Project Summary / Abstract Targeted therapy for the treatment of non-small cell lung cancer (NSCLC) has greatly improved patient outcomes compared to traditional chemotherapy. However, it is estimated that up to 20% of lung cancer patients receive first-line therapy prior to EGFR mutational analysis, necessitating the need for a rapid and cheap blood-based circulating tumor (ct) DNA test for efficient therapy selection. Additionally, ctDNA can be useful for rebiopsy and monitoring of response to therapy or acquired resistance to therapy. We have successfully developed a highly multiplexable magneto-nanosensor assay for the detection of “hot spot” mutation panel for lung cancer [targeted therapy selection or therapy monitoring and prognosis.] Specific aims of the project are: 1) Validate ctDNA EGFR magneto-nanosensor assay accuracy and correlations to clinical outcome on expanded cohort. We hypothesize that our magneto-nanosensor assay can detect “hot spot” EGFR mutations Exon19 deletion, L858R, and T790M with high sensitivity and specificity in a large cohort of NSCLC patients. Additionally, this assay can be well suited for prognosis and therapy response monitoring. A follow-up blood draw 2 weeks after initiating TKI therapy can be highly predictive of progression free survival. Additionally, we hypothesize that frequent testing (every 2-3 months) on the magneto-nanosensor assay could be used in lieu of radiographic imaging if patient maintains favorable response, and that the magneto-nanosensor assay could detect progression of disease prior to imaging. 2) Develop an extensive ctDNA mutation panel assay on magneto-nanosensor arrays [for NSCLC treatment selection, monitoring, and prognosis.] We hypothesize that our previously developed EGFR ctDNA magneto- nanosensor mutation assay can be further expanded to include mutations relevant to third-generation TKI resistance mechanisms and mutations in other genes to better aid in NSCLC [treatment selection, monitoring, and prognosis.] In this aim, our goal is to develop a ctDNA mutation panel that can help identify treatment options for NSCLC patients, not only EGFR mutations but also including KRAS, BRAF V600E, ALK-EML4 fusions, and ROS1 fusions. [We aim to monitor patients throughout their treatment course for responsiveness and prognosis.] The expected outcomes of this project are clinical validation of a ctDNA EGFR magneto-nanosensor assay, expanding and multiplexing the existing EGFR assay to include KRAS, BRAF V600E, ALK-EML4 fusions, and ROS1 fusions, and utilizing the assay [for efficient therapy selection, prognosis, and/or response monitoring.]