Eµ-Myc grows similarly in the left and right cervical and inguinal lymph nodes.

Abstract DLBCL gene expression signatures from single-nucleus analyses have recently identified new tumor microenvironment (TME) archetypes (LymphoMAPs) linked to RCHOP and CAR-T cell sensitivity. While some archetypes are enriched at diagnosis or relapse, their stability during disease progression remains unclear. We analyzed longitudinal DLBCL cases using LymphoMAP signatures and histologic annotation to assess archetype switching after RCHOP. Methods: We utilized two independent well-annotated DLBCL cohorts with paired diagnosis/relapse biopsies:23 in-house (46 samples) and 44 published cases (88 samples). Gene expression in our cohort was profiled using the NanoString Cancer Immune Panel on fixed (FFPE) specimens. The external dataset comprised whole-transcriptome from FFPE or fresh-frozen samples pre- and post-RCHOP. LymphoMapR predicted the archetypes from normalized counts: enriched in fibroblast and tumor-associated macrophages (FMAC), naïve and memory T cell (LN), activated macrophages and exhausted T cell (TEX). Samples with classification probability <0.7 were excluded. Additionally, we correlated the archetypes with histologic annotation from our cohort. Immunohistochemistry (IHC) supported T cell markers (CD3, CD4, CD8) and PDL1 expression. Multiplex OPAL imaging quantified macrophage markers (CD68, CD163) with immune checkpoint (CD47), and chemokines (CXCL9 and CXCL10) with receptor (CXCR3). Results: In the in-house cohort, 8 patients relapsed following only RCHOP, others received ≥1 relapse regimen. One patient per cohort was excluded for low LymphoMAP confidence. Notably, archetype switching occurred in 13 of 22 patients (59%, 95% CI: 36-79%) in-house and 18 of 43 patients (42%, 95% CI: 28-57%) externally. To gain statistical power, the cohorts were combined, yielding an overall switch rate of 48% (95% CI: 36-60%). Switching was less frequent for patients with LN at diagnosis (38%, 95% CI: 23-57%) than FMAC (59%, 95% CI: 36-78%) or TEX (57%, 95% CI: 33-79%), but no consistent direction of switch or correlation with relapse timing was observed. Germinal center B-cell-like (GCB) DLBCLs were predominantly LN archetype (58%), whereas non-GCB encompassed all LymphoMAP archetypes, suggesting an association with cell-of-origin. Histology was consistent with LymphoMAP prediction: T cell abundance (CD3, CD4, CD8) and CXCR3 expression were highest in TEX, followed by LN and FMAC (p<0.05). IHC also demonstrated a significant decrease in CD3 and CD8 T cells at relapse compared with diagnosis. Conclusions: Based on our observations, the immune environment of DLBCL is dynamic. Although relapse samples exhibited more T cell-depleted environments, changes in TME archetypes frequently occurred, without a dominant pattern. In the context of patient selection for CAR-T cell therapy, relapsed TME should not be inferred from TME at diagnosis. Citation Format: Raoul Santiago, Raquel Aloyz, Stéphanie Bianco, Svetlana Dmitrienko, Nathalie Johnson, Andreas I. Papadakis, Naciba Benlimame, Cynthia Guilbert, Alan Spatz, François E. Mercier, Madelyne Abraham, Laura Hilton, David W. Scott, Koren K. Mann, Sarit Assouline. Tumor immune micro-environment (TME) dynamics in longitudinal diffuse large B-cell lymphoma (DLBCL) cases from diagnosis to RCHOP relapse [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 7944.

Organophosphate esters (OPEs), widely used as flame retardants and plasticizers, are environmental toxicants known to disrupt lipid metabolism. Although most studies have focused on individual OPEs, environmental exposures typically involve complex mixtures. Our previous studies demonstrated that a representative OPE mixture from Canadian household dust promotes cholesterol and lipid droplet accumulation in THP-1 macrophages. However, the molecular mechanisms underlying this lipid dysregulation remain unclear. Here, we employed tandem mass tag (TMT)-based quantitative proteomics to investigate how OPE mixtures alter protein expression and lipid regulation in macrophages. THP-1 macrophages were exposed to vehicle or environmentally relevant dilutions of the OPE mixture for 48 h. Lysates were subjected to TMT labeling and mass spectrometry. Bioinformatic analyses using STRING and Ingenuity Pathway Analysis identified 162 differentially expressed proteins, with unsupervised clustering highlighting cholesterol biosynthesis as a key pathway. Further validation via qPCR and upstream analysis implicated the sterol regulatory element-binding protein 2 (SREBP2) signaling axis in OPE-induced cholesterol biosynthesis. Functional assays revealed that atorvastatin-mediated HMG-CoA reductase inhibition, the rate limiting enzyme in cholesterol biosynthesis, prevents cholesterol buildup and lipid droplet formation in macrophages. These findings provide the first evidence that an environmentally relevant OPE mixture can induce cholesterol biosynthesis in human macrophages. These studies provide mechanistic evidence that an environmentally relevant mixture of organophosphate esters induces cholesterol biosynthesis in macrophages. These findings link real-world exposure to lipid pathways implicated in metabolic disease and support the need for updated regulatory standards that protect human health.

Tungsten is an emerging environmental contaminant, highlighting the urgent need to elucidate its toxicological characteristics and assess long-term health risks. Our previous investigations show that tungsten deposition in the bone is associated with stalled pre-B lymphocyte differentiation, inhibition of osteogenesis, and increased intervertebral disc degeneration and fibrosis. To delineate the underlying molecular mechanisms, we employed CRISPR-based genomics on NALM-6 cells and identified Solute Carrier Family 26 Member 2 (SLC26A2), a sulfate/chloride antiporter, as a pivotal mediator of tungsten-induced toxicity. SLC26A2 deletion reduced tungsten-induced growth inhibition and intercellular tungsten levels. Functional impairment of SLC26A2 is associated with chondrodysplasias, thus, we hypothesized that tungsten would impair the development of cartilage and bone tissues. Indeed, tungstate exposure impaired chondrogenesis and osteogenesis in murine limb cultures, which was reversible by sulfate supplementation. Our study demonstrates that tungsten exploits SLC26A2 for cellular entry and correlates with bone development disruption through proteoglycan and collagen depletion.

Relapsed and refractory diffuse large B-cell lymphoma (rrDLBCL) presents a significant challenge in hematology-oncology, with approximately 30% to 40% of patients with DLBCL experiencing relapse or resistance to treatment. This underscores the urgent need to better understand the molecular mechanisms governing therapeutic resistance. Signal transducer and activator of transcription 6 (STAT6) has been previously identified as a gene with recurrent D419 gain-of-function mutations in rrDLBCL. We demonstrated previously that when STAT6D419 mutations are present in DLBCL tumor cells, transcription of the chemokine CCL17 (aka TARC) is increased, and tumors have increased infiltration of CD4+ T cells. However, the implication of increased T-cell infiltration has not been reported previously. In the present study, we developed a mouse model of STAT6D419N mutant DLBCL that recapitulates the critical features of human STAT6D419-mutant DLBCL, including increased expression of phospho-STAT6, increased CD4+ T-cell invasion, and resistance to doxorubicin treatment. We report CD4+ T cells in STAT6D419N tumors exhibit higher expression of the receptor for CCL17, CCR4. Using ex vivo functional assays, we demonstrated that STAT6D419N tumor cells are directly chemoattractive to CCR4+ CD4+ T cells, and CCR4 inhibition using a small-molecule antagonist reduced CD4+ T-cell infiltration into STAT6D419N tumors and made STAT6D419N tumors regain therapeutic sensitivity to doxorubicin. Using PhenoCycler imaging of human rrDLBCL samples, we found that STAT6D419 tumors have increased expression of phospho-STAT6 and increased cellular interactions between phospho-STAT6+ tumor cells and CD4+/CCR4+ CD4+ T cells. Thus, our data identify CCR4 as a therapeutic target in STAT6D419-mutant rrDLBCL.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a growing global health burden and a major contributor to chronic liver disease. Evidence suggests arsenic (As) and/or cadmium (Cd) exposure may influence MASLD development, yet the effects of chronic low-dose metal mixtures on hepatic inflammation and immune responses remain unclear. Using the apolipoprotein E-knockout mouse model, we examined low-dose As and Cd exposure in male and female mice. We focused on hepatic steatosis and inflammation. Steatosis-related changes were assessed via lipid metabolism gene expression and PLIN2 levels. No significant changes were observed in males; however, females exposed to the metal mixture showed increased PLIN2 expression. In contrast, males exhibited an inflammatory phenotype following combined exposure. High-plex single-cell imaging (PhenoCycler) in male livers revealed increased Ki67+ hepatocytes, enhanced β-catenin signal, and elevated CD8+ T-cell infiltration, indicating enhanced proliferation and immune activation. These findings suggest sex-dependent responses to low-dose As and Cd, with females showing subtle steatotic changes and males a pronounced inflammatory signature. Collectively, combined metal exposure induces hepatic priming in both sexes; males show an increased inflammatory response with cellular proliferation in the absence of overt steatosis or fibrosis, whereas females demonstrate increased steatosis without inflammation or fibrosis.

Exposure to individual organophosphate esters (OPEs) has been linked to immune dysfunction. However, the effect of OPEs as environmentally relevant mixtures on the immune system remains poorly understood. This study examines how parental exposure to an OPE mixture impacts the immune status of Sprague-Dawley rats and their offspring. Sprague-Dawley rats were fed a control or OPE-supplemented diet containing 13 OPEs detected in >85% of Canadian homes. Only male offspring of OPE-exposed animals showed a significant reduction in CD43lowHis48hi splenic monocyte-macrophages. There were no significant changes in CD43lowHis48hi splenic monocyte-macrophages in the F0 generation or female offspring. However, the OPE mixture significantly altered serum cytokine levels in both sexes and generations, with females and offspring experiencing more pronounced changes. Notably, female progeny had elevated levels of chemokines associated with monocyte recruitment. In vitro follow-up studies revealed that the OPE mixture delays monocyte-to-macrophage transition and monocyte migration in both sexes. These results indicate that an environmentally relevant OPE mixture disrupts immune function by affecting monocyte recruitment and differentiation but does not reveal clear sex differences. However, when combined with cytokine findings, these results support a hypothesis that OPE exposure causes male-specific decreases in CD43lowHis48hi monocyte-macrophages that are absent in females due to compensatory inflammation. These studies demonstrate that an environmentally relevant mixture of OPEs can alter basal immune status in the offspring of exposed animals. This work will be useful for risk assessment studies and regulations protecting human health.

OBJECTIVE:To examine the association of low-level urine As with incident stroke, acute myocardial infarction (AMI), and heart failure (HF) among never smokers. METHODS:Between 1993 and 1997, the Danish Diet, Cancer and Health (DCH) cohort enrolled 19,394 participants, ages 50-64 yrs, who reported never smoking. Among these never smokers with baseline urine samples available for arsenic speciation, we identified incident cases of stroke (N = 457), AMI (N = 696), and HF (N = 803) using the Danish National Patient Registry. We also randomly selected a sub-cohort of 541 men and 548 women. We used modified Cox proportional hazards models to estimate adjusted hazard ratios (HR) for each incident CVD outcome in relation to urine As adjusted for creatinine. RESULTS:We did not find an association with stroke, AMI, or HF across unadjusted and adjusted models using our primary exposure metric, sum of As species (iAs + MMA + DMA), AsB-corrected. We also did not find evidence of an association of secondary metrics (%DMA, %MMA) with AMI or HF, although HRs were consistently elevated for upper quartile %MMA. We observed associations for stroke with upper quartile %DMA (HR = 0.57; 95 % CI: 0.40-0.82), and with third quartile %MMA (HR = 1.52; 95 % CI: 1.07-2.20) but not the upper quartile %MMA (HR = 1.20; 95 % CI: 0.85-1.70). CONCLUSIONS:We found little evidence of an association between low-level urine As species and AMI, HF, or stroke among Danish never smokers, although further research is needed to disentangle a potential cardiovascular role of %MMA.

Millions of people are exposed to concentrations of arsenic that exceed the World Health Organization's limit of 10 μg/L. Inorganic arsenic (iAsIII) is metabolized by arsenic 3 methyltransferase (As3mt) that converts it to methylated arsenicals, mono and dimethyl arsonous acid (MMA and DMA). Chronic arsenic exposure is linked to an increased risk of chronic kidney disease (CKD), however, the effects of arsenic exposure on kidney development remain unclear. We hypothesized that exposure to arsenicals impairs nephron formation during mouse kidney development. Mouse embryonic kidney explants were treated with iAsIII and MMAIII (1.5 μM or 200 μg/L). iAsIII inhibited growth of kidney explants and ureteric bud branching morphogenesis at embryonic day 11.5 (E11.5) and E12.5, but not at E13.5. Similar effects were observed when kidney explants were treated with MMAIII. Additionally, iAsIII exposure increased apoptosis in the metanephric mesenchyme of E11.5 explants and decreased Gdnf transcription. To assess the impact of iAS exposure in utero and early postnatal life, female mice harboring a humanized version of AS3MT and wild-type mice with murine As3mt were exposed to iAsIII throughout gestation and weaning and their offspring were analyzed for kidney defects. Pups with human AS3MT exposed to 1.5 μM iAsIII in utero, showed a 20 % reduction in kidney weight normalized to body weight and a 28 % reduction in nephron number, compared to kidneys of wild-type mice. In conclusion, exposure to arsenicals during embryonic development impairs ureteric bud branching morphogenesis and decreases nephron endowment, which may predispose to CKD in adulthood.

BACKGROUND:Millions worldwide are exposed to elevated levels of arsenic that significantly increase their risk of developing atherosclerosis, a pathology primarily driven by immune cells. While the impact of arsenic on immune cell populations in atherosclerotic plaques has been broadly characterized, cellular heterogeneity is a substantial barrier to in-depth examinations of the cellular dynamics for varying immune cell populations. OBJECTIVES:This study aimed to conduct single-cell multi-omics profiling of atherosclerotic plaques in apolipoprotein E knockout (ApoE-/-) mice to elucidate transcriptomic and epigenetic changes in immune cells induced by arsenic exposure. METHODS:The ApoE-/- mice were fed a high-fat diet and were exposed to either 200 ppb arsenic in drinking water or a tap water control, and single-cell multi-omics profiling was performed on atherosclerotic plaque-resident immune cells. Transcriptomic and epigenetic changes in immune cells were analyzed within the same cell to understand the effects of arsenic exposure. RESULTS:Our data revealed that the transcriptional profile of macrophages from arsenic-exposed mice were significantly different from that of control mice and that differences were subtype specific and associated with cell-cell interaction and cell fates. Additionally, our data suggest that differences in arsenic-mediated changes in chromosome accessibility in arsenic-exposed mice were statistically more likely to be due to factors other than random variation compared to their effects on the transcriptome, revealing markers of arsenic exposure and potential targets for intervention. DISCUSSION:These findings in mice provide insights into how arsenic exposure impacts immune cell types in atherosclerosis, highlighting the importance of considering cellular heterogeneity in studying such effects. The identification of subtype-specific differences and potential intervention targets underscores the significance of understanding the molecular mechanisms underlying arsenic-induced atherosclerosis. Further research is warranted to validate these findings and explore therapeutic interventions targeting immune cell dysfunction in arsenic-exposed individuals. https://doi.org/10.1289/EHP14285.

Tungsten, a transition metal with widespread applications, is increasingly recognized as an environmental contaminant with potential health impacts. While tungsten exposure has been previously associated with increased DNA damage, its specific effects on DNA repair mechanisms remain poorly understood. Here, we found that tungsten alone did not induce DNA damage in vitro, as assessed by γ-H2AX phosphorylation. However, tungsten exacerbated DNA double-strand breaks induced by genotoxic agents, delayed γ-H2AX resolution, and induced cell cycle arrest. Mechanistically, GFP-based reporter assays revealed that tungsten impairs both homologous recombination (HR) and non-homologous end joining (NHEJ), which coincided with reduced recruitment of critical repair proteins, including BRCA1, 53BP1, and ATM at DSB sites. To assess functional consequences of impaired DNA repair due to tungsten exposure, we examined immunoglobulin (Ig) class switch recombination (CSR), a critical antibody diversification process reliant on the repair of DSBs. Using the CH12F3 B cell line, we showed that tungsten significantly impaired cytokine stimulated-CSR from IgM to IgA without affecting cell proliferation or activation-induced cytidine deaminase (Aicda) expression. Furthermore, ex vivo activation of splenic B cells confirmed that tungsten exposure inhibits CSR from IgM to IgG1, independent of cell proliferation. Together, these data indicate that tungsten exposure impairs canonical DSB repair pathways and CSR, highlighting the potential consequences for immune function upon environmental or occupational exposure to tungsten.

Cardiovascular diseases (CVDs), the leading cause of arsenic-related morbidity and mortality, are a major public health concern in many countries, including Bangladesh. Understanding the mediators of increased CVD risk associated with arsenic exposure is decisive. Fractalkine (CX3CL1), a CX3C chemokine with both chemotactic and adhesive functions, acts by interacting with its specific receptor, CX3CR1. Fractalkine is implicated in atherosclerosis, leading to CVDs. However, the association between chronic arsenic exposure and serum fractalkine levels and its implication in CVDs have not yet been documented. The present study aimed to investigate the relationship between arsenic exposure and serum fractalkine levels, especially regarding the risk of CVDs. The study included 413 participants from low- and high-arsenic exposure rural areas in Bangladesh. Fractalkine levels in serum were measured by immunoassay, and the risk of CVDs was assessed by measuring the serum high-density lipoprotein cholesterol (HDL-C) and hypertension. We found that participants from high-exposure areas had nearly twice the serum fractalkine levels compared to those in low-exposure area (p < 0.001). A one-unit increase of log2-transformed water, hair, and nail arsenic showed significantly higher propensity score adjusted odd ratios (aORs) in the second and third tertiles of serum fractalkine in comparison to the first tertile. Serum fractalkine levels were inversely associated with HDL-C levels and positively linked to blood pressure and ORs of hypertension. Finally, the associations between arsenic exposure and hypertension were found to be mediated by serum fractalkine. These results suggest that arsenic-exposure-related increases in fractalkine levels may be implicated in the pathogenesis of CVDs.

Background: Diet is known to impact cardiovascular disease (CVD) risk, but evidence for the essential minerals of magnesium (Mg), calcium (Ca), and potassium (K) is inconsistent. Methods: We conducted a case–cohort study within a non-smoking subgroup of the Danish Diet, Cancer and Health cohort, a prospective study of 50–64-year-olds recruited between 1993–1997. We identified incident heart failure (HF), acute myocardial infarction (AMI) and stroke cases through 2015 with an 1135-member subcohort. We measured the dietary intake of minerals, also known as elements, and calculated a combined dietary intake (CDI) score based on joint Ca, Mg and K intakes (mg/d) from Food Frequency Questionnaires. We estimated adjusted hazard ratios (HRs) with Cox proportional hazard models. Results: Most HRs examining associations between CDI score and CVD were null. However, the third quartile of CDI was associated with a lower risk for heart failure (HR: 0.89; 95% CI: 0.67, 1.17), AMI (HR: 0.79; 95% CI: 0.60, 1.04), and stroke (HR: 0.63; 95% CI: 0.44, 0.88). Conclusions: We did not find consistent evidence to suggest that higher levels of essential minerals are associated with incident HF, AMI, and stroke, though results suggest a potential U-shaped relationship between select minerals and CVD outcomes.

Worldwide, there is an increasing prevalence of hepatic diseases. The most common diseases include alcoholic-associated liver disease (ALD), metabolic dysfunction-associated fatty liver disease/ metabolic dysfunction-associated steatohepatitis (MAFLD/MASH) and viral hepatitis. While there are many important mediators of these diseases, there is increasing recognition of the importance of the inflammatory immune response in hepatic disease pathogenesis. Hepatic inflammation triggers the onset and progression of liver diseases. Chronic and sustained inflammation can lead to fibrosis, then cirrhosis and eventually end-stage cancer, hepatocellular carcinoma. Importantly, growing evidence suggest that metal exposure plays a role in hepatic disease pathogenesis. While in recent years, studies have linked metal exposure and hepatic steatosis, studies emphasizing metal-induced hepatic inflammation are limited. Hepatic inflammation is an important hallmark of fatty liver disease. This review aims to summarize the mechanisms of arsenic (As), cadmium (Cd) and chromium (Cr)-induced hepatic inflammation as they contribute to hepatic toxicity and to identify data gaps for future investigation.

Multi-omics datasets present unrivaled opportunities to understand complex biological processes from comprehensive perspectives, but the distinct feature spaces associated with different modalities pose challenges in data integration and downstream analyses. Here we report scCross, an algorithm based on the variational that can integrate the features from different modalities to learn joint cell embeddings. We applied scCross to integrate dualomics and triple-omics data. scCross is able to map all data modalities to a shared latent space, allowing for various downstream data analytic tasks. scCross outperforms existing methods for data integration and is able to generate single-cell data across modalities. Moreover, scCross can perform multi-omics simulation and infer intra-modal and inter-modal perturbations to manipulate the cellular states, which can be leveraged to discover interventions and therapeutics against various diseases.

Metal exposure is linked to numerous pathological outcomes including cancer, cardiovascular disease, and diabetes. Over the past decades, we have made significant progress in our understanding of how metals are linked to disease, but there is still much to learn. In October 2022, experts studying the consequences of metal exposures met in Montréal, Québec, to discuss recent advances and knowledge gaps for future research. Here, we present a summary of presentations and discussions had at the meeting.

Despite being designed for smoking cessation, e-cigarettes and their variety of flavors have become increasingly attractive to teens and young adults. This trend has fueled concerns regarding the potential role of e-cigarettes in advancing chronic diseases, notably those affecting the cardiovascular system. E-cigarettes contain a mixture of metals and chemical compounds, some of which have been implicated in cardiovascular diseases like atherosclerosis. Our laboratory has optimized in vivo exposure regimens to mimic human vaping patterns. Using these established protocols in an inducible (AAV-PCSK9) hyperlipidemic mouse model, this study tests the hypothesis that a chronic exposure to e-cigarette aerosols will increase atherosclerotic plaques. The exposures were conducted using the SCIREQ InExpose™ nose-only inhalation system and STLTH or Vuse products for 16 weeks. We observed that only male mice exposed to STLTH or Vuse aerosols had significantly increased plasma total cholesterol, triglycerides, and LDL cholesterol levels compared to mice exposed to system air. Moreover, these male mice also had a significant increase in aortic and sinus plaque area. Male mice exposed to e-cigarette aerosol had a significant reduction in weight gain over the exposure period. Our data indicate that e-cigarette use in young hyperlipidemic male mice increases atherosclerosis in the absence of significant pulmonary and systemic inflammation. These results underscore the need for extensive research to unravel the long-term health effects of e-cigarettes.

Background The tumour microenvironment (TME) consists of tumour-supportive immune cells, endothelial cells, and fibroblasts. PhenoCycler, a high-plex single cell spatial biology imaging platform, is used to characterize the complexity of the TME. Researchers worldwide harvest and bank tissues from mouse models which are employed to model a plethora of human disease. With the explosion of interest in spatial biology, these panoplies of archival tissues provide a valuable resource to answer new questions. Here, we describe our protocols for developing tunable PhenoCycler multiplexed imaging panels and describe our open-source data analysis pipeline. Using these protocols, we used PhenoCycler to spatially resolve the TME of 8 routinely employed pre-clinical models of lymphoma, breast cancer, and melanoma preserved as FFPE. Results Our data reveal distinct TMEs in the different cancer models that were imaged and show that cell-cell contacts differ depending on the tumour type examined. For instance, we found that the immune infiltration in a murine model of melanoma is altered in cellular organization in melanomas that become resistant to αPD-1 therapy, with depletions in a number of cell-cell interactions. Conclusions This work presents a valuable resource study seamlessly adaptable to any field of research involving murine models. The methodology described allows researchers to address newly formed hypotheses using archival materials, bypassing the new to perform new mouse studies.