Results of LentiMutate on EGFRdel746-750 cDNA packaged using M-RT and osimertinib in the HCC827 cell line. Values shown are the averages of two biological replicates. The fold enrichment of "All nucleotides" is the sum of substitution rates for all nucleotides in the with doxycycline replicates divided by the sum of substitution rates for all nucleotides without doxycycline (parental) at one nucleotide position. Nucleotide w/ max error refers to the highest substitution rate for a single nucleotide at one nucleotide position.
Cancer Prevention and Research Institute of Texas (CPRIT)
National Cancer Institute (NCI)
United States Department of Health and Human ServicesFind out more...
Howard Hughes Medical Institute (HHMI)
ARTICLE ABSTRACTIdentifying resistance mutations in a drug target provides crucial information. Lentiviral transduction creates multiple types of mutations due to the error-prone nature of the HIV-1 reverse transcriptase (RT). Here we optimized and leveraged this property to identify drug resistance mutations, developing a technique we term LentiMutate. This technique was validated by identifying clinically relevant EGFR resistance mutations, then applied to two additional clinical anticancer drugs: imatinib, a BCR-ABL inhibitor, and AMG 510, a KRAS G12C inhibitor. Novel deletions in BCR-ABL1 conferred resistance to imatinib. In KRAS-G12C or wild-type KRAS, point mutations in the AMG 510 binding pocket or oncogenic non-G12C mutations conferred resistance to AMG 510. LentiMutate should prove highly valuable for clinical and preclinical cancer-drug development.
LentiMutate can evaluate a drug's on-target activity and can nominate resistance mutations before they occur in patients, which could accelerate and refine drug development to increase the survival of patients with cancer.