TY - JOUR
T1 - Assessment of translation rate in leukemic cells and immune cells of the microenvironment by OPP protein synthesis assay
AU - Klapp, Vanessa
AU - Gumustekin, Ozgu
AU - Paggetti, Jerome
AU - Moussay, Etienne
AU - Largeot, Anne
N1 - Acknowledgments
This work was supported by grants from the Luxembourg National Research Fund (FNR) and Fondation Cancer to VK, OG, EM, JP and AL (PRIDE19/14254520/i2TRON, C20/BM/14582635, C20/BM/14592342, and C23/BM/17987391).
Publisher Copyright:
© 2024 Elsevier Inc.
PY - 2024/7/2
Y1 - 2024/7/2
N2 - Despite being tightly regulated, messenger RNA (mRNA) translation, a manner in which cells control expression of genes and rapidly respond to stimuli, is highly dysfunctional and plastic in pathologies including cancer. Conversely, the investigation of molecular mechanisms whereby mRNA translation becomes aberrant in cancer, as well as inhibition thereof, become critical in developing novel therapeutic approaches. More specifically, in malignancies such as chronic lymphocytic leukemia in which aberrant global and transcript specific translation has been linked with poorer patient outcomes, targeting translation is a relevant approach, with various translation inhibitors under development. Here we elaborate on a protein synthesis assay by flow cytometry, O-propargyl-puromycin, demonstrating global mRNA translation rate with a variety of different applications including cell lines, primary cells or co-culture systems in vitro. This method provides a comprehensive tool in quantifying the rate of global mRNA translation in cancer cells, as well as that of the tumor microenvironment cells, or in response to inhibitory therapeutic agents while offering the possibility to simultaneously assess other cellular markers.
AB - Despite being tightly regulated, messenger RNA (mRNA) translation, a manner in which cells control expression of genes and rapidly respond to stimuli, is highly dysfunctional and plastic in pathologies including cancer. Conversely, the investigation of molecular mechanisms whereby mRNA translation becomes aberrant in cancer, as well as inhibition thereof, become critical in developing novel therapeutic approaches. More specifically, in malignancies such as chronic lymphocytic leukemia in which aberrant global and transcript specific translation has been linked with poorer patient outcomes, targeting translation is a relevant approach, with various translation inhibitors under development. Here we elaborate on a protein synthesis assay by flow cytometry, O-propargyl-puromycin, demonstrating global mRNA translation rate with a variety of different applications including cell lines, primary cells or co-culture systems in vitro. This method provides a comprehensive tool in quantifying the rate of global mRNA translation in cancer cells, as well as that of the tumor microenvironment cells, or in response to inhibitory therapeutic agents while offering the possibility to simultaneously assess other cellular markers.
KW - FL3
KW - Flavagline
KW - Immunotherapy
KW - mRNA translation inhibition
KW - PBMC
UR - http://www.scopus.com/inward/record.url?scp=85197385917&partnerID=8YFLogxK
U2 - 10.1016/bs.mcb.2024.06.006
DO - 10.1016/bs.mcb.2024.06.006
M3 - Article
AN - SCOPUS:85197385917
SN - 0091-679X
VL - 189
JO - Methods in Cell Biology
JF - Methods in Cell Biology
ER -