Metabolic Pathways in T-Cell Acute Lymphoblastic Leukemia (T-ALL)
- Determine the metabolic status and regulation of primary T-cell acute lymphoblastic
leukemia (T-ALL) relative to control resting peripheral T cells.
- Establish the effects of metabolic inhibition on metabolic stress pathways and
- Determine how metabolic inhibition interacts with chemotherapy or targeted therapy
drugs to kill T-ALL cells.
OUTLINE: T-ALL samples cultured alone or with gamma secretase inhibitors (GSI) or PI3K
inhibitors are analyzed for metabolic characteristics including glucose transporter 1
(Glut1) expression, mitochondrial mass, phospho-flow for 5' adenosine
monophosphate-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), and mammalian
target of rapamycin (mTOR) by flow cytometry. T-ALL samples and normal CD4+ T cells
(control) are also exposed to ± 2-deoxyglucose or ± the glutaminolysis inhibitor media and
analyzed for metabolic stress responses over time in particular, AMPK activation, autophagy
(immunofluorescence for LC3-II processing), and BCL2-associated X protein (Bak) and Bax
activation to indicate apoptosis. These cells (T-ALL and control) are then cultured with
cyclophosphamide, dexamethasone, or the B-cell CLL/lymphoma 2 (Bcl-2) inhibitor, ABT-737, to
determine cell death over time.
Metabolic status of primary T-ALL
Jeffrey C. Rathmell, PhD
Duke Cancer Institute
United States: Federal Government