title: A Highly Sensitive and Quantitative Test Platform for Detection of NSCLC EGFR Mutations in Urine and Plasma.
abstract: In approximately 60% of patients with NSCLC who are receiving EGFR tyrosine kinase inhibitors, resistance develops through the acquisition of EGFR T790M mutation. We aimed to demonstrate that a highly sensitive and quantitative next-generation sequencing analysis of EGFR mutations from urine and plasma specimens is feasible.
  
  
  
  Short footprint mutation enrichment next-generation sequencing assays were used to interrogate EGFR activating mutations and the T790M resistance mutation in urine or plasma specimens from patients enrolled in TIGER-X (NCT01526928), a phase 1/2 clinical study of rociletinib in previously treated patients with EGFR mutant-positive advanced NSCLC.
  
  
  
  Of 63 patients, 60 had evaluable tissue specimens. When the tissue result was used as a reference, the sensitivity of EGFR mutation detection in urine was 72% (34 of 47 specimens) for T790M, 75% (12 of 16) for L858R, and 67% (28 of 42) for exon 19 deletions. With specimens that met a recommended volume of 90 to 100 mL, the sensitivity was 93% (13 of 14 specimens) for T790M, 80% (four of five) for L858R, and 83% (10 of 12) for exon 19 deletions. A comparable sensitivity of EGFR mutation detection was observed in plasma: 93% (38 of 41 specimens) for T790M, 100% (17 of 17) for L858R, and 87% (34 of 39) for exon 19 deletions. Together, urine and plasma testing identified 12 additional T790M-positive cases that were either undetectable or inadequate by tissue test. In nine patients monitored while receiving treatment with rociletinib, a rapid decrease in urine T790M levels was observed by day 21.
  
  
  
  DNA derived from NSCLC tumors can be detected with high sensitivity in urine and plasma, enabling diagnostic detection and monitoring of therapeutic response from these noninvasive "liquid biopsy" samples.
authors.affiliation: City of Hope Comprehensive Cancer Center, Duarte, California. Electronic address: KRechamp@coh.org.
authors.forename: Karen L
authors.initials: KL
authors.lastname: Reckamp
authors.affiliation: Trovagene, Inc., San Diego, California.
authors.forename: Vladislava O
authors.initials: VO
authors.lastname: Melnikova
authors.affiliation: Clovis Oncology, Inc., Boulder, Colorado.
authors.forename: Chris
authors.initials: C
authors.lastname: Karlovich
authors.affiliation: Massachusetts General Hospital, Boston, Massachusetts.
authors.forename: Lecia V
authors.initials: LV
authors.lastname: Sequist
authors.affiliation: University of Colorado, Denver, Colorado.
authors.forename: D Ross
authors.initials: DR
authors.lastname: Camidge
authors.affiliation: Stanford University Medical Center, Stanford, California.
authors.forename: Heather
authors.initials: H
authors.lastname: Wakelee
authors.affiliation: Hospices Civils de Lyon, Lyon, France.
authors.forename: Maurice
authors.initials: M
authors.lastname: Perol
authors.affiliation: Dana Farber Cancer Institute, Boston, Massachusetts.
authors.forename: Geoffrey R
authors.initials: GR
authors.lastname: Oxnard
authors.affiliation: Trovagene, Inc., San Diego, California.
authors.forename: Karena
authors.initials: K
authors.lastname: Kosco
authors.affiliation: Trovagene, Inc., San Diego, California.
authors.forename: Peter
authors.initials: P
authors.lastname: Croucher
authors.affiliation: Trovagene, Inc., San Diego, California.
authors.forename: Errin
authors.initials: E
authors.lastname: Samuelsz
authors.affiliation: Trovagene, Inc., San Diego, California.
authors.forename: Cecile Rose
authors.initials: CR
authors.lastname: Vibat
authors.affiliation: Trovagene, Inc., San Diego, California.
authors.forename: Shiloh
authors.initials: S
authors.lastname: Guerrero
authors.affiliation: Clovis Oncology, Inc., Boulder, Colorado.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Geis
authors.affiliation: Beverly Hills Cancer Center, Beverly Hills, CA; City of Hope, Duarte, California.
authors.forename: David
authors.initials: D
authors.lastname: Berz
authors.affiliation: Clovis Oncology, Inc., Boulder, Colorado.
authors.forename: Elaina
authors.initials: E
authors.lastname: Mann
authors.affiliation: Clovis Oncology, Inc., Boulder, Colorado.
authors.forename: Shannon
authors.initials: S
authors.lastname: Matheny
authors.affiliation: Clovis Oncology, Inc., Boulder, Colorado.
authors.forename: Lindsey
authors.initials: L
authors.lastname: Rolfe
authors.affiliation: Clovis Oncology, Inc., Boulder, Colorado.
authors.forename: Mitch
authors.initials: M
authors.lastname: Raponi
authors.affiliation: Trovagene, Inc., San Diego, California.
authors.forename: Mark G
authors.initials: MG
authors.lastname: Erlander
authors.affiliation: Barbara Karmanos Cancer Institute, Detroit, Michigan.
authors.forename: Shirish
authors.initials: S
authors.lastname: Gadgeel
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics* / Q000473:pathology; D004311:Double-Blind Method; D066246:ErbB Receptors / Q000097:blood* / Q000652:urine*; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D009154:Mutation; D012189:Retrospective Studies
chemical_list: D066246:ErbB Receptors
keywords: Circulating tumor DNA; EGFR mutations; NSCLC; T790M; Urine