Citation
- Authors: Bodineau C. et al.
- Year: 2021
- Journal: Nat Commun 12 4814
- Applications: in vitro / DNA, siRNA / INTERFERin, jetPEI
- Cell type: U-2 OS
Description: Human bone osteosarcoma
Known as: U2OS
Method
The siRNA mediated knockdown was performed using 10 nM of siRNA transfected with INTERFERin® siRNA transfection reagent (POLYplus transfection) following the manufacturer’s instructions. Cells were incubated in contact with the transfection agent for 48 h and then treated with the indicated conditions for 24, 48, 72, or 96 h.
The plasmid transfections were carried out using Jetpei (Polyplus Transfection) according to the manufacturer’s instructions. 70% confluent cells were transfected with the indicated amount of plasmid, 24 h later cells were treated as indicated.
Abstract
Glutamoptosis is the induction of apoptotic cell death as a consequence of the aberrant activation of glutaminolysis and mTORC1 signaling during nutritional imbalance in proliferating cells. The role of the bioenergetic sensor AMPK during glutamoptosis is not defined yet. Here, we show that AMPK reactivation blocks both the glutamine-dependent activation of mTORC1 and glutamoptosis in vitro and in vivo. We also show that glutamine is used for asparagine synthesis and the GABA shunt to produce ATP and to inhibit AMPK, independently of glutaminolysis. Overall, our results indicate that glutamine metabolism is connected with mTORC1 activation through two parallel pathways: an acute alpha-ketoglutarate-dependent pathway; and a secondary ATP/AMPK-dependent pathway. This dual metabolic connection between glutamine and mTORC1 must be considered for the future design of therapeutic strategies to prevent cell growth in diseases such as cancer.