Arginine methylation coordinates with phosphorylation to regulate NADP+synthesis

ObjectiveWe sought to characterize how different compositions of the diet can alter feeding behavior and energy expenditure in mice, ultimately driven by changes in sympathetic activation and thermogenesis.HypothesisMice on a high fat diet (HFD) will consume more calories/day compared to high carbohydrate diet (HCD) and standard chow fed mice. Mice on HFD will have decreased expression of UCP1 in both brown and white adipose tissue. There will be decreased expression for several lipolytic enzymes in the white adipose of HFD mice.MethodsA total of 21 mice were divided into three ad libitum fed diet groups: HFD, HCD, and standard chow. Each group was fed their respective diet for 7 weeks. Changes in weight and percent body fat were recorded by NMR. At week 5, glucose tolerance tests were performed on all mice. Both a 24‐hour heat production and respiratory exchange ratio were collected on each group using a metabolic cage system. Mice were sacrificed at the conclusion of week 7 and organs were collected for molecular analysis.ResultsWe show that mice on both ad libitum HFD and HCD consume significantly more calories/day when compared to standard chow. Both HFD and HCD fed mice have a significantly increased body weight and percent body fat when compared to standard chow; however, the HFD mice had a far greater increase in weight compared to HCD fed mice despite no difference in calorie consumption. Mice on a HFD had a lower 24‐hour average heat production per body mass when compared to standard chow and HCD mice and a significantly impaired glucose tolerance test. Furthermore, UCP1 mRNA expression is significantly elevated in brown adipose tissue (BAT) of HCD mice compared to that of HFD and standard chow mice. There were no differences in UCP1 expression in subcutaneous or visceral white adipose tissue (WAT) among all groups. There were also no differences in tyrosine hydroxylase (TH) expression in BAT between all mice. Three markers of lipolysis were decreased in visceral WAT of HFD mice compared to HCD and standard chow counterparts.DiscussionMice consumed significantly more calories when fed a HFD or HCD. These mice gain significantly more weight compared to chow with HFD mice gaining weight far more than the HCD group. This major difference in weight gain despite no significant difference in calorie consumption between HFD and HCD suggests differences may be determined by energy expenditure. This evidence is further supported by the fact that HFD fed mice had a significantly lower daily heat production compared to HCD and standard chow mice. We also show that HCD increases thermogenesis in mouse BAT which may be protective against weight gain. Finally, lipolysis signaling seems to be impaired in HFD mice, but no data supports that this is due to differences in sympathetic activation. Further work will include analyzing the same tissues in isocaloric, restricted diet fed mice.

Accumulating studies have shown that methylation of tumor suppressor genes plays an important role in tumorigenesis. Deuterosome assembly protein 1 (DEUP1) has been implicated as a suppressor gene in some tumors and promoter methylation led to silencing of its expression. However, the roles of DEUP1 promoter methylation and expression in hepatocellular carcinoma (HCC) are not clear. In the present study, the expression and methylation of the DEUP1 promoter in HCC was investigated and the correlations with HCC occurrence and development were explored. A total of 60 HCC tumor and adjacent non-tumor tissues were included in this study. Reverse transcription-polymerase chain reaction, bisulfite PCR sequencing, immunohistochemistry and western blotting were applied to detect the methylation status of the DEUP1 promoter and its expression, and to analyze their associations with clinicopathological data. The results showed that the mRNA and protein expression of DEUP1 in adjacent non-tumor tissues was significantly increased compared with in the HCC tissues. DEUP1 promoter methylation was detected in 46/60 (76.7%) tumor tissues and there was a negative correlation between promoter methylation and DEUP1 protein expression (P<0.05). Analysis of the clinicopathological data revealed that the mRNA and protein expression of DEUP1, and its promoter methylation status, was associated with tumor node metastasis stage and tumor differentiation. Taken together, the results of the present study suggested that methylation of the DEUP1 promoter maybe an important mechanism for gene inactivation and has a critical role in the occurrence and development of liver cancer.

Recently, the prevention of cardiovascular events has become one of most important purpose of diabetes care. We have previously reported that GLP-1 receptor agonist exendin-4 (BBRC 2011) and DPP-4 inhibitor linagliptin (Cardiovasc Diabetol 2014) attenuate neiontima formation after vascular injury. On the other hand, PPAR agonist is also reported its vascular protective effect. Then, we examined whether selective PPAR alpha agonist pemafibrate could attenuate neointima formation after vascular injury and vascular smooth muscle cell (VSMC) proliferation. We performed endothelial denudation injury in mice treated with high fat diet (HFD) or normal chow. Orally administered pemafibrate significantly attenuated neointima formation after vascular injury in both HFD and normal chow mice. Interestingly, pemafibrate increased serum FGF-21 concentration and decreased serum insulin concentration in HFD mice. In addition, body weight was slightly but significantly decreased by pemafibrate in HFD mice. In vitro experiments, pemafibrate attenuated VSMC proliferation in growth curve in a dose dependent manner, but not another PPAR alpha agonist bezafibrate. This suggested that the attenuation of VSMC proliferation by pemafibrate could be the drug effect, not class effect of PPAR alpha agonist. BrdU assay revealed that pemafibrate inhibited DNA synthesis in VSMC dose-dependently. In cell cycle distribution, G1 to S phase entry was significantly inhibited by pemafibrate in flow cytometry analysis. However, apoptosis was not induced by pemafibrate in VSMC in TUNEL assay. These data suggested that pemafibrate attenuates neiontima formation after vascular injury and VSMC proliferation inhibiting cell cycle progression. Disclosure T. Horikawa: None. T. Nomiyama: None. H. Takahashi: None. T. Kawanami: None. Y. Hamaguchi: None. T. Tanaka: None. T. Yanase: None.

Objective Non-alcoholic fatty liver disease (NAFLD) is an increasingly common disease, which can lead to hepatocellular carcinoma (HCC). It is associated with an increased portal pressure, which can alter the intestinal barrier, increase the translocation of bacterial products, and further worsen NAFLD. We hypothesized that this vicious circle can be broken by surgical porto-systemic vein shunting (PSVS), and previously demonstrated that PSVS can decrease the histological features of NAFLD in a high-fat diet (HFD) mouse model. We now test whether PSVS can also impact de-novo hepatocarcinogenesis. Methods C57BL/6 mice received HFD starting from 4 weeks of age. HCC was induced by intraperitoneal injection of DEN at 25mg/kg on week 2 and PSVS (n = 18) (or sham surgery (n = 18)) are created at 8 weeks. HCC burden was assessed by MRI and, finally, by macroscopic and histomorphology assessments. HCC features of aggressiveness, including solid growth pattern and fat component have been also evaluated. Results At 40 weeks of HFD feeding, tumors were identified in all the animals. Shunted HFD mice showed a reduced number of tumor nodules compared to sham (median nodules 8 vs 14, -42.9%; p = 0.0471) while associated to a greater average total tumor volume (709.3 vs 197 mm3, +258,6%; p = 0.0245). This correlated with an increased median tumor volume in shunted mice (16.30 vs 72.45 mm3, +344,5%; p = 0.0011). Notably, HCC histology of shunted mice was hallmarked by accentuated trend concerning HCC fatty change combined to a less pronounced solid growth pattern (p = 0.193). Conclusion PSVS leads to the presence of larger HCCs, potentially linked to the proportionally increased arterial supply of the liver. However, it demonstrates a protective effect on HCC carcinogenesis (&amp;lt; number of tumors). Collectively, this data suggests that portal pressure could represent a potential therapeutic target to attenuate liver steatosis and NAFLD-related HCC carcinogenesis.

This study was performed to identify crucial oncogenes modulated by DNA methylation in hepatocellular carcinoma (HCC) and look for new drugs for HCC treatment. The data of TCGA-LIHC cohort were obtained from UCSC database. Weighted gene co-expression network analysis and multiple machine learning algorithms were applied to screen the crucial prognosis-related genes in HCC. Then these genes were further screened by DNA methylation status. Ten-eleven translocation 1 (TET1) was overexpressed in HCC cell lines, and its biological functions and regulatory effects on the oncogenes were explored by qPCR, methylation-specific polymerase chain reaction, cell viability assay, Western blot, etc. Molecular docking was applied to evaluate the binding affinity between atractylenolide I (AT-I) and TET1, and the tumor-suppressive functions of AT-I were examined with both in vitro and in vivo models. In this work, 12 crucial genes related to HCC prognosis were obtained, among which six genes were with differential methylation status in HCC tissues, including AKR1B10, ALPK3, NQO1, NT5DC2, SFN, and SPP1. The expression levels of ALPK3 and NT5DC2 were positively regulated by TET1, the crucial mediator of demethylation. TET1 overexpression increased the viability and stemness of HCC cells. AT-I had good binding affinity with TET1, and repressed its activity. AT-I promoted the methylation of ALPK3 and NT5DC2 promoter regions, and reduced their expression, and repressed the growth of HCC cells. In summary, DNA methylation contributes to HCC progression, and AT-I represses the malignancy of HCC cells by inhibiting TET1-mediated abnormal DNA demethylation.

Lead (Pb) is a widespread toxic endocrine disruptor that could cause liver damage and gut microbiota dysbiosis. However, the causal relationship and underlying mechanisms between the gut microbiota and Pb-induced liver injury are unclear. In this study, we investigated the metabolic toxicity caused by Pb exposure in normal chow (Chow) and high-fat diet (HFD) mice and confirmed the causal relationship by fecal microbial transplantation (FMT) and antibiotic cocktail experiments. The results showed that Pb exposure exacerbated HFD-induced hepatic lipid deposition, fibrosis, and inflammation, but it had no significant effect on Chow mice. Pb increased serum lipopolysaccharide (LPS) levels and induced intestinal inflammation and barrier damage by activating TLR4/NFκB/MLCK in HFD mice. Furthermore, Pb exposure disrupted the gut microbiota, reduced short-chain fatty acid (SCFA) concentrations and the colonic SCFA receptors, G protein-coupled receptor (GPR) 41/43/109A, in HFD mice. Additionally, Pb significantly inhibited the hepatic GPR109A-mediated adenosine 5'-monophosphate-activated protein kinase (AMPK) pathway, resulting in hepatic lipid accumulation. FMT from Pb-exposed HFD mice exacerbated liver damage, disturbed lipid metabolic pathways, impaired intestinal barriers, and altered the gut microbiota and metabolites in recipient mice. However, mice exposed to HFD + Pb and HFD mice had similar levels of these biomarkers in microbiota depleted by antibiotics. In conclusion, our study provides new insights into gut microbiota dysbiosis as a potential novel mechanism for human health related to liver function impairment caused by Pb exposure.

The genomic bases of the adipose tissue abnormalities induced by chronic positive calorie excess have been only partially elucidated. We adopted a genome-wide approach to directly test whether long-term high-fat diet (HFD) exposure affects the DNA methylation profile of the mouse adipose tissue and to identify the functional consequences of these changes. We have used epididymal fat of mice fed either high-fat (HFD) or regular chow (STD) diet for 5 months and performed genome-wide DNA methylation analyses by methylated DNA immunoprecipitation sequencing (MeDIP-seq). Mouse Homeobox (Hox) Gene DNA Methylation PCR, RT-qPCR and bisulphite sequencing analyses were then performed. Mice fed the HFD progressively expanded their adipose mass accompanied by a significant decrease in glucose tolerance (P<0.001) and insulin sensitivity (P<0.05). MeDIP-seq data analysis revealed a uniform distribution of differentially methylated regions (DMR) through the entire adipocyte genome, with a higher number of hypermethylated regions in HFD mice (P<0.005). This different methylation profile was accompanied by increased expression of the Dnmt3a DNA methyltransferase (Dnmt; P<0.05) and the methyl-CpG-binding domain protein Mbd3 (P<0.05) genes in HFD mice. Gene ontology analysis revealed that, in the HFD-treated mice, the Hox family of development genes was highly enriched in differentially methylated genes (P=0.008). To validate this finding, Hoxa5, which is implicated in fat tissue differentiation and remodeling, has been selected and analyzed by bisulphite sequencing, confirming hypermethylation in the adipose tissue from the HFD mice. Hoxa5 hypermethylation was associated with downregulation of Hoxa5 mRNA and protein expression. Feeding animals previously exposed to the HFD with a standard chow diet for two further months improved the metabolic phenotype of the animals, accompanied by return of Hoxa5 methylation and expression levels (P<0.05) to values similar to those of the control mice maintained under standard chow. HFD induces adipose tissue abnormalities accompanied by epigenetic changes at the Hoxa5 adipose tissue remodeling gene.

In the present study, Potentilla fulgens L. methanolic root extract (PF) was examined to evaluate its comparative ultrastructure effect on the liver and kidney of diabetic and high-fat diet (HFD) mice models. The diabetic mice and HFD mice were treated with PF and separately with their respective standard drugs, metformin, and orlistat respectively. The kidney and liver were isolated on the last day for transmission electron microscope (TEM) study. TEM results showed a remarkable change in the nucleus with distorted shape in the liver and kidney of diabetic and HFD mice. The endoplasmic reticulum tubules were found to be deteriorated, fragmented, and dilated in both models. Mitochondria membranes were disrupted and were found to be less in number in the liver and kidney of both the mice models when compared to their respective control groups. The presence of lipid droplets was observed only in HFD mice, which is a pathological hallmark of a metabolic disease. Treatment with PF restored these ultrastructural abnormalities observed in diabetic and HFD mice.

Introduction: A recent Phase 1/2 clinical trial evaluating the efficacy of the inducible nitric oxide synthase (iNOS, NOS2) inhibitor L-NMMA in combination with taxanes for patients with chemorefractory, locally advanced breast cancer or metastatic triple-negative breast cancer (TNBC) reported an overall response rate (ORR) of 45.8%. Notably, stratification by body mass index (BMI) revealed that obese patients had a significantly higher ORR (85.6%) compared to non-obese patients (46.7%), correlating with elevated iNOS expression in tumor biopsies from obese patients. While obesity typically worsens disease-free and overall survival in TNBC, these findings suggest a potential role for iNOS inhibition in overcoming obesity-driven tumor progression. To investigate further, we studied the impact of iNOS inhibition on tumor growth and treatment response in an obese mouse model and examined the role of the gut microbiome. Obesity was induced in C57BL/6 mice using a high-fat diet (HFD) for 10 weeks prior to orthotopic implantation of E0771 TNBC cells in the mammary fat pad of female mice. Control mice were fed a normal diet (ND). Results:- HFD-fed mice developed larger, more aggressive tumors, with detectable burden occurring ∼3 weeks earlier. Tumors in HFD mice reached humane endpoints ∼17 days sooner than ND mice, whose tumors grew to 80% of HFD tumor size in the same period. Lung metastasis was significantly higher in HFD mice. - L-NMMA combined with docetaxel delayed tumor growth in both diet groups, though the untreated/treated tumor size ratio was higher in HFD mice (6.5) than ND mice (2.1). - Gut microbiome analysis revealed diet- and tumor-associated compositional changes. FMT (fecal microbiota transplantation) from ND mice to HFD mice reduced tumor size and improved therapeutic response, while FMT from HFD mice to ND mice increased tumor size and impaired response. Immunohistochemistry showed elevated iNOS levels in tumors of HFD mice, which normalized following ND-FMT. Conversely, ND mice receiving HFD-FMT displayed increased iNOS expression. Serum cytokine profiles reflected these tumor and treatment differences. Conclusion: These findings suggest that HFD-induced gut microbiome alterations create a pro-inflammatory environment driving increased iNOS expression, promoting tumor aggressiveness and influencing therapeutic response. Targeting the gut microbiota, alongside iNOS inhibition, could provide novel strategies for managing obesity-associated TNBC. Citation Format: Ivonne Uzair, Ann C. Anselme, Wei Qian, Jianying Zhou, Adriana E. Rosato, Roberto R. Rosato, Jenny C. Chang. Unraveling the microbiome-iNOS connection in obesity-driven triple-negative breast cancer [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 1606.

The current study investigated preventive responses to Houttuynia cordata (HC) standardized extract in obesity induced by a high-fat diet (HFD) in mice. High-performance liquid chromatography was carried out to analyze bioactive components in HC. Mice were divided into HFD groups as a control and HFD groups fed with 1 and 4 % of HC supplementation diet. Changes of the body weight of mice, blood biological factors, and histopathological markers were measured. Bioactive components from HC were characterized to contain chlorogenic acid, caffeic acid, rutin, and quercitrin. HC supplementation reduced body weight, glucose, and total cholesterol levels, but it did not affect aspartic acid transaminase, alanine transaminase, or triglyceride amounts in plasma from mice induced by HFD. A 4 % HC supplementation showed a remarkable decrease in the number of large adipocyte in subcutaneous and abdominal adipose tissue. Histopathological evaluation of liver injury induced by HFD was observed to be improved by HC supplementation, leading to attenuated levels of cytoplasmic vacuolation of hepatocytes. From these results, we explored that standardized extract of HC could be a potential natural product to be directly associated with the control of weight.

Rett syndrome (RTT) is an X-linked neurodevelopmental disorder caused by mutation of the methyl-CpG-binding protein 2 (MECP2) gene. Although RTT has been associated with obesity, the underlying mechanism has not yet been elucidated. In this study, female heterozygous Mecp2-null mice (Mecp2+/- mice), a model of RTT, were fed a normal chow diet or high-fat diet (HFD), and the changes in molecular signaling pathways were investigated. Specifically, we examined the expression of genes related to the hypothalamus and dopamine reward circuitry, which represent a central network of feeding behavior control. In particular, dopamine reward circuitry has been shown to regulate hedonic feeding behavior, and its disruption is associated with HFD-related changes in palatability. The Mecp2+/- mice that were fed the normal chow showed normal body weight and food consumption, whereas those fed the HFD showed extreme obesity with hyperphagia, an increase of body fat mass, glucose intolerance, and insulin resistance compared with wild-type mice fed the HFD (WT-HFD mice). The main cause of obesity in Mecp2+/--HFD mice was a remarkable increase in calorie intake, with no difference in oxygen consumption or locomotor activity. Agouti-related peptide mRNA and protein levels were increased, whereas proopiomelanocortin mRNA and protein levels were reduced in Mecp2+/--HFD mice with hyperleptinemia, which play an essential role in appetite and satiety in the hypothalamus. The conditioned place preference test revealed that Mecp2+/- mice preferred the HFD. Tyrosine hydroxylase and dopamine transporter mRNA levels in the ventral tegmental area, and dopamine receptor and dopamine- and cAMP-regulated phosphoprotein mRNA levels in the nucleus accumbens were significantly lower in Mecp2+/--HFD mice than those of WT-HFD mice. Thus, HFD feeding induced dysregulation of food intake in the hypothalamus and dopamine reward circuitry, and accelerated the development of extreme obesity associated with addiction-like eating behavior in Mecp2+/- mice.

Background and aimsAberrant DNA methylation of cyclin-dependent kinase-like 2 (CDKL2) had been observed in several types of tumors. Herein, the present study was aimed to explore the epigenetic and expression status of CDKL2 and evaluate the diagnostic potential of CDKL2 methylation in hepatocellular carcinoma (HCC). MethodsThe methylation status of CDKL2 was detected by methylation-sensitive restriction enzyme based quantitative PCR (MSRE-qPCR) and bisulfite genomic sequencing (BGS). The mRNA expression of CDKL2 was measured using real-time quantitative PCR (qPCR). The correlations between the methylation of CDKL2 and mRNA expression, clinicopathological features were evaluated. ResultsCompared with normal liver tissues, the methylation levels of CDKL2 were significantly increased in the HCC tissues and cell lines (All p < 0.05). And the receiver operating characteristic (ROC) analysis showed that the hypermethylation of CDKL2 had a high specificity and sensitivity to distinguish adjacent non-tumor tissues from HCC tissues. Additionally, the mRNA expression levels of CDKL2 were decreased both in HCC tissues and cell lines than those in normal liver tissues (All p < 0.05), and the expression could be upregulated by 5-aza-2′-deoxycytidine treatment in HCC cell lines. Furthermore, the methylation of CDKL2 was negatively correlated with its mRNA expression (p < 0.001, rs = −0.513), and was associated with gender (p = 0.023), age (p = 0.001) and tumor size (p = 0.016). ConclusionsOur results showed that CDKL2 promoter hypermethylation played an important role in hepatocarcinogenesis and might be a valuable biomarker for HCC diagnosis.

Obesity in postmenopausal women is a risk factor for breast cancer. Since 25-30% of women in the USA are obese, the public health implications of obesity and breast cancer are immense. Activation of inflammatory pathways is one plausible mechanism underlying the association between obesity and increased breast cancer risk. We have used a novel immunocompetent mouse model of luminal breast cancer to demonstrate that luminal tumor progression is enhanced in ovariectomized mice fed a high fat diet (HFD). The obese mammary fat pads are proinflammatory as measured by an increase in M1 polarized macrophages, increased TNF-α production and increased matrix metalloproteinase (MMP)-2 and -9 expression. To evaluate activatable cell penetrating peptide (ACPP) targeting of mammary tumors in the obese fat pad we utilized a version of the MMP cleavable ACPP that has a Cy5 fluorophore attached to the polycationic domain of the peptide and a Cy7 conjugated to the polyanionic domain. Upon cleavage of the peptide by MMPs, the fluorescence resonance energy transfer (FRET) between the two dyes is disrupted and the Cy5 emission increases while the Cy7 re-emission diminishes. This change in emission spectra is measured and the ratio of Cy5 to Cy7 fluorescence is used as a measure of peptide cleavage. The ACPP was administered at a dose of 0.4nmol/g and allowed to circulate for 2 hours prior to harvesting of the fat pads and imaging. The Cy5/Cy7 ratio was 24% higher in the obese fat pads, with a 45% increase in fluorescence when examining just the Cy5 emission. To determine if tumors could be identified in the obese fat pads with the higher background resulting from increased macrophage recruitment and MMP activity, normal chow (NC) and HFD fed C57Bl/6 mice were injected with syngeneic luminal Py230 and claudin low EMT Py8119 cell lines derived from spontaneous polyomavirus middle T (PyVmT) tumor homogenates. Mice harboring both Py230 and Py8119 tumors in their thoracic mammary fat pads were dosed with either the MMP cleavable FRET ACPP or an uncleavable control ACPP. Two hours after peptide injection, mice were sacrificed and the skin was removed to image the tumors in the context of the fat pad. In concordance with our previous results, tumor growth of the Py230 cells was significantly greater in the HFD mice. Interestingly, growth of the more aggressive Py8119 tumors was not enhanced in the HFD microenvironment. For both cell lines, the tumors were clearly delineated using the Cy5/Cy7 ratio. The Py8119 tumors had a ratio of 6.1±0.7 in NC mice and 5.4±0.3 in the HFD mice, and a similar trend was observed in the less aggressive luminal Py230 tumors, with ratios of 4.9±0.8 for NC and 3.8±0.4 for HFD fed mice. This was consistent with analysis showing increased macrophage recruitment in the Py8119 tumors compared with the Py230 tumors, but no difference in recruitment between tumors of either type in mice fed NC or HFD. Our results indicate that although inflammation associated with obesity results in higher background fluorescence in the mammary fat pad, this is not a barrier to visualizing the tumor tissue in the fat pad. Since a large amount of adipose tissue can be problematic during surgery, MMP FRET ACPPs may be useful for identifying and resecting breast cancers in obese patients leading to improved survival. Citation Format: Jessica L. Crisp, Elamprakash N. Savariar, Jonathan P. Hasselmann, Heekyung Chung, Roger Y. Tsien, Lesley G. Ellies. MMP FRET ACPP imaging of mammary tumors in a mouse model of postmenopausal obesity. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research: Genetics, Biology, and Clinical Applications; Oct 3-6, 2013; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2013;11(10 Suppl):Abstract nr A005.

Hepatocellular carcinoma (HCC) is a liver disease with a complex underlying mechanism, and patients with HCC have low survival rates. Iron metabolism plays a crucial role in the pathogenesis of HCC; however, the prognostic value of iron metabolism-related and methylated genes for HCC needs to be further explored. In the present study, we identified differentially expressed genes (DEGs) that play a role in iron metabolism and DNA methylation in HCC from The Cancer Genome Atlas. Four of these DEGs, whose expression levels are correlated with HCC prognosis, namely, RRM2, FTCD, CYP2C9, and ATP6V1C1, were further used to construct a prognostic model for HCC, wherein the risk score was calculated using the gene expression of the four DEGs. This could be used to predict the overall survival of HCC patients for 1, 3, and 5 years. Results of a multivariate Cox regression analysis further indicated that the risk score was an independent variable correlated with the prognosis of HCC patients. The identified gene signature was further validated using an independent cohort of HCC patients from the International Cancer Genome Consortium. Weighted gene co-expression network analysis and gene set enrichment analysis were performed to identify potential regulatory mechanisms of the gene signature in HCC. Taken together, we identified key prognostic factors of iron metabolism-related and methylated genes for HCC, providing a potential treatment strategy for HCC.

Obesity is associated with overactivation of the renin‐angiotensin system hormone angiotensin (Ang) II, which increases blood pressure via multiple mechanisms. Conversely, Ang‐(1‐7), a vasodilatory hormone that opposes the actions of Ang II, is deficient in animal models of obesity. Our previous data show that in obese mice chronic systemic Ang‐(1‐7) administration can attenuate weight gain by increasing energy expenditure as well as improve insulin sensitivity. In this study, we hypothesized that Ang‐(1‐7) would prevent the hypertension and increased sympathetic tone also associated with this mouse model of obesity. To test this, adult male C57BL/6J mice received 12‐week Ang‐(1‐7) (400 ng/kg/min) or saline infusion via subcutaneous osmotic mini‐pumps. Immediately following mini‐pump implantation, mice were placed on a 60% high fat diet (HFD) or a standard chow diet. The three groups of mice in this study were chow diet saline (chow; n=6), HFD saline (HFD; n=3) and HFD Ang‐(1‐7) [n=8]. Blood pressure was measured during the last week of treatment via a carotid artery catheter connected to a strain‐gauge transducer and blood pressure analyzer. Cardiac sympathetic and parasympathetic tone and sympathovagal balance were calculated from the blood pressure signal using spectral analysis frequency domain methods. Consistent with our previous studies, Ang‐(1‐7) attenuated weight gain in HFD mice (36±2 chow vs. 44±2, HFD vs. 38±1 grams HFD Ang‐(1‐7); P=0.013). Ang‐(1‐7) also prevented the rise in systolic blood pressure with HFD (105±6 chow vs. 134±9 HFD vs. 111±5 mm Hg HFD Ang‐(1‐7); P=0.031), with a similar trend for diastolic blood pressure (P=0.084). There was no significant effect of Ang‐(1‐7) on heart rate (P=0.973). The Ang‐(1‐7)‐mediated blood pressure lowering in HFD mice was not associated with significant changes in cardiac sympathetic tone (P=0.303), parasympathetic tone (P=0.115), or sympathovagal balance (P=0.521). These data suggest that Ang‐(1‐7) prevents obesity hypertension in mice perhaps through autonomic‐independent mechanisms. Overall, these findings provide rationale for targeting Ang‐(1‐7) for treatment of obesity‐related hypertension.Support or Funding InformationHL122507This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.