High-fat High-sucrose Research Articles

OR18-03-23 Maternal Methyl Donor Nutrients Supplementation to a High-Fat High-Sucrose Diet Modulates Systemic Vitamin D Signaling and Renal Inflammation Among Adult Offspring

OR18-03-23 Maternal Methyl Donor Nutrients Supplementation to a High-Fat High-Sucrose Diet Modulates Systemic Vitamin D Signaling and Renal Inflammation Among Adult Offspring

1544-P: Circadian Timing of SGLT2 Inhibitor Administration Modulates Therapeutic Effects on Pancreatic Islet Function, Composition, and Gene Expression

Timing of drug administration based on endogenous circadian cycles (i.e., chronotherapy) has been shown to enhance treatment efficacy and improve outcomes in type 2 diabetes (T2DM). Sodium-Glucose Co-transporter 2 inhibitors (SGLT2i) are potent glucose lowering agents which improve cardiovascular health and exert systemic benefits on metabolic organs (e.g., pancreatic islets). Systemic benefits of SGLT2i are attributed to their ability to mimic a fasting state thus raising a question whether chronotherapeutic exposure to SGLT2i timed to the fasting vs. feeding circadian cycle will produce divergent benefits in T2DM. To address this, we generated a diet-induced T2DM mouse model by exposing male C57BL/6 mice to 14 weeks of high fat/high sucrose (HFHS) diet. Following induction of diabetes, mice were assigned into 3 groups for 4 weeks: 1) DAPAFAST: mice with daily Dapagliflozin (DAPA) oral gavage (1 mg/kg) given at the onset of the fasting (FAST) circadian cycle; 2) DAPAFED: daily DAPA oral gavage at the onset of the feeding (FED) cycle; 3) DAPAVEH: daily vehicle (VEH) gavage. DAPAFAST and DAPAFED groups demonstrated comparable lowering of basal and OGTT glucose versus DAPAVEH (~30%, p<0.05). In contrast, whereas improved glucose tolerance in DAPAFAST was associated with enhanced insulin secretion (p<0.05 vs. DAPAVEH), no changes in β-cell function was noted in DAPAFED mice. Consistently, RNAseq analysis of DAPAFAST isolated islets revealed attenuated expression of genes enriched for key pathways regulating β-cell failure such as p53, FOXO, and TGFβ signaling (all p<0.05). We also noted divergent effects on islet morphology with DAPAFAST group uniquely characterized by increased α-cell mass compared to DAPAFED and DAPAVEH (~70% increase, p<0.05). These data demonstrate divergent metabolic and islet effects of SGLT2i therapy highlighting the importance of chronotherapeutic approaches for optimal treatment of T2DM. Disclosure M.Dhanasekaran: None. K.Rakshit: None. T.K.Her: None. A.Matveyenko: None.

1561-P: Dietary Proteins Impact Glucose Tolerance and Hepatic Insulin Resistance in Mice through Modulation of the Urea Cycle

We recently reported that dietary proteins (DP) modulate diet-induced obesity and insulin resistance in mice (Nat Commun 12: 3377, 2021). We posit that these metabolic alterations are caused by differential activation of the hepatic urea cycle in response to different amino acid (AA) compositions and total nitrogen load. Objective: To test whether DP sources alter hepatic metabolism and insulin resistance through the urea cycle. Methods: Metabolic phenotypes, hepatic urea cycle and insulin resistance were measured in mice fed low-fat, low-sucrose (LFLS) or high-fat, high-sucrose (HFHS) diets containing either pork (PP), soy (SP) or casein (CP) protein for 12 weeks. Based on their AA composition, PP had the highest nitrogen load, followed by SP and CP. Results: When compared to SP and CP, feeding PP to HFHS mice induced more glucose intolerance which was correlated with higher post-prandial AA-derived nitrogen load in the portal vein. PP feeding in HFHS mice induced hepatic insulin resistance, revealed by impaired insulin-induced Akt phosphorylation on Ser473. Mice fed the LFLS diet were, however, protected from PP-induced glucose intolerance and insulin resistance, and this was associated with robust increase in the protein levels of urea cycle enzymes (CPS1, OTC) in the liver. In starked contrast, induction of urea cycle enzymes was blunted in the liver of PP-fed HFHS mice. PP feeding also increased the expression of inducible nitric oxide synthase (iNOS) and reactive oxygen species (ROS) as showed by DHE staining of liver sections. Conclusion: Dietary PP increases total nitrogen load activating the urea cycle, which may protect against metabolic dysfunction and hepatic insulin resistance in LFLS mice. This protective mechanism is lost in PP-fed HFHS mice, likely by downregulation of urea cycle activity, when the liver is overloaded with dietary fat. iNOS induction could be acting as an alternative nitrogen handling pathway, causing a build-up of mitochondrial ROS. Disclosure F.Bégin: None. A.Caron: None. A.Marette: Advisory Panel; Valbiotis, Amancia, Plexus, Acasti.

1664-P: High Fat High Sucrose Exposure during Lactation but Not during the Post-Weaning Period Programs Impaired Glucose Homeostasis

Nutrition during critical developmental windows plays a pivotal role in shaping long-term metabolic health. Exposure to a Western diet in early life may lead to altered metabolic programing, potentially influencing the risk to develop obesity and type 2 diabetes. In particular, an early exposure to an obesogenic diet may induce a metabolic program that only emerges upon dietary challenges later in life. Accordingly, understanding the long-term impact of nutritional insults in different stages of early life may provide strategies for primary prevention. Therefore, the present study aimed to determine which time window in the post-natal period is most critical for long-term metabolic health. To this end, pups born to lean mouse dams were exposed to a high-fat-high-sucrose (HFHS) diet for 3 weeks during lactation or in the post-weaning period. Thereafter, mice were either fed a regular chow diet until the age of 30 weeks or exposed to a second bout of HFHS diet for the last 12 weeks. Metabolic health and development of obesity was assessed by regular monitoring of glucose homeostasis and body weight gain. At the age of 12 weeks, mice with lactational exposure to a HFHS diet revealed a significantly impaired glucose tolerance compared to control mice, while glucose tolerance of post-weaning exposed mice was similar to the control group. Moreover, a second bout of HFHS diet impaired glucose tolerance and increased body and adipose tissue weight in mice with lactational exposure to a significant greater extent than in post-weaning exposed mice. Preliminary data suggests that increased adipose tissue inflammation might be mechanistically involved in impaired glucose tolerance of lactation exposed mice. Hence, exposure to HFHS during lactation but not during the post-weaning period programs impaired glucose homeostasis, suggesting that the lactational window may be the most critical in terms of metabolic programing. Disclosure A.Goergen: None. S.Wueest: None. D.Konrad: Research Support; Novo Nordisk A/S.

1673-P: Metabolic Outcomes and Changes in Innate Immunity Induced by Diesel Exhaust Particles Airway Exposure and High-Fat High-Sucrose Diet

Epidemiological studies have shown that exposure to diesel exhaust particles (DEP) is associated with metabolic diseases. We used mice with glucose intolerance and nonalcoholic fatty liver disease (NAFLD) caused by a high-fat, high-sucrose diet (HFHSD), which mimics a Western diet, to investigate the mechanism of metabolic diseases exacerbation via changes in innate immunity in the lungs by airway exposure to DEP. Six-week-old C57BL6/J male mice were fed HFHSD, and DEP was administered endotracheally once a week for eight weeks. Chronic exposure to DEP in HFHSD-fed mice increased inflammatory cells involved in innate immunity, such as type1 innate lymphoid cells (ILC1s), ILC2s, ILC3s, and M1 macrophages in the lungs and raised local inflammatory cytokine levels. This inflammation spread throughout the body, suggesting the association with the progression of NAFLD via increased inflammatory cells involved in innate immunity, such as ILC1s, ILC3s, M1 macrophages, and natural killer cells in the liver, and inflammatory cytokine levels in the liver which in turn exacerbated glucose intolerance. These findings contribute to a better understanding of the role of innate immunity in air pollution-related systemic diseases, especially metabolic diseases. Disclosure Y.Hasegawa: None. T.Okamura: Research Support; Yamada Bee Company, Inc. M.Hamaguchi: Research Support; Yamada Bee Company, Inc.. M.Fukui: None. H.Takano: None. Funding Japan Science and Technology Agency (JPMJCR19H3)

Voluntary physical activity modulates self-selection of a high-caloric choice diet in male Wistar rats

Physical exercise training has been positioned as a behavioral strategy to prevent or alleviate obesity via promotion of energy expenditure as well as modulation of energy intake resulting from changes in dietary preference. Brain adaptations underlying the latter process are incompletely understood. Voluntary wheel running (VWR) is a self-reinforcing rodent paradigm that mimics aspects of human physical exercise training. Behavioral and mechanistic insight from such fundamental studies can help optimize therapies that improve body weight and metabolic health based on physical exercise training in humans.To assess the effects of VWR on dietary self-selection, male Wistar rats were given access to a two-component “no-choice” control diet (CD; consisting of prefabricated nutritionally complete pellets and a bottle with tap water) or a four-component free-choice high-fat high-sucrose diet (fc-HFHSD; consisting of a container with prefabricated nutritionally complete pellets, a dish with beef tallow, a bottle with tap water, and a bottle with 30% sucrose solution). Metabolic parameters and baseline dietary self-selection behavior during sedentary (SED) housing were measured for 21 days, after which half of the animals were allowed to run on a vertical running wheel (VWR) for another 30 days. This resulted in four experimental groups (SEDCD, SEDfc-HFHSD, VWRCD, and VWRfc-HFHSD). Gene expression of opioid and dopamine neurotransmission components, which are associated with dietary self-selection, was assessed in the lateral hypothalamus (LH) and nucleus accumbens (NAc), two brain regions involved in reward-related behavior, following 51 and 30 days of diet consumption and VWR, respectively.Compared to CD controls, consumption of fc-HFHSD before and during VWR did not alter total running distances. VWR and fc-HFHSD had opposite effects on body weight gain and terminal fat mass. VWR transiently lowered caloric intake and increased and decreased terminal adrenal and thymus mass, respectively, independent of diet. VWR during fc-HFHSD consumption consistently increased CD self-selection, had an acute negative effect on fat self-selection, and a delayed negative effect on sucrose solution self-selection compared to SED controls. Gene expression of opioid and dopamine neurotransmission components in LH and NAc were unaltered by fc-HFHSD or VWR. We conclude that VWR modulates fc-HFHSD component self-selection in a time-dependent manner in male Wistar rats.

Effects of risk factors on evoked pain patterns in rat models of experimental knee osteoarthritis.

Pain experiences in patients with knee osteoarthritis (OA) may be influenced differently by OA risk factors, reducing the translatability of preclinical research into the clinic. Our objective was to contrast evoked pain patterns after exposure to different OA risk factors including acute joint trauma, chronic instability, or obesity/metabolic syndrome using rat models of experimental knee OA. We tested longitudinal patterns of evoked pain behaviors (knee pressure pain threshold and hindpaw withdrawal threshold) in young male rats exposed to different OA-inducing risk factors including (1) nonsurgical joint trauma (impact-induced anterior cruciate ligament (ACL) rupture); (2) surgical joint destabilization (ACL + medial meniscotibial ligament transection); and (3) high fat/sucrose (HFS) diet-induced obesity. Histopathology for synovitis, cartilage damage, and subchondral bone morphology was performed. Pressure pain threshold was reduced (more pain) most, and earlier by joint trauma (Week 4-12) and HFS (Week 8-28) than by joint destabilization (Week 12). Hindpaw withdrawal threshold was reduced transiently after joint trauma (Week 4), with smaller and later reductions after joint destabilization (Week 12), but not with HFS. Synovial inflammation occurred at Week 4 after joint trauma and instability but only coincided with pain behaviors after joint trauma. Cartilage and bone histopathology were most severe after joint destabilization and least severe with HFS. The pattern, intensity, and timing of evoked pain behaviors varied due to OA risk factor exposure and were inconsistently associated with histopathological OA features. These findings may help to explain the challenges with translating preclinical OA pain research to multimorbid clinical OA contexts.

Oral Carnosine Supplementation Restores Insulin Sensitivity but not Glucose Tolerance in Mice With Glutathione Peroxidase-4 Deficiency

Obesity presents in about 90% of the diabetic patients, a risk factor that makes them more susceptible to cardiometabolic disorders, particularly insulin resistance and ultimately T2 diabetes. Increased oxidative stress and specifically lipid peroxidation are known to be major pathogenic factors driving these disorders in obesity. We recently reported that diabetic patients were found to have significantly lower level of glutathione peroxidase 4 (GPx4)- an antioxidant selenoenzyme which neutralizes lipid peroxides. Carnosine, a histidine-containing dipeptide which is highly concentrated in muscle and brain, has shown a remarkable therapeutic potential for preventing diabetes and its complications when administered orally in experimental and clinical studies of obesity. The extent to which carnosine can mitigate these disorders in the presence of GPx4 deficiency is not known however, and this is critical to determining its efficacy in diabetes patients. To address this question, wild-type (WT) and GPx4+/− male were randomly assigned to either standard chow (CNTL) or high fat high sucrose diet (HFHS) for 16 weeks. Seven weeks after starting the diet, half of the HFHS diet groups received 80 mM carnosine oral carnosine supplementation in their drinking water. Glucose and insulin tolerance tests performed 8 weeks after starting the diet revealed that carnosine significantly improved glucose tolerance in obese WT (P<0.05) but not obese GPx4+/− mice. Interestingly, carnosine treatment did significantly improve insulin sensitivity in the GPx4+/− mice, and to a lesser extent the WT mice. At the molecular level, we observed that HFHS diet up-regulated GPx4 protein in cardiac tissue from WT but not GPx4+/− mice, and protein oxidation was significantly reduced with carnosine treatment in both genotypes. Collectively, these findings indicate that carnosine supplementation could potentially mitigate insulin resistance even in patients with compromised antioxidant capacity (e.g., diabetes). Further analysis is in progress to evaluate the effectiveness of carnosine supplementation on additional cardiometabolic complications that are associated with diet induced obesity. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Select human milk oligosaccharide supplementation in post‐weanling rats affects metabolism and gut microbiota into adulthood

Feeding infants with human milk versus formula can produce long-lasting benefits, including reduced risk of inflammatory diseases. Most infant formulas do not contain human milk oligosaccharides (HMOs), which are important carbohydrates in human breast milk displaying prebiotic properties. The study's aim was to examine the effect of select HMOs in the post-weaning period. Metabolic and microbial outcomes were measured in female and male rats whose post-weaning diet was supplemented with 3'sialyllactose and 2'-O-fucosyllactose (low dose, 0.75% of each, or high dose, 2% of each). It was determined whether exposure to the HMOs would attenuate metabolic dysfunction associated with a high fat/sucrose (HFS) diet. HMO supplementation resulted in dose-dependent and sex-dependent effects, with the high HMO female group displaying reduced food intake and percentage body fat and an increase in Blautia abundance. Early life HMO supplementation did not prevent the effects of an HFS diet on metabolic outcomes; however, rats that received high dose HMOs followed by the HFS diet were the only HFS group that maintained a measurable abundance of Blautia. This study highlights the potential for HMOs in follow-up infant formulas. Further investigation should include variable HMO mixtures and doses to optimize infant nutrition at this critical stage.

Reactive Oxygen Species in the Aorta and Perivascular Adipose Tissue Precedes Endothelial Dysfunction in the Aorta of Mice with a High-Fat High-Sucrose Diet and Additional Factors.

Metabolic syndrome (Mets) is the major contributor to the onset of metabolic complications, such as hypertension, type 2 diabetes mellitus (DM), dyslipidemia, and non-alcoholic fatty liver disease, resulting in cardiovascular diseases. C57BL/6 mice on a high-fat and high-sucrose diet (HFHSD) are a well-established model of Mets but have minor endothelial dysfunction in isolated aortas without perivascular adipose tissue (PVAT). The purpose of this study was to evaluate the effects of additional factors such as DM, dyslipidemia, and steatohepatitis on endothelial dysfunction in aortas without PVAT. Here, we employed eight-week-old male C57BL/6 mice fed with a normal diet (ND), HFHSD, steatohepatitis choline-deficient HFHSD (HFHSD-SH), and HFHSD containing 1% cholesterol and 0.1% deoxycholic acid (HFHSD-Chol) for 16 weeks. At week 20, some HFHSD-fed mice were treated with streptozocin to develop diabetes (HFHSD-DM). In PVAT-free aortas, the endothelial-dependent relaxation (EDR) did not differ between ND and HFHSD (p = 0.25), but in aortas with PVAT, the EDR of HFHSD-fed mice was impaired compared with ND-fed mice (p = 0.005). HFHSD-DM, HFHSD-SH, and HFHSD-Chol impaired the EDR in aortas without PVAT (p < 0.001, p = 0.019, and p = 0.009 vs. ND, respectively). Furthermore, tempol rescued the EDR in those models. In the Mets model, the EDR is compromised by PVAT, but with the addition of DM, dyslipidemia, and SH, the vessels themselves may result in impaired EDR.

Environmental Enrichment and Metformin Improve Metabolic Functions, Hippocampal Neuron Survival, and Hippocampal-Dependent Memory in High-Fat/High-Sucrose Diet-Induced Type 2 Diabetic Rats

Background: The Western-style diet-induced type 2 diabetes mellitus (T2D) may eventually trigger neurodegeneration and memory impairment. Thus, it is essential to identify effective therapeutic strategies to overcome T2D complications. This study aimed to investigate the effects of environmental enrichment (EE) and metformin interventions on metabolic dysfunctions, hippocampal neuronal death, and hippocampal-dependent memory impairments in high-fat/high-sucrose (HFS) diet-induced T2D rats. Methods: Thirty-two male rats (200-250 g) were divided into four groups: C group (standard diet + conventional cage); D group (HFS diet + conventional cage); DE group (HFS diet + EE cage/6hr daily); and DM group (HFS diet + metformin + conventional cage). Body weight was measured every week. T-maze tasks, anthropometric, biochemical, histological, and morphometric parameters were measured. The expression changes of hippocampal genes were also analyzed. Results: The anthropometric and biochemical parameters were improved in DE and DM groups compared with the D group. DE and DM groups had significantly higher T-maze percentages than the D group. These groups also had better histological and morphometric parameters than the D group. The interventions of EE and metformin enhanced the expression of hippocampal genes related to neurogenesis and synaptic plasticity (BDNF/TrkB binding, PI3K-Akt, Ras-MAPK, PLCγ-Ca2+, and LTP). Conclusion: Environmental enrichment (EE) and metformin improved metabolic functions, hippocampal neuron survival, and hippocampal-dependent memory in HFS diet-induced T2D rats. The underlying mechanisms of these interventions involved the expression of genes that regulate neurogenesis and synaptic plasticity.

Combined Intake of Fish Oil and D-Fagomine Prevents High-Fat High-Sucrose Diet-Induced Prediabetes by Modulating Lipotoxicity and Protein Carbonylation in the Kidney.

Obesity has been recognized as a major risk factor for chronic kidney disease, insulin resistance being an early common metabolic feature in patients suffering from this syndrome. This study aims to investigate the mechanism underlying the induction of kidney dysfunction and the concomitant onset of insulin resistance by long-term high-fat and sucrose diet feeding in Sprague Dawley rats. To achieve this goal, our study analyzed renal carbonylated protein patterns, ectopic lipid accumulation and fatty acid profiles and correlated them with biometrical and biochemical measurements and other body redox status parameters. Rats fed the obesogenic diet developed a prediabetic state and incipient kidney dysfunction manifested in increased plasma urea concentration and superior levels of renal fat deposition and protein carbonylation. An obesogenic diet increased renal fat by preferentially promoting the accumulation of saturated fat, arachidonic, and docosahexaenoic fatty acids while decreasing oleic acid. Renal lipotoxicity was accompanied by selectively higher carbonylation of proteins involved in the blood pH regulation, i.e., bicarbonate reclamation and synthesis, amino acid, and glucose metabolisms, directly related to the onset of insulin resistance. This study also tested the combination of antioxidant properties of fish oil with the anti-diabetic properties of buckwheat D-Fagomine to counteract diet-induced renal alterations. Results demonstrated that bioactive compounds combined attenuated lipotoxicity, induced more favorable lipid profiles and counteracted the excessive carbonylation of proteins associated with pH regulation in the kidneys, resulting in an inhibition of the progression of the prediabetes state and kidney disease.

Prospective insight into the role of benzyl propylene glycoside as a modulator of the cGAS-STING signaling pathway in the management of nonalcoholic fatty pancreas animal model

BackgroundNonalcoholic fatty pancreatitis (NAFP) is one of the metabolic syndrome manifestations that need further studies to determine its molecular determinants and find effective medications. We aimed to investigate the potential effect of benzyl propylene glycoside on NAFP management via targeting the pancreatic cGAS-STING pathway-related genes (DDX58, NFκB1 & CHUK) and their upstream regulator miRNA (miR-1976) that were retrieved from bioinformatics analysis.MethodsThe rats were fed either normal chow or a high-fat high-sucrose diet (HFHS), as a nutritional model for NAFP. After 8 weeks, the HFHS-fed rats were subdivided randomly into 4 groups; untreated HFHS group (NAFP model group) and three treated groups which received 3 doses of benzyl propylene glycoside (10, 20, and 30 mg/kg) daily for 4 weeks, parallel with HFHS feeding.ResultsThe molecular analysis revealed that benzyl propylene glycoside could modulate the expression of the pancreatic cGAS-STING pathway-related through the downregulation of the expression of DDX58, NFκB1, and CHUK mRNAs and upregulation of miR-1976 expression. Moreover, the applied treatment reversed insulin resistance, inflammation, and fibrosis observed in the untreated NAFP group, as evidenced by improved lipid panel, decreased body weight and the serum level of lipase and amylase, reduced protein levels of NFκB1 and caspase-3 with a significant reduction in area % of collagen fibers in the pancreatic sections of treated animals.Conclusionbenzyl propylene glycoside showed a potential ability to attenuate NAFP development, inhibit pancreatic inflammation and fibrosis and reduce the pathological and metabolic disturbances monitored in the applied NAFP animal model. The detected effect was correlated with modulation of the expression of pancreatic (DDX58, NFκB1, and CHUK mRNAs and miR-1976) panel.

Intestinal cell type-specific communication networks underlie homeostasis and response to Western diet.

The small intestine plays a key role in immunity and mediates inflammatory responses to high fat diets. We have used single-cell RNA-sequencing (scRNA-seq) and statistical modeling to examine gaps in our understanding of the dynamic properties of intestinal cells and underlying cellular mechanisms. Our scRNA-seq and flow cytometry studies of different layers of intestinal cells revealed new cell subsets and modeled developmental trajectories of intestinal intraepithelial lymphocytes, lamina propria lymphocytes, conventional dendritic cells, and enterocytes. As compared to chow-fed mice, a high-fat high-sucrose (HFHS) "Western" diet resulted in the accumulation of specific immune cell populations and marked changes to enterocytes nutrient absorption function. Utilizing ligand-receptor analysis, we profiled high-resolution intestine interaction networks across all immune cell and epithelial structural cell types in mice fed chow or HFHS diets. These results revealed novel interactions and communication hubs among intestinal cells, and their potential roles in local as well as systemic inflammation.

Abstract PD12-05: PD12-05 Adipocyte directed vaccination to reduce the risk of breast cancer

Abstract Background: Obesity is an important risk factor for breast cancer and women with metabolic syndrome may be at the highest risk. Infiltration of CD8 T-cells into fat is an early event in obesity. Type I cytokines secreted by CD8 T-cells upregulate costimulatory molecules on enlarged adipocytes. The adipocytes, now antigen presenting cells, further stimulate Type I T-cell activation. The resulting T-cells compete for glucose and fatty acids which leads to metabolic dysfunction in both the adipose tissue and the T-cells themselves. The T-cells are not able to maintain tumor immune surveillance and secretion of adipokines promotes malignant transformation. Immunologic memory prevents inflammation from resolving even if an individual becomes normal weight. Strategies to increase Type II (anti-inflammatory) T-cells in inflamed adipose could have clinical benefit. Methods: We developed a method of CD4 epitope identification that includes functional screening for Th1 or Th2 epitopes. We use a multi-algorithm approach to ensure responsiveness across diverse HLA alleles. We identified Th2 selective epitopes associated with high IL-10 secretion for 6 adipocyte associated antigens that become overexpressed in inflamed adipocytes (IGF-IR, HIF-1a, DUSP1, FABP4, PAI-1 and ATGL). The epitopes were highly homologous between mouse and man (median 100% (range-82-100%). When the epitopes were used to immunize mice, all antigens generated a significant IL-10 response compared to control (p&amp;lt; 0.05). We questioned whether our “adipocyte directed” vaccine (ADVac) could prevent the development of breast cancer in obese mice. Results: First, C57BL/6 mice were fed a high fat high sucrose (HFHS) diet or normal chow. When mice became obese, vaccination with ADVac or adjuvant alone (Alum) was initiated. Four weeks after the final vaccine, visceral adipose tissue showed significantly fewer CD8 T-cells in the obese mice immunized with ADVac as compared to the control, p=0.0011. The decrease in CD8 T-cells was specific for adipose tissue as no change was observed in matched spleen. There was a significant increase in T-regulatory cells in the adipose tissue of mice immunized with ADVac as compared to control, p=0.031. Two weeks after the final vaccine, a glucose tolerance test (GTT) and insulin tolerance (ITT) showed blood glucose concentrations were significantly lower at all time points for the ADVac-immunized obese mice as compared to the control obese mice (p&amp;lt; 0.01 for all). TgMMTV-neu develop aggressive breast cancer when made obese. Ten-week old TgMMTV-neu mice were fed a HFHS diet for 4 weeks, then randomized into 2 cohorts when obese, one cohort receiving the adjuvant only (Alum) and one receiving ADVac. Mice were sacrificed at 31 weeks of age when all controls had developed tumor. ITT showed the glucose levels in the blood were significantly lower in the ADVac group as compared to control (p&amp;lt; 0.0001). Fewer CD8 T-cells were observed in mammary adipose tissue of AdVac immunized mice compared to control (p=0.001). There was significantly less leptin detected in the serum of ADVac vaccinated mice compared to Alum immunized, p=0.024. The median age of tumor development was 25 weeks in controls and 29 weeks in the immunized group (p=0.009). Sixty percent (9/15) of the vaccinated mice were tumor free at study termination, whereas 100% of the control mice had developed tumor. Conclusions: ADVac represents the first vaccine to lower breast cancer risk in obesity. Vaccination corrected metabolic dysfunction as evidenced by reversal of diabetes and prevented breast cancer in the majority of obese ADVac immunized mice. Further studies are ongoing evaluating the systemic distribution of ADVac specific T-cells and the safety of the approach. Citation Format: Mary Disis, Lauren Corulli, Erin R. Rodmaker, Denise Cecil. PD12-05 Adipocyte directed vaccination to reduce the risk of breast cancer [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr PD12-05.

Once a week consumption of Western diet over twelve weeks promotes sustained insulin resistance and non-alcoholic fat liver disease in C57BL/6 J mice

The Western diet (high in fat and sucrose) consumption is a highly prevalent feature in the whole world, mainly due to the increasing consumption of ultra-processed foods (UPF), which are cheaper and easier-to-eat, as compared to fresh and highly nutritive meals. Epidemiological studies have associated UPF consumption with development of obesity, non-alcoholic fat liver disease (NAFLD) and insulin resistance. For molecular studies, mice fed with Western diets have been used to characterize signaling pathways involved in these diet-induced pathologies. However, these studies fed mice continuously with the diets, which is not compatible with what occurs in real life, when consumption is occasional. Here, we fed mice once-a-week with a high fat, high sucrose (HFHS) diet and compared these animals with those fed continuously with HFHS diet or with a standard diet. Our results show that after a single day of consuming HFHS, animals presented impaired oral glucose tolerance test (oGTT) as compared to control group. Although this impairment was reversed after 24 h consuming regular diet, repetition of HFHS consumption once-a-week aggravated the picture such as after 12-weeks, oGTT impairment was not reversed after 6 days under control diet. Liver steatosis, inflammation, impaired insulin signaling pathway and endoplasmic reticulum stress are similar comparing animals that consumed HFHS once-a-week with those that continuously consumed HFHS, though weekly-fed animals did not gain as much weight. Therefore, we conclude that regimen of one day HFHS plus 6 days normal diet over 12 weeks is sufficient to induce insulin resistance and NAFLD in mice.

Knockout of murine Lyplal1 confers sex-specific protection against diet-induced obesity.

Human genome-wide association studies found single-nucleotide polymorphisms (SNPs) near LYPLAL1 (Lysophospholipase-like protein 1) that have sex-specific effects on fat distribution and metabolic traits. To determine whether altering LYPLAL1 affects obesity and metabolic disease, we created and characterized a mouse knockout (KO) of Lyplal1. We fed the experimental group of mice a high-fat, high-sucrose (HFHS) diet for 23 weeks, and the controls were fed regular chow diet. Here, we show that CRISPR-Cas9 whole-body Lyplal1 KO mice fed an HFHS diet showed sex-specific differences in weight gain and fat accumulation as compared to chow diet. Female, not male, KO mice weighed less than WT mice, had reduced body fat percentage, had white fat mass, and had adipocyte diameter not accounted for by changes in the metabolic rate. Female, but not male, KO mice had increased serum triglycerides, decreased aspartate, and decreased alanine aminotransferase. Lyplal1 KO mice of both sexes have reduced liver triglycerides and steatosis. These diet-specific effects resemble the effects of SNPs near LYPLAL1 in humans, suggesting that LYPLAL1 has an evolutionary conserved sex-specific effect on adiposity. This murine model can be used to study this novel gene-by-sex-by-diet interaction to elucidate the metabolic effects of LYPLAL1 on human obesity.

SGLT2 inhibitor ertugliflozin decreases elevated intracellular sodium, and improves energetics and contractile function in diabetic cardiomyopathy

BackgroundElevated myocardial intracellular sodium ([Na+]i) was shown to decrease mitochondrial calcium ([Ca2+]MITO) via mitochondrial sodium/calcium exchanger (NCXMITO), resulting in decreased mitochondrial ATP synthesis. The sodium-glucose co-transporter 2 inhibitor (SGLT2i) ertugliflozin (ERTU) improved energetic deficit and contractile dysfunction in a mouse model of high fat, high sucrose (HFHS) diet-induced diabetic cardiomyopathy (DCMP). As SGLT2is were shown to lower [Na+]i in isolated cardiomyocytes, we hypothesized that energetic improvement in DCMP is at least partially mediated by a decrease in abnormally elevated myocardial [Na+]i. MethodsForty-two eight-week-old male C57BL/6J mice were fed a control or HFHS diet for six months. In the last month, a subgroup of HFHS-fed mice was treated with ERTU. At the end of the study, left ventricular contractile function and energetics were measured simultaneously in isolated beating hearts by 31P NMR (Nuclear Magnetic Resonance) spectroscopy. A subset of untreated HFHS hearts was perfused with vehicle vs. CGP 37157, an NCXMITO inhibitor. Myocardial [Na+]i was measured by 23Na NMR spectroscopy. ResultsHFHS hearts showed diastolic dysfunction, decreased contractile reserve, and impaired energetics as reflected by decreased phosphocreatine (PCr) and PCr/ATP ratio. Myocardial [Na+]i was elevated > 2-fold in HFHS (vs. control diet). ERTU reversed the impairments in HFHS hearts to levels similar to or better than control diet and decreased myocardial [Na+]i to control levels. CGP 37157 normalized the PCr/ATP ratio in HFHS hearts. ConclusionsElevated myocardial [Na+]i contributes to mitochondrial and contractile dysfunction in DCMP. Targeting myocardial [Na+]i and/or NCXMITO may be an effective strategy in DCMP and other forms of heart disease associated with elevated myocardial [Na+]i.

High-fat/high-sucrose diet worsens metabolic outcomes and widespread hypersensitivity following early-life stress exposure in female mice.

Exposure to stress early in life has been associated with adult-onset comorbidities such as chronic pain, metabolic dysregulation, obesity, and inactivity. We have established an early-life stress model using neonatal maternal separation (NMS) in mice, which displays evidence of increased body weight and adiposity, widespread mechanical allodynia, and hypothalamic-pituitary-adrenal axis dysregulation in male mice. Early-life stress and consumption of a Western-style diet contribute to the development of obesity; however, relatively few preclinical studies have been performed in female rodents, which are known to be protected against diet-induced obesity and metabolic dysfunction. In this study, we gave naïve and NMS female mice access to a high-fat/high-sucrose (HFS) diet beginning at 4 wk of age. Robust increases in body weight and fat were observed in HFS-fed NMS mice during the first 10 wk on the diet, driven partly by increased food intake. Female NMS mice on an HFS diet showed widespread mechanical hypersensitivity compared with either naïve mice on an HFS diet or NMS mice on a control diet. HFS diet-fed NMS mice also had impaired glucose tolerance and fasting hyperinsulinemia. Strikingly, female NMS mice on an HFS diet showed evidence of hepatic steatosis with increased triglyceride levels and altered glucocorticoid receptor levels and phosphorylation state. They also exhibited increased energy expenditure as observed via indirect calorimetry and expression of proinflammatory markers in perigonadal adipose. Altogether, our data suggest that early-life stress exposure increased the susceptibility of female mice to develop diet-induced metabolic dysfunction and pain-like behaviors.

Gestational Exercise Antagonises the Impact of Maternal High-Fat High-Sucrose Diet on Liver Mitochondrial Alterations and Quality Control Signalling in Male Offspring.

Maternal high-caloric nutrition and related gestational diabetes mellitus (GDM) are relevant modulators of the intrauterine environment, increasing the risk of liver metabolic alterations in mothers and offspring. In contrast, as a non-pharmacological approach against metabolic disorders, exercise is highly recommended in GDM treatment. We analysed whether gestational exercise (GE) protects mothers from diet-induced GDM metabolic consequences and mitigates liver mitochondrial deleterious alterations in their 6-week-old male offspring. Female Sprague Dawley rats were fed with control or high-fat high-sucrose (HFHS) diet and kept sedentary or submitted to GE. Male offspring were sedentary and fed with control diet. Sedentary HFHS mothers and their offspring showed impaired hepatic mitochondrial biogenesis and morphological evidence of mitochondrial remodelling. In contrast, GE-related beneficial effects were demonstrated by upregulation of mitochondrial biogenesis signalling markers and mitochondrial fusion proteins and downregulation of mitochondrial fission protein. Alterations in miR-34a, miR-130b, and miR-494, associated with epigenetic regulation of mitochondrial biogenesis, suggested that GE is a more critical modulator of intergenerational changes in miRs expression than the maternal diet. Our data showed that GE positively modulated the altered hepatic mitochondrial biogenesis and dynamics markers and quality control signalling associated with maternal HFHS-diet-related GDM in mothers and offspring.