Search

BACKGROUND This study assesses different technologies for detecting epidermal growth factor receptor (EGFR) mutations from circulating tumor DNA in patients with EGFR T790M-positive advanced non-small cell lung cancer (NSCLC) from the AURA3 study (NCT02151981), and it evaluates clinical responses to osimertinib and platinum-pemetrexed according to the plasma T790M status. METHODS Tumor tissue biopsy samples were tested for T790M during screening with the cobas EGFR Mutation Test (cobas tissue). Plasma samples were collected at screening and at the baseline and were retrospectively analyzed for EGFR mutations with the cobas EGFR Mutation Test v2 (cobas plasma), droplet digital polymerase chain reaction (ddPCR; Biodesix), and next-generation sequencing (NGS; Guardant360, Guardant Health). RESULTS With cobas tissue test results as a reference, the plasma T790M positive percent agreement (PPA) was 51% (110 of 215 samples) by cobas plasma, 58% (110 of 189) by ddPCR, and 66% (136 of 207) by NGS. Plasma T790M detection was associated with a larger median baseline tumor size (56 mm for T790M-positive vs 39 mm for T790M-negative; P

Purpose: To assess the utility of the cobas EGFR Mutation Test, with tissue and plasma, for first-line osimertinib therapy for patients with EGFR-mutated (EGFRm; Ex19del and/or L858R) advanced or metastatic non–small cell lung cancer (NSCLC) from the FLAURA study (NCT02296125). Experimental Design: Tumor tissue EGFRm status was determined at screening using the central cobas tissue test or a local tissue test. Baseline circulating tumor (ct)DNA EGFRm status was retrospectively determined with the central cobas plasma test. Results: Of 994 patients screened, 556 were randomized (289 and 267 with central and local EGFR test results, respectively) and 438 failed screening. Of those randomized from local EGFR test results, 217 patients had available central test results; 211/217 (97%) were retrospectively confirmed EGFRm positive by central cobas tissue test. Using reference central cobas tissue test results, positive percent agreements with cobas plasma test results for Ex19del and L858R detection were 79% [95% confidence interval (CI), 74–84] and 68% (95% CI, 61–75), respectively. Progression-free survival (PFS) superiority with osimertinib over comparator EGFR-TKI remained consistent irrespective of randomization route (central/local EGFRm-positive tissue test). In both treatment arms, PFS was prolonged in plasma ctDNA EGFRm-negative (23.5 and 15.0 months) versus -positive patients (15.2 and 9.7 months). Conclusions: Our results support utility of cobas tissue and plasma testing to aid selection of patients with EGFRm advanced NSCLC for first-line osimertinib treatment. Lack of EGFRm detection in plasma was associated with prolonged PFS versus patients plasma EGFRm positive, potentially due to patients having lower tumor burden.

Summary Background The Wee1 (WEE1hu) inhibitor adavosertib and gemcitabine have shown preclinical synergy and promising activity in early phase clinical trials. We aimed to determine the efficacy of this combination in patients with ovarian cancer. Methods In this double-blind, randomised, placebo-controlled, phase 2 trial, women with measurable recurrent platinum-resistant or platinum-refractory high-grade serous ovarian cancer were recruited from 11 academic centres in the USA and Canada. Women were eligible if they were aged 18 years or older, had an Eastern Cooperative Oncology Group performance status of 0–2, a life expectancy of more than 3 months, and normal organ and marrow function. Women with ovarian cancer of non-high-grade serous histology were eligible for enrolment in a non-randomised exploratory cohort. Eligible participants with high-grade serous ovarian cancer were randomly assigned (2:1), using block randomisation (block size of three and six) and no stratification, to receive intravenous gemcitabine (1000 mg/m2 on days 1, 8, and 15) with either oral adavosertib (175 mg) or identical placebo once daily on days 1, 2, 8, 9, 15, and 16, in 28-day cycles until disease progression or unacceptable toxicity. Patients and the team caring for each patient were masked to treatment assignment. The primary endpoint was progression-free survival. The safety and efficacy analysis population comprised all patients who received at least one dose of treatment. The trial is registered with ClinicalTrials.gov , NCT02151292 , and is closed to accrual. Findings Between Sept 22, 2014, and May 30, 2018, 124 women were enrolled, of whom 99 had high-grade serous ovarian cancer and were randomly assigned to adavosertib plus gemcitabine (65 [66%]) or placebo plus gemcitabine (34 [34%]). 25 women with non-high-grade serous ovarian cancer were enrolled in the exploratory cohort. After randomisation, five patients with high-grade serous ovarian cancer were found to be ineligible (four in the experimental group and one in the control group) and did not receive treatment. Median age for all treated patients (n=119) was 62 years (IQR 54–67). Progression-free survival was longer with adavosertib plus gemcitabine (median 4·6 months [95% CI 3·6–6·4] with adavosertib plus gemcitabine vs 3·0 months [1·8–3·8] with placebo plus gemcitabine; hazard ratio 0·55 [95% CI 0·35–0·90]; log-rank p=0·015). The most frequent grade 3 or worse adverse events were haematological (neutropenia in 38 [62%] of 61 participants in the adavosertib plus gemcitabine group vs ten [30%] of 33 in the placebo plus gemcitabine group; thrombocytopenia in 19 [31%] of 61 in the adavosertib plus gemcitabine group vs two [6%] of 33 in the placebo plus gemcitabine group). There were no treatment-related deaths; two patients (one in each group in the high-grade serous ovarian cancer cohort) died while on study medication (from sepsis in the experimental group and from disease progression in the control group). Interpretation The observed clinical efficacy of a Wee1 inhibitor combined with gemcitabine supports ongoing assessment of DNA damage response drugs in high-grade serous ovarian cancer, a TP53-mutated tumour type with high replication stress. This therapeutic approach might be applicable to other tumour types with high replication stress; larger confirmatory studies are required. Funding US National Cancer Institute Cancer Therapy Evaluation Program, Ontario Institute for Cancer Research, US Department of Defense, Princess Margaret Cancer Foundation, and AstraZeneca.

Introduction Osimertinib is an oral, central nervous system–active, EGFR tyrosine kinase inhibitor (TKI) for the treatment of EGFR T790M–positive advanced NSCLC. Here we have evaluated EGFR mutation frequencies in two phase II studies of osimertinib (AURA extension and AURA2). Methods After progression while receiving their latest line of therapy, patients with EGFR mutation–positive advanced NSCLC provided tumor samples for mandatory central T790M testing for the study selection criteria. Tumor tissue mutation analysis for patient selection was performed with the Roche cobas EGFR Mutation Test (European Conformity–in vitro diagnostic, labeled investigational use only) (Roche Molecular Systems, Pleasanton, CA). Patients should not have been prescreened for T790M mutation status. The cobas test results were compared with those of the MiSeq next-generation sequencing system (Illumina, San Diego, CA), which was used as a reference method. Results Samples from 324 and 373 patients screened for AURA extension and AURA2, respectively, produced valid cobas test results. The T790M detection rates were similar between AURA extension and AURA2 (64% and 63%, respectively). The pooled T790M rate was 63%, with no difference by ethnicity (63% for Asian and non-Asian patients alike) or immediately prior treatment with an EGFR TKI (afatinib, 69%; erlotinib, 69%; and gefitinib, 63%). A higher proportion of patients had T790M detected against a background of exon 19 deletions versus L858R mutation (73% versus 58% [ p = 0.0002]). In both trials the cobas test demonstrated high sensitivity (positive percent agreement) and specificity (negative percent agreement) for T790M detection when compared with the next-generation sequencing reference method: positive percent agreement of 91% versus 89% and negative percent agreement of 97% versus 98%. Conclusions In both trials, the rate of detection of T790M mutation in patients with advanced NSCLC was approximately 63% and was unaffected by immediately prior treatment with an EGFR TKI or ethnicity.

Objectives Reliable epidermal growth factor receptor (EGFR) mutation testing techniques are required to identify eligible patients with EGFR mutation/T790M positive advanced non-small cell lung cancer (NSCLC), for treatment with osimertinib (AZD9291), an oral, potent, irreversible EGFR tyrosine kinase inhibitor (TKI) selective for EGFR-TKI-sensitizing and T790M resistance mutations over wild-type EGFR. There is no current consensus regarding the best method to detect EGFR T790M mutations. The aim of this study was to describe the concordance between local testing, which used a variety of methods, and central testing, using the cobas ® EGFR Mutation Test, for EGFR-sensitizing mutations and the T790M resistance mutation. Materials and methods Tumor samples were obtained from all patients screened for inclusion onto the osimertinib Phase I expansion component of the AURA Phase I/II study (NCT01802632). Samples underwent central laboratory testing for EGFR-sensitizing mutations and T790M resistance mutation using the cobas ® EGFR Mutation Test. Results were compared with local laboratory test results, based on other testing methodologies including Sanger sequencing, therascreen ® , PNAClamp™, and Sequenom MassARRAY ® . Results Central laboratory testing was successful in 99% of samples passing histopathology review and testing success rates were comparable across the three central laboratories. Concordance between central and local testing for common sensitizing mutations was high (>98%) and concordance for the T790M mutation was also high (>90%). Tumor heterogeneity, along with other technical factors may have influenced this result. Conclusions Within the osimertinib AURA Phase I study, EGFR mutation testing across three centralized laboratories using the cobas ® EGFR Mutation Test was feasible and successful, with strong concordance between local and central laboratory results, including for T790M. The cobas ® EGFR Mutation Test has subsequently been approved as the companion diagnostic test for osimertinib in the USA and Japan.

Osimertinib (TAGRISSO) is an irreversible epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, selective for sensitizing and resistance mutations in the receptor. It was initially developed for second and later line use in patients whose disease had progressed on an earlier generation inhibitor due to the acquisition of the T790M resistance mutation. To identify patients whose tumor had acquired T790M-positive resistance, a companion diagnostic was codeveloped alongside the small molecule. The Roche cobas® EGFR Mutation Test v2 was developed and clinical utility of T790M mutation detection was demonstrated for osimertinib; the test was initially approved for use with formalin-fixed paraffin-embedded tissue and later with circulating tumor DNA (ctDNA) from plasma samples. Several challenges were overcome to enable simultaneous FDA approval of the companion diagnostic and the new drug application as well as the approval of a plasma ctDNA-based companion diagnostic the following year; these are discussed.

Objectives: In AURA3 (NCT02151981), osimertinib treatment provided significant clinical benefit compared with platinum-pemetrexed in patients with epidermal growth factor receptor (EGFR) T790M-positive advanced non-small-cell lung cancer (NSCLC), whose tumors had progressed on previous EGFR-tyrosine kinase inhibitor therapy. This retrospective analysis investigated detection rates for T790M, common (exon 19 deletion and L858R), and rare EGFR mutations in tissue samples from the screened population of AURA3. Materials and methods: In AURA3, eligible patients were randomized 2:1 to receive oral osimertinib 80 mg once daily or intravenous platinum-pemetrexed every 3 weeks for up to six cycles. Tumor tissue samples were centrally tested for EGFR mutations using the cobas® EGFR Mutation Test (Version 2). T790M-positive status was a key inclusion criteria. Results: A total of 820 screened patients had a valid EGFR mutation test result, of whom 452 (55%) were T790M-positive. Detection rates were similar by ethnicity (Asian versus non-Asian) for T790M (53% versus 58%) and exon 19 deletions (56% versus 63%). Conversely, the L858R rate was higher among Asian patients versus non-Asian patients (39% versus 28%; p = 0.0017). In the overall population, a higher proportion of patients had T790M detected against a background of exon 19 deletion versus L858R mutations (64% versus 47%; p Conclusion: Among AURA3 screened patients with EGFR mutation-positive advanced NSCLC, approximately half had detectable T790M in their tumor tissue, a rate unaffected by ethnicity. Results are consistent with previous reports of T790M detection rate in this patient population.

Objectives To assess the ability of different technology platforms to detect epidermal growth factor receptor (EGFR) mutations, including T790M, from circulating tumor DNA (ctDNA) in advanced non-small cell lung cancer (NSCLC) patients. Materials and methods A comparison of multiple platforms for detecting EGFR mutations in plasma ctDNA was undertaken. Plasma samples were collected from patients entering the ongoing AURA trial (NCT01802632), investigating the safety, tolerability, and efficacy of AZD9291 in patients with EGFR-sensitizing mutation-positive NSCLC. Plasma was collected prior to AZD9291 dosing but following clinical progression on a previous EGFR-tyrosine kinase inhibitor (TKI). Extracted ctDNA was analyzed using two non-digital platforms (cobas ® EGFR Mutation Test and therascreen™ EGFR amplification refractory mutation system assay) and two digital platforms (Droplet Digital™ PCR and BEAMing digital PCR [dPCR]). Results Preliminary assessment (38 samples) was conducted using all four platforms. For EGFR-TKI-sensitizing mutations, high sensitivity (78–100%) and specificity (93–100%) were observed using tissue as a non-reference standard. For the T790M mutation, the digital platforms outperformed the non-digital platforms. Subsequent assessment using 72 additional baseline plasma samples was conducted using the cobas ® EGFR Mutation Test and BEAMing dPCR. The two platforms demonstrated high sensitivity (82–87%) and specificity (97%) for EGFR-sensitizing mutations. For the T790M mutation, the sensitivity and specificity were 73% and 67%, respectively, with the cobas ® EGFR Mutation Test, and 81% and 58%, respectively, with BEAMing dPCR. Concordance between the platforms was >90%, showing that multiple platforms are capable of sensitive and specific detection of EGFR-TKI-sensitizing mutations from NSCLC patient plasma. Conclusion The cobas ® EGFR Mutation Test and BEAMing dPCR demonstrate a high sensitivity for T790M mutation detection. Genomic heterogeneity of T790M-mediated resistance may explain the reduced specificity observed with plasma-based detection of T790M mutations versus tissue. These data support the use of both platforms in the AZD9291 clinical development program.

Background Adavosertib (AZD1775) is a first-in-class, selective, small-molecule inhibitor of Wee1. Previously we reported safety and efficacy of adavosertib monotherapy in patients (pts) with advanced solid tumors (NCT02482311; Bauer et al Cancer Res 2016;76[14 Suppl]); here we report safety and efficacy data from expansion cohorts based on tumor type. Methods A total of 80 pts grouped into 6 biomarker-matched cohorts (Table) received adavosertib (175 mg PO bid; days 1–3 and 8–10 per 21-day cycle). Eligible pts had: confirmed diagnosis of ovarian cancer (OC), small-cell lung cancer (SCLC) or triple-negative breast cancer (TNBC); prior treatment (Tx) for metastatic/recurrent disease (≥3 prior Tx for pts with BRCAwt OC; progression following PARPi Tx for BRCA1/2m OC pts; ≤1 chemotherapy-based Tx for SCLC and ≥1 chemotherapy-based Tx for TNBC); measurable disease. Primary objective assessments: ORR; DCR; PFS (RECIST v1.1); safety. Tumor assessments were performed every 6 weeks in year 1 and every 12 weeks thereafter until disease progression or intolerable toxicity. Blood and tumor samples were collected for correlative biomarker and pharmacokinetic (PK) analyses. Results Median total Tx duration was 2.4 months. Most frequently reported adverse events (AEs) were diarrhea (61%), nausea (50%) and fatigue (43%). Most commonly reported grade ≥3 AEs were diarrhea (7.5%), nausea, fatigue and small intestine obstruction (6%). AEs leading to dose interruptions (22.5%), reductions (11.3%) or discontinuations (16.3%) were reported. The study showed preliminary antitumor activity, particularly in BRCAwt as well as PARPi-failure BRCAm OC pts (Table). PK and biomarker analyses will be presented. Conclusions Adavosertib was generally well tolerated and showed preliminary antitumor activity. DCR was modest across all patient cohorts. Patient cohortParameterOC, BRCAwtOC, BRCAm, PARPi failureTNBC, CCNE1/MYC/MYCL1/ MYCN non-amplifiedTNBC, CCNE1/MYC/MYCL1/ MYCN amplifiedSCLC, CCNE1/MYC/MYCL1/ MYCN non-amplifiedSCLC, CCNE1/MYC/MYCL1/MYCN amplifiedTotalPatients who received treatment, n163013612380Median age, years (range)62.5 (47–83)59.5 (44–73)58.0 (35–81)54.0 (43–78)64.5 (54–74)63.0 (56–69)60.0(35–83)Prior regimens, n (%) 1 2 3 4 5 6 >6 Median0 0 1 (6) 3 (19) 2 (13) 1 (6) 9 (56) 70 0 2 (7) 3 (10) 7 (23) 8 (27) 10 (33) 60 3 (23) 2 (15) 1 (8) 2 (15) 3 (23) 2 (15) 50 0 1 (17) 2 (33) 2 (33) 1 (17) 0 4.53 (25) 5 (42) 3 (25) 0 1 (8) 0 0 20 1 (33) 2 (67) 0 0 0 0 33 (4) 9 (11) 11 (14) 9 (11) 14 (18) 13 (16) 21 (26) 5ECOG PS, n (%) 0 18 (50) 8 (50)10 (33) 20 (67)4 (31) 9 (69)1 (17) 5 (83)2 (17) 10 (83)1 (33) 2 (67)26 (33)54 (68)ORR, n (%) CR* PR* SD ≥5 weeks PD NE1 (6) 0 1 (6) 10 (63) 4 (25) 1 (6)1 (3) 0 1 (3) 22 (73) 5 (17) 2 (7)0 0 0 9 (69) 3 (23) 1 (8)0 0 0 3 (50) 3 (50) 01 (8) 0 1 (8) 3 (25) 7 (58) 1 (8)0 0 0 1 (33) 2 (67) 03 (4) 0 3 (4) 48 (60) 24 (30) 5 (6)DCR, n (%)†11 (69)23 (77)9 (69)3 (50)4 (33)1 (33)51 (64)Median PFS, months4.53.93.12.01.31.23.0*Confirmed CR/PR; †DCR determined by CR + PR + SD. BRCA1/2m, BRCA1 or BRCA2 mutated; BRCAwt, BRCA1 and BRCA2 wild type; CR, complete response; DCR, disease control rate; ECOG PS, Eastern Cooperative Oncology Group performance status; NE, not evaluable; ORR, objective response rate; PARPi, PARP inhibitor; PD, progressive disease; PFS, progression-free survival; PR, partial response; SD, stable disease Citation Format: Todd M. Bauer, Kathleen Moore, Janet S. Rader, Fiona Simpkins, Alain Mita, J Thaddeus Beck, Lowell Hart, Quincy Chu, Amit Oza, Anna V. Tinker, Karen So, Esteban Rodrigo Imedio, Sanjeev Kumar, Ganesh M. Mugundu, Suzanne Jenkins, Juliann Chmielecki, Suzanne Jones, David R. Spigel, Siqing Fu. Open-label, multicenter, Phase Ib study to assess safety, tolerability and efficacy of adavosertib monotherapy in patients with advanced solid tumors: Expansion cohorts [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr CT012.

Introduction Tumor biopsies for detecting EGFR mutations in advanced NSCLC are invasive, costly, and not always feasible for patients with late-stage disease. The clinical utility of the cobas EGFR Mutation Test v2 (Roche Molecular Systems, Inc., Pleasanton, CA) with plasma samples from patients with NSCLC at disease progression after previous EGFR tyrosine kinase inhibitor therapy was investigated to determine eligibility for osimertinib treatment. Methods Matched tumor tissue and plasma samples from patients screened for the AURA extension and AURA2 phase II studies were tested for EGFR mutations by using tissue- and plasma-based cobas EGFR mutation tests. Plasma test performance was assessed by using the cobas tissue test and a next-generation sequencing method (MiSeq [Illumina Inc., San Diego, CA]) as references. The objective response rate, measured by blinded independent central review, was assessed in patients receiving osimertinib with a plasma T790M mutation–positive status. Results During screening, 551 patients provided matched tumor tissue and plasma samples. Pooled analysis of the positive and negative percent agreements between the cobas plasma and tissue tests for detection of T790M mutation were 61% and 79%, respectively. Comparing cobas plasma test with next-generation sequencing demonstrated positive and negative percent agreements of 90% or higher. The objective response rate was 64% (95% confidence interval: 57–70) in T790M mutation–positive patients by both cobas tissue and plasma tests (evaluable for response). Conclusions The cobas plasma test detected the T790M mutation in 61% of tumor tissue T790M mutation–positive patients. To mitigate the risk of false-negative plasma results, patients with a negative plasma result should undergo a tissue test where feasible.

Introduction: Increase in targeted therapies has resulted in the need for a single assay capable of detecting diverse biomarkers indicated for these agents. Comprehensive genomic profiling (CGP) provides such a solution, but due to the complexity and number of assays available today, standardization of validation has become critically important. We present FoundationOne CDx, the first NGS-based comprehensive companion diagnostics (CDx) platform developed and performed in compliance with FDA 21 CFR part 820. The assay interrogates 324 genes, and has CDx indications in five tumor types associated with 17 targeted therapies (Table 1). The versatile assay design will facilitate streamlined development of future CDx indications. Methods: DNA extracted from FFPE tumor tissue underwent whole-genome shotgun library construction and hybridization-based capture, followed by sequencing using Illumina HiSeq 4000. Sequence data were processed using a proprietary analysis pipeline designed to detect base substitutions, indels, copy number alterations, rearrangements, microsatellite instability (MSI), and tumor mutational burden (TMB). Results: Clinical validity was established such that the concordance between CGP and approved CDx were statistically non-inferior to that of two runs of approved CDx. For analytical validity, limit of detection (LoD) was at allele frequency 4% for known substitutions and indels. LoD was 16% tumor content for copy number amplifications, 30% for homozygous deletions, 11% for rearrangements, 12% for MSI, and 20% for TMB. Concordance with an orthogonal NGS platform was 94.6% for substitutions and indels. Within-assay reproducibility had PPA 99.4%. Conclusion: Rapid expansion of targeted therapies and CDx has necessitated a new approach and urgency to defining performance standards. We developed a comprehensive CDx assay and demonstrated clinical and analytical validity to support and accelerate using CGP for routine clinical care. Table 1. Companion Diagnostic IndicationsIndicationBiomarkerTherapyNon-small cell lung cancer (NSCLC)EGFR exon 19 deletions and EGFR exon 21 L858R alterationsafatinib, gefitinib, or erlotinibEGFR exon 20 T790M alterationsosimertinibALK rearrangementsalectinib, crizotinib, or ceritinibBRAF V600Edabrafenib in combination with trametinibMelanomaBRAF V600Edabrafenib, vemurafenibBRAF V600E and V600Ktrametinib, cobimetinib, in combination with vemurafenibBreast cancerERBB2 (HER2) amplificationtrastuzumab, ado-trastuzumab-emtansine, or pertuzumabColorectal cancerKRAS wild-type (absence of mutations in codons 12 and 13)cetuximabKRAS and NRAS wild-type (absence of mutations in exons 2, 3, and 4)panitumumabOvarian cancerBRCA1/2 alterationsrucaparib Citation Format: James X. Sun, Yali Li, Coren Milbury, Joel Skoletsky, Christine Burns, Wai-ki Yip, Jun Luo, Ninad Dewal, Adrienne Johnson, Kyle Gowen, Jing Tong, Yuting He, Jie He, Pei Ma, Jared White, Steve Roels, John Truesdell, Eric Peters, Houston Gilbert, Charlie Wu, Erica Schleifman, Johannes Noe, Carl Barrett, Kenneth Thress, Suzanne Jenkins, Julia Elvin, Geoff Otto, Doron Lipson, Jeffrey Ross, Vincent Miller, Philip Stephens, Michael Doherty, Christine Vietz. A clinically validated comprehensive companion diagnostic platform for care of patients with advanced cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4757.

Whole genome approaches using single nucleotide polymorphism (SNP) markers have the potential to transform complex disease genetics and expedite pharmacogenetics research. This has led to a requirement for high-throughput SNP genotyping platforms. Development of a successful high-throughput genotyping platform depends on coupling reliable assay chemistry with an appropriate detection system to maximise efficiency with respect to accuracy, speed and cost. Current technology platforms are able to deliver throughputs in excess of 100 000 genotypes per day, with an accuracy of >99%, at a cost of 20–30 cents per genotype. In order to meet the demands of the coming years, however, genotyping platforms need to deliver throughputs in the order of one million genotypes per day at a cost of only a few cents per genotype. In addition, DNA template requirements must be minimised such that hundreds of thousands of SNPs can be interrogated using a relatively small amount of genomic DNA. As such, it is predicted that the next generation of high-throughput genotyping platforms will exploit large-scale multiplex reactions and solid phase assay detection systems.

This study has assessed the ability of five hand-washing agents, Hibisol, Hibiscrub, Betadine, 70% isopropyl alcohol and bar soap, to decontaminate the surfaces of Biogel D, Surgikos Microtouch and Ansell Gammex latex medical gloves after repeated inoculation with Staphylococcus aureus. The glove and hand-washing agents were compared in a Latin Square experiment, and a standardized handwashing regime followed. Each of the gloves used in the study was later tested electrically for micropuncture formation. On the basis of microbial recovery, no specific combination of glove and hand washing agent proved superior, the marker organism being isolated on a sporadic basis. In addition, 17% of the gloves showed evidence of micropunctures after five sequential inoculations and washings. These results suggest that, even under strictly controlled conditions, glove surfaces cannot be reliably and consistently cleaned of microbial contamination, nor can they be guaranteed to remain puncture-free. Ideally, therefore, a new pair of gloves should be worn for each patient.

One of the major goals of pharmacogenetics is to elucidate mechanisms and identify patients at increased risk of adverse events (AEs). To date, however, there have been only a few successful examples of this type of approach. In this paper, we describe a retrospective case-control pharmacogenetic study of an AE of unknown mechanism, characterized by elevated levels of serum alanine aminotransferase (ALAT) during long-term treatment with the oral direct thrombin inhibitor ximelagatran. The study was based on 74 cases and 130 treated controls and included both a genome-wide tag single nucleotide polymorphism and large-scale candidate gene analysis. A strong genetic association between elevated ALAT and the MHC alleles DRB1(*)07 and DQA1(*)02 was discovered and replicated, suggesting a possible immune pathogenesis. Consistent with this hypothesis, immunological studies suggest that ximelagatran may have the ability to act as a contact sensitizer, and hence be able to stimulate an adaptive immune response.

Objective AZD9291 is an oral, selective, irreversible EGFR-TKI, effective against EGFR-TKI-sensitizing (EGFRm) and resistance T790M mutations. Pre-study tumor samples from pts enrolled into expansion cohorts of a PhI study (AURA, NCT01802632) were tested centrally for T790M status. Pts with T790M positive tumor samples had higher objective response rate (ORR) than pts with no detectable T790M (T790M ‘negative’): 78/127 (61%) and 13/61 (21%), respectively (Jänne et al 2015). Exploratory analyses to characterize AZD9291 activity in pts whose tumors were T790M negative by central test were performed. Methods Pts with EGFRm aNSCLC and acquired resistance to EGFR-TKIs were enrolled into expansion cohorts of the AURA study. The T790M status of pre-study tumor samples was determined locally (any method allowed) and/or centrally using the cobasTM EGFR mutation test (Roche Molecular Systems, Inc.). Presence of T790M in plasma derived ctDNA was determined by digital PCR (BEAMing). Pts with a centrally determined tumor based T790M negative status were then classified into one of nine groups according to local tumor status (T790M positive, negative, or unknown) and ctDNA status (T790M positive, negative, or unknown). Clinical outcome is reported for each group. Results 69 pts with T790M negative tumor status were determined centrally, and received AZD9291; 35 and 21 were also negative by local test and ctDNA, respectively and 7 were negative by all 3 tests; despite a negative central test, T790M positive tumor status was identified in 11 pts by prior local test and 26 by ctDNA. Confirmed responses by RECIST 1.1, n/N (%) in centrally tested tissue samples with no detectable T790M (Data cut-off 01 May 2015)T790M status of plasma derived ctDNA (Method: BEAMing)PositiveNegativeUnknownTotalT790M status of tumor tissue determined by a local test (Method: Any)Positive2/6 (33)0/1 (0)4/4 (100)6/11 (55)Negative2/14 (14)0/7 (0)2/14 (14)4/35 (11)Unknown2/6 (33)3/13 (23)2/4 (50)7/23 (26)Total6/26 (23)3/21 (14)8/22 (36)17/69 (25) 17 pts with a central T790M negative status responded to AZD9291 treatment. ORR was 55% (6/11) in pts with a central T790M negative/local positive test result. No objective responses (0/7) were observed in pts with T790M negative tumor status by all three tests. Additional analysis of the sub-groups, by immediate prior TKI, EGFRm status, and duration of response will be available for presentation. Discussion In 69 pts with centrally identified T790M negative tumor status, 17 achieved a confirmed response. Of these, 12 were T790M positive by local test, ctDNA, or both. Different sensitivity and specificity of assays, tumor heterogeneity, and ctDNA shedding may explain positive results in this small group of pts classified as central T790M negative. The data support the hypothesis that T790M directed inhibitors are active against T790M driven resistance. Citation Format: Pasi A. Jänne, Enriqueta Felip, Dong-Wan Kim, Thomas John, Daniel E. Haggstrom, Myung-Ju Ahn, Kenneth S. Thress, Suzanne C. Jenkins, Helen Mann, Mireille V. Cantarini, Simon P. Dearden, James Chih-Hsin Yang. Characterization of the activity of AZD9291 in patients (pts) with T790M ‘negative’ advanced non-small cell lung cancer (aNSCLC). [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr B105.

Aim: AZD9291 is in clinical development to address T790M-mediated drug resistance associated with treatment of EGFR-mutant NSCLC. Genotyping of circulating plasma DNA may provide an option to identify patients (pts) unable to provide tissue biopsies. We aimed to study plasma levels of EGFR T790M in relation to AZD9291 drug activity. Methods: 101 pts with advanced NSCLC consented to plasma analysis as part of the Phase I study of AZD9291(NCT01802632). Central tumour genotyping was performed when feasible using the cobas™ assay. After pre-amplification of plasma DNA, levels of pre-dose T790M, L858R and exon 19 del variants were quantified using an emulsion PCR assay (Sysmex). Cases with no detectable plasma-sensitising mutation were omitted from T790M analysis. Response rate (RR) as of 16 Jan 2014 was calculated as the proportion of pts with ≥1 follow-up tumour measurement having a diameter decrease ≥30%. Results: Plasma assays for L858R and 19 del were studied in 44 pts with 19 del and 28 pts with L858R centrally confirmed in their tumour. A threshold of 0.05% mutant had 0% false positive rate (FPR) and 83% sensitivity for these mutations. In 64 pts with central T790M results, the same threshold applied to plasma T790M had 50% FPR, 89% sensitivity. Notably, of 6 cases falsely T790M+ in plasma, 5 were also T790M+ using a second plasma assay (cobas™). RR was 61% (25/41) and 24% (5/21) in 62 pts with central T790M+ and T790M- tumour genotyping (p = 0.008). Similarly, RR was 65% (33/51) and 38% (8/21) in 72 pts with T790M+ and T790M- plasma genotyping (p Conclusions: Plasma levels of EGFR-TKI-sensitising mutations correlate more closely with tumour genotype than T790M, likely due to the greater genomic heterogeneity of resistance. RR is higher in T790M+ pts than in T790M- pts, either using tumour or plasma genotyping. Plasma T790M genotyping using emulsion PCR may be an attractive alternative to rebiopsy for tumour genotyping of NSCLC pts with acquired EGFR resistance. Disclosure: K. Thress: Employment and stock ownership: AstraZeneca; J.C. Yang: Advisory boards: Boehringer Ingelheim, Eli Lilly, Pfizer, Novartis, Roche/Genentech, AstraZeneca, Merck, Bayer, Clovis Oncology. Corporate-sponsored research: Boehringer Ingelheim; D. Planchard: Advisory boards: AstraZeneca, Boehringer Ingelheim, Lilly, Novartis, Pfizer, Roche, BMS; R. Brant: Employment and stock ownership: AstraZeneca; H. Carr: Employment and stock ownership: AstraZeneca; S. Dearden: Employment and stock ownership: AstraZeneca; S. Jenkins: Employment and stock ownership: AstraZeneca; M. Cantarini: Employment and stock ownership: AstraZeneca; S. Ghiorghiu: Employment and stock ownership: AstraZeneca; J.C. Barrett: Employment and stock ownership: AstraZeneca; P.A. Janne: Consultant or advisory role: AstraZeneca, Boehringer Ingelheim, Clovis Oncology, Pfizer, Merrimack Pharmaceuticals, Chugai, Immunogen. Stock ownership: Gatekeeper. Other: Lab Corp; G.R. Oxnard: Compensated consultancy: Astra-Zeneca, Boehringer Ingelheim, Clovis, Genentech, Novartis, Sanofi. All other authors have declared no conflicts of interest.

e19051 Background: EGFR tyrosine kinase inhibitors (TKIs) have demonstrated clinical benefit in patients (pts) with advanced EGFR mutation-positive NSCLC. Eventually pts develop acquired resistance...

8092 Background: EGFR tyrosine kinase inhibitors are highly effective in treating EGFR mutant+ NSCLC. Patients invariably relapse and resistance is most often associated with an EGFR T790M mutation...

The influence of nursing on the decision-making process of selecting healthcare information systems (HISs) is increasing. Systems planning and implementation are based not only on the capabilities of information technology but also on forces in the environment that impact health care. Computer decisions in the 1970s were primarily financially oriented. Hospitals saw the need to automate their patient billing process and other key systems, such as general ledger and accounts payable. Advanced technology led to increased sophistication of hardware and software capabilities, and concurrently the information needs of the hospital increased. More extensive reporting requirements, as well as the need for management reporting on productivity, became the norm.

Emerging evidence suggests that circulating tumor DNA (ctDNA) provides an alternative diagnostic sample when surgical biopsies are inaccessible, and could enable disease diagnosis, frequent assessment of disease progression, and almost real-time monitoring of the emergence of resistance mutations. We hypothesized that novel highly sensitive technologies enable EGFR T790M mutation detection in ctDNA from patients that had previously been treated with and become resistant to EGFR-TKIs. We have used 3 technologies to assess T790M mutation detection rate. Samples comprised patients with sensitizing EGFR mutations and wild type EGFR patients who had been previously treated with EGFR TKI therapies. In approximately 50% of EGFR mutation positive cases, acquired resistance to EGFR-TKIs is mediated by an EGFR T790M mutation. Reference data for T790M mutations from tumor material was unavailable.We undertook a comprehensive cross-technology comparison of three technology platforms: ARMS based detection using the Roche cobas EGFR mutation detection kit; digital droplet PCR using the BioRad ddPCR instrument (by MolecularMD) and bead based digital PCR using the Inostics BEAMing technology for the detection of T790M mutations. In total, 135 frozen plasma samples, obtained from 2 clinical studies, were used to estimate T790M detection rates in ctDNA. Within the study group, 72 samples were taken from EGFR mutation positive patients, of which around 36 (expected range 28-44) would be expected to have T790M mutations, the remaining samples would be expected to be T790M negative. Each individual patient plasma sample was split and evaluated across multiple platforms. In addition, the suitability of 3 ctDNA preparation kits (cobas Circulating DNA Isolation kit, QIAamp Circulating Nucleic Acid kit and QIAamp DNA Mini kit) in terms of DNA yield was evaluated. The QIAamp Circulating Nucleic Acid kit and cobas Circulating DNA Isolation kit were equally effective in preparing ctDNA from plasma samples. Overall, concordance of EGFR T790M mutation status was high between all 3 methods. In addition, the false positive rate in the known EGFR mutation negative cases was low for all 3 methods. However, differences were seen in the rate of false negative results (assay sensitivity) between methods. DNA preparation kits specifically designed for use with ctDNA samples are preferable to other methods. Current technologies for the detection of T790M mutations in ctDNA, where patient tumor material is not available, still require further development to increase detection rates in these samples. Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C196. Citation Format: Helen Brown, Roz Brant, Simon Dearden, Suzanne Jenkins, Kenneth Thress, Alain Horvais, Ruth March, J. Carl Barrett. EGFR mutation detection in ctDNA isolated from NSCLC patient plasma; a cross-platform comparison of leading technologies. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C196.

Summary “ligan Stew,” a nutrition, education program consisting of films and supplementary learning materials, was given to 140 fourth-grade students. To evaluate the effectiveness of the method, changes in nutrition knowledge were measured by a written test given before and after the program. In addition, the change in dietary intake of the children were assessed using two 3-day food records. Children who viewed the films and participated in the activities increased their knowledge of nutrition significantly more than did control groups. While only some of the fourth-graders involved improved their diets when compared with controls, it should be noted that these were the children who showed the largest increase in nutrition knowledge. Since the objective of nutrition education is to change dietary intakes in such a way as to improve health, the results are encouraging. The “Mulligan Stew” program is especially valuable to nutrition educators because it is suitable for use on television, in or out of school.

• An acute gastrointestinal tract illness affected 213 (52%) of 407 campers and 64 (53%) of 121 staff members attending a boys' camp in the Catoctin Mountains of Maryland during the summer of 1981. Nausea was the predominant symptom for ill campers and staff members (73%), but more staff members experienced diarrhea (49%) than did campers (9%). Twenty-three individuals had more than one episode of illness compatible with the case definition. Eight of nine paired blood specimens from ill staff members showed a fourfold increase in antibody titer to Norwalk virus by radioimmunoassay. Environmental inspections and laboratory tests failed to implicate a common source of exposure. ( AJDC 1985;139:787-789)