Exosome-Based Detection of EGFR T790M in Plasma from Non–Small Cell Lung Cancer Patients

Purpose: About 60% of non–small cell lung cancer (NSCLC) patients develop resistance to targeted epidermal growth factor receptor (EGFR) inhibitor therapy through the EGFR T790M mutation. Patients with this mutation respond well to third-generation tyrosine kinase inhibitors, but obtaining a tissue biopsy to confirm the mutation poses risks and is often not feasible. Liquid biopsies using circulating free tumor DNA (cfDNA) have emerged as a noninvasive option to detect the mutation; however, sensitivity is low as many patients have too few detectable copies in circulation. Here, we have developed and validated a novel test that overcomes the limited abundance of the mutation by simultaneously capturing and interrogating exosomal RNA/DNA and cfDNA (exoNA) in a single step followed by a sensitive allele-specific qPCR.

Experimental Design: ExoNA was extracted from the plasma of NSCLC patients with biopsy-confirmed T790M-positive (N = 102) and T790M-negative (N = 108) samples. The T790M mutation status was determined using an analytically validated allele-specific qPCR assay in a Clinical Laboratory Improvement Amendment laboratory.

Results: Detection of the T790M mutation on exoNA achieved 92% sensitivity and 89% specificity using tumor biopsy results as gold standard. We also obtained high sensitivity (88%) in patients with intrathoracic disease (M0/M1a), for whom detection by liquid biopsy has been particularly challenging.

Conclusions: The combination of exoRNA/DNA and cfDNA for T790M detection has higher sensitivity and specificity compared with historical cohorts using cfDNA alone. This could further help avoid unnecessary tumor biopsies for T790M mutation testing.

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Zeitschriftenbeiträge (peer-reviewed)
Exosome-Based Detection of EGFR T790M in Plasma from Non–Small Cell Lung Cancer Patients
Clinical Cancer Research
Elena Castellanos-Rizaldos, Prof. Dr. Dominik Grimm , Vasisht R. Tadigotla, James Hurley, John Healy, Patricia L. Neal, Mia Sher, Raajdeep Venkatesan, Chris Karlovich, Mitch Raponi, Anne Krug, Mikkel Noerholm, Jihane Tannous, Bakhos A. Tannous, Luis E. Raez, Johan K. Skog
Castellanos-Rizaldos, Elena; Grimm, Dominik; Tadigotla, Vasisht R.; Hurley, James; Healy, John; Neal, Patricia L.; Sher, Mia; Venkatesan, Raajdeep; Karlovich, Chris; Raponi, Mitch; Krug, Anne; Noerholm, Mikkel; Tannous, Jihane; Tannous, Bakhos A.; Raez, Luis E.; Skog, Johan K. (2018): Exosome-Based Detection of EGFR T790M in Plasma from Non–Small Cell Lung Cancer Patients. Clinical Cancer Research 24 (12), S. 2944-2950. DOI: 10.1158/1078-0432.CCR-17-3369