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Background  Inhibition of the epidermal growth factor receptor (EGFR) has shown clinical success in patients with advanced non-small cell lung cancer (NSCLC). Somatic mutations of EGFR were found in lung adenocarcinoma that lead to exquisite dependency on EGFR signaling; thus patients with EGFR-mutant tumors are at high chance of response to EGFR inhibitors. However, imaging approaches affording early identification of tumor response in EGFR-dependent carcinomas have so far been lacking.    Methodology/Principal Findings  We performed a systematic comparison of 3′-Deoxy-3′-[ 18 F]-fluoro-L-thymidine ([ 18 F]FLT) and 2-[ 18 F]-fluoro-2-deoxy-D-glucose ([ 18 F]FDG) positron emission tomography (PET) for their potential to identify response to EGFR inhibitors in a model of EGFR-dependent lung cancer early after treatment initiation. While erlotinib-sensitive tumors exhibited a striking and reproducible decrease in [ 18 F]FLT uptake after only two days of treatment, [ 18 F]FDG PET based imaging revealed no consistent reduction in tumor glucose uptake. In sensitive tumors, a decrease in [ 18 F]FLT PET but not [ 18 F]FDG PET uptake correlated with cell cycle arrest and induction of apoptosis. The reduction in [ 18 F]FLT PET signal at day 2 translated into dramatic tumor shrinkage four days later. Furthermore, the specificity of our results is confirmed by the complete lack of [ 18 F]FLT PET response of tumors expressing the T790M erlotinib resistance mutation of EGFR.    Conclusions  [ 18 F]FLT PET enables robust identification of erlotinib response in EGFR-dependent tumors at a very early stage. [ 18 F]FLT PET imaging may represent an appropriate method for early prediction of response to EGFR TKI treatment in patients with NSCLC.

Early Detection of Erlotinib Treatment Response in NSCLC by 3′-Deoxy-3′-[18F]-Fluoro-L-Thymidine ([18F]FLT) Positron Emission Tomography (PET)