New insights into adipose tissue VEGF-A actions in the control of obesity and insulin resistance

Altres ajuts: The work in our lab relevant to this commentary was supported by grants from EUMODIC (LSHG-CT-2006-037188).Vascular endothelial growth factor A (VEGF-A) is classically viewed as a key factor in angiogenesis and tissue remodeling. However, recent evidence suggests a potential role of this growth factor in the control of energy metabolism and adipose tissue function. In this regard, we and others have described the effects of the up and downregulation of VEGF-A in adipose tissue on the control of energy homeostasis. VEGF-A overexpression protects against diet-induced obesity and insulin resistance. The observation that VEGF-A overexpression leads to an increase in brown adipose tissue (BAT) thermogenesis and also promotes a "BAT-like" phenotype in white adipose tissue depots is of particular relevance for the understanding of the mechanisms underlying obesity development. In addition, VEGF-A may not only have pro-inflammatory but also anti-inflammatory properties, with a chemotactic activity specific for M2 anti-inflammatory macrophages. This new scientific evidence highlights the importance that VEGF-A actions on metabolism could have on the design of new treatments for obesity, insulin resistance and obesity-related disorders

PADAMOT : project overview report

Background and relevance to radioactive waste management International consensus confirms that placing radioactive wastes and spent nuclear fuel deep underground in a geological repository is the generally preferred option for their long-term management and disposal. This strategy provides a number of advantages compared to leaving it on or near the Earth’s surface. These advantages come about because, for a well chosen site, the geosphere can provide: • a physical barrier that can negate or buffer against the effects of surface dominated natural disruptive processes such as deep weathering, glaciation, river and marine erosion or flooding, asteroid/comet impact and earthquake shaking etc. • long and slow groundwater return pathways from the facility to the biosphere along which retardation, dilution and dispersion processes may operate to reduce radionuclide concentration in the groundwater. • a stable, and benign geochemical environment to maximise the longevity of the engineered barriers such as the waste containers and backfill in the facility. • a natural radiation shield around the wastes. • a mechanically stable environment in which the facility can be constructed and will afterwards be protected. • an environment which reduces the likelihood of the repository being disturbed by inadvertent human intrusion such as land use changes, construction projects, drilling, quarrying and mining etc. • protection against the effects of deliberate human activities such as vandalism, terrorism and war etc. However, safety considerations for storing and disposing of long-lived radioactive wastes must take into account various scenarios that might affect the ability of the geosphere to provide the functionality listed above. Therefore, in order to provide confidence in the ability of a repository to perform within the deep geological setting at a particular site, a demonstration of geosphere “stability” needs to be made. Stability is defined here to be the capacity of a geological and hydrogeological system to minimise the impact of external influences on the repository environment, or at least to account for them in a manner that would allow their impacts to be evaluated and accounted for in any safety assessments. A repository should be sited where the deep geosphere is a stable host in which the engineered containment can continue to perform according to design and in which the surrounding hydrogeological, geomechanical and geochemical environment will continue to operate as a natural barrier to radionuclide movement towards the biosphere. However, over the long periods of time during which long-lived radioactive wastes will pose a hazard, environmental change at the surface has the potential to disrupt the stability of the geosphere and therefore the causes of environmental change and their potential consequences need to be evaluated. As noted above, environmental change can include processes such as deep weathering, glaciation, river and marine erosion. It can also lead to changes in groundwater boundary conditions through alternating recharge/discharge relationships. One of the key drivers for environmental change is climate variability. The question then arises, how can geosphere stability be assessed with respect to changes in climate? Key issues raised in connection with this are: • What evidence is there that 'going underground' eliminates the extreme conditions that storage on the surface would be subjected to in the long term? • How can the additional stability and safety of the deep geosphere be demonstrated with evidence from the natural system? As a corollary to this, the capacity of repository sites deep underground in stable rock masses to mitigate potential impacts of future climate change on groundwater conditions therefore needs to be tested and demonstrated. To date, generic scenarios for groundwater evolution relating to climate change are currently weakly constrained by data and process understanding. Hence, the possibility of site-specific changes of groundwater conditions in the future can only be assessed and demonstrated by studying groundwater evolution in the past. Stability of groundwater conditions in the past is an indication of future stability, though both the climatic and geological contexts must be taken into account in making such an assertion

Methodology for Hydrogeochemical Sampling to Characterise Groundwaters in Crystalline Bedrock: Developments Made within the Swedish Radwaste Programme

The search by SKB (Swedish Nuclear Fuel and Waste Management Co.) for a site to locate the deep geological repository for spent nuclear fuel in Sweden has involved geoscientific investigations at several locations since the 1970s. The objectives were to characterise geologically a bedrock volume as well as its hydrogeology and hydrochemistry. To acquire high-quality hydrogeochemical data, a complete system for groundwater sampling and analysis, as well as for interpretation strategies, has been developed through a continuous process of modification and refinement. Since the largest part of the Swedish bedrock is composed of granitoids, the site investigations had to adapt to the special difficulties of fractured crystalline rocks. This paper discusses the problems with groundwater sampling that are specific to fractured crystalline rocks and describes the solutions adopted and methods developed by SKB since the early 2000s during the site investigations. The methodology described in this paper for the characterisation of deep groundwaters in crystalline rocks is not only applicable in the context of radioactive waste disposal but also useful when sampling groundwaters for any purpose in such rocks. Sampling of groundwaters in fractured rocks at depth, often down to approximately 1, 000 m, involves special challenges since the natural conditions of the groundwater are easily disturbed, especially by the initial drilling, but also by every subsequent activity performed in the borehole, including the actual groundwater sampling. The sampling strategy presented in this paper shows that planning of the sampling preferably starts already when the drilling procedure is decided. Each following step is described in detail and includes tracing the drilling fluid, selecting the best borehole sections to sample, procedures for the actual sampling, and selection of analytical protocol; all this with the goal of taking representative samples. Although the evaluation of the sampling uncertainties is not a straightforward procedure, an adequate categorisation routine has been established to classify groundwater samples regarding sample quality, representativeness, and suitability for further interpretations and modelling

AAV-mediated Sirt1 overexpression in skeletal muscle activates oxidative capacity but does not prevent insulin resistance

Type 2 diabetes is characterized by triglyceride accumulation and reduced lipid oxidation capacity in skeletal muscle. SIRT1 is a key protein in the regulation of lipid oxidation and its expression is reduced in the skeletal muscle of insulin resistant mice. In this tissue, Sirt1 up-regulates the expression of genes involved in oxidative metabolism and improves mitochondrial function mainly through PPARGC1 deacetylation. Here we examined whether Sirt1 overexpression mediated by adeno-associated viral vectors of serotype 1 (AAV1) specifically in skeletal muscle can counteract the development of insulin resistance induced by a high fat diet in mice. AAV1- Sirt1 -treated mice showed up-regulated expression of key genes related to β-oxidation together with increased levels of phosphorylated AMP protein kinase. Moreover, SIRT1 overexpression in skeletal muscle also increased basal phosphorylated levels of AKT. However, AAV1- Sirt1 treatment was not enough to prevent high fat diet-induced obesity and insulin resistance. Although Sirt1 gene transfer to skeletal muscle induced changes at the muscular level related with lipid and glucose homeostasis, our data indicate that overexpression of SIRT1 in skeletal muscle is not enough to improve whole-body insulin resistance and that suggests that SIRT1 has to be increased in other metabolic tissues to prevent insulin resistance

Expression of the TGF-beta1 system in human testicular pathologies

In non-obstructive azoospermia, histological patterns of Sertoli cell-only Syndrome (SCO) and hypospermatogenesis (H) are commonly found. In these pathologies, Leydig cell hyperplasia (LCH) is detected in some patients. Since TGF-β1 is involved in cellular proliferation/development, the aim of this work was to analyze the expression of TGF-β1, its receptors TGFBRII, TGFBRI (ALK-1 and ALK-5), and the co-receptor endoglin in human biopsies from patients with idiopathic infertilityFil: Gonzalez, Candela Rocio. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina; Argentina. Universidad de Buenos Aires. Facultad de Medicina; ArgentinaFil: Matzkin, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina; Argentina. Universidad de Buenos Aires. Facultad de Medicina; ArgentinaFil: Frungieri, Monica Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina; Argentina. Universidad de Buenos Aires. Facultad de Medicina; ArgentinaFil: Terradas, Claudio. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos"Carlos G. Durand"; Argentina. Instituto Médico IPREFER; ArgentinaFil: Ponzio, Roberto . Universidad de Buenos Aires. Facultad de Medicina; ArgentinaFil: Puigdomenech, Elisa. Instituto Médico IPREFER;; ArgentinaFil: Levalle, Oscar. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos"Carlos G. Durand"; ArgentinaFil: Calandra, Ricardo Saul. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina; ArgentinaFil: Gonzalez Calvar, Silvia Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina; Argentina. Universidad de Buenos Aires. Facultad de Medicina; Argentin

BMP4 gene therapy enhances insulin sensitivity but not adipose tissue browning in obese mice

Bone morphogenetic protein 4 (BMP4) adeno-associated viral vectors of serotype 8 (AAV8) gene therapy targeting the liver prevents the development of obesity in initially lean mice by browning the large subcutaneous white adipose tissue (WAT) and enhancing energy expenditure. Here, we examine whether this approach could also reduce established obesity. Dietary-induced obese C57BL6/N mice received AAV8 BMP4 gene therapy at 17-18 weeks of age. They were kept on a high-fat diet and phenotypically characterized for an additional 10-12 weeks. Following termination, the mice underwent additional characterization in vitro. Surprisingly, we observed no effect on body weight, browning of WAT, or energy expenditure in these obese mice, but whole-body insulin sensitivity and glucose tolerance were robustly improved. Insulin signaling and insulin-stimulated glucose uptake were increased in both adipose cells and skeletal muscle. BMP4 also decreased hepatic glucose production and reduced gluconeogenic enzymes in the liver, but not in the kidney, in addition to enhancing insulin action in the liver. Our findings show that BMP4 prevents, but does not reverse, established obesity in adult mice, while it improves insulin sensitivity independent of weight reduction. The BMP antagonist Noggin was increased in WAT in obesity, which may account for the lack of browning

HMGA1 overexpression in adipose tissue impairs adipogenesis and prevents diet-induced obesity and insulin resistance

High-Mobility-Group-A1 (HMGA1) proteins are non-histone proteins that regulate chromatin structure and gene expression during embryogenesis, tumourigenesis and immune responses. In vitro studies suggest that HMGA1 proteins may be required to regulate adipogenesis. To examine the role of HMGA1 in vivo, we generated transgenic mice overexpressing HMGA1 in adipose tissues. HMGA1 transgenic mice showed a marked reduction in white and brown adipose tissue mass that was associated with downregulation of genes involved in adipogenesis and concomitant upregulation of preadipocyte markers. Reduced adipogenesis and decreased fat mass were not associated with altered glucose homeostasis since HMGA1 transgenic mice fed a regular-chow diet exhibited normal glucose tolerance and insulin sensitivity. However, when fed a high-fat diet, overexpression of HMGA1 resulted in decreased body-weight gain, reduced fat mass, but improved insulin sensitivity and glucose tolerance. Although HMGA1 transgenic mice exhibited impaired glucose uptake in adipose tissue due to impaired adipogenesis, the increased glucose uptake observed in skeletal muscle may account for the improved glucose homeostasis. Our results indicate that HMGA1 plays an important function in the regulation of white and brown adipogenesis in vivo and suggests that impaired adipocyte differentiation and decreased fat mass is not always associated with impaired whole-body glucose homeostasis

Adipose Tissue Overexpression of Vascular Endothelial Growth Factor Protects Against Diet-Induced Obesity and Insulin Resistance

During the expansion of fat mass in obesity, vascularization of adipose tissue is insufficient to maintain tissue normoxia. Local hypoxia develops and may result in altered adipokine expression, proinflammatory macrophage recruitment, and insulin resistance. We investigated whether an increase in adipose tissue angiogenesis could protect against obesity-induced hypoxia and, consequently, insulin resistance. Transgenic mice overexpressing vascular endothelial growth factor (VEGF) in brown adipose tissue (BAT) and white adipose tissue (WAT) were generated. Vessel formation, metabolism, and inflammation were studied in VEGF transgenic mice and wild-type littermates fed chow or a high-fat diet. Overexpression of VEGF resulted in increased blood vessel number and size in both WAT and BAT and protection against high-fat diet-induced hypoxia and obesity, with no differences in food intake. This was associated with increased thermogenesis and energy expenditure. Moreover, whole-body insulin sensitivity and glucose tolerance were improved. Transgenic mice presented increased macrophage infiltration, with a higher number of M2 anti-inflammatory and fewer M1 proinflammatory macrophages than wild-type littermates, thus maintaining an anti-inflammatory milieu that could avoid insulin resistance. These studies suggest that overexpression of VEGF in adipose tissue is a potential therapeutic strategy for the prevention of obesity and insulin resistance

Long-Term Efficacy and Safety of Insulin and Glucokinase Gene Therapy for Diabetes : 8-Year Follow-Up in Dogs

Diabetes is a complex metabolic disease that exposes patients to the deleterious effects of hyperglycemia on various organs. Achievement of normoglycemia with exogenous insulin treatment requires the use of high doses of hormone, which increases the risk of life-threatening hypoglycemic episodes. We developed a gene therapy approach to control diabetic hyperglycemia based on co-expression of the insulin and glucokinase genes in skeletal muscle. Previous studies proved the feasibility of gene delivery to large diabetic animals with adeno-associated viral (AAV) vectors. Here, we report the long-term (∼8 years) follow-up after a single administration of therapeutic vectors to diabetic dogs. Successful, multi-year control of glycemia was achieved without the need of supplementation with exogenous insulin. Metabolic correction was demonstrated through normalization of serum levels of fructosamine, triglycerides, and cholesterol and remarkable improvement in the response to an oral glucose challenge. The persistence of vector genomes and therapeutic transgene expression years after vector delivery was documented in multiple samples from treated muscles, which showed normal morphology. Thus, this study demonstrates the long-term efficacy and safety of insulin and glucokinase gene transfer in large animals and especially the ability of the system to respond to the changes in metabolic needs as animals grow older