Glucose Metabolism Genes Research Articles

Gegen Qinlian Decoction Coordinately Regulates PPARγ and PPARα to Improve Glucose and Lipid Homeostasis in Diabetic Rats and Insulin Resistance 3T3-L1 Adipocytes.

Gegen Qinlian Decoction (GQD), a well-documented traditional Chinese Medicine (TCM) formula, was reported with convincing anti-diabetic effects in clinical practice. However, the precise antidiabetic mechanism of GQD remains unknown. In this study, the anti-hyperglycemic and/or lipid lowering effects of GQD were demonstrated in high-fat diet with a low dose of streptozotocin induced diabetic Sprague-Dawley rats and insulin resistance (IR)-3T3-L1 adipocytes. GQD treatment increased expression and activity levels of both PPARγ and PPARα in adipocytes, which transcriptionally affected an ensemble of glucose and lipid metabolic genes in vivo and in vitro. The results clearly indicated that GQD treatment intervened with multiple pathways controlled by concomitantly downstream effects of adipocytic PPARγ and PPARα, to influence two opposite lipid pathways: fatty acid oxidation and lipid synthesis. Antagonist GW9662 decreased the mRNA expression of Pparγ and target genes Adpn and Glut4 whereas GW6471 decreased the mRNA expression of Pparα and target genes Cpt-1α, Lpl, Mcad, Lcad, Acox1, etc. Nuclear location and activity experiments showed that more PPARγ and PPARα shuttled into nuclear to increase its binding activities with target genes. GQD decreased the phosphorylation level of ERK1/2 and/or CDK5 to elevate PPARγ and PPARα activities in IR-3T3-L1 adipocytes through post-translational modification. The increase in p-p38MAPK and SIRT1 under GQD treatment may be attributed to partially reduce PPARγ adipogenesis activity and/or activate PPARα activity. Compared with the rosiglitazone-treated group, GQD elevated Cpt-1α expression, decreased diabetic biomarker Fabp4 expression, which produced an encouraging lipid profile with triglyceride decrease partially from combined effects on upregulated adipocytic PPARγ and PPARα activities. These results suggested that GQD improved diabetes by intervening a diverse array of PPARγ and PPARα upstream and downstream signaling transduction cascades, which jointly optimized the expression of target gene profiles to promote fatty acid oxidation and accelerate glucose uptake and utilization than PPARγ full agonist rosiglitazone without stimulating PPARα activity. Thus, GQD showed anti-diabetic/or antihyperglycemic effects, partially through regulating adipocytic PPARα and PPARγ signaling systems to maintaining balanced glucose and lipid metabolisms. This study provides a new insight into the anti-diabetic effect of GQD as a PPARα/γ dual agonist to accelerate the clinical use.

213-OR: Obesity-Induced Astrogliosis Is Regulated by the Diabesity Factor HMG20A

It is well established that systemic glucose homeostasis is imparted by an intricate cross-talk among several organs including the central nervous system (CNS) and pancreatic islets. In analogy to islet beta cells, hypothalamic astrocytes are glucose sensors that facilitate the CNS response to changing metabolic environment. Therefore, astrocytes and beta cells may share a common genetic signature implicated in coupling glucose metabolism to cellular output. We recently demonstrated that the diabesity factor HMG20A is essential for beta-cell functional maturity and adaptation to physiological stress (obesity, type 2 diabetes and pregnancy). As this chromatin remodelling factor also dictates CNS development, we sought to determine whether HMG20A is expressed in astrocytes and whether it potentiates astrocyte function in response to environmental cues. We found that HMG20A is expressed in hypothalamic astrocytes and is upregulated in diet-induced obesity and glucose-intolerant mice, correlating with increased transcript levels of GFAP and IL-1β, indicative of reactive astrocytes (astrogliosis). High glucose, but not lipids, transiently decreased HMG20A expression in isolated mouse primary astrocytes. HMG20A silencing in astrocytes resulted in repression of pro-inflammatory as well as lipid and glucose metabolism genes. These pathways are up-regulated in activated astrocytes (astrogliosis) to resolve stress conditions. As such astrocyte depleted of HMG20A were more susceptible to cell death. Furthermore, motor neuron viability in astrocyte conditioned media was significantly hindered after HMG20A depletion in astrocytes. We posit that HMG20A is involved in the astrocyte polarization state. Under physiological pressure such as obesity, HMG20A expression may be transiently decreased by high glucose but long-term it is increased to induce astrogliosis in an attempt to preserve the neuronal network and glucose homeostasis. Disclosure J.M. Mellado-Gil: None. E. Fuente-Martin: None. P.I. Lorenzo: None. N. Cobo-Vuilleumier: None. J.A. Guerrero-Martínez: None. S.Y. Romero-Zerbo: None. J.M. Franco: None. J. Perez-Cabello: None. M. Alvarez-Dolado: None. D. Pozo: None. F.J. Bermudez-Silva: None. J.C. Reyes: None. B.R. Gauthier: None. Funding ISCIII (PI13/00593 to B.R.G.), (PI13/00309 to F.J.B-S.); Ministerio de Economia y Competidividad/Fondos FEDER (PI10/00871, PI13/00593, BFU2017-83588-P to B.R.G.); BFU2017-85420-R (to J.C.R.); (JCI-2012-12491, IJCI-2015-26238 to E.F-M.); Consejería de Salud Junta de Andalucía (PI-0006-2016 to E.F-M.)

Interleukin 6-independent metabolic reprogramming as a driver of cancer-related fatigue

Fatigue is a common and debilitating symptom of cancer with few effective interventions. Cancer-related fatigue (CRF) is often associated with increases in inflammatory cytokines, however inflammation may not be requisite for this symptom, suggesting other biological mediators also play a role. Because tumors are highly metabolically active and can amplify their energetic toll via effects on distant organs, we sought to determine whether CRF could be explained by metabolic competition exacted by the tumor. We used a highly metabolically active murine E6/E7/hRas model of head and neck cancer for this purpose. Mice with or without tumors were submitted to metabolic constraints in the form of voluntary wheel running or acute overnight fasting and their adaptive behavioral (home cage activity and fasting-induced wheel running) and metabolic responses (blood glucose, ketones, and liver metabolic gene expression) were monitored. We found that the addition of running wheel was necessary to measure activity loss, used as a surrogate for fatigue in this study. Tumor-bearing mice engaged in wheel running showed a decrease in blood glucose levels and an increase in lactate accumulation in the skeletal muscle, consistent with inhibition of the Cori cycle. These changes were associated with gene expression changes in the livers consistent with increased glycolysis and suppressed gluconeogenesis. Fasting also decreased blood glucose in tumor-bearing mice, without impairing glucose or insulin tolerance. Fasting-induced increases in wheel running and ketogenesis were suppressed by tumors, which was again associated with a shift from gluconeogenic to glycolytic metabolism in the liver. Blockade of IL-6 signaling with a neutralizing antibody failed to recover any of the behavioral or metabolic outcomes. Taken together, these data indicate that metabolic competition between the tumor and the rest of the organism is an important component of fatigue and support the hypothesis of a central role for IL-6-independent hepatic metabolic reprogramming in the pathophysiology of CRF.

Expression profiling and analysis of some miRNAs in subcutaneous white adipose tissue during development of obesity

BackgroundMicroRNAs are emerging as new mediators in the regulation of adipocyte physiology and have been approved to play a role in obesity. Despite several studies have focused on microRNA expression profiles and functions in different metabolic tissues, little is known about their response to nutritional interventions in white adipose tissue during obesity stages, and whether they differ in this response to weight-reduction strategy is poorly understood. Our objectives were to study the dysregulation of some miRNAs in subcutaneous inguinal white adipose tissue during weight change, expansion/reduction; in response to both a high-fat diet and switching to a normal diet feeding, and to evaluate them as potential biomarkers and therapeutic targets for early obesity managementMethodA hundred 6-week-old male Wister rats were randomly divided into a normal diet group (N.D), a high-fat diet group (H.F.D), and a switched to a normal diet group (H.F.D/N.D). At the beginning and at intervals 2 weeks, serum lipid, hormone levels, total body fat mass, and inguinal subcutaneous white adipose tissue mass (WAT) measurements were recorded using dual-energy X-ray absorptiometry (DEXA). The expression levels of microRNAs were evaluated using real-time PCR.ResultsSignificant alterations were observed in serum glucose, lipid profile, and adipokine hormones during the early stages of obesity development. Alteration in rno-mir 30a-5p, rno-mir 133a-5p, and rno-mir 107-5p expression levels were observed at more than one time point. While rno-let-7a-5p, rno-mir 193a-5p, and rno-mir125a-5p were downregulated and rno-mir130a-5p was upregulated at all time points within 2 to 4 weeks in response to H.F.D feeding for 10 weeks. The impact of switching to normal diet has a reversed effect on lipid profile, adipokine hormone levels, and some miRNAs. The bioinformatics results have identified a novel and important pathway related to inflammatory signalling.ConclusionOur research demonstrated significant alterations in some adipocyte-expressed miRNAs after a short time of high caloric diet consumption. This provides further evidence of the significant role of nutrition as an epigenetic factor in regulation of lipid and glucose metabolism genes by modulating of related key miRNAs. Therefore, we suggest that miRNAs could be used as biomarkers for adiposity during diet-induced obesity. Perhaps limitation in calories intake is a way to manipulate obesity and associated metabolic disorders. Further studies are needed to fully elucidate the role of microRNAs in the development of obesity

Fat Distribution in Women Is Associated With Depot-Specific Transcriptomic Signatures and Chromatin Structure.

BackgroundPreferential accumulation of fat in the upper body (apple shape) is associated with higher risk of developing metabolic syndrome relative to lower body fat (pear shape). We previously discovered that chromatin openness partially defined the transcriptome of preadipocytes isolated from abdominal and gluteofemoral fat. However, the molecular mechanisms underlying interindividual variation in body shape are unknown.MethodsAdipocyte fraction was isolated from abdominal and gluteofemoral fat biopsies of premenopausal women (age and body mass index matched) segregated initially only by their waist-to-hip ratio. We evaluated transcriptomic and chromatin accessibility using RNA sequencing and assay for transposase-accessible chromatin using sequencing (ATAC-seq) along with key clinical parameters.ResultsOur data showed that higher lower body fat mass was associated with better lipid profile and free fatty acid decrease after glucose administration. Lipid and glucose metabolic pathways genes were expressed at higher levels in gluteofemoral adipocyte fraction in pears, whereas genes associated with inflammation were higher both in abdominal and gluteofemoral apple adipocyte fraction. Gluteofemoral adipocyte chromatin from pear-shaped women contained a significantly higher number of differentially open ATAC-seq peaks relative to chromatin from the apple-shaped gluteofemoral adipocytes. In contrast, abdominal adipocyte chromatin openness showed few differences between apple- and pear-shaped women. We revealed a correlation between gene transcription and open chromatin at the proximity of the transcriptional start site of some of the differentially expressed genes.ConclusionsIntegration of data from all 3 approaches suggests that chromatin openness partially governs the transcriptome of gluteofemoral adipocytes and may be involved in the early metabolic syndrome predisposition associated with body shape.

COMP-Ang-1 Improves Glucose Uptake in db/db Mice with Type 2 Diabetes.

Cartilage oligomeric matrix protein (COMP)-Angiopoietin-1 is a potent angiopoietin-1 (Ang-1) variant that possesses therapeutic potential in angiogenesis and vascular endothelial dysfunction. Noteworthy, we have shown that COMP-Ang-1 improves hyperglycemia and neuroregeneration in ob/ob mice. However, the mechanism of the antidiabetic effect of COMP-Ang-1 is completely unknown. Therefore, we elucidated the diabetes protective molecular mechanisms of COMP-Ang-1 in diabetic db/db mouse model. COMP-Ang-1 (0.5 ng/g body weight) or aqueous NaCl solution was injected intraperitoneally per day in 21 consecutive days into 3-month old, male db/db mice (n=10 per group). Blood glucose and HbA1c levels were determined at baseline and 21 days after COMP-Ang-1 or NaCl treatment. The effect of COMP-Ang-1 on glucose uptake was investigated by euglycemic-hyperinsulinemic clamp studies and key genes of glucose metabolism were studied by Western blot analysis. Our findings indicate that COMP-Ang-1 improves glucose metabolism in a tissue specific manner by regulating HIF-1α transcriptional genes of GLUT-1 expression.

Gene Expression in LEW.1WR1 Rats Indicate Insulin Resistance in Muscle and Liver Tissue

FAT10 is a ubiquitin‐like protein and type 1 diabetes (T1D) susceptibility gene that may play a role in age‐related inflammation, adiposity, cancer, and kidney disease. The young T1D susceptible, LEW.1WR1 rat overexpresses FAT10 and has higher fasting concentrations of blood insulin. It is unclear how the overexpression of FAT10 directly affects insulin sensitivity. It is also unclear if the initial insulin sensitivity of this animal model plays a role in disease susceptibility. We measured the relative expression of lipid and glucose metabolism genes in skeletal muscle and liver tissue in LEW.1WR1 and LEW/ssNHsd rats to determine if 12‐week old LEW.1WR1 rats were insulin resistant. We also tested the gene expression in pancreatic islets to determine if there were changes in endocrine pancreas cell health and hormone production. The results indicate that gene expression increased in the islets for glucagon, insulin, and PLIN5 (Perilipin‐5), and cyclin D expression decreased in LEW.1WR1 rats, suggesting reduced compensation and a potentially lipotoxic environment for beta cells. The increase in glucagon and insulin also supports peripheral insulin resistance. In liver tissue, UBD (Ubiquitin‐D), a gene suspected to regulate beta‐oxidation, was increased. INSIG (Insulin‐induced gene 1) and GLUT2 (Glucose transporter 2) gene expression decreased, indicating decreased expression of insulin‐regulated gene and glucose transporters, respectively. In muscle tissues, ATGL (Adipose triglyceride lipase), GAPDH (Glyceraldehyde‐3‐Phosphate Dehydrogenase), and PNPLA2 (Patatin Like Phospholipase Domain Containing 2) gene expression decreased, indicating a reduced ability to burn fat as energy and glucose intolerance in muscle tissue. These results, in tandem with previous work, suggest that fat storage in organs may account for the difference in weight previously observed between LEW.1WR1 and LEW/ssNHsd rats, and that LEW.1WR1 rats were developing insulin resistance which further suggests these rats are a novel animal model for pre‐diabetes.Support or Funding InformationThe authors would like to thank the UAH Office of the Vice President for Research and Economic Development, The Alabama Space Grant Consortium, ALSAMP, Adriel D. Johnson Sr. Fellowship, College of Science, Department of Chemistry, Department of Biology, Honors College, and Office of the Provost for the financial support for the resources, supplies, and student salaries.

Effects of Intermittent Hypoxia on Expression of Glucose Metabolism Genes in MCF7 Breast Cancer Cell Line.

Hypoxic condition induces molecular alterations which affect the survival rate and chemo-resistant phenotype of cancer cells. The aim of this study is to investigate the influence of intermittent hypoxic conditions on the expression of glucose metabolism genes in breast cancer MCF7 cell line. The gene expression was analyzed using a polymerase chain reaction-array method. In addition, the cell resistance, survival and migration rates were examined to assure the hypoxic influence on the cells. 30 hypoxic episodes induced the Warburg effect through significant (p-value < 0.05) upregulation of the expression of PCK2, PHKG1, ALDOC, G6PC, GYS2, ALDOB, HK3, PKLR, PGK2, PDK2, ACO1 and H6PD genes that are involved in glycolysis, were obtained. Furthermore, the expression of the major gluconeogenesis enzyme genes was significantly (ANOVA, p-value < 0.05) downregulated. These molecular alterations were associated with increased MCF7 cell division and migration rate. However, molecular and phenotypic changes induced after 30 episodes were normalized in MCF7 cells exposed to 60 hypoxic episodes. It is concluded, from this study, that 30 intermitted hypoxic episodes increased the survival rate of MCF7 breast cancer cells and induced the Warburg effect through upregulation of the expression of genes involved in the glycolysis pathway. These results may increase our understanding of the molecular alterations of breast cancer cells under hypoxic conditions.

Reduced caloric intake and periodic fasting independently contribute to metabolic effects of caloric restriction.

Caloric restriction (CR) has positive effects on health and longevity. CR in mammals implements time‐restricted (TR) feeding, a short period of feeding followed by prolonged fasting. Periodic fasting, in the form of TR or mealtime, improves metabolism without reduction in caloric intake. In order to understand the relative contribution of reduced food intake and periodic fasting to the health benefits of CR, we compared physiological and metabolic changes induced by CR and TR (without reduced food intake) in mice. CR significantly reduced blood glucose and insulin around the clock, improved glucose tolerance, and increased insulin sensitivity (IS). TR reduced blood insulin and increased insulin sensitivity, but in contrast to CR, TR did not improve glucose homeostasis. Liver expression of circadian clock genes was affected by both diets while the mRNA expression of glucose metabolism genes was significantly induced by CR, and not by TR, which is in agreement with the minor effect of TR on glucose metabolism. Thus, periodic fasting contributes to some metabolic benefits of CR, but TR is metabolically different from CR. This difference might contribute to differential effects of CR and TR on longevity.

Glucose homeostasis and glucose tolerance were impaired with elevated lipid to starch ratios in practical diets for the omnivorous genetically improved farmed tilapia Oreochromis niloticus

The present study was performed to ascertain if glucose metabolism and glucose tolerance could be regulated by dietary lipid to starch ratios in the omnivorous genetically improved farmed tilapia Oreochromis niloticus. Three isonitrogenous (ca 34.5% protein) and isocaloric (ca 14.5 kJ/g) practical diets were formulated with elevated lipid at the cost of starch provision, being designated as diets L6S23 (5.55% lipid and 22.5% starch), L9S18 (8.77% lipid and 18.1% starch) and L12S13 (12.0% lipid and 13.8% starch). Tilapia juveniles (initial mean body weight: 23.0 g/fish) were allocated to 12 rectangular tanks (250 L) with 20 fish per tank, and were fed experimental diets to apparent satiation for 8 weeks. At the end of the trial, approximately 24 h after the last feeding, fish in each tank were bulk-weighed and counted. Nine fish per tank were sampled to analyze the whole-body composition (3 fish), blood chemistry (3 fish) and the expression of glucose-metabolic genes (3 fish). Thirty-six of the remaining fish per treatment were subjected to an acute glucose tolerance test. The results showed that the growth (weight gain) and feed utilization (feed and protein efficiency ratios) of tilapia were not affected by different treatments. Intraperitoneal fat ratio (1.32%) and lipid percentages in the liver (9.69%) and whole-body (10.2%) of the L12S13 fish were higher than those of the L6S23 fish (0.75%, 6.61% and 8.17%, respectively). The L9S18 (5.24 mmol/L) and L12S13 fish (5.59 mmol/L) obtained higher plasma glucose levels than the L6S23 fish (4.10 mmol/L). In the liver, the mRNA levels of representative genes involved in glycolysis (glucokinase) and glucose release (glucose-6-phosphatase catalytic subunit a2) were simultaneously up-regulated in the L12S13 fish as compared with the L6S23 fish, indicating that a futile cycle between glucose and glucose-6-phosphate was associated with elevated dietary lipid to starch ratios. In the white muscle, the mRNA levels of glucose transporter 1a (glut1a), glut4, hexokinase 1b, phosphofructokinase muscle type a (pfkma), pfkmb and glycogen synthase 1 were 0.44-, 0.71-, 0.58-, 0.51-, 0.72- and 0.53-fold lower in the L12S13 fish than in the L6S23 fish, suggesting that muscular glucose transport and utilization were depressed with elevated dietary lipid to starch ratios. Although it took similar time (3 h) to restore plasma glucose for all treatments, plasma glucose was lower in the L6S23 fish than in the L12S13 fish during 1–3 h after the glucose injection. Additionally, plasma glucose of the L9S18 and L12S13 fish decreased continually along the injection time (during 7–10 h) as compared with the L6S23 fish, which further proved that the regulatory mechanisms of glucose homeostasis were impaired in tilapia that were fed elevated lipid to starch ratios. Our data confirmed that the impairment of glucose homeostasis was also associated with high fat diet reception in the omnivorous tilapia.

High-Dose Dexamethasone Manipulates the Tumor Microenvironment and Internal Metabolic Pathways in Anti-Tumor Progression.

High-dose dexamethasone (DEX) is used to treat chemotherapy-induced nausea and vomiting or to control immunotherapy-related autoimmune diseases in clinical practice. However, the underlying mechanisms of high-dose DEX in tumor progression remain unaddressed. Therefore, we explored the effects of high-dose DEX on tumor progression and the potential mechanisms of its anti-tumor function using immunohistochemistry, histological examination, real-time quantitative PCR (qPCR), and Western blotting. Tumor volume, blood vessel invasion, and levels of the cell proliferation markers Ki67 and c-Myc and the anti-apoptotic marker Bcl2 decreased in response to high-dose DEX. However, the cell apoptosis marker cleaved caspase 3 increased significantly in mice treated with 50 mg/kg DEX compared with controls. Some genes associated with immune responses were significantly downregulated following treatment with 50 mg/kg DEX e.g., Cxcl9, Cxcl10, Cd3e, Gzmb, Ifng, Foxp3, S100a9, Arg1, and Mrc1. In contrast, the M1-like tumor-associated macrophages (TAMs) activation marker Nos2 was shown to be increased. Moreover, the expression of peroxisome proliferator-activated receptors α and γ (Pparα and Pparg, respectively) was shown to be significantly upregulated in livers or tumors treated with DEX. However, high-dose DEX treatment decreased the expression of glucose and lipid metabolic pathway-related genes such as glycolysis-associated genes (Glut1, Hk2, Pgk1, Idh3a), triglyceride (TG) synthesis genes (Gpam, Agpat2, Dgat1), exogenous free fatty acid (FFA) uptake-related genes (Fabp1, Slc27a4, and CD36), and fatty acid oxidation (FAO) genes (Acadm, Acaa1, Cpt1a, Pnpla2). In addition, increased serum glucose and decreased serum TG and non-esterified fatty acid (NEFA) were observed in DEX treated-xenografted tumor mice. These findings indicate that high-dose DEX-inhibited tumor progression is a complicated process, not only activated by M1-like TAMs, but also decreased by the uptake and consumption of glucose and lipids that block the raw material and energy supply of cancer cells. Activated M1-like TAMs and inefficient glucose and lipid metabolism delayed tumor cell growth and promoted apoptosis. These findings have important implications for the application of DEX combined with drugs that target key metabolism pathways for tumor therapy in clinical practice.

P.209 Reward sensitivity, glucose metabolism, and candidate genes: variants of the translocatior protein (TSPO) and TFAP2B genes in a cohort study

This paper represents a sequel to Jureczka and Ochal (2019) where numerical solution of a quasistatic contact problem is considered for an elastic body in frictional contact with a moving foundation. The model takes into account wear of the contact surface of the body caused by the friction. Some preliminary error analysis for a fully discrete approximation of the contact problem was provided in Jureczka and Ochal (2019). In this paper, we consider a more general fully discrete numerical scheme for the contact problem, derive optimal order error bounds and present computer simulation results showing that the numerical convergence orders match the theoretical predictions.

Low oxygen tension activates glucose metabolism, improves antioxidant capacity and augment developmental potential of ovine embryos in vitro

Context Oxygen (O2) is one of the most powerful regulators of embryo function. Nevertheless, most in vitro embryo production studies do not consider O2 as a determining factor. Aim The present study was designed to assess the effect of different O2 (5 and 20%) concentrations on the developmental ability and expression of genes related to cellular antioxidant functions and glucose metabolism in the in vitro produced ovine embryos. Methods In vitro sheep embryos were produced at different O2 (5 and 20%) concentrations as per the laboratory protocol. Developmental stages of embryos at different O2 concentrations were compared. Messenger RNA abundance of antioxidant and glucose metabolism genes in embryos produced at different O2 concentrations were compared. Key results No significant (P &amp;lt; 0.05) effect of different O2 concentrations on oocyte maturation and cleavage rate was observed. In contrast, significantly (P &amp;lt; 0.05) more number of morula and blastocysts were observed at 5 compared with 20%O2. The expression level of the genes related to antioxidant functions (GPX, SOD1, SOD2 and CAT) and glucose metabolism (G6PD and HPRT) were found significantly (P &amp;lt; 0.05) greater in the embryos generated with 5 compared with 20% O2. In contrast, the expression of GAPDH did not differ significantly (P &amp;lt; 0.05) between the groups. Conclusions Ovine embryos at 5%O2 generated low ROS and synthesised more GSH due to the activation of G6PD and GPX that in turn increased the antioxidant capability and developmental potential of the embryos. Implications Embryos at higher O2 concentration (20%) generated more reactive oxygen species (ROS) that caused oxidative damage to the embryos and in turn reduced their developmental ability and alter gene expression.

Identification of a Subtype of Hepatocellular Carcinoma with Poor Prognosis Based on Expression of Genes within the Glucose Metabolic Pathway.

Hepatocellular carcinoma (HCC) is the most prevalent primary cancer and a highly aggressive liver malignancy. Liver cancer cells reprogram their metabolism to meet their needs for rapid proliferation and tumor growth. In the present study, we investigated the alterations in the expression of the genes involved in glucose metabolic pathways as well as their association with the clinical stage and survival of HCC patients. We found that the expressions of around 30% of genes involved in the glucose metabolic pathway are consistently dysregulated with a predominant down-regulation in HCC tumors. Moreover, the differentially expressed genes are associated with an advanced clinical stage and a poor prognosis. More importantly, unsupervised clustering analysis with the differentially expressed genes that were also associated with overall survival (OS) revealed a subgroup of patients with a worse prognosis including reduced OS, disease specific survival, and recurrence-free survival. This aggressive subtype had significantly increased expression of stemness-related genes and down-regulated metabolic genes, as well as increased immune infiltrates that contribute to a poor prognosis. Collectively, this integrative study indicates that expressions of the glucose metabolic genes could be used as potential prognostic markers and/or therapeutic targets, which might be helpful in developing precise treatment for patients with HCC.

Effects of acute hyperglycaemia stress on indices of glucose metabolism and glucose transporter genes expression in cobia ( Rachycentron canadum )

The objective of the present study is to preliminarily clarify the mechanism of carbohydrates metabolism in cobia (Rachycentron canadum) (85 ± 3 g) receiving injection of glucose solution. We examined plasma glucose (GLU), total protein (TP), triglyceride (TG), cholesterol (CHOL), insulin, liver glycogen and muscle glycogen, activities of hepatic hexokinase (HK), phosphofructokinase (PFK) and pyruvate kinase (PK), as well as relative expressions of glucose transporter 1 (GLUT1), GLUT2, GLUT3, GLUT4, GLUT5 and GLUT9 in hemocyte, liver and muscle of R. canadum when fish were injected with 200 μl of glucose solution (255 mg/ml) after 0, 1, 2, 4, 8, 12, 24 and 48 hr. Fish received injection of 0.68% saline served as control. Results indicated that the plasma GLU, TG and CHOL increased and reached peak at 1, 8 and 48 hr postinjection (hpi) respectively. The hepatic glycogen increased from 1 hpi, and reached peak at 8 hpi, plasma insulin increased at 1 hpi, and reached peak at 2 hpi, and activity of hepatic PK peaked at 8 hpi. Furthermore, the relative expressions of GLUT1, GLUT2, GLUT3, GLUT4 and GLUT5 in hemocytes reached peak at 1,4, 8, 4 and 8 hpi, respectively, relative expressions of GLUT2, GLUT3, GLUT5 and GLUT9 in liver reached peak at 24, 24, 12 and 24 hpi, respectively, and relative expressions of GLUT1 and GLUT3 in muscle were significantly higher at 2 and 2–4 hpi, respectively compared with those in controls. In conclusion, low ability of utilizing glucose in R. canadum may be attributed to insufficient insulin secretion, low activities of key glycolytic enzymes (HK, PFK and PK) regulated by glucose injection and slow increase of GLUTs.

Localisation of PGK1 determines metabolic phenotype to balance metastasis and proliferation in patients with SMAD4-negative pancreatic cancer

ObjectivePancreatic ductal adenocarcinoma (PDAC) is the most aggressive type of GI tumour, and it possesses deregulated cellular energetics. Although recent advances in PDAC biology have led to the discovery of...

Coopted temporal patterning governs cellular hierarchy, heterogeneity and metabolism in Drosophila neuroblast tumors.

It is still unclear what drives progression of childhood tumors. During Drosophila larval development, asymmetrically-dividing neural stem cells, called neuroblasts, progress through an intrinsic temporal patterning program that ensures cessation of divisions before adulthood. We previously showed that temporal patterning also delineates an early developmental window during which neuroblasts are susceptible to tumor initiation (Narbonne-Reveau et al., 2016). Using single-cell transcriptomics, clonal analysis and numerical modeling, we now identify a network of twenty larval temporal patterning genes that are redeployed within neuroblast tumors to trigger a robust hierarchical division scheme that perpetuates growth while inducing predictable cell heterogeneity. Along the hierarchy, temporal patterning genes define a differentiation trajectory that regulates glucose metabolism genes to determine the proliferative properties of tumor cells. Thus, partial redeployment of the temporal patterning program encoded in the cell of origin may govern the hierarchy, heterogeneity and growth properties of neural tumors with a developmental origin.

Abstract 4517: Tumor-associated macrophages are reprogrammed by pancreatic ductal adenocarcinoma cells through tumor-induced DNA methylation on metabolic genes

Abstract Metabolism is shifted toward glycolysis with increased glucose uptake in anti-tumor M1 macrophages, and toward oxidative phosphorylation with significant decreased glucose uptake in pro-tumor M2 macrophages in Pancreatic ductal adenocarcinoma (PDA). Current study aims to investigate the mechanism by which tumor cells reprogram tumor associated macrophages metabolically and functionally. Using mouse bone marrow derived macrophages and human macrophages derived from peripheral blood mononuclear cells for contact and non-contact co-culture experiments with PDA tumor cells, we showed PDA tumor cells, through direct cell-cell contact confirmed by Lucifer yellow dye-coupling assay, induced DNA methylation and downregulation of a panel of glucose metabolism and oxidative phosphorylation genes selectively in M1 but not M2 macrophages by methylation specific PCR, methylation microarray analysis and qPCR, RNA-seq analysis. Glucose-response genes such as IL10 were subsequently activated. Through IL10 and its receptor IL-10R on tumor cells, macrophages enhanced the in vitro migration of tumor cells in trans-well migration assay verified by shRNA knockdown of IL10 and IL10R respectively. Exogenous infusion of both M1 and M2 macrophages promoted PDA metastasis in orthotopic mouse model after resident macrophage depletion, although pretreating with DNA demethylating agent Decitabine prevented M1, but not M2 macrophages in promoting metastasis, suggesting M1 macrophages were reprogrammed metabolically and functionally by tumor cells to become pro-cancerous. To interrogate the mechanism that regulates the tumor-induced DNA methylation in macrophages, we first examined chromatin remodeling complex Polycomb Repressive Complex 2 (PRC2), with its major component enhancer of zeste homolog 2 (EZH2) previously shown to recruit DNA methyltransferases to specific chromatin regions for epigenetic gene regulation. We found EZH2 protein was most significantly increased in M1 macrophages while Ser21 phosphorylated EZH2 (pSer21-EZH2) remained constant, indicating a decreased percentage of pSer21-EZH2/EZH2 in M1 macrophages co-cultured with tumor cells. Further identification of potential F-box containing ubiquitin ligases that interact with pSer21-EZH2 by Co-immunoprecipitation suggested EZH2 may be stabilized in M1 macrophages by tumor cells through inhibiting the ubiquitin-dependent proteolysis of pSer21-EZH2 in a phosphorylation dependent manner. This study thus far reveals a novel mechanism that reprograms M1 macrophages phenotypically into M2-like macrophages and functionally into pro-cancerous macrophages. Identification of cell surface receptors that mediate the direct tumor-macrophage contact are actively being sought, which will lead to the development of therapeutic blockades to reverse the macrophage reprogramming. Citation Format: Xingyi Pan, Mengwen Zhang, Kenji Fujiwara, Noelle R. Jurcak, Stephen Muth, Lei Zheng. Tumor-associated macrophages are reprogrammed by pancreatic ductal adenocarcinoma cells through tumor-induced DNA methylation on metabolic genes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4517.

Abstract 4938: Identification of a subtype of hepatocellular carcinoma with poor prognosis based on expression of genes within the glucose metabolic pathway

Abstract Hepatocellular carcinoma (HCC) is the most prevalent primary cancer and a highly aggressive liver malignancy. Liver cancer cells reprogram their metabolic pathways to meet their needs for rapid proliferation and generation of the biomass for tumor growth. The accelerated aerobic glycolysis in tumor has been considered one of the hallmarks that distinguish cancer cells from normal cells. In the present study, liver hepatocellular cancer dataset of the Cancer Genome Atlas (LIHC TCGA) and datasets from Gene Expression Omnibus (GEO) were used to investigate the alterations in expressions of the genes involved in the glucose metabolic pathways as well as their association with the patient clinical stage and survival in liver cancer. In this study, 97 genes including glucose and lactate transporters, the enzymes involved in glucose metabolism (glycolysis, gluconeogenesis, regulation of glucose metabolism, tricarboxylic acid cycle (TCA), pentose phosphate pathway), and glycogen metabolism (glycogen synthesis, glycogen degradation, regulation of glycogen metabolism) were included. The results showed that the expressions of around 50% of genes involved in the glucose metabolic pathway are altered in liver tumors compared to the corresponding normal tissues (p-values&amp;lt;0.005 and fold change≥1.5). More importantly, the differentially expressed genes are associated with advanced AJCC (American Joint Committee on Cancer) clinical stage (p-values&amp;lt;0.05), and reduced overall survival (OS) and recurrence-free survival (RFS) as determined by multivariate Cox proportional hazard models (p-values&amp;lt;0.05). Unexpectedly but interestingly, supervised clustering with the differentially expressed genes of the LIHC TCGA data identified a subgroup of patients with worse OS compared to those patients that clustered with the normal samples (p-value=0.00015, and 5-year median survival time (months) is 25.50 with 95% CI (20.90, 42.37) vs. 47.43 with 95% CI (40.33, NA)). This group of patients had decreased activation (p-values&amp;lt;0.0001) of FXR/RXR and LXR/RXR involved in lipid, cholesterol and glucose metabolism, but had increased atherosclerosis and leukocyte extravasation signaling (p-values&amp;lt;0.0001) activated during inflammation. This indicates those patients had decreased metabolism and increased inflammation which may contribute to the worse OS. Collectively, this study indicates that expressions of the glucose metabolic genes could be used as potential prognostic markers as well as therapeutic targets for liver cancer. Citation Format: Xiaoli Zhang, Kalpana Ghoshal, Lang Li. Identification of a subtype of hepatocellular carcinoma with poor prognosis based on expression of genes within the glucose metabolic pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4938.

Abstract # 3188 Spinal cord injury perturbs circadian rhythms

Spinal cord injury (SCI) can disrupt many physiological functions. The circadian system helps maintain homeostasis throughout the body by regulating daily rhythms in physiology and behavior. Circadian rhythms likely exist in all cell throughout the body, yet whether SCI alters daily rhythms remains under-studied. Here, we hypothesized that SCI in rats would disrupt several prominent circadian outputs including glucocorticoids, core temperature, activity, and circadian/inflammatory gene expression. Female and male rats received clinically relevant thoracic (T-) 8 moderate-to-severe contusion SCI (or laminectomy sham surgery). Circadian measures – including rhythms of plasma corticosterone, body temperature and activity (using small implanted transmitters), plasma glucose, and gene expression – were studied before and after surgery. First, we found that SCI disrupted intraspinal and peripheral rhythms of clock and inflammatory gene expression. Circadian rhythms in peripheral cells are entrained by “oscillatory factors”, including glucocorticoids, body temperature, and activity. SCI increased and disrupted rhythms of the major rodent glucocorticoid, corticosterone. Pre-surgery and sham rats displayed expected rhythms in activity and body temperature, whereas SCI rats had blunted daily rhythms in activity and body temperature. In parallel, SCI increased plasma glucose and liver expression of glucose metabolism genes. Our data show that moderate SCI in rats causes wide-ranging circadian dysfunction that is severe at acute time points and gradually recovers. Normalizing post-SCI circadian rhythms could enhance recovery of homeostasis and quality-of-life.