3D high-content screening for the identification of compounds that target cells in dormant tumor spheroid regions.
Cancer cells in poorly vascularized tumor regions need to adapt to an unfavorable metabolic microenvironment. As distance from supplying blood vessels increases, oxygen and nutrient concentrations decrease and cancer cells react by stopping cell cycle progression and becoming dormant. As cytostatic drugs mainly target proliferating cells, cancer cell dormancy is considered as a major resistance mechanism to this class of anti-cancer drugs. Therefore, substances that target cancer cells in poorly vascularized tumor regions have the potential to enhance cytostatic-based chemotherapy of solid tumors. With three-dimensional growth conditions, multicellular tumor spheroids (MCTS) reproduce several parameters of the tumor microenvironment, including oxygen and nutrient gradients as well as the development of dormant tumor regions. We here report the setup of a 3D cell culture compatible high-content screening system and the identification of nine substances from two commercially available drug libraries that specifically target cells in inner MCTS core regions, while cells in outer MCTS regions or in 2D cell culture remain unaffected. We elucidated the mode of action of the identified compounds as inhibitors of the respiratory chain and show that induction of cell death in inner MCTS core regions critically depends on extracellular glucose concentrations. Finally, combinational treatment with cytostatics showed increased induction of cell death in MCTS. The data presented here shows for the first time a high-content based screening setup on 3D tumor spheroids for the identification of substances that specifically induce cell death in inner tumor spheroid core regions. This validates the approach to use 3D cell culture screening systems to identify substances that would not be detectable by 2D based screening in otherwise similar culture conditions.
Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin, Germany.
Carsten
C
Wenzel
Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin, Germany.
Björn
B
Riefke
Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin, Germany.
Stephan
S
Gründemann
Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin, Germany.
Alice
A
Krebs
Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin, Germany.
Sven
S
Christian
Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin, Germany.
Florian
F
Prinz
Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin, Germany.
Marc
M
Osterland
Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin, Germany.
Sven
S
Golfier
Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin, Germany.
Sebastian
S
Räse
Physical Biology Group, Buchmann Institute for Molecular Life Sciences (BMLS), Goethe University Frankfurt, Germany.
Nariman
N
Ansari
Max Planck Institute of Molecular Cell Biology and Genetics, High-Throughput Technology Development Studio (TDS), Dresden, Germany.
Milan
M
Esner
Max Planck Institute of Molecular Cell Biology and Genetics, High-Throughput Technology Development Studio (TDS), Dresden, Germany.
Marc
M
Bickle
Physical Biology Group, Buchmann Institute for Molecular Life Sciences (BMLS), Goethe University Frankfurt, Germany.
Francesco
F
Pampaloni
Physical Biology Group, Buchmann Institute for Molecular Life Sciences (BMLS), Goethe University Frankfurt, Germany.
Christian
C
Mattheyer
Physical Biology Group, Buchmann Institute for Molecular Life Sciences (BMLS), Goethe University Frankfurt, Germany.
Ernst H
EH
Stelzer
Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin, Germany.
Karsten
K
Parczyk
Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin, Germany.
Stefan
S
Prechtl
Bayer Pharma AG, Global Drug Discovery, Muellerstrasse 178, 13353 Berlin, Germany. Electronic address: Patrick.Steigemann@bayer.com.
Patrick
P
Steigemann
D000970:Antineoplastic Agents / Q000302:isolation & purification* / Q000494:pharmacology; D018929:Cell Culture Techniques; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D004579:Electron Transport / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology; D005260:Female; D005947:Glucose / Q000378:metabolism; D006801:Humans; D018874:Spheroids, Cellular / Q000187:drug effects*; D019311:Staurosporine / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D059016:Tumor Microenvironment / Q000502:physiology
D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D019311:Staurosporine; D005947:Glucose
3D cell culture; 3D high-content screening; Advanced light microscopy; Multicellular tumor spheroids; Tumor dormancy