Abstract Obesity is associated with gut dysbiosis, chronic inflammation, and insulin resistance. We assessed the proportional change in fecal microbial populations in a pilot study (n = 34) of peri/postmenopausal women with a body mass index ≥28 kg/m2 who were randomized to receive either 3.25 g/day of omega-3 fatty acids or a placebo during a weight loss intervention. Body composition was assessed using dual X-ray absorptiometry, and fecal and blood samples were collected. The median weight change was −10%. Among participants who lost ≥10% of their weight, those assigned to omega-3 fatty acids showed the greatest decrease in the Firmicutes:Bacteroidetes ratio and displayed favorable changes in systemic biomarkers. Notable increases in the proportional abundance of short-chain fatty acid (SCFA)–producing microbes including Phocaeicola vulgatus and Alistipes putredinis were observed in women receiving omega-3, which correlated with improvements in breast cancer biomarkers such as bioavailable estradiol, adiponectin:leptin ratio, and C-reactive protein levels. Women administered omega-3 fatty acids displayed increased % change in plasma SCFA propionate and decreased butyrate, suggesting that intervention differentially modulated circulating bacteria-derived SCFA metabolites. High-dose omega-3 fatty acids, when added to a behavioral weight loss intervention, promoted beneficial shifts in the gut microbiome and associated with improved breast cancer risk factor biomarkers. Prevention Relevance: Obesity is a modifiable risk factor for breast cancer, characterized by chronic inflammation and altered adipokines. This trial addresses the need to enhance weight loss by targeting underlying metabolic and inflammatory drivers. We show that omega-3 polyunsaturated fatty acids (eicosapentaenoic acid/docosahexaenoic acid) shift SCFA-producing microbiota, increase propionate, and correlate with improved breast cancer risk factor biomarkers.

Objective This study aimed to assess the spectrum and frequency of adverse events (AEs) linked to glucagon-like peptide-1 receptor agonists (GLP-1RAs) using the US FDA Adverse Event Reporting System (FAERS). Emphasis was placed on emerging safety concerns in context-specific use.Methods A retrospective analysis of FAERS reports between 2012 and 2025 was conducted. Five commonly prescribed FDA-approved GLP-1RAs were included. Disproportionality analyses were applied to detect AE signals. Subgroup analyses evaluated associations by indication, GLP-1RAs compared to other drugs, and AEs specific to individual GLP-1RAs.Results From over 18 million FAERS reports, 137,451 involved GLP-1RAs. The most frequent AEs were gastrointestinal, nutritional and metabolic, and psychiatric disorders, occurring at higher rates compared to other drugs. In diabetes use, GLP-1RAs were associated with retinopathy, hearing loss, and cataracts. In contrast, when prescribed for weight management/obesity, nutritional, metabolic, and psychiatric AEs predominated. We also developed an open-access portal for AE exploration, available at http://glp1.tanlab.org.Conclusions GLP-1RAs are linked to a broad range of AEs across indications. These findings stress the need for careful clinical monitoring and long-term safety evaluation. This study also illustrates how real-world evidence can inform safety communications, as well as hypothesis generation for research on next-generation GLP-1RAs.

Supplementary Table S2. Representativeness of Study Participants

A discovery in mice reveals why fasting enhances a type of breast cancer treatment — a hormone-signalling pathway and gene-expression changes have key roles. A discovery in mice reveals why fasting enhances a type of breast cancer treatment — a hormone-signalling pathway and gene-expression changes have key roles.

Supplementary Figure S4. α and β diversity indexes in patients by menopause and hormone replacement therapy use in the 6-month fecal samples. A. Shannon index B. Bray-Curtis β-diversity principal coordinate analysis (PCoA).

Obesity alters systemic metabolism and immune function, yet how obesity and tumor progression regulate extracellular vesicle (EV) composition and function within the tumor microenvironment remains unclear. Using a preclinical model of diet-induced obesity (DIO) and triple-negative breast cancer (TNBC), we investigated how obesity and tumor stage shape the proteomic composition of EVs from visceral adipose tissue (VAT-EVs) and mammary tumors (tumor-EVs), and how these EVs regulate immune and tumor cell metabolism. Orthotopically transplanted metM-Wnt lung tumors were classified as early (∼0.5 cm³) or late (∼1.0 cm³), and EV proteomes were analyzed by mass spectrometry. At early stages, tumor-EVs from DIO mice, compared with control lean mice, were depleted in immune-related proteins, whereas VAT-EVs were enriched in mitochondrial and fatty acid oxidation proteins. In contrast, at later stages, tumor-EVs from DIO mice were enriched in lipid metabolism and oxidative stress-associated proteins, while VAT-EVs exhibited loss of mitochondrial proteins consistent with metabolic dysfunction. Functionally, tumor-EVs and VAT-EVs differentially regulated CD8 T cell mitochondrial activity and cytokine production and induced distinct, stage-dependent metabolic reprogramming in non-aggressive epithelial-like (E-Wnt) versus mesenchymal-like (M-Wnt) tumor cells. These findings suggest that obesity and tumor progression dynamically reshapes EV cargo, enabling EV-mediated metabolic reprogramming that may contribute to immune suppression and TNBC progression.

Supplementary Table S1. Baseline characteristics for women in the weight loss intervention who had 6- month fecal samples available for analysis. Mean ± SD; (range).

Chronic choline insufficiency reprograms hepatic metabolism and drives insulin resistance independent of obesity. While complete choline deficiency causes liver injury, the metabolic consequences of sustained, suboptimal intake, observed in ∼90% of US adults, remain poorly defined. Here, we used integrated lipidomic, metabolomic, and transcriptomic profiling to determine how graded choline intake (0.5, 1.4, or 6.3 g/kg) regulates hepatic metabolism during a control (Con) or high-fat (HF) diet-induced obesity regimen. Under Con diets, low choline intake induced a distinct metabolic state characterized by remodeled hepatic lipid architecture, particularly within triglyceride and glycerolipid species, without altering bulk triglyceride accumulation. Mechanistically, low choline disrupted phospholipid balance and induced a coordinated, sex-dependent transcriptional response, identifying ethanolamine-phosphate phospho-lyase (ETNPPL) and the fatty acid transporter CD36 as top choline-responsive genes. These metabolic effects were unique to the Con low choline group, as a high-fat diet masked all choline-dependent variations. Specifically, ETNPPL protein abundance increased under low choline Con conditions in males but not females. Functionally, this sustained restriction led to progressive hyperglycemia and insulin resistance exclusively in male mice, whereas females remained metabolically protected. Together, these findings demonstrate that chronic choline restriction remodels hepatic lipid metabolism in the absence of obesity and define a CD36-ETNPPL axis linking choline availability to sex-specific insulin resistance.

Supplementary Figure S3. α and β diversity indexes in patients with matched baseline and 6-month fecal samples. A. Shannon index B. Bray-Curtis β-diversity principal coordinate analysis (PCoA).

This current framework (version 3) updates the previous one published within the WCRF Gateway on Health Open Research (publication data of version 2: 23 September 2025). The purpose of the Framework is to provide guidance on how to: identify, review, and summarise the biological mechanisms that underpin the associations between diet, nutrition, body weight, and physical activity and cancer incidence found within CUP Global assess the strength of evidence that a given biological mechanism operates between an exposure and outcome of interest These biological plausibility investigations apply to 1) exposures with an established link to cancer (strong evidence in CUP/CUP Global) and for which there are strong mechanistic data, but further work is needed (for example, a poorer understanding of the evidence for certain cancer sites); 2) exposures with a strong epidemiological link to cancer (strong evidence in CUP/CUP Global) but with limited mechanistic understanding; 3) exposures with a putative epidemiological link to cancer (limited suggestive evidence in the CUP/CUP Global) for which more robust mechanistic data would strengthen conclusions. Note that mechanisms evaluated using this Framework are defined as biological processes linking diet, nutrition, body weight, and physical activity and cancer incidence. The focus is primarily on molecular mechanisms but may expand to physiological mechanisms depending on the research question.

Obesity affects over 40% of women in the US and increases the risk and progression of several cancers, including triple-negative breast cancer (TNBC), in part through chronic low-grade inflammation and impaired antitumor immunity. While weight loss can reverse obesity-driven cancer risk, cost and other factors limit the accessibility of effective weight loss interventions. This study investigated whether sulindac, a nonsteroidal anti-inflammatory drug (NSAID), could mitigate obesity-driven TNBC progression. Using multiple preclinical models, we demonstrate that sulindac treatment abrogates obesity-accelerated tumor growth and metastasis without affecting body weight or composition. Bulk transcriptomic profiling revealed obesity-driven suppression of immune-related gene signatures in the tumor microenvironment (TME)—including antigen presentation—while sulindac treatment restored these signatures. Single-cell RNA sequencing identified sulindac-mediated reprogramming of tumoral metabolism toward oxidative phosphorylation and restoration of antigen presentation machinery in tumor-associated macrophages. Sulindac also reversed obesity-driven reduction in T cell receptor diversity within the TME. We conclude that sulindac treatment remodels the TME and restores obesity-associated impairments of immunosurveillance, offering a potentially accessible intervention to limit obesity-driven TNBC progression. We demonstrate that NSAIDs, which are generally safe, cheap, and readily available, limit the burden of obesity-driven TNBC preclinically, warranting further evaluation as a targeted clinical intervention.

Metabolomic profiles are increasingly being used to identify responders to dietary interventions. Advances using this approach are particularly needed to personalize and enhance the effectiveness of dietary weight loss interventions. Using obese Diversity Outbred (DO) mice that model genetic and phenotypic heterogeneity of human populations, we aimed to identify urinary metabolite signatures associated with responsiveness to calorie restriction (CR)-mediated weight loss. DO mice (150 males, 150 females) were fed a high-fat diet for 12 weeks to induce obesity, then urine was collected and an 8-week CR regimen (30% decrease in energy intake) initiated. At study completion, mice were rank-ordered according to their percent body weight change, with mice in the extreme quartiles deemed CR responders (n = 67) versus nonresponders (n = 67). Targeted semi-quantitative metabolomics identified elevated glutamic acid and hydroxyproline as key urinary metabolites that distinguish CR responders from CR nonresponders, independent of sex. Three urinary metabolites (glutamic acid, hydroxyproline, and putrescine) distinguished male CR responders from nonresponders. Six metabolites (glutamic acid, hydroxyproline, dopamine, histamine, lysine, and spermine) distinguished female CR responders from nonresponders. Multivariate receiver operating characteristic analyses integrated these metabolites to reveal potential sex specific and sex-independent associations of CR-mediated weight loss. Further, pathway analysis identified several metabolic pathways, including arginine and proline metabolism, and alanine, aspartate, and glutamate biosynthesis, that distinguished CR responders from nonresponders and could be indicative of metabolic reprogramming to enhance insulin sensitivity and energy metabolism.

BACKGROUND:Palmitoleic acid (POA) is an n-7 monounsaturated fatty acid. Preclinical studies suggest cis-POA lowers inflammation and improves metabolism. However, the impact of POA supplementation on inflammatory/metabolic biomarkers in humans is not well understood. OBJECTIVES:The primary aim was to investigate if cis-POA lowers circulating high-sensitivity C-reactive protein (hs-CRP) relative to placebo. Secondary endpoints were interleukin-6, tumor necrosis factor-α, fasting glucose, insulin, glycosylated hemoglobin, low-density lipoprotein cholesterol, red blood cell (RBC), and plasma fatty acid concentration. Exploratory endpoints were total cholesterol, high-density lipoprotein cholesterol, triglycerides, leptin, ghrelin, peptide YY, and adiponectin. METHODS:The randomized double-blinded parallel arm trial enrolled 123 participants with hs-CRP concentrations of 2 mg/L or higher. Participants consumed 500 mg/d or 1000 mg/d of marine-source POA or placebo for 12 wk. Fasting blood draws were used to quantify plasma inflammatory/metabolic biomarkers at baseline and 12-wk by multiplex immunoassays. Plasma and RBC POA concentrations were quantified with gas chromatography. Dietary intake was assessed with the Nutrition Data System for Research. Analysis of covariance analysis was used to investigate if there is an effect of either baseline or dosage on inflammatory/metabolic biomarkers. RESULTS:At baseline, all 3 groups had similar hs-CRP concentrations [geometric mean (standard deviation) = 0.57 (0.17), 0.54 (0.20), and 0.53 (0.23)] at 1000 mg/d, 500 mg/d, and placebo, respectively. There were no changes in hs-CRP concentration within and between the 3 groups in response to the supplementation. Analysis of covariance analysis showed there was no significant influence of baseline hs-CRP or POA dosage on changes in hs-CRP. There were also no changes in secondary or exploratory endpoints in response to placebo or POA. Significant changes were measured in select plasma and RBC fatty acids that were significantly related to POA dosage (P < 0.001) but not baseline. CONCLUSIONS:Compared to placebo, supplementation for 12 wk with 500 mg/d or 1000 mg/d POA did not significantly lower hs-CRP or change other biomarkers.

Breast cancer is the most common cancer among women, and metastasis is the leading cause of mortality. It is still unknown how breast cancer cells metabolically adapt to successfully metastasize to different organs to survive adverse conditions, including varying nutrient availability. The purpose of this study is to elucidate the metabolic characteristics and glucose adaptation mechanisms of breast cancer cells that preferentially metastasize to the lungs or the liver. Using a Wnt-driven breast cancer model with preferential metastasis to lung (metM-WntLung) or liver (metM-WntLiver), we measured 14C-glucose uptake, 13C6-glucose metabolic flux, metabolic enzyme levels, and cell viability under normal (5 mM), high (25 mM), and low (1 or 0 mM) glucose conditions. Under normal glucose conditions, metM-WntLung cells were more glycolytic, exhibiting greater flux of 13C6-glucose-derived carbons into glycolytic intermediates, such as pyruvate and lactate. In contrast, metM-WntLiver cells favored oxidative phosphorylation, with higher levels of 13C6-glucose-derived carbons in tricarboxylic acid (TCA) cycle metabolites such as oxaloacetate indicative of higher pyruvate carboxylase (PC) activity. Exposure to high glucose reduced metM-WntLiver cell viability, with no effect on metM-WntLung cells, suggesting better adaptability of metM-WntLung cells to glucose excess. This was accompanied by increased PC activity and oxidative phosphorylation in metM-WntLung cells, whereas metM-WntLiver cells shifted to a more glycolytic phenotype. Under glucose deprivation, metM-WntLung cells were more viable than metM-WntLiver cells, suggesting that metM-WntLung cells have better adaptability to glucose deprivation. Inhibiting phosphoenolpyruvate carboxykinase, a key enzyme in gluconeogenesis, reduced metM-WntLung cell viability compared to metM-WntLiver cells. Similarly, inhibiting catabolism of glutamine, a gluconeogenic substrate, decreased metM-WntLung cell viability compared to metM-WntLiver cells, indicating that metM-WntLung cells rely on more on gluconeogenesis and glutamine metabolism under glucose deprivation. Our findings reveal that metM-WntLung cells exhibit greater metabolic flexibility to glucose than metM-WntLiver cells by shifting from glycolysis to oxidative phosphorylation under high glucose conditions while utilizing gluconeogenesis and glutamine under glucose deprivation conditions.

INTRODUCTION:Triple-negative breast cancer (TNBC), which tends to be more advanced when diagnosed and more aggressive than other breast cancer subtypes, is accelerated by obesity. Hypertrophic adipocytes and cancer cells exhibit increased oxidative stress and altered redox homeostasis, influencing therapeutic outcomes. Enzymes implicated in both redox regulation and TNBC include glutathione peroxidase 4 (GPX4; reduces lipid peroxides) and pyruvate carboxylase (PC; essential in oxidative stress protection). Using preclinical models, we characterized interactions between GPX4, PC, and oxidative stress in TNBC cells, and established effects of GPX4 suppression on TNBC progression. In TNBC cells, PC knockdown increased GPX4 expression, while GPX4 knockdown increased PC expression. GPX4 inhibition by erastin or RSL3 enhanced TNBC cell death in vitro, and antioxidants mitigated the cytotoxicity. In obese mice, GPX4 knockdown, versus scramble control: (i) reduced tumor burden following orthotopic transplantation of TNBC cells; and (ii) reduced lung metastasis following tail vein injection of TNBC cells in combination with chemotherapy (carboplatin) but not immunotherapy (anti-CTLA4 plus anti-PD1). We conclude that GPX4 and PC expression are inversely related in TNBC cells, and GPX4 and obesity interact to impact TNBC progression and treatment responses. Moreover, GPX4-mediated redox defense, alone or in combination with chemotherapy, is a targetable vulnerability for treating TNBC, including obesity-related TNBC. IMPLICATION:GPX4 suppression, alone or with current TNBC therapies, impacts outcomes in preclinical TNBC models with or without obesity and offers a new, plausible mechanistic target for TNBC treatment.

Metastasis is the primary cause of breast cancer-related deaths. We investigated the metabolic adaptations of glucose, a major energy substrate, in breast cancer cells that metastasize to lung versus liver to determine the metabolic adaptations that may support site-specific metastasis. Utilizing a breast cancer model that preferentially metastasizes to the lung (metM-WntLung cells; MLg) or liver (metM-WntLiver cells; MLr), we measured 14C-glucose uptake, 13C6-glucose flux into metabolic pathways, and viability under varying glucose concentrations. Results show that in 5 mM glucose (normal), MLg is more glycolytic with greater flux of 13C6-glucose-derived carbons into the glycolytic intermediates pyruvate and lactate in comparison to MLr. In contrast, MLr showed higher levels of 13C6-glucose-derived carbons in the tricarboxylic acid cycle metabolites including oxaloacetate and malate indicative of higher oxidative phosphorylation and higher pyruvate carboxylase (PC) activity. Interestingly, high glucose (25 mM) exposure reduced viability of MLr by 39%, suggesting MLg’s adaptability to varying glucose levels. Although glucose uptake similarly increased in all cell lines under high glucose, flux results showed greater oxidative phosphorylation and increased PC activity in MLg, while MLr shifted to a more glycolytic phenotype. These results were consistent with improved oxidative stress protection in high glucose in MLg, but not MLr. Further, MLg deprived of glucose were more viable (9%) compared to MLr, suggesting a potential alternate use of other substrates to overcome glucose deprivation. Inhibiting the rate-limiting enzyme for gluconeogenesis, phosphoenolpyruvate carboxykinase, reduced viability of MLg by 9% compared to MLr. Further, inhibiting the catabolism of glutamine, a gluconeogenic substrate, reduced viability of MLg by 13% compared to MLr, suggesting that MLg utilizes the contribution of glutamine and gluconeogenesis in conditions of low glucose. Overall, breast cancer cells with preferential metastasis to lung are more adaptable to varying glucose concentrations than breast cancer cells with preferential metastasis to liver by shifting metabolism from glycolysis to oxidative phosphorylation under high glucose conditions and utilizing glutamine and gluconeogenesis under low glucose conditions. Marjorie Anne A. Layosa, Mike Wendt, Stephen Hursting, Dorothy Teegarden. Breast cancer cells that preferentially metastasize to lung are more adaptable to varying glucose levels compared to those that metastasize to the liver [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 5392.

BACKGROUND:Predictors of premature death and cancer development are needed to more precisely identify individuals who may warrant preventive intervention. Circulating insulin-like growth factor (IGF)-binding protein-7 (IGFBP7) and, to a lesser extent, the IGFBP7/IGF-1 ratio are emerging biomarkers of renal and cardiovascular morbidity. However, their relationships with aging, obesity, mortality, and cancer risk remain unclear. METHODS:This hypothesis-generating study investigated plasma IGFBP7, IGF-1, and their ratio as predictors of all-cause mortality and the incidence of any cancer (excluding nonmelanoma skin cancer), obesity-related cancer (composite of 13 cancer types), and breast cancer in a large longitudinal cohort of postmenopausal women. We assessed the relationships of each biomarker with age, body mass index, and each outcome (bivariately and controlling for age, body mass index, race, physical activity, education, income, marital status, alcohol intake, smoking, diabetes, and hormone therapy) in 793 Women's Health Initiative Observational Study participants (mean, 19.4-year follow-up). RESULTS:In adjusted analyses, IGFBP7 increased with age and obesity and was positively associated with risks of all-cause mortality [HR = 2.42 (95% confidence interval, 1.37-4.26); P = 0.002], any cancer [HR = 2.04 (1.05-3.94); P = 0.035], and obesity-related cancer [HR = 1.58 (0.99-2.51); P = 0.053]. Also in adjusted analyses, the IGFBP7/IGF-1 ratio increased with age and was positively associated with all-cause mortality [HR = 1.51 (1.14-1.99); P = 0.004] and any cancer incidence [HR = 5.44 (1.13-26.1); P = 0.034]. CONCLUSIONS:Plasma IGFBP7 and the IGFBP7/IGF-1 ratio are positively associated with age, obesity (IGFBP7 only), mortality, and cancer in postmenopausal women. IMPACT:Plasma IGFBP7 may represent an age- and obesity-sensitive biomarker of increased risk of developing cancer and/or dying prematurely.