Quantifying cell cycle-dependent drug sensitivities in cancer using a high throughput synchronisation and screening approach.


Type
Article
Change log
Authors
Johnson, Timothy I 
Minteer, Christopher J 
Kottmann, Daniel 
Dunlop, Charles R 
Fernández, Sandra Bernaldo de Quirós 
Abstract

BACKGROUND: Chemotherapy and targeted agent anti-cancer efficacy is largely dependent on the proliferative state of tumours, as exemplified by agents that target DNA synthesis/replication or mitosis. As a result, cell cycle specificities of a number of cancer drugs are well known. However, they are yet to be described in a quantifiable manner. METHODS: A scalable cell synchronisation protocol used to screen a library of 235 anti-cancer compounds exposed over six hours in G1 or S/G2 accumulated AsPC-1 cells to generate a cell cycle specificity (CCS) score. FINDINGS: The synchronisation method was associated with reduced method-related cytotoxicity compared to nocodazole, delivering sufficient cell cycle purity and cell numbers to run high-throughput drug library screens. Compounds were identified with G1 and S/G2-associated specificities that, overall, functionally matched with a compound's target/mechanism of action. This annotation was used to describe a synergistic schedule using the CDK4/6 inhibitor, palbociclib, prior to gemcitabine/AZD6738 as well as describe the correlation between the CCS score and published synergistic/antagonistic drug schedules. INTERPRETATION: This is the first highly quantitative description of cell cycle-dependent drug sensitivities that utilised a tractable and tolerated method with potential uses outside the present study. Drug treatments such as those shown to be G1 or S/G2 associated may benefit from scheduling considerations such as after CDK4/6 inhibitors and being first in drug sequences respectively. FUNDING: Cancer Research UK (CRUK) Institute core grants C14303/A17197 and C9545/A29580. The Li Ka Shing Centre where this work was performed was generously funded by CK Hutchison Holdings Limited, the University of Cambridge, CRUK, The Atlantic Philanthropies and others.

Description
Keywords
Cancer, cell cycle, combination treatment, high throughput screening, synchronisation, Cell Culture Techniques, Cell Cycle, Cell Line, Tumor, Cell Proliferation, Cell Survival, Deoxycytidine, Drug Screening Assays, Antitumor, Drug Synergism, HeLa Cells, High-Throughput Screening Assays, Humans, MCF-7 Cells, Neoplasms, Nocodazole, Piperazines, Pyridines, Small Molecule Libraries, Time Factors, Tubulin Modulators, Gemcitabine
Journal Title
EBioMedicine
Conference Name
Journal ISSN
2352-3964
2352-3964
Volume Title
68
Publisher
Elsevier BV
Sponsorship
Pancreatic Cancer UK (FLF2015_03_Cambridge)