Peroxisome Proliferator-activated Receptor-gamma Coactivator-1alpha Research Articles

Prevalence of Gly482Ser polymorphism of PPARGC1A in Cuban Americans

Type 2 Diabetes (T2D) has grown to be a major public health issue worldwide. The fundamental clinical characteristic of T2D is high blood glucose levels which may be caused by anomalies in one or more different molecular pathways regulated by certain genes. The Gly482Ser polymorphism in human peroxisome proliferator‐activated receptor gamma coactivator‐1 alpha (PPARGC1A) has been associated with T2D. We genotyped Gly482Ser polymorphism in the PPARGC1A gene from 315 Cuban Americans (164 without T2D / 151 with T2D) using ABI TaqMan assays. Descriptive statistical analysis was performed using SPSS software. The genotype distribution for rs8192678 (Gly482Ser) polymorphism was found to be 0.42 for Gly/Gly, 0.47 for Gly/Ser and 0.10 for Ser/Ser with T2D and 0.47, 0.41 and 0.11 for Gly/Gly, Gly/Ser and Ser/Ser respectively without T2D. The findings in individuals without T2D demonstrate presence of homozygous genotype Gly/Gly to be highest. However, the prevalence of heterozygous genotype Gly/Ser was found to be relatively higher in individuals with T2D than without T2D. The clinical importance of the results has to be established in further studies. Funding was provided by an NIH grant.

Does DNA Methylation of PPARGC1A Influence Insulin Action in First Degree Relatives of Patients with Type 2 Diabetes?

Epigenetics may play a role in the pathophysiology of type 2 diabetes (T2D), and increased DNA methylation of the metabolic master regulator peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PPARGC1A) has been reported in muscle and pancreatic islets from T2D patients and in muscle from individuals at risk of T2D. This study aimed to investigate DNA promoter methylation and gene expression of PPARGC1A in skeletal muscle from first degree relatives (FDR) of T2D patients, and to determine the association with insulin action as well as the influence of family relation. We included 124 Danish FDR of T2D patients from 46 different families. Skeletal muscle biopsies were excised from vastus lateralis and insulin action was assessed by oral glucose tolerance tests. DNA methylation and mRNA expression levels were measured using bisulfite sequencing and quantitative real-time PCR, respectively. The average PPARGC1A methylation at four CpG sites situated 867-624 bp from the transcription start was associated with whole-body insulin sensitivity in a paradoxical positive manner (β = 0.12, P = 0.03), supported by a borderline significant inverse correlation with fasting insulin levels (β = −0.88, P = 0.06). Excluding individuals with prediabetes and overt diabetes did not affect the overall result. DNA promoter methylation was not associated with PPARGC1A gene expression. The familiality estimate of PPARGC1A gene expression was high (h2 = 79±27% (h2±SE), P = 0.002), suggesting genetic regulation to play a role. No significant effect of familiality on DNA methylation was found. Taken together, increased DNA methylation of the PPARGC1A promoter is unlikely to play a major causal role for the development of insulin resistance in FDR of patients with T2D.

High-intensity interval training increases intrinsic rates of mitochondrial fatty acid oxidation in rat red and white skeletal muscle

High-intensity interval training (HIIT) can increase mitochondrial volume in skeletal muscle. However, it is unclear whether HIIT alters the intrinsic capacity of mitochondrial fatty acid oxidation, or whether such changes are associated with changes in mitochondrial FAT/CD36, a regulator of fatty acid oxidation, or with reciprocal changes in the nuclear receptor coactivator (peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1α)) and the corepressor (receptor-interacting protein 140 (RIP140)). We examined whether HIIT alters fatty acid oxidation rates in the isolated subsarcolemmal (SS) and intermyofibrillar (IMF) mitochondria of red and white skeletal muscle and (or) induces changes in muscle PGC-1α and RIP140 proteins and mitochondrial FAT/CD36 protein content. Rats were divided into untrained or HIIT-trained groups. HIIT animals performed 10 bouts of 1-min high-intensity treadmill running (30-55 m·min(-1)), separated by 2 min of rest, for 5 days a week for 4 weeks. As expected, after the training period, HIIT increased mitochondrial enzymes (citrate synthase, COXIV, and β-hydroxyacyl CoA dehydrogenase) in red and white muscle, indicating that muscle mitochondrial volume had increased. HIIT also increased the rates of palmitate oxidation in mitochondria of red (37% for SS and 19% for IMF) and white (36% for SS and 12% for IMF) muscle. No changes occurred in SS and IMF mitochondrial FAT/CD36 proteins, despite increasing FAT/CD36 at the whole-muscle level (27% for red and 22% for white). Concurrently, muscle PGC-1α protein was increased in red (22%) and white (16%) muscle, but RIP140 was not altered. These results indicate that increases in SS and IMF mitochondrial fatty acid oxidation induced by HIIT are accompanied by an increase in PGC-1α, but not RIP140 or FAT/CD36.

Maternal exercise during pregnancy affects mitochondrial enzymatic activity and biogenesis in offspring brain

The present study addresses whether exercise during pregnancy in mouse alters mitochondrial function in the brains of the resultant offspring. We divided pregnant mice into four groups: a control group and groups of mice that exercised for 20 (E20m), 30 (E30m) and 40 min/d (E40m). The pregnant mice ran on a treadmill at 12 m/min, 5 d/week for a duration of 3 weeks. The protein expression of cytochrome c oxidase subunit Va (CVa) was downregulated in the offspring of the E20m group, unlike that in the control animals, whereas CVa expression was reserved in the E40m neonates. The F1-ATPase catalytic core (Core) protein expression levels were the highest in the E40m group neonates. Complex I, IV and ATPase activities were significantly lower in the E20m group than that in the control group neonates and were reserved in the E30m and E40m group neonates. The activities of citrate synthase and pyruvate dehydrogenase were consistent with those of complex I, IV and ATPase. Peroxisome proliferator-activated receptor-gamma coactivator 1-alpha, mitochondrial transcription factor A, nuclear respiratory factor-1 and mitochondrial DNA showed high levels of expression in the E40m neonates compared with the other groups. Malondialdehyde (MDA) levels in E40m neonates were higher than that in the controls but were lower than that in the E20m neonates. Finally, 40 min/d of maternal exercise improved mitochondrial function in the resultant pups and was concomitant with brain-derived trophic factor induction in the hippocampus, thereby functionally improving short-term memory.

Regulation of Mitochondrial Oxidative Metabolism by Tumor Suppressor FLCN

Birt-Hogg-Dubé (BHD) syndrome is a hereditary hamartoma syndrome that predisposes patients to develop hair follicle tumors, lung cysts, and kidney cancer. Genetic studies of BHD patients have uncovered the causative gene, FLCN, but its function is incompletely understood. Mice with conditional alleles of FLCN and/or peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PPARGC1A), a transcriptional coactivator that regulates mitochondrial biogenesis, were crossbred with mice harboring either muscle creatine kinase (CKM) -Cre or myogenin (MYOG) -Cre transgenes to knock out FLCN and/or PPARGC1A in muscle, or cadherin 16 (CDH16)- Cre transgenes to knock out FLCN and/or PPARGC1A in kidney. Real-time polymerase chain reaction, immunoblotting, electron microscopy, and metabolic profiling assay were performed to evaluate mitochondrial biogenesis and function in muscle. Immunoblotting, electron microscopy, and histological analysis were used to investigate expression and the pathological role of PPARGC1A in FLCN-deficient kidney. Real-time polymerase chain reaction, oxygen consumption measurement, and flow cytometry were carried out using a FLCN-null kidney cancer cell line. All statistical analyses were two-sided. Muscle-targeted FLCN knockout mice underwent a pronounced metabolic shift toward oxidative phosphorylation, including increased mitochondrial biogenesis (FLCN ( f/f ) vs FLCN ( f/f ) /CKM-Cre: % mitochondrial area mean = 7.8% vs 17.8%; difference = 10.0%; 95% confidence interval = 5.7% to 14.3%; P < .001), and the observed increase in mitochondrial biogenesis was PPARGC1A dependent. Reconstitution of FLCN-null kidney cancer cells with wild-type FLCN suppressed mitochondrial metabolism and PPARGC1A expression. Kidney-targeted PPARGC1A inactivation partially rescued the enlarged kidney phenotype and abrogated the hyperplastic cells observed in the FLCN-deficient kidney. FLCN deficiency and subsequent increased PPARGC1A expression result in increased mitochondrial function and oxidative metabolism as the source of cellular energy, which may give FLCN-null kidney cells a growth advantage and drive hyperplastic transformation.

Salvianolic Acid A Protects the Peripheral Nerve Function in Diabetic Rats through Regulation of the AMPK-PGC1α-Sirt3 Axis

Salvianolic acid A (SalA) is one of the main efficacious, water-soluble constituents of Salvia miltiorrhiza Bunge. This study investigated the protective effects of SalA on peripheral nerve in diabetic rats. Administration of SalA (0.3, 1 and 3 mg/kg, ig) was started from the 5th week after strepotozotocin (STZ60 mg/kg) intraperitoneal injection and continued for 8 weeks. Paw withdrawal mechanical threshold (PWMT) and motor nerve conduction velocity (MNCV) were used to assess peripheral nerve function. The western blot methods were employed to test the expression levels of serine-threonine liver kinase B1 (LKB1), AMP-activated protein kinase (AMPK), peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1α), silent information regulator protein3 (sirtuin 3/Sirt3) and neuronal nitric oxide synthase (nNOS) in sciatic nerve. Results showed that SalA administration could increase PWMT and MNCV in diabetic rats; reduce the deterioration of sciatic nerve pathology; increase AMPK phosphorylation level, up-regulate PGC-1α, Sirt3 and nNOS expression, but had no influence on LKB1. These results suggest that SalA has protective effects against diabetic neuropathy. The beneficial effects of SalA on peripheral nerve function in diabetic rats might be attributed to improvements in glucose metabolism through regulation of the AMPK-PGC1α-Sirt3 axis.

Weight Phenotype Diagnostic Test Method: Body Mass Index or Body Fat Percent for Gene Expression

Obesity continues increasing at epidemic levels worldwide, as does the number of genetic studies that focus on obesity. Body mass index (BMI) is often used to characterize weight phenotypes and obesity status due to its simplicity. Refined measurements of body composition may be needed to understand variations in gene expression. This study explores gene expression when individuals are characterized as overweight based on BMI versus body fat percent. Individuals were recruited to a natural history protocol at the National Institutes of Health. Twelve Caucasian participants with the highest and lowest BMI were included. Whole-body air displacement plethysmography was performed to calculate body fat percent, and BMI was calculated. Fasting whole blood was collected and RNA extracted. Quantitative real time PCR array was used to determine expression of 96 obesity related genes. The PCR array from participants with high BMI compared to low BMI showed dysregulation of four genes: peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PPARGC1A), pro-opiomelanocortin (POMC), growth hormone secretagogue receptor (GHSR), and leptin (LEP), whereas participants with high body fat compared to low body fat showed dysregulation of one gene: PPARGC1A. This research showed differential gene expression and clinical indices depending on method of weight classification.

Abnormality in Expression Levels of Gluconeogenesis-Related Genes by High-Dose Supplementation with Pyridoxamine in Mice

Pyridoxamine supplementation caused the alteration of the expression of genes encoding six gluconeogenesis-related proteins. The expression levels of phosphoenolpyruvate carboxykinase, pyruvate kinase, and pyruvate dehydrogenase kinase 4 in the pyridoxamine-supplemented mice were higher than those in the control mice. In contrast, the pyridoxamine supplementation caused lower expression levels of peroxisome proliferator-activated receptor-gamma coactivator-1alpha, carbohydrate response element-binding protein, glucocorticoid receptor, and glucose-6-phosphatase. The pyridoxamine-supplemented mice showed significantly low glucose clearance in a glucose tolerance test, but they showed no symptoms of diabetes, which was estimated according to the levels of hemoglobin A1c and blood glucose. Pyruvate challenge testing suggested that pyridoxamine supplementation enhanced gluconeogenic activity from pyruvate. The results showed that a high-dose of pyridoxamine may require a careful inquiry concerning its validity.

ACE I/D, ACTN3 R577X, PPARD T294C and PPARGC1A Gly482Ser polymorphisms and physical fitness in Taiwanese late adolescent girls

Physical performance of youth is influenced by various factors, including body composition, biological maturity status, level of habitual physical activity, and muscular strength. Muscular strength has been largely attributed to genetic effects. To exclude possible confounding effects from various acquired factors, this study examined the relationships between polymorphisms of the angiotensin-converting enzyme (ACE), α-actinin-3 (ACTN3), peroxisome proliferator-activated receptor delta (PPARD), and peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PPARGC1A) genes and performance as measured by six fitness tests (handgrip strength of dominant hand, 30- and 60-s sit-ups, standing long jump, 60-m dash, and 800-m run) in 170 sedentary adolescent girls with the adjustment of anthropometric characteristics. We found that subjects with the ACE DD genotype were significantly heavier than those with I allele, while those with the ACTN3 RR genotype had higher fat-free mass percentage (FFM%) than those with the XX genotype. In addition, those with the PPARD TT genotype were significantly taller, heavier, and had a greater FFM than those with the CC genotype. Subjects with the ACE DD, ACTN3 RR and PPARD TC genotype had better performance in handgrip strength, 30- and 60-s sit-up tests, and standing long jump, respectively, when individual gene was analyzed independently after adjusting anthropometric characteristics. In the gene combination analysis, subjects with ACE DD, ACTN3 RR and PPARD TT genotype had significantly greater performance in handgrip strength. Overall, the results indicate that the genes studied have a modest influence on individual performance as assessed by specific fitness and strength tests in female late adolescents.

Regulatory roles for L-arginine in reducing white adipose tissue.

As the nitrogenous precursor of nitric oxide, L-arginine regulates multiple metabolic pathways involved in the metabolism of fatty acids, glucose, amino acids, and proteins through cell signaling and gene expression. Specifically, arginine stimulates lipolysis and the expression of key genes responsible for activation of fatty acid oxidation to CO2 and water. The underlying mechanisms involve increases in the expression of peroxisome proliferator-activated receptor-gamma coactivator-1 alpha (PGC-1 alpha), mitochondrial biogenesis, and the growth of brown adipose tissue growth. Furthermore, arginine regulates adipocyte-muscle crosstalk and energy partitioning via the secretion of cytokines and hormones. In addition, arginine enhances AMP-activated protein kinase (AMPK) expression and activity, thereby modulating lipid metabolism and energy balance toward the loss of triacylglycerols. Growing evidence shows that dietary supplementation with arginine effectively reduces white adipose tissue in Zucker diabetic fatty rats, diet-induced obese rats, growing-finishing pigs, and obese patients with type II diabetes. Thus, arginine can be used to prevent and treat adiposity and the associated metabolic syndrome.

Modulation of Longevity and Tissue Homeostasis by the Drosophila PGC-1 Homolog

In mammals, the PGC-1 transcriptional coactivators are key regulators of energy metabolism, including mitochondrial biogenesis and respiration, which have been implicated in numerous pathogenic conditions, including neurodegeneration and cardiomyopathy. Here, we show that overexpression of the Drosophila PGC-1 homolog (dPGC-1/spargel) is sufficient to increase mitochondrial activity. Moreover, tissue-specific overexpression of dPGC-1 in stem and progenitor cells within the digestive tract extends life span. Long-lived flies overexpressing dPGC-1 display a delay in the onset of aging-related changes in the intestine, leading to improved tissue homeostasis in old flies. Together, these results demonstrate that dPGC-1 can slow aging both at the level of cellular changes in an individual tissue and also at the organismal level by extending life span. Our findings point to the possibility that alterations in PGC-1 activity in high-turnover tissues, such as the intestine, may be an important determinant of longevity in mammals.

HDACs confer glucagon responsiveness to FoxO for gluconeogenesis in the liver

In mammals deprived of food, induction of hepatic glucose production (HGP) is important to ensure energy homeostasis, including prevention of hypoglycemia in response to the energy demands [1], and is especially critical in sites that predominantly utilize glucose as an energy source, such as the brain, red blood cells, and renal medulla. Glucose homeostasis is maintained through tight regulation of glucose production (i.e., glycogenolysis and gluconeogenesis) in the liver during the state of fasting and uptake of glucose in peripheral tissues under the fed condition. The pancreatic hormones glucagon and insulin are central to this regulation. During fasting, glucagon enhances HGP to maintain euglycemia. Conversely, when nutrients are increased after a meal, insulin blunts HGP and increases glucose uptake in the skeletal muscle and adipose tissue to prevent hyperglycemia. However, these processes are dysregulated in type 2 diabetes, resulting in the development of hyperglycemia [2]. The aberrant increase of HGP during the postabsorptive state in diabetic patients is primarily due to suppression failure by insulin, resulting primarily from excessive gluconeogenesis rather than glycogenolysis [3]. Gluconeogenesis is mainly regulated at the transcriptional level of rate-limiting gluconeogenic enzymes, such as glucose-6-phosphatase (G6pc; G6Pase) and phosphoenolpyruvate carboxykinase (Pck1; PEPCK) through hormone-dependent modulation of transcriptional regulators, including the transcription factors for FoxO (forkhead box O), CREB (cAMP response element binding protein), HNF4ahepatocytes nuclear factor 4a and glucocorticoid receptor, and transcriptional coactivators, such as PGC-1a (peroxisome proliferator-activated receptor gamma coactivator-1 alpha), CRTC2 (CREB-regulated transcription coactivator 2), and the histone acetyltransferase CBP (CREB-binding protein) [4]. Glucagon stimulates the induction of gluconeogenic genes through the following signaling pathway: cAMP, a second messenger of the glucagon receptor, leads cAMPdependent protein kinase (PKA)-mediated activation of the CRTC2/CREB complex to induce PGC-1a, which in turn coactivates the FoxO and HNF4a transcription factors on the G6pc and Pck1 promoters (Fig. 1). Insulin blunts gluconeogenic gene induction by affecting the same pathway. This is mediated mainly through the serine/threonine kinase Akt, which inactivates FoxO and PGC-1a via direct phosphorylation, as well as CRTC2 via SIK (salt-inducible kinase) 1/2 [5] (Fig. 1). The FoxO family of transcription factors, mainly FoxO1 and FoxO3 in mammalian liver [6], plays pivotal roles in regulating gluconeogenesis, as shown by studies on in vivo loss of and gain of function study; FoxO1 loss of function in the liver reduces HGP by inhibiting gluconeogenesis and glycogenolysis, resulting in neonatal and fasting-induced hypoglycemia [6], whereas its gain of function impairs hormonal regulation of HGP, including gluconeogenesis [7]. FoxO is regulated by insulin; Akt-dependent phosphorylation promotes its nuclear exclusion and inactivates FoxO1, leading to the reduction of gluconeogenic genes [8]. FoxO is also controlled by cAMP, which promotes its dephosphorylation and nuclear retention in cultured hepatocytes [8]. In vivo, FoxO1 loss of function in the liver attenuates gluconeogenic gene induction and HGP during fasting [6]. Although these findings indicate that FoxO is indispensable for cAMP-dependent induction of gluconeogenic genes, the M. Matsumoto (&) Department of Molecular Metabolic Regulation, Diabetes Research Center, National Center for Global Health and Medicine, 1-21-1 Toyama, Shinjuku-ku, Tokyo 162-8655, Japan e-mail: mmatsumoto@ri.ncgm.go.jp

Novel thromboxane A2 analog-induced IUGR mouse model.

Rodents, particularly rats, are used in the majority of intrauterine growth restriction (IUGR) research. An important tool that is lacking in this field is the ability to impose IUGR on transgenic mice. We therefore developed a novel mouse model of chronic IUGR using U-46619, a thromboxane A2 (TXA2) analog, infusion. TXA2 overproduction is prevalent in human pregnancies complicated by cigarette smoking, diabetes mellitus and preeclampsia. In this model, U-46619 micro-osmotic pump infusion in the last week of C57BL/6J mouse gestation caused maternal hypertension. IUGR pups weighed 15% less, had lighter brain, lung, liver and kidney weights, but had similar nose-to-anus lengths compared with sham pups at birth. Metabolically, IUGR pups showed increased essential branched-chain amino acids. They were normoglycemic yet hypoinsulinemic. They showed decreased hepatic mRNA levels of total insulin-like growth factor-1 and its variants, but increased level of peroxisome proliferator-activated receptor-gamma coactivator-1 alpha. IUGR offspring were growth restricted from birth (P1) through postnatal day 21 (P21). IUGR males caught up with sham males in weight by P28, whereas IUGR females caught up with sham females by P77. IUGR males surpassed sham males in weight by P238. In summary, we have a non-brain sparing IUGR mouse model that has a relative ease of surgical IUGR induction and exhibits features similar to the chronic IUGR offspring of humans and other animal models. As transgenic technology predominates in mice, this model now permits the imposition of IUGR on transgenic mice to interrogate mechanisms of fetal origins of adult disease.

A follow‐up case–control association study of tractable (druggable) genes in recurrent major depression

The High-Throughput Disease-specific target Identification Program (HiTDIP) aimed to study case-control association samples for 18 common diseases. Here we present the results of a follow-up case-control association study of HiTDIP in major depressive disorder (MDD). The HiTDIP in MDD was conducted in a sample of 974 cases of recurrent MDD of white German origin collected at the Max-Planck Institute (MP-GSK) and 968 ethnically matched controls screened for lifetime absence of depression. Six genes were identified as of interest for a follow-up, based on the strength of the association and based on the interest as potential candidate target for developing new treatment for depression: Solute Carrier Family 4 Member 10 (SLC4A10), Dipeptidyl Peptidase IV (DPP4), Dopamine Receptor D3 (DRD3), Zinc Finger Protein 80 (ZNF80), Nitric Oxide Synthase 2A (NOS2A) and Peroxisome Proliferator-Activated Receptor-Gamma, Coactivator 1, Alpha (PPARGC1A). Within the current study, we attempted to follow-up these findings in a sample from the UK, the Depression Case Control (DeCC) sample consisting of 1,196 cases and 842 screened controls, phenotyped using exactly the same methods as the MP-GSK sample. Performing Cochran-Mantel-Haenzel statistics to test for genotypic and/or allelic differences between the DeCC and MP-GSK samples, we found no significant differences, thus being able to combine the two samples for association testing. In the combined sample of 2,170 MDD cases and 1,810 controls, there were positive findings in the Nitric Oxide Synthase 2A (NOS2A) gene both using single SNP analysis and haplotype analysis.

Abstract LB-121: Polymorphic variants in PPARGC1A and PCMTD1 associate with earlier age of onset in ER-negative breast cancer

Abstract Background: Breast cancer is the most frequent cancer and leading cause of cancer death among women worldwide. As &amp;lt;10% of breast cancers are due to mutations in BRCA1 and BRCA2, other perturbations such as single nucleotide polymorphisms (SNPs) may be genetic contributors. Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PPARGC1A) enhances estrogen receptor (ER) and estrogen related receptor gamma (ERRγ) activity by acting as a coactivator. ERRγ has been demonstrated to regulate vascular endothelial growth factor mRNA expression in both ER-positive and ER-negative breast cancer cell lines, indicating that it may accelerate angiogenesis and metastasis. PCMTD1 encodes protein-L-isoaspartate O-methyltransferase domain-containing protein 1, a S-adenosylmethionine-dependent methyltransferase whose biological relevance is largely unknown to present. SNPs in PPARGC1A have been correlated with metabolic diseases, but within the three studies evaluating cancer phenotypes for this gene, only Thr612Met (76873 C→T) revealed a significant association with familial breast cancers. Experimental Procedure: SNPs in PPARGC1A and PCMTD1 were selected using a bioinformatics approach based on SNP associations with drug response in NCI-60 cell lines and evidence of selection bias for haplotype frequencies. Allelic discrimination assays were performed on genomic DNA isolated from whole blood from over 900 Caucasian breast cancer patients with ductal carcinomas. Clinical information was linked to genotypes to search for genotype-phenotype correlations, and statistical analyses were carried out. Results: Analysis of PPARGC1A rs4619879 revealed a mean age of diagnosis of ER-negative cases for CC and AA carriers at 50.1 and 43.8 years, respectively (p=0.019). The difference was less striking in ER-positive ductal carcinomas, with only a 2.7year earlier onset for AA carriers (p=0.013). For PCMTD1 rs9298462, the mean age at diagnosis of ER-negative cases for CC carriers was 41.2, compared to 48.8 for the TT carriers (p=0.004). No significant difference was seen in ER-positive ductal carcinomas (p=0.15). Conclusions: Our results indicate that polymorphisms in PPARGC1A and PCMTD1, two transcriptional regulators, may play a role in age at diagnosis in breast cancer, particularly ER-negative ductal carcinomas. We are performing functional studies to further evaluate these findings and to elucidate the molecular mechanisms underlying these observations. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-121. doi:10.1158/1538-7445.AM2011-LB-121

Association of peroxisome proliferator‐activated receptor gamma coactivator 1 alpha (PPARGC1A) gene polymorphisms and type 2 diabetes mellitus: a meta‐analysis

The association between peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PPARGC1A) polymorphisms and type 2 diabetes mellitus (T2DM) has been investigated in several studies, but these studies yielded contradictory results. We conducted a meta-analysis to assess the association between three polymorphisms (Gly482Ser, Thr394Thr and Thr612Met) in PPARGC1A and T2DM. A literature-based search was conducted to collect data. The additive model was chosen to investigate the association between the three polymorphisms and T2DM. The random effects model was used if there was heterogeneity between studies. In addition, subgroup meta-analyses were made according to the ethnic groups. Twenty-three studies were enrolled in this meta-analysis (7539 cases and 9562 controls for Gly482Ser, 1818 cases and 2376 controls for Thr394Thr, 2042 cases and 1289 controls for Thr612Met). In the combined analysis of all eligible studies, a significant association was found between Gly482Ser, Thr394Thr and T2DM with pooled odds ratios 1.19 [95% confidence interval (CI) 1.05-1.34] and 1.33 (95% CI 1.04-1.70), respectively, but great heterogeneity was found between studies. In the subgroup meta-analyses, we found that Gly482Ser and Thr394Thr polymorphisms were associated with the risk of T2DM, and the pooled odds ratios were 1.66 (95% CI 1.28-2.15) and 1.72 (95% CI 1.45-2.03), respectively, in the Indian population; no significant evidence was found in the Caucasian and East Asian populations. This meta-analysis indicated that Gly482Ser and Thr394Thr polymorphisms of PPARGC1A gene were significantly associated with the risk of T2DM, especially in the Indian population. No relationship was found between the Thr612Met and risk of T2DM.

Mitochondrial defect and PGC-1α dysfunction in parkin-associated familial Parkinson's disease

Mutations in the parkin gene are expected to play an essential role in autosomal recessive Parkinson's disease. Recent studies have established an impact of parkin mutations on mitochondrial function and autophagy. In primary skin fibroblasts from two patients affected by an early onset Parkinson's disease, we identified a hitherto unreported compound heterozygous mutation del exon2-3/del exon3 in the parkin gene, leading to the complete loss of the full-length protein. In both patients, but not in their heterozygous parental control, we observed severe ultrastructural abnormalities, mainly in mitochondria. This was associated with impaired energy metabolism, deregulated reactive oxygen species (ROS) production, resulting in lipid oxidation, and peroxisomal alteration. In view of the involvement of parkin in the mitochondrial quality control system, we have investigated upstream events in the organelles' biogenesis. The expression of the peroxisome proliferator-activated receptor gamma-coactivator 1-alpha (PGC-1α), a strong stimulator of mitochondrial biogenesis, was remarkably upregulated in both patients. However, the function of PGC-1α was blocked, as revealed by the lack of its downstream target gene induction. In conclusion, our data confirm the role of parkin in mitochondrial homeostasis and suggest a potential involvement of the PGC-1α pathway in the pathogenesis of Parkinson's disease. This article is part of a Special Issue entitled: Translating nuclear receptors from health to disease.

Peroxisome Proliferator-activated Receptor-γ Coactivator-1α (PGC-1α) Stimulates VLDL Assembly through Activation of Cell Death-inducing DFFA-like Effector B (CideB)

Abnormalities in very low density lipoprotein (VLDL) assembly and secretion impact intrahepatic lipid homeostasis, plasma lipoprotein profile, and energy metabolism of distal peripheral tissues. We have evaluated the role of the transcriptional coactivator, the peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha), in VLDL assembly and secretion. PGC-1alpha overexpression in HepG2 cells led to diminished rates of triglyceride (TG) synthesis but strongly stimulated VLDL-TG secretion, markedly increasing the efficiency of secretion of newly synthesized TG. PGC-1alpha overexpression increased the rate of secretion of apoB100 and promoted secretion of larger, less dense VLDL particles. PGC-1alpha overexpression in intact mouse liver also stimulated rates of VLDL TG secretion and attenuated hepatic TG accumulation resulting from high fat diet feeding. To determine the molecular mechanisms mediating the effect of PGC-1alpha on VLDL assembly, we evaluated the expression of several candidate mediators known to be involved in VLDL assembly or hepatic lipid homeostasis. Cell death-inducing DFFA-like effector B (CideB) expression was greatly induced by PGC-1alpha, and siRNA against CideB reversed the effects of PGC-1alpha on the secretion of TG and VLDL-sized particles by HepG2 cells, indicating that CideB is a critical mediator of stimulatory effects of PGC-1alpha on VLDL secretion. Collectively, these data suggest that PGC-1alpha plays an important role in partitioning cytoplasmic TG toward the VLDL secretory compartments and promoting VLDL secretion via transcriptional induction of CideB.

Sirtuin 3, a New Target of PGC-1α, Plays an Important Role in the Suppression of ROS and Mitochondrial Biogenesis

BackgroundSirtuin 3 (SIRT3) is one of the seven mammalian sirtuins, which are homologs of the yeast Sir2 gene. SIRT3 is the only sirtuin with a reported association with the human life span. Peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) plays important roles in adaptive thermogenesis, gluconeogenesis, mitochondrial biogenesis and respiration. PGC-1α induces several key reactive oxygen species (ROS)-detoxifying enzymes, but the molecular mechanism underlying this is not well understood.ResultsHere we show that PGC-1α strongly stimulated mouse Sirt3 gene expression in muscle cells and hepatocytes. Knockdown of PGC-1α led to decreased Sirt3 gene expression. PGC-1α activated the mouse SIRT3 promoter, which was mediated by an estrogen-related receptor (ERR) binding element (ERRE) (−407/−399) mapped to the promoter region. Chromatin immunoprecipitation and electrophoretic mobility shift assays confirmed that ERRα bound to the identified ERRE and PGC-1α co-localized with ERRα in the mSirt3 promoter. Knockdown of ERRα reduced the induction of Sirt3 by PGC-1α in C2C12 myotubes. Furthermore, Sirt3 was essential for PGC-1α-dependent induction of ROS-detoxifying enzymes and several components of the respiratory chain, including glutathione peroxidase-1, superoxide dismutase 2, ATP synthase 5c, and cytochrome c. Overexpression of SIRT3 or PGC-1α in C2C12 myotubes decreased basal ROS level. In contrast, knockdown of mSIRT3 increased basal ROS level and blocked the inhibitory effect of PGC-1α on cellular ROS production. Finally, SIRT3 stimulated mitochondrial biogenesis, and SIRT3 knockdown decreased the stimulatory effect of PGC-1α on mitochondrial biogenesis in C2C12 myotubes.ConclusionOur results indicate that Sirt3 functions as a downstream target gene of PGC-1α and mediates the PGC-1α effects on cellular ROS production and mitochondrial biogenesis. Thus, SIRT3 integrates cellular energy metabolism and ROS generation. The elucidation of the molecular mechanisms of SIRT3 regulation and its physiological functions may provide a novel target for treating ROS-related disease.

Exercise intensity-dependent regulation of peroxisome proliferator-activated receptor γ coactivator-1α mRNA abundance is associated with differential activation of upstream signalling kinases in human skeletal muscle

Skeletal muscle contraction increases intracellular ATP turnover, calcium flux, and mechanical stress, initiating signal transduction pathways that modulate peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1alpha)-dependent transcriptional programmes. The purpose of this study was to determine if the intensity of exercise regulates PGC-1alpha expression in human skeletal muscle, coincident with activation of signalling cascades known to regulate PGC-1alpha transcription. Eight sedentary males expended 400 kcal (1674 kj) during a single bout of cycle ergometer exercise on two separate occasions at either 40% (LO) or 80% (HI) of . Skeletal muscle biopsies from the m. vastus lateralis were taken at rest and at +0, +3 and +19 h after exercise. Energy expenditure during exercise was similar between trials, but the high intensity bout was shorter in duration (LO, 69.9 +/- 4.0 min; HI, 36.0 +/- 2.2 min, P < 0.05) and had a higher rate of glycogen utilization (P < 0.05). PGC-1alpha mRNA abundance increased in an intensity-dependent manner +3 h after exercise (LO, 3.8-fold; HI, 10.2-fold, P < 0.05). AMP-activated protein kinase (AMPK) (2.8-fold, P < 0.05) and calcium/calmodulin-dependent protein kinase II (CaMKII) phosphorylation (84%, P < 0.05) increased immediately after HI but not LO. p38 mitogen-activated protein kinase (MAPK) phosphorylation increased after both trials (2.0-fold, P < 0.05), but phosphorylation of the downstream transcription factor, activating transcription factor-2 (ATF-2), increased only after HI (2.4-fold, P < 0.05). Cyclic-AMP response element binding protein (CREB) phosphorylation was elevated at +3 h after both trials (80%, P < 0.05) and class IIa histone deacetylase (HDAC) phosphorylation increased only after HI (2.0-fold, P < 0.05). In conclusion, exercise intensity regulates PGC-1alpha mRNA abundance in human skeletal muscle in response to a single bout of exercise. This effect is mediated by differential activation of multiple signalling pathways, with ATF-2 and HDAC phosphorylation proposed as key intensity-dependent mediators.