Multifactorial resistance to aminopeptidase inhibitor prodrug CHR2863 in myeloid leukemia cells: down-regulation of carboxylesterase 1, drug sequestration in lipid droplets and pro-survival activation ERK/Akt/mTOR.
Aminopeptidase inhibitors are receiving attention as combination chemotherapeutic agents for the treatment of refractory acute myeloid leukemia. However, the factors determining therapeutic efficacy remain elusive. Here we identified the molecular basis of acquired resistance to CHR2863, an orally available hydrophobic aminopeptidase inhibitor prodrug with an esterase-sensitive motif, in myeloid leukemia cells. CHR2863 enters cells by diffusion and is retained therein upon esterase activity-mediated conversion to its hydrophilic active metabolite drug CHR6768, thereby exerting amino acid depletion. Carboxylesterases (CES) serve as candidate prodrug activating enzymes given CES1 expression in acute myeloid leukemia specimens. We established two novel myeloid leukemia sublines U937/CHR2863(200) and U937/CHR2863(5uM), with low (14-fold) and high level (270-fold) CHR2863 resistance. The latter drug resistant cells displayed: (i) complete loss of CES1-mediated drug activation associated with down-regulation of CES1 mRNA and protein, (ii) marked retention/sequestration of the prodrug, (iii) a substantial increase in intracellular lipid droplets, and (iv) a dominant activation of the pro-survival Akt/mTOR pathway. Remarkably, the latter feature coincided with a gain of sensitivity to the mTOR inhibitor rapamycin. These finding delineate the molecular basis of CHR2863 resistance and offer a novel modality to overcome this drug resistance in myeloid leukemia cells.
Department of Rheumatology, Amsterdam Rheumatology and Immunology Center, VU University Medical Center, Amsterdam, The Netherlands.|Present address: Department of Clinical Chemistry, UMCU, Utrecht, The Netherlands.
Sue Ellen
SE
Verbrugge
Department of Rheumatology, Amsterdam Rheumatology and Immunology Center, VU University Medical Center, Amsterdam, The Netherlands.
Marjon
M
Al
The Fred Wyszkowsky Cancer Research Laboratory, Faculty of Biology, The Technion-Israel Institute of Technology, Haifa, Israel.
Yehuda G
YG
Assaraf
Department of Rheumatology, Amsterdam Rheumatology and Immunology Center, VU University Medical Center, Amsterdam, The Netherlands.|Department of Medical Oncology, VU University Medical Center, Amsterdam, The Netherlands.|Present address: Institute of Biophysics, Medical University of Graz, Graz, Austria.
Sarah
S
Kammerer
Department of Rheumatology, Amsterdam Rheumatology and Immunology Center, VU University Medical Center, Amsterdam, The Netherlands.|Department of Medical Oncology, VU University Medical Center, Amsterdam, The Netherlands.
Durga M S H
DM
Chandrupatla
Department of Medical Oncology, VU University Medical Center, Amsterdam, The Netherlands.
Richard
R
Honeywell
Department of Physiology, VU University, Amsterdam, The Netherlands.
Rene P J
RP
Musters
Department of Medical Oncology, VU University Medical Center, Amsterdam, The Netherlands.
Elisa
E
Giovannetti
Department of Molecular Cell Biology, VU University, Amsterdam, The Netherlands.
Tom
T
O'Toole
Departments of Pathology and Cardiac Surgery, ICaR-VU, VU University Medical Center, Amsterdam, The Netherlands.
George L
GL
Scheffer
Chroma Therapeutics Ltd, Abingdon, United Kingdom.|Present address: Immunocore Ltd, Oxford, UK.
David
D
Krige
Department of Medical Oncology, VU University Medical Center, Amsterdam, The Netherlands.
Tanja D
TD
de Gruijl
Departments of Pathology and Cardiac Surgery, ICaR-VU, VU University Medical Center, Amsterdam, The Netherlands.
Hans W M
HW
Niessen
Department of Rheumatology, Amsterdam Rheumatology and Immunology Center, VU University Medical Center, Amsterdam, The Netherlands.
Willem F
WF
Lems
Isala Hospital, Zwolle, The Netherlands.
Pieternella A
PA
Kramer
Departments of Pathology and Cardiac Surgery, ICaR-VU, VU University Medical Center, Amsterdam, The Netherlands.
Rik J
RJ
Scheper
Department of Pediatric Oncology/Hematology, VU University Medical Center, Amsterdam, The Netherlands.
Jacqueline
J
Cloos
Department of Hematology, VU University Medical Center, Amsterdam, The Netherlands.
Gert J
GJ
Ossenkoppele
Department of Medical Oncology, VU University Medical Center, Amsterdam, The Netherlands.
Godefridus J
GJ
Peters
Department of Rheumatology, Amsterdam Rheumatology and Immunology Center, VU University Medical Center, Amsterdam, The Netherlands.
Gerrit
G
Jansen
D000626:Aminopeptidases / Q000378:metabolism*; D017209:Apoptosis; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D015536:Down-Regulation; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D006801:Humans; D066292:Lipid Droplets; D010766:Phosphorylation; D011355:Prodrugs / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism*
D011355:Prodrugs; C546842:MTOR protein, human; D051057:Proto-Oncogene Proteins c-akt; D058570:TOR Serine-Threonine Kinases; D002265:Carboxylic Ester Hydrolases; D000626:Aminopeptidases
aminopeptidase; carboxylesterase; lipid droplets; mTOR; rapamycin