In vivo fuel utilization and metabolic regulation by brown adipose tissue

Información del proyecto

GLUCOBAT

Identificador del acuerdo de subvención: 101029456

DOI

10.3030/101029456

Proyecto cerrado

Fecha de la firma de la CE 6 Abril 2021

Fecha de inicio 1 Octubre 2021

Fecha de finalización 30 Septiembre 2024

Financiado con arreglo a

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Coste total

€ 286 921,92

Aportación de la UE

€ 286 921,92

Coordinado por

KOBENHAVNS UNIVERSITET
Denmark

Periodic Reporting for period 2 - GLUCOBAT (In vivo fuel utilization and metabolic regulation by brown adipose tissue)

Período documentado: 2023-10-01 hasta 2024-09-30

Resumen del contexto y de los objetivos generales del proyecto

Brown adipose tissue (BAT) thermogenesis has gained significant clinical interest as a strategy against obesity and diabetes because of its potential to expend excess calories as heat. However, our understanding of BAT metabolism on an organismal level is still rudimentary, thus, to advance BAT-targeted therapies, we must first comprehensively define how BAT utilizes its key fuels, such as glucose in vivo. It is widely assumed that the major role of glucose during thermogenesis is to provide energy for the TCA cycle and subsequent uncoupling by the key thermogenic protein in BAT, uncoupling protein 1 (UCP1). However, recent literature reveal that this assumption is oversimplified since glucose has many fates once it enters cells, feeding into additional metabolic pathways beyond the TCA cycle, which are necessary for optimum thermogenesis.
The overall goal of this project is to investigate if and how glucose supports multiple metabolic pathways, beyond providing energy for the TCA cycle, that are required for thermogenesis, and moreover, how the fate of glucose-derived metabolites potentially varies depending on the thermogenic stimulus and duration. In addition, the role of cold-regulated solute carrier (SLC)-transporters in regulating glucose metabolism and thermogenesis will be investigated.
This project will expand our understanding of how thermogenic adipocytes utilize glucose and will potentially reveal novel therapeutic strategies to enhance thermogenesis.

Trabajo realizado desde el comienzo del proyecto hasta el final del período abarcado por el informe y los principales resultados hasta la fecha

In this project, I have used state-of-the-art glucose-tracing technology that utilizes mass spectrometry, combined with novel CRISPR/Cas9 genome editing strategies to precisely define how thermogenic adipose tissue utilizes glucose in vivo.
I have performed tracing experiments in brown adipocytes in vitro to investigate the importance of glucose for thermogenesis, to validate findings performed in vivo. Glucose tracing in mice acclimated to different temperatures (TN (30 °C), RT (22 °C) or to SC (6 °C)) and acutely to cold showed and confirmed my hypothesis that glucose carbons are rapidly incorporated into glycolysis and TCA cycle, as well as several auxiliary pathways, including NADPH, nucleotide, and phospholipid synthesis pathways. Furthermore, I have investigated with other carbon tracers if other carbon sources could be utilized by the BAT as energy for the TCA to drive thermogenesis. I have optimized and performed stable isotopic infusion tracing experiments to assess minimum-perturbative glucose tracing in mice acclimated to different temperatures and in UCP1-loss-of-function mice.
In this project I have investigated the role of solute carrier transporters of the SCL family for glucose utilization and thermogenesis in BAT. To investigate the most cold-regulated SLC-transporters in BAT, I assessed the expression of one of the transporters after acclimation to different temperatures and in different tissues. The mRNA expression was induced in BAT and in inguinal white adipose tissue in cold, which indicate a function of this transporter for thermogenesis. I furthermore investigated the loss-of-function by performing siRNA transfections in mature brown adipocytes and observed changed oxygen consumption measured on the Seahorse XF analyser. CRISPR/Cas9 gene editing technology has been used to investigate the loss-of-function in vivo.

Avances que van más allá del estado de la técnica e impacto potencial esperado (incluida la repercusión socioeconómica y las implicaciones sociales más amplias del proyecto hasta la fecha)

Since data analysis is still ongoing, the final impact of the project remains to be determined, but by increasing our knowledge on the glucose utilization in BAT, this project could provide bases for the development of targeted therapies.

Figure of substrate utilization in BAT.