TY - JOUR
T1 - Lipid droplet-mediated scavenging as novel intrinsic and adaptive resistance factor against the multikinase inhibitor ponatinib
AU - Englinger, Bernhard
AU - Laemmerer, Anna
AU - Moser, Patrick
AU - Kallus, Sebastian
AU - Röhrl, Clemens
AU - Pirker, Christine
AU - Baier, Dina
AU - Mohr, Thomas
AU - Niederstaetter, Laura
AU - Meier-Menches, Samuel M.
AU - Gerner, Christopher
AU - Gabler, Lisa
AU - Gojo, Johannes
AU - Timelthaler, Gerald
AU - Senkiv, Julia
AU - Jäger, Walter
AU - Kowol, Christian R.
AU - Heffeter, Petra
AU - Berger, Walter
PY - 2020/9/15
Y1 - 2020/9/15
N2 - Ponatinib is a small molecule multi-tyrosine kinase inhibitor clinically approved for anticancer therapy. Molecular mechanisms by which cancer cells develop resistance against ponatinib are currently poorly understood. Likewise, intracellular drug dynamics, as well as potential microenvironmental factors affecting the activity of this compound are unknown. Cell/molecular biological and analytical chemistry methods were applied to investigate uptake kinetics/subcellular distribution, the role of lipid droplets (LDs) and lipoid microenvironment compartments in responsiveness of FGFR1-driven lung cancer cells toward ponatinib. Selection of lung cancer cells for acquired ponatinib resistance resulted in elevated intracellular lipid levels. Uncovering intrinsic ponatinib fluorescence enabled dissection of drug uptake/retention kinetics in vitro as well as in mouse tissue cryosections, and revealed selective drug accumulation in LDs of cancer cells. Pharmacological LD upmodulation or downmodulation indicated that the extent of LD formation and consequent ponatinib incorporation negatively correlated with anticancer drug efficacy. Co-culturing with adipocytes decreased ponatinib levels and fostered survival of cancer cells. Ponatinib-selected cancer cells exhibited increased LD levels and enhanced ponatinib deposition into this organelle. Our findings demonstrate intracellular deposition of the clinically approved anticancer compound ponatinib into LDs. Furthermore, increased LD biogenesis was identified as adaptive cancer cell-defense mechanism via direct drug scavenging. Together, this suggests that LDs represent an underestimated organelle influencing intracellular pharmacokinetics and activity of anticancer tyrosine kinase inhibitors. Targeting LD integrity might constitute a strategy to enhance the activity not only of ponatinib, but also other clinically approved, lipophilic anticancer therapeutics.
AB - Ponatinib is a small molecule multi-tyrosine kinase inhibitor clinically approved for anticancer therapy. Molecular mechanisms by which cancer cells develop resistance against ponatinib are currently poorly understood. Likewise, intracellular drug dynamics, as well as potential microenvironmental factors affecting the activity of this compound are unknown. Cell/molecular biological and analytical chemistry methods were applied to investigate uptake kinetics/subcellular distribution, the role of lipid droplets (LDs) and lipoid microenvironment compartments in responsiveness of FGFR1-driven lung cancer cells toward ponatinib. Selection of lung cancer cells for acquired ponatinib resistance resulted in elevated intracellular lipid levels. Uncovering intrinsic ponatinib fluorescence enabled dissection of drug uptake/retention kinetics in vitro as well as in mouse tissue cryosections, and revealed selective drug accumulation in LDs of cancer cells. Pharmacological LD upmodulation or downmodulation indicated that the extent of LD formation and consequent ponatinib incorporation negatively correlated with anticancer drug efficacy. Co-culturing with adipocytes decreased ponatinib levels and fostered survival of cancer cells. Ponatinib-selected cancer cells exhibited increased LD levels and enhanced ponatinib deposition into this organelle. Our findings demonstrate intracellular deposition of the clinically approved anticancer compound ponatinib into LDs. Furthermore, increased LD biogenesis was identified as adaptive cancer cell-defense mechanism via direct drug scavenging. Together, this suggests that LDs represent an underestimated organelle influencing intracellular pharmacokinetics and activity of anticancer tyrosine kinase inhibitors. Targeting LD integrity might constitute a strategy to enhance the activity not only of ponatinib, but also other clinically approved, lipophilic anticancer therapeutics.
KW - cancer
KW - drug sequestration
KW - lipid droplets
KW - Ponatinib
KW - resistance
KW - Cell Proliferation
KW - Signal Transduction
KW - Receptor, Fibroblast Growth Factor, Type 1/antagonists & inhibitors
KW - Humans
KW - Tumor Microenvironment
KW - Drug Resistance, Neoplasm
KW - Pyridazines/pharmacokinetics
KW - Protein Kinase Inhibitors/pharmacokinetics
KW - Xenograft Model Antitumor Assays
KW - Animals
KW - Imidazoles/pharmacokinetics
KW - Lung Neoplasms/drug therapy
KW - Cell Line, Tumor
KW - Mice
KW - Lipid Droplets/metabolism
UR - http://www.scopus.com/inward/record.url?scp=85081268004&partnerID=8YFLogxK
U2 - 10.1002/ijc.32924
DO - 10.1002/ijc.32924
M3 - Article
C2 - 32064608
AN - SCOPUS:85081268004
SN - 0020-7136
VL - 147
SP - 1680
EP - 1693
JO - International Journal of Cancer
JF - International Journal of Cancer
IS - 6
ER -
