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Background Approximately 30% of tumor biopsies from patients with advanced-stage lung adenocarcinomas yield insufficient tissue for successful molecular subtyping. We have analyzed the clinical utility of next-generation sequencing (NGS) of cell-free circulating tumor DNA (ctDNA) in patients with inadequate tumor samples for tissue genotyping. Patients and methods We conducted the study in a multi-institutional prospective cohort of clinically unselected patients with advanced-stage lung adenocarcinomas with insufficient tissue for EGFR, ALK or ROS1 genotyping across 12 Spanish institutions (n  =   93). ctDNA NGS was carried out by Guardant Health (Guardant360, Redwood City, CA), using a hybrid-capture-based 73-gene panel. Variants were deemed actionable if they were part of the OncoKB precision oncology knowledge database and classified in four levels of actionability based on their clinical or preclinical evidence for drug response. Results Eighty-three out of 93 patients (89%) had detectable levels of ctDNA. Potentially actionable level 1–4 genomic alterations were detected in 53 cases (57%), of which 13 (14%) had level 1–2A alterations (Food and Drug Administration-approved and standard-care biomarkers according to lung cancer guidelines). Frequencies of each genomic alteration in ctDNA were consistent with those observed in unselected pulmonary adenocarcinomas. The majority of the patients (62%), particularly those with actionable alterations (87%), had more than one pathogenic variant in ctDNA. The median turnaround time to genomic results was 13 days. Twelve patients (13%) received genotype-matched therapies based on ctDNA results, deriving the expected clinical benefit. Patients with co-occurring pathogenic alterations had a significantly shorter median overall survival as compared with patients without co-occurring pathogenic alteration (multivariate hazard ratio = 5.35, P  =   0.01). Conclusion Digital NGS of ctDNA in lung cancers with insufficient tumor samples for tissue sequencing detects actionable variants that frequently co-occur with other potentially clinically relevant genomic alterations, allowing timely initiation of genotype-matched therapies. [ABSTRACT FROM AUTHOR]

Background Colorectal cancer (CRC) has been shown to acquire RAS and EGFR ectodomain mutations as mechanisms of resistance to epidermal growth factor receptor (EGFR) inhibition (anti-EGFR). After anti-EGFR withdrawal, RAS and EGFR mutant clones lack a growth advantage relative to other clones and decay; however, the kinetics of decay remain unclear. We sought to determine the kinetics of acquired RAS/EGFR mutations after discontinuation of anti-EGFR therapy. Patients and methods We present the post-progression circulating tumor DNA (ctDNA) profiles of 135 patients with RAS / BRAF wild-type metastatic CRC treated with anti-EGFR who acquired RAS and/or EGFR mutations during therapy. Our validation cohort consisted of an external dataset of 73 patients with a ctDNA profile suggestive of prior anti-EGFR exposure and serial sampling. A separate retrospective cohort of 80 patients was used to evaluate overall response rate and progression free survival during re-challenge therapies. Results Our analysis showed that RAS and EGFR relative mutant allele frequency decays exponentially (r 2=0.93 for RAS ; r 2=0.94 for EGFR) with a cumulative half-life of 4.4 months. We validated our findings using an external dataset of 73 patients with a ctDNA profile suggestive of prior anti-EGFR exposure and serial sampling, confirming exponential decay with an estimated half-life of 4.3 months. A separate retrospective cohort of 80 patients showed that patients had a higher overall response rate during re-challenge therapies after increasing time intervals, as predicted by our model. Conclusion These results provide scientific support for anti-EGFR re-challenge and guide the optimal timing of re-challenge initiation. [ABSTRACT FROM AUTHOR]

Background: To identify predictive markers for responders in lapatinib-treated patients and to demonstrate molecular changes during lapatinib treatment via cell-free genomics. Patients and methods: We prospectively evaluated the efficacy of combining lapatinib with capecitabine and oxaliplatin as first line neoadjuvant therapy in patients with previously untreated, HER2-overexpressing advanced gastric cancer. A parallel biomarker study was conducted by simultaneously performing immunohistochemistry and next-generation sequencing (NGS) with tumor and blood samples. Results: Complete response was confirmed in 7/32 patients (21.8%), 2 of whom received radical surgery with pathologicconfirmed complete response. Fifteen partial responses (46.8%) were observed, resulting in a 68.6% overall response rate. NGS of the 16 tumor specimens demonstrated that the most common co-occurring copy number alteration was CCNE1 amplification, which was present in 40% of HER2þ tumors. The relationship between CCNE1 amplification and lack of response to HER2-targeted therapy trended toward statistical significance (66.7% of non-responders versus 22.2% of responders harbored CCNE1 amplification; P=0.08). Patients with high level ERBB2 amplification by NGS were more likely to respond to therapy, compared with patients with low level ERBB2 amplification (P=0.02). Analysis of cfDNA showed that detectable ERBB2 copy number amplification in plasma was predictive to the response (100%, response rate) and changes in plasma-detected genomic alterations were associated with lapatinib sensitivity and/or resistance. The follow-up cfDNA genomics at disease progression demonstrated that there are emergences of other genomic aberrations such as MYC, EGFR, FGFR2 and MET amplifications. Conclusions: The present study showed that HER2þGC patients respond differently according to concomitant genomic aberrations beyond ERBB2, high ERBB2 amplification by NGS or cfDNA can be a positive predictor for patient selection, and tumor genomic alterations change significantly during targeted agent therapy. [ABSTRACT FROM AUTHOR]

Background: While deregulation of the cyclin D1-CDK4/6-retinoblastoma pathway is common in hormone receptor positive (HR+) breast cancer, Rb is usually intact in HR+breast cancer, and targeted CDK 4/6 inhibitors that act upstream of Rb, are routinely being utilized in clinical practice. However, factors that can lead to clinical resistance to CDK 4/6 inhibitors are not known. Patients and methods: We identified patients who had pre- and post-genotyping in tissue and peripheral blood samples after receiving CDK 4/6 inhibitors. Genotyping was carried out in tumor tissue or blood collected before start of CDK 4/6 inhibitor and after disease progression on CDK 4/6 inhibitor, covering more than 90% of the coding region in RB1. Results: We identified detectable acquired RB1 mutations in circulating tumor DNA (ctDNA) after exposure to CDK4/6 inhibitor (palbociclib, palbociclib, ribociclib) for 5, 8, and 13 months, respectively, in three patients. The RB1 mutations included substitution in donor splicing site of exon 8 of the RB1 gene in patient #1; substitution in donor splicing site of exon 22 of RB1 gene, exon 19 deletion, exon 3 insertion in patient #2; and RB1 exon 16 H483Y mutation in patient #3. None of these RB1 mutations were present in the pre-CDK 4/6 specimen highlighting these molecular alterations, which lead to functional loss of Rb1, likely emerged under selective pressure from the CDK4/6 inhibitor potentially confering therapeutic resistance. Conclusion: This is the first clinical report to describe the emergence of somatic RB1 mutations after exposure to palbociclib or ribociclib, in patients with metastatic breast cancer. Further research is needed to validate these findings, identify how these mutations temporally emerge under selective pressure of CDK 4/6 inhibitor, and develop rational therapeutic strategies. [ABSTRACT FROM AUTHOR]

MET amplification (amp) is rare in untreated metastatic colorectal cancer (mCRC), but cell-free DNA (cfDNA) analysis identifies it in ∼20% with anti-EGFR monoclonal antibody (mAb)-refractory disease. CfDNA analysis reveals both focal gene amp and gene copy number gain due to chromosomal aneuploidy (non-focal MET amp). We analyzed cfDNA from Japanese (JPN) and United States (US) patients (pts) with anti-EGFR mAb-refractory mCRC to investigate the prevalence of focal and non-focal MET amp. We studied MET amp in pts with mCRC in the Nationwide Cancer Genome Screening Project in JPN (GOZILA; UMIN000029315; JPN cohort) and in a phase I/II trial of cabozantinib (C) +/- panitumumab (P) in pts with RAS wild-type mCRC (NCT02008383; US cohort). MET amp was detected with the Guardant360 assay. Focal MET amp was defined as either the only amp on chromosome (chr) 7q or MET copy number ³4. Non-focal MET amp was defined as co-amp with ³1 other genes (CDK6 or BRAF) located on chr 7q and MET copy number <4. 244 JPN cohort pts were analyzed from 2/2018-12/2018, and 81 pts in the US cohort, from 8/2014-2/2018. In pts with prior anti-EGFR mAb (JPN: n = 184; US: n = 70), focal and non-focal MET amp were detected in 8 (4.3%) and 30 (16.3%) pts in the JPN cohort, and in 9 (12.9%) and 4 (5.7%) US pts. Without prior anti-EGFR mAb (JPN: n = 60; US: n = 11), focal and non-focal MET amp were detected in 1 (1.7%) and 4 (6.7%) pts in the JPN cohort, and 0 (0%) and 1 (9.1%) US pts. The focal MET amp rate combined was 6.7% (17/254) with prior anti-EGFR mAb. In the US cohort, 4/6 pts with focal MET amp treated with C or C+P had a reduction in RECIST lesions as best response, with one PR, while 2 pts with non-focal MET amp had tumor growth as best response. The prevalence of focal MET amp detected by cfDNA analysis in mCRC pts was similar in the JPN and US cohorts. Given preliminary efficacy results from the US trial, focal MET amp may be a [A1] predictor of benefit from a MET inhibitor. A clinical trial evaluating a MET inhibitor in mCRC pts with focal MET amp is underway. The authors. Has not received any funding. A. Kawazoe: Honoraria (self), Research grant / Funding (institution): Ono Oharmaceutical; Honoraria (self), Research grant / Funding (institution): Taiho Pharmaceutical; Research grant / Funding (institution): Dainippon Sumitomo Pharma; Research grant / Funding (institution): Takeda Pharmaceutical; Research grant / Funding (institution): Astellas Pharmaceutical; Research grant / Funding (institution): MSD. Y. Nakamura: Research grant / Funding (institution): Ono Pharmaceutical; Research grant / Funding (institution): Taiho Pharmaceutical. J.I. Odegaard: Full / Part-time employment: Guardant Health. M.I. Lefterova: Full / Part-time employment: Guardant Health. R. Lanman: Full / Part-time employment: Guardant Health. T. Yoshino: Research grant / Funding (institution): Novartis Pharma K.K.; Research grant / Funding (institution): MSD.K.K.; Research grant / Funding (institution): Sumitomo Dainippon Pharma Co., Ltd.; Research grant / Funding (institution): CHUGAI PHARMACEUTICAL CO., LTD.; Research grant / Funding (institution): Sanofi K.K.; Research grant / Funding (institution): DAIICHI SANKYO COMPANY, LIMITED; Research grant / Funding (institution): PAREXEL International Inc.; Research grant / Funding (institution): ONO PHARMACEUTICAL CO., LTD.. J.H. Strickler: Honoraria (self), Advisory / Consultancy, Research grant / Funding (institution): Amgen; Honoraria (self), Advisory / Consultancy: Bayer; Honoraria (self): Chugai; Honoraria (institution), Advisory / Consultancy, Research grant / Funding (institution): Seattle Genetics; Advisory / Consultancy, Research grant / Funding (institution): Genentech/Roche; Advisory / Consultancy: AstraZeneca; Advisory / Consultancy, Leadership role, Research grant / Funding (institution): OncoMed; Advisory / Consultancy: Celgene; Advisory / Consultancy: Proteus Digital Health; Advisory / Consultancy: Chengdu Kanghong Biotechnology Co., Ltd; Research grant / Funding (institution): Exelixis; Research grant / Funding (institution): Sanofi; Research grant / Funding (institution): Nektar; Research grant / Funding (institution): Abbvie; Research grant / Funding (institution): Gilead Sciences; Research grant / Funding (institution): Macrogenics; Research grant / Funding (institution): Leap Therapeutics; Research grant / Funding (institution): MedImmune. All other authors have declared no conflicts of interest. [ABSTRACT FROM AUTHOR]