize that such distinct arginine metabolism in M1- versus M2-TAMs is attributed to different availability of BH4 (NO synthase cofactor) and that its replenishment would reprogram M2-TAMs to M1-TAMs. Recently,we reported that sepiapterin (SEP),the endogenous BH4 precursor,elevates the expression of M1-TAM markers within HER2+ tumors. Here,we show that SEP restores BH4 levels in M2-like macrophages,which then redirects arginine metabolism to NO synthesis and converts M2 type to M1 type. The reprogrammed macrophages exhibit full-fledged capabilities of antigen presentation and induction of effector T cells to trigger immunogenic cell death of HER2+ cancer cells. This study substantiates the utility of SEP in the metabolic shift of the HER2+ breast tumor microenvironment as a novel immunotherapeutic strategy. View Publication

IntroductionChronic inflammation of the gastrointestinal tissues underlies gastrointestinal inflammatory disorders,leading to tissue damage and a constellation of painful and debilitating symptoms. These disorders include inflammatory bowel diseases (Crohn’s disease and ulcerative colitis),and eosinophilic disorders (eosinophilic esophagitis and eosinophilic duodenitis). Gastrointestinal inflammatory disorders can often present with overlapping symptoms necessitating the use of invasive procedures to give an accurate diagnosis.MethodsThis study used peripheral blood mononuclear cells from individuals with Crohn’s disease,ulcerative colitis,eosinophilic esophagitis,and eosinophilic duodenitis to better understand the alterations to the transcriptome of individuals with these diseases and identify potential markers of active inflammation within the peripheral blood of patients that may be useful in diagnosis. Single-cell RNA-sequencing was performed on peripheral blood mononuclear cells isolated from the blood samples of pediatric patients diagnosed with gastrointestinal disorders,including Crohn’s disease,ulcerative colitis,eosinophilic esophagitis,eosinophilic duodenitis,and controls with histologically healthy gastrointestinal tracts.ResultsWe identified 730 (FDR < 0.05) differentially expressed genes between individuals with gastrointestinal disorders and controls across eight immune cell types.DiscussionThere were common patterns among GI disorders,such as the widespread upregulation of MTRNR2L8 across cell types,and many differentially expressed genes showed distinct patterns of dysregulation among the different gastrointestinal diseases compared to controls,including upregulation of XIST across cell types among individuals with ulcerative colitis and upregulation of Th2-associated genes in eosinophilic disorders. These findings indicate both overlapping and distinct alterations to the transcriptome of individuals with gastrointestinal disorders compared to controls,which provide insight as to which genes may be useful as markers for disease in the peripheral blood of patients. View Publication

Crohn’s disease (CD) is a complex chronic inflammatory disorder with both gastrointestinal and extra-intestinal manifestations associated immune dysregulation. Analyzing 202,359 cells from 170 specimens across 83 patients,we identify a distinct epithelial cell type in both terminal ileum and ascending colon (hereon as ‘LND’) with high expression of LCN2,NOS2,and DUOX2 and genes related to antimicrobial response and immunoregulation. LND cells,confirmed by in-situ RNA and protein imaging,are rare in non-IBD controls but expand in active CD,and actively interact with immune cells and specifically express IBD/CD susceptibility genes,suggesting a possible function in CD immunopathogenesis. Furthermore,we discover early and late LND subpopulations with different origins and developmental potential. A higher ratio of late-to-early LND cells correlates with better response to anti-TNF treatment. Our findings thus suggest a potential pathogenic role for LND cells in both Crohn’s ileitis and colitis. Crohn’s disease (CD) is a complex disease associated with immune dysregulation. Here the authors use multimodal data to identify and characterize an epithelial cell population,termed ‘LND’ cells,in both terminal ileum and ascending colon,with LND interacting locally with immune cells and potentially contributing to CD pathology. View Publication

Platelets are key mediators of atherothrombosis,yet,limited tools exist to identify individuals with a hyperreactive platelet phenotype. In this study,we investigate the association of platelet hyperreactivity and cardiovascular events,and introduce a tool,the Platelet Reactivity ExpreSsion Score (PRESS),which integrates platelet aggregation responses and RNA sequencing. Among patients with peripheral artery disease (PAD),those with a hyperreactive platelet response (>60% aggregation) to 0.4 µM epinephrine had a higher incidence of the 30 day primary cardiovascular endpoint (37.2% vs. 15.3% in those without hyperreactivity,adjusted HR 2.76,95% CI 1.5–5.1,p = 0.002). PRESS performs well in identifying a hyperreactive phenotype in patients with PAD (AUC [cross-validation] 0.81,95% CI 0.68 –0.94,n = 84) and in an independent cohort of healthy participants (AUC [validation] 0.77,95% CI 0.75 –0.79,n = 35). Following multivariable adjustment,PAD individuals with a PRESS score above the median are at higher risk for a future cardiovascular event (adjusted HR 1.90,CI 1.07–3.36; p = 0.027,n = 129,NCT02106429). This study derives and validates the ability of PRESS to discriminate platelet hyperreactivity and identify those at increased cardiovascular risk. Future studies in a larger independent cohort are warranted for further validation. The development of a platelet reactivity expression score opens the possibility for a personalized approach to antithrombotic therapy for cardiovascular risk reduction. Platelet hyperreactivity is associated with cardiovascular events in patients with PAD. Here the authors derive and validate a circulating platelet genetic signature to discriminate platelet hyperreactivity and cardiovascular risk. View Publication