A screen to identify drug resistant variants to target-directed anti-cancer agents.
The discovery of oncogenes and signal transduction pathways important for mitogenesis has triggered the development of target-specific small molecule anti-cancer compounds. As exemplified by imatinib (Gleevec), a specific inhibitor of the Chronic Myeloid Leukemia (CML)-associated Bcr-Abl kinase, these agents promise impressive activity in clinical trials, with low levels of clinical toxicity. However, such therapy is susceptible to the emergence of drug resistance due to amino acid substitutions in the target protein. Defining the spectrum of such mutations is important for patient monitoring and the design of next-generation inhibitors. Using imatinib and BCR/ABL as a paradigm for a drug-target pair, we recently reported a retroviral vector-based screening strategy to identify the spectrum of resistance-conferring mutations. Here we provide a detailed methodology for the screen, which can be generally applied to any drug-target pair.
Azam
Mohammad
M
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School and Division of Pediatric Hematology/Oncology, Children's Hospital and Dana Farber Cancer Institute, Boston, MA 02115 USA. Children's Hospital, 300 Longwood Ave, Boston, MA 02115. USA.
Raz
Tal
T
Nardi
Valentina
V
Opitz
Sarah L.
SL
Daley
George Q.
GQ