T. Ferre - Academia.edu (original) (raw)

Papers by T. Ferre

The FASEB Journal, 1996

To study the role of glucokinase (GK) in the control of glucose metabolism in the liver, transgen... more To study the role of glucokinase (GK) in the control of glucose metabolism in the liver, transgenic mice were generated in which GK was overexpressed under control of the P-enolpyruvate carboxykinase gene promoter. Whereas the expression of the GK gene in starved control mice was blocked, this promoter was able to direct the expression of the enzyme to the liver of starved transgenic mice. Furthermore, starved transgenic mice showed levels of GK activity fourfold higher than those of starved control and similar to those of fed control. This activation of GK led to an increase in the intracellular concentration of glucose 6-phosphate, which was also related to an induction of glycogen accumulation. In addition, L-pyruvate kinase (L-PK) activity increased in transgenic mice, which when starved showed similar levels of activity to control fed mice. The induction of L-PK caused an increase in the hepatic lactate concentration. Furthermore, hepatocytes in primary culture from transgenic mice incubated with 20 mM glucose produced levels of lactate threefold higher than controls, but no difference was noted when the hepatocytes from control and transgemc mice were incubated with 2 mM glucose. These results demonstrated in vivo that the activation of GK is a rate-limiting step in the induction of glycolysis and glycogen synthesis. These changes in liver glucose metabolism led to a marked reduction in blood glucose (30%) and insulin (40%) concentrations. Furthermore, transgenic mice showed lower levels of blood glucose after an intraperitoneal glucose tolerance test, indicating that GK overexpression caused an increase in blood glucose disposal by the liver. All these findings show the key role of liver GK in the control of whole-body glucose homeostasis.

Scientific Reports, 2015

High-Mobility-Group-A1 (HMGA1) proteins are non-histone proteins that regulate chromatin structur... more 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 m...

The FASEB Journal, 2003

Alterations in hepatic glucose metabolism play a key role in the development of the hyperglycemia... more Alterations in hepatic glucose metabolism play a key role in the development of the hyperglycemia observed in type 2 diabetes. Because the transcription factor c-Myc induces hepatic glucose uptake and utilization and blocks gluconeogenesis, we examined whether hepatic overexpression of c-myc counteracts the insulin resistance induced by a high-fat diet. After 3 months on this diet, control mice became obese, hyperglycemic, and hyperinsulinemic, indicating that they had developed insulin resistance. In contrast, transgenic mice remained lean and showed improved glucose disposal and normal levels of blood glucose and insulin, indicating that they had developed neither obesity nor insulin resistance. These findings were concomitant with normalization of hepatic glucokinase and pyruvate kinase gene expression and enzyme activity, which led to normalization of intrahepatic glucose-6-phosphate and glycogen content. In the liver of control mice fed a high-fat diet, the expression of genes encoding proteins that control energy metabolism, such as sterol receptor element binding protein 1-c, peroxisome proliferator activated receptor α, and uncoupling protein-2, was altered. In contrast, in the liver of transgenic mice fed a high-fat diet, the expression of these genes was normal. These results suggest that c-myc overexpression counteracted the obesity and insulin resistance induced by a high-fat diet by modulating the expression of genes that regulate hepatic metabolism.

Human Gene Therapy, 2000

Chronic hyperglycemia is responsible for diabetes-specific microvascular and macrovascular compli... more Chronic hyperglycemia is responsible for diabetes-specific microvascular and macrovascular complications. To reduce hyperglycemia, key tissues may be engineered to take up glucose. To determine whether an increase in skeletal muscle glucose phosphorylation leads to increased glucose uptake and to normalization of diabetic alterations, the liver enzyme glucokinase (GK) was expressed in muscle of transgenic mice. GK has a high K m for glucose and its activity is not inhibited by glucose 6-phosphate. The presence of GK activity in skeletal muscle resulted in increased concentrations of glucose 6-phosphate and glycogen. These mice showed lower glycemia and insulinemia, increased serum lactate levels, and higher blood glucose disposal after an intraperitoneal glucose tolerance test. Furthermore, transgenic mice were more sensitive to injection of low doses of insulin, which led to increased blood glucose disposal. In addition, streptozotocin (STZ)-treated transgenic mice showed lower levels of blood glucose than STZ-treated controls and maintained body weight. Moreover, injection of insulin to STZ-treated transgenic mice led to normoglycemia, while STZ-treated control mice remained highly hyperglycemic. Thus, these results are consistent with a key role of glucose phosphorylation in regulating glucose metabolism in skeletal muscle. Furthermore, this study suggests that engineering skeletal muscle to express GK may be a new approach to the therapy of diabetes mellitus.

Human Gene Therapy, 2002

Type 1 diabetic patients depend on insulin replacement therapy. However, chronic hyperglycemia du... more Type 1 diabetic patients depend on insulin replacement therapy. However, chronic hyperglycemia due to failure to maintain proper glycemic control leads to microvascular, macrovascular, and neurological complications. Increased glucose disposal by tissues engineered to overexpress key regulatory genes in glucose transport or phosphorylation can reduce diabetic hyperglycemia. Here we report that differentiated myoblast cells expressing the glucose-phosphorylating enzyme glucokinase (GK) showed a glucose-dependent increase in glucose uptake and utilization in vitro. Transplantation of GK-expressing myotubes into healthy mice did not alter blood glucose levels and recipient mice maintained normoglycemia. After streptozotocin treatment, mice transplanted with GK-expressing myotubes counteracted hyperglycemia, polydipsia, and polyphagia, whereas mice transplanted with control myotubes developed diabetes. Similarly, diabetic mice transplanted with control myotubes remained hyperglycemic. In contrast, transplantation of GK-expressing myotubes into diabetic mice lowered hyperglycemia. These results suggest that the use of genetically engineered muscle cells to express glucokinase may provide a glucose-regulated approach to reduce diabetic hyperglycemia.

Diabetologia, 2003

Aims/hypothesis. Glucokinase overexpression in the liver increases glucose uptake and utilization... more Aims/hypothesis. Glucokinase overexpression in the liver increases glucose uptake and utilization, and improves glucose tolerance in young transgenic mice. Here, we examined the long-term effects of hepatic overexpression of glucokinase on glucose homeostasis. Moreover, we determined whether glucokinase overexpression counteracted high-fat diet-induced insulin resistance. Methods. Transgenic mice overexpressing glucokinase in liver under the control of the phosphoenolpyruvate carboxykinase promoter, fed either a standard diet or a high-fat diet, were studied. We used non-transgenic littermates as controls. Results. Transgenic mice over 6 months old developed impaired glucose tolerance. In addition, at 12 months of age, transgenic mice showed mild hyperglycaemia, hyperinsulinaemia and hypertriglyceridaemia. In spite of increased glucokinase activity, the liver of these mice accumulated less glycogen and increased triglyceride deposition. When 2-month-old glucose-tolerant mice were fed a high-fat diet, transgenic mice gained more body weight and became hyperglycaemic and hyperinsulinaemic. This was concomitant to glucose intolerance, liver steatosis and whole-body insulin resistance. Conclusion/interpretation. Long-term overexpression of glucokinase increases hepatic lipogenesis and circulating lipids, which lead to insulin resistance. Our results also suggest that the liver plays a key role in the onset of diabetes.

Diabetologia, 2008

Aims/hypothesis IL-6 is released by the adipose tissue and increased circulating levels in obesit... more Aims/hypothesis IL-6 is released by the adipose tissue and increased circulating levels in obesity are associated with hyperinsulinaemia and insulin resistance. Short-term experiments suggest that increased IL-6 release by the skeletal muscle following exercise may improve insulin sensitivity. Methods In order to examine the effect of chronically elevated IL-6 levels, we overexpressed Il6 in skeletal muscle in mice using an electro-transfer procedure. Results Circulating IL-6 levels were increased and the animals rapidly lost both weight and body fat, but food intake was unchanged, which is consistent with the finding that IL-6 increased energy expenditure. Insulin levels were inappropriately elevated and combined with hypoglycaemia in spite of reduced 2-deoxy-D-glucose uptake by skeletal muscle. Insulin-stimulated glucose uptake by skeletal muscles ex vivo was reduced, probably due to the decreased amounts of glucose transporter (GLUT)-4. Beta cell insulin content was increased, while apparent beta cell mass was unchanged. Circulating serum amyloid A cluster levels were increased tenfold due to a pronounced proinflammatory state in the liver with infiltration of inflammatory cells. However, no liver steatosis was found, which may be accounted for by concomitant AMP kinase activation. Conclusions/interpretation Chronically elevated IL-6 levels lead to inappropriate hyperinsulinaemia, reduced body weight, impaired insulin-stimulated glucose uptake by the skeletal muscles and marked inflammation in the liver. Thus, the pleiotrophic effects of chronically elevated IL-6 levels preclude any obvious usefulness in treating obesity or its associated metabolic complications in man, despite the fact that weight reduction may be expected.

Diabetologia, 2010

Aims/hypothesis In adipocytes, triacylglycerol synthesis depends on the formation of glycerol 3-p... more Aims/hypothesis In adipocytes, triacylglycerol synthesis depends on the formation of glycerol 3-phosphate, which originates either from glucose, through glycolysis, or from lactate, through glyceroneogenesis. However, glucose is traditionally viewed as the main precursor of the glycerol backbone and thus, enhanced glucose uptake would be expected to result in increased triacylglycerol synthesis and contribute to obesity. Methods To further explore this issue, we generated a mouse model with chronically increased glucose uptake in adipose tissue by expressing Gck, which encodes the glucokinase enzyme. Results Here we show that the production of high levels of glucokinase led to increased adipose tissue glucose uptake and lactate production, improved glucose tolerance and higher whole-body and skeletal muscle insulin sensitivity. There was no parallel increase in glycerol 3-phosphate synthesis in vivo, fat accumulation or obesity. Moreover, at high glucose concentrations, in cultured fat cells overproducing glucokinase, glycerol 3-phosphate synthesis from pyruvate decreased, while glyceroneogenesis increased in fat cells overproducing hexokinase II. Conclusions/interpretations These findings indicate that the absence of glucokinase inhibition by glucose 6phosphate probably led to increased glycolysis and blocked S. Muñoz and S. Franckhauser contributed equally to this study.

Diabetes, 2002

Insulin replacement therapy in type 1 diabetes is imperfect because proper glycemic control is no... more Insulin replacement therapy in type 1 diabetes is imperfect because proper glycemic control is not always achieved. Most patients develop microvascular, macrovascular, and neurological complications, which increase with the degree of hyperglycemia. Engineered muscle cells continuously secreting basal levels of insulin might be used to improve the efficacy of insulin treatment. Here we examined the control of glucose homeostasis in healthy and diabetic transgenic mice constitutively expressing mature human insulin in skeletal muscle. Fed transgenic mice were normoglycemic and normoinsulinemic and, after an intraperitoneal glucose tolerance test, showed increased glucose disposal. When treated with streptozotocin (STZ), transgenic mice showed increased insulinemia and reduced hyperglycemia when fed and normoglycemia and normoinsulinemia when fasted. Injection of low doses of soluble insulin restored normoglycemia in fed STZ-treated transgenic mice, while STZ-treated controls remained ...

Diabetes, 2006

Obesity and insulin resistance are associated with increased serum free fatty acids (FFAs). Thus,... more Obesity and insulin resistance are associated with increased serum free fatty acids (FFAs). Thus, a reduction in circulating FFAs may increase insulin sensitivity. This could be achieved by increasing FFA reesterification in adipose tissue. Transgenic mice with increased adipose tissue glyceroneogenesis, caused by overexpression of phosphoenolpyruvate carboxykinase (PEPCK), show increased FFA reesterification and develop obesity but are insulin sensitive. Here, we examined whether these transgenic mice were protected from diet-induced insulin resistance. Surprisingly, when fed a high-fat diet for a short period (6 weeks), transgenic mice developed severe obesity and were more hyperinsulinemic, glucose intolerant, and insulin resistant than controls. The high triglyceride accumulation prevented white adipose tissue from buffering the flux of lipids in circulation and led to increased serum triglyceride levels and fat deposition in liver. Furthermore, circulating leptin and FFA concen...

Diabetes, 2013

Adipose tissue is pivotal in the regulation of energy homeostasis through the balance of energy s... more Adipose tissue is pivotal in the regulation of energy homeostasis through the balance of energy storage and expenditure and as an endocrine organ. An inadequate mass and/or alterations in the metabolic and endocrine functions of adipose tissue underlie the development of obesity, insulin resistance, and type 2 diabetes. To fully understand the metabolic and molecular mechanism(s) involved in adipose dysfunction, in vivo genetic modification of adipocytes holds great potential. Here, we demonstrate that adeno-associated viral (AAV) vectors, especially serotypes 8 and 9, mediated efficient transduction of white (WAT) and brown adipose tissue (BAT) in adult lean and obese diabetic mice. The use of short versions of the adipocyte protein 2 or uncoupling protein-1 promoters or micro-RNA target sequences enabled highly specific, long-term AAV-mediated transgene expression in white or brown adipocytes. As proof of concept, delivery of AAV vectors encoding for hexokinase or vascular endothe...

Biochemical Journal, 2002

Overexpression of the c-Myc transcription factor in liver induces glucose uptake and utilization.... more Overexpression of the c-Myc transcription factor in liver induces glucose uptake and utilization. Here we examined the effects of c-myc overexpression on the expression of hepatocyte-specific transcription factor genes which regulate the expression of genes controlling hepatic metabolism. At 4 months after streptozotocin (STZ) treatment, most diabetic control mice were highly hyperglycaemic and died, whereas in STZ-treated transgenic mice hyperglycaemia was markedly lower, the serum levels of β-hydroxybutyrate, triacylglycerols and non-esterified fatty acids were normal, and they had greater viability in the absence of insulin. Furthermore, long-term STZ-treated transgenic mice showed similar glucose utilization and storage to healthy controls. This was consistent with the expression of glycolytic genes becoming normalized. In addition, restoration of gene expression of the transcription factor, sterol receptor element binding protein 1c, was observed in the livers of these transgen...

Diabetes, 2012

During the expansion of fat mass in obesity, vascularization of adipose tissue is insufficient to... more 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...

Scientific Reports, 2015

Proceedings of the National Academy of Sciences, 1996

Hyperglycemia is a common feature of diabetes mellitus. It results from a decrease in glucose uti... more Hyperglycemia is a common feature of diabetes mellitus. It results from a decrease in glucose utilization by the liver and peripheral tissues and an increase in hepatic glucose production. Glucose phosphorylation by glucokinase is an initial event in glucose metabolism by the liver. However, glucokinase gene expression is very low in diabetic animals. Transgenic mice expressing the P-enolpyruvate carboxykinase/glucokinase chimeric gene were generated to study

Human Gene Therapy, 2000

Chronic hyperglycemia is responsible for diabetes-specific microvascular and macrovascular compli... more Chronic hyperglycemia is responsible for diabetes-specific microvascular and macrovascular complications. To reduce hyperglycemia, key tissues may be engineered to take up glucose. To determine whether an increase in skeletal muscle glucose phosphorylation leads to increased glucose uptake and to normalization of diabetic alterations, the liver enzyme glucokinase (GK) was expressed in muscle of transgenic mice. GK has a high Km for glucose and its activity is not inhibited by glucose 6-phosphate. The presence of GK activity in skeletal muscle resulted in increased concentrations of glucose 6-phosphate and glycogen. These mice showed lower glycemia and insulinemia, increased serum lactate levels, and higher blood glucose disposal after an intraperitoneal glucose tolerance test. Furthermore, transgenic mice were more sensitive to injection of low doses of insulin, which led to increased blood glucose disposal. In addition, streptozotocin (STZ)-treated transgenic mice showed lower levels of blood glucose than STZ-treated controls and maintained body weight. Moreover, injection of insulin to STZ-treated transgenic mice led to normoglycemia, while STZ-treated control mice remained highly hyperglycemic. Thus, these results are consistent with a key role of glucose phosphorylation in regulating glucose metabolism in skeletal muscle. Furthermore, this study suggests that engineering skeletal muscle to express GK may be a new approach to the therapy of diabetes mellitus.

Diabetologia, 2009

Aims/hypothesis Due to their ability to regulate various signalling pathways (cytokines, hormones... more Aims/hypothesis Due to their ability to regulate various signalling pathways (cytokines, hormones, growth factors), the suppressor of cytokine signalling (SOCS) proteins are thought to be promising therapeutic targets for metabolic and inflammatory disorders. Hence, their role in vivo has to be precisely determined. Methods We generated transgenic mice constitutively producing SOCS-3 in skeletal muscle to define whether the sole abundance of SOCS-3 is sufficient to induce metabolic disorders and whether SOCS-3 is implicated in physiological roles distinct from metabolism. Results We demonstrate here that chronic expression of SOCS-3 in skeletal muscle leads to overweight in mice and worsening of high-fat diet-induced systemic insulin resistance. Counter-intuitively, insulin sensitivity in muscle of transgenic mice appears to be unaltered. However, following constitutive SOCS-3 production, several genes had deregu-P. Lebrun and E. Cognard contributed equally to this work.

Diabetologia, 2010

In adipocytes, triacylglycerol synthesis depends on the formation of glycerol 3-phosphate, which ... more In adipocytes, triacylglycerol synthesis depends on the formation of glycerol 3-phosphate, which originates either from glucose, through glycolysis, or from lactate, through glyceroneogenesis. However, glucose is traditionally viewed as the main precursor of the glycerol backbone and thus, enhanced glucose uptake would be expected to result in increased triacylglycerol synthesis and contribute to obesity. To further explore this issue, we generated a mouse model with chronically increased glucose uptake in adipose tissue by expressing Gck, which encodes the glucokinase enzyme. Here we show that the production of high levels of glucokinase led to increased adipose tissue glucose uptake and lactate production, improved glucose tolerance and higher whole-body and skeletal muscle insulin sensitivity. There was no parallel increase in glycerol 3-phosphate synthesis in vivo, fat accumulation or obesity. Moreover, at high glucose concentrations, in cultured fat cells overproducing glucokinase, glycerol 3-phosphate synthesis from pyruvate decreased, while glyceroneogenesis increased in fat cells overproducing hexokinase II. These findings indicate that the absence of glucokinase inhibition by glucose 6-phosphate probably led to increased glycolysis and blocked glyceroneogenesis in the mouse model. Furthermore, this study suggests that under physiological conditions, when blood glucose increases, glyceroneogenesis may prevail over glycolysis for triacylglycerol formation because of the inhibition of hexokinase II by glucose 6-phosphate. Together these results point to the indirect pathway (glucose to lactate to glycerol 3-phosphate) being key for fat deposition in adipose tissue.

Diabetes, 2012

Animals. Transgenic mice expressing the aP2/Vegf chimeric gene were obtained by microinjection of... more Animals. Transgenic mice expressing the aP2/Vegf chimeric gene were obtained by microinjection of oocytes from C57Bl6/SJL mice. Mice were kept in a pathogen-free facility (Servei d'Estabulació de Ratolins-Centre de Biotecnologia Animal i Teràpia Gènica) and maintained under a 12-h light-dark cycle at 22°C. Mice were fed ad libitum for 15 weeks with a chow diet (2018S Harlan Teklad, Madison, WI) or an HFD (TD88137 Harlan Teklad). When stated, mice were fasted for 16 h.

Diabetes, 2013

Diabetes is associated with severe secondary complications, largely caused by poor glycemic contr... more Diabetes is associated with severe secondary complications, largely caused by poor glycemic control. Treatment with exogenous insulin fails to prevent these complications completely, leading to significant morbidity and mortality. We previously demonstrated that it is possible to generate a "glucose sensor" in skeletal muscle through coexpression of glucokinase and insulin, increasing glucose uptake and correcting hyperglycemia in diabetic mice. Here, we demonstrate long-term efficacy of this approach in a large animal model of diabetes. A one-time intramuscular administration of adeno-associated viral vectors of serotype 1 encoding for glucokinase and insulin in diabetic dogs resulted in normalization of fasting glycemia, accelerated disposal of glucose after oral challenge, and no episodes of hypoglycemia during exercise for .4 years after gene transfer. This was associated with recovery of body weight, reduced glycosylated plasma proteins levels, and long-term survival without secondary complications. Conversely, exogenous insulin or gene transfer for insulin or glucokinase alone failed to achieve complete correction of diabetes, indicating that the synergistic action of insulin and glucokinase is needed for full therapeutic effect. This study provides the first proof-of-concept in a large animal model for a gene transfer approach to treat diabetes.

The FASEB Journal, 1996

Scientific Reports, 2015

The FASEB Journal, 2003

Human Gene Therapy, 2000

Human Gene Therapy, 2002

Diabetologia, 2003

Diabetologia, 2008

Diabetologia, 2010

Diabetes, 2002

Diabetes, 2006

Diabetes, 2013

Biochemical Journal, 2002

Diabetes, 2012

Scientific Reports, 2015

Proceedings of the National Academy of Sciences, 1996

Human Gene Therapy, 2000

Chronic hyperglycemia is responsible for diabetes-specific microvascular and macrovascular compli... more Chronic hyperglycemia is responsible for diabetes-specific microvascular and macrovascular complications. To reduce hyperglycemia, key tissues may be engineered to take up glucose. To determine whether an increase in skeletal muscle glucose phosphorylation leads to increased glucose uptake and to normalization of diabetic alterations, the liver enzyme glucokinase (GK) was expressed in muscle of transgenic mice. GK has a high Km for glucose and its activity is not inhibited by glucose 6-phosphate. The presence of GK activity in skeletal muscle resulted in increased concentrations of glucose 6-phosphate and glycogen. These mice showed lower glycemia and insulinemia, increased serum lactate levels, and higher blood glucose disposal after an intraperitoneal glucose tolerance test. Furthermore, transgenic mice were more sensitive to injection of low doses of insulin, which led to increased blood glucose disposal. In addition, streptozotocin (STZ)-treated transgenic mice showed lower levels of blood glucose than STZ-treated controls and maintained body weight. Moreover, injection of insulin to STZ-treated transgenic mice led to normoglycemia, while STZ-treated control mice remained highly hyperglycemic. Thus, these results are consistent with a key role of glucose phosphorylation in regulating glucose metabolism in skeletal muscle. Furthermore, this study suggests that engineering skeletal muscle to express GK may be a new approach to the therapy of diabetes mellitus.

Diabetologia, 2009

Diabetologia, 2010

Diabetes, 2012

Diabetes, 2013