Next-generation sequencing for tumor mutation quantification using liquid biopsies

2020 ◽  
Vol 58 (2) ◽  
pp. 306-313 ◽  
Author(s):  
Mariano Provencio ◽  
Clara Pérez-Barrios ◽  
Miguel Barquin ◽  
Virginia Calvo ◽  
Fabio Franco ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Next Generation Sequencing ◽  
Correlation Coefficient ◽  
Resistance Mutation ◽  
Response To Treatment ◽  
Clinical Samples ◽  
Next Generation ◽  
Liquid Biopsies ◽  
Nsclc Patients ◽  
Generation Sequencing

AbstractBackgroundNon-small cell lung cancer (NSCLC) patients benefit from targeted therapies both in first- and second-line treatment. Nevertheless, molecular profiling of lung cancer tumors after first disease progression is seldom performed. The analysis of circulating tumor DNA (ctDNA) enables not only non-invasive biomarker testing but also monitoring tumor response to treatment. Digital PCR (dPCR), although a robust approach, only enables the analysis of a limited number of mutations. Next-generation sequencing (NGS), on the other hand, enables the analysis of significantly greater numbers of mutations.MethodsA total of 54 circulating free DNA (cfDNA) samples from 52 NSCLC patients and two healthy donors were analyzed by NGS using the Oncomine™ Lung cfDNA Assay kit and dPCR.ResultsLin’s concordance correlation coefficient and Pearson’s correlation coefficient between mutant allele frequencies (MAFs) assessed by NGS and dPCR revealed a positive and linear relationship between the two data sets (ρc = 0.986; 95% confidence interval [CI] = 0.975–0.991; r = 0.987; p < 0.0001, respectively), indicating an excellent concordance between both measurements. Similarly, the agreement between NGS and dPCR for the detection of the resistance mutation p.T790M was almost perfect (K = 0.81; 95% CI = 0.62–0.99), with an excellent correlation in terms of MAFs (ρc = 0.991; 95% CI = 0.981–0.992 and Pearson’s r = 0.998; p < 0.0001). Importantly, cfDNA sequencing was successful using as low as 10 ng cfDNA input.ConclusionsMAFs assessed by NGS were highly correlated with MAFs assessed by dPCR, demonstrating that NGS is a robust technique for ctDNA quantification using clinical samples, thereby allowing for dynamic genomic surveillance in the era of precision medicine.

scholarly journals Next Generation Sequencing for Gene Fusion Analysis in Lung Cancer: A Literature Review

Diagnostics ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 521
Author(s):  
Rossella Bruno ◽  
Gabriella Fontanini
Keyword(s):  
Lung Cancer ◽  
Next Generation Sequencing ◽  
Gene Fusion ◽  
Repetitive Sequences ◽  
Clinical Samples ◽  
Gene Fusions ◽  
Next Generation ◽  
Hybrid Capture ◽  
Fusion Analysis ◽  
Generation Sequencing

Gene fusions have a pivotal role in non-small cell lung cancer (NSCLC) precision medicine. Several techniques can be used, from fluorescence in situ hybridization and immunohistochemistry to next generation sequencing (NGS). Although several NGS panels are available, gene fusion testing presents more technical challenges than other variants. This is a PubMed-based narrative review aiming to summarize NGS approaches for gene fusion analysis and their performance on NSCLC clinical samples. The analysis can be performed at DNA or RNA levels, using different target enrichment (hybrid-capture or amplicon-based) and sequencing chemistries, with both custom and commercially available panels. DNA sequencing evaluates different alteration types simultaneously, but large introns and repetitive sequences can impact on the performance and it does not discriminate between expressed and unexpressed gene fusions. RNA-based targeted approach analyses and quantifies directly fusion transcripts and is more accurate than DNA panels on tumor tissue, but it can be limited by RNA quality and quantity. On liquid biopsy, satisfying data have been published on circulating tumor DNA hybrid-capture panels. There is not a perfect method for gene fusion analysis, but NGS approaches, though still needing a complete standardization and optimization, present several advantages for the clinical practice.

Impact of next-generation sequencing on clinical outcomes in advanced EGFR-mutant lung cancer patients after resistance to osimertinib.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. e20726-e20726
Author(s):  
Jia-Tao Cheng ◽  
Jin-Ji Yang ◽  
Yi-Long Wu
Keyword(s):  
Lung Cancer ◽  
Next Generation Sequencing ◽  
Clinical Outcomes ◽  
Objective Response ◽  
Next Generation ◽  
Significant Difference ◽  
Egfr Mutant ◽  
Nsclc Patients ◽  
Ngs Data ◽  
Generation Sequencing

e20726 Background: Osimertinib is used to treat EGFR-mutant non–small-cell lung cancer (NSCLC) with acquired T790M mutation. Next-generation sequencing (NGS) is helpful to understand mechanisms of resistance to osimertinib. However, whether NGS after resistance to osimertinib has an impact on clinical outcomes of patients treated with subsequent treatments has been elusive. Methods: We retrospectively identified advanced, EGFR-mutant T790M positive NSCLC patients treated with the 2nd or further-line osimertinib from January 27th, 2015 to January 31th, 2019 at our institute. Genetic profiles and clinical outcomes were analyzed. These patients were divided into 2 groups based on NGS data after resistance to osimertinib. Progression-free survival1 (PFS1) was calculated from the start of osimertinib to progression or death. PFS2 was calculated from the start of subsequent-line treatment to progression or death. Objective response rate (ORR) of subsequent-line treatments was evaluated by RECIST1.1. Results: Among 187 patients treated with osimertinib, 66 had NGS data and 27 had no NGS data after progression. Maintained EGFR T790M was detected in 23 patients (34.8%), and loss of T790M was seen in 43 patients (65.2%). Mutations of EGFR C797S were detected in 12 patients (18.1% overall; 52.2% of those with retained T790M), 11 in cis with a maintained T790M, 1 in trans with a maintained T790M. There was no significant difference in median PFS1 between the maintained T790M group and the loss of T790M group (10.8 vs. 7.0 months, P = 0.085).The NGS group was treated with TKIs according to the results of NGS strictly (n = 36), the non-NGS group received chemotherapy or best supportive care (n = 11).There was a significant difference in median PFS2 between the NGS and non-NGS groups (5.4 vs. 2.9 months, P = 0.043). The ORR of the NGS group was significantly superior to that of the non-NGS group (16.2% vs 11.1%, P < 0.001). Conclusions: NGS after resistance to osimertinib might favor clinical outcomes of advanced EGFR-mutant NSCLC patients. Further more investigations are warranted.

TP53 fusions with next-generation sequencing (NGS) in Chinese non-small cell lung cancer (NSCLC): A multicenter study.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. e21742-e21742
Author(s):  
Huafei Chen ◽  
Wen xian Wang ◽  
Chunwei Xu ◽  
You-cai Zhu ◽  
Wu Zhuang ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Next Generation Sequencing ◽  
Multicenter Study ◽  
Lung Tumor ◽  
Next Generation ◽  
Entire Cohort ◽  
Detection Techniques ◽  
Nsclc Patients ◽  
Next Generation Sequencing Ngs ◽  
Generation Sequencing

e21742 Background: Currently, with the advances in detection techniques, such as next generation sequencing (NGS), more and more rare or atypical TP53 fusions had been identified. Such as TP53-PSMD14, the importance of EGFR signaling in the pathogenesis of lung cancer and the efficacy of EGFR-TKI treatment had been demonstrated. The aim of this study was to evaluate the prevalence of TP53 fusions in Chinese NSCLC populations, which had not been reported earlier, and to describe targeting potential in Chinese NSCLC populations. Methods: A multicenter study in China was initiated from February 2014, and NSCLC patients have been enrolled as of December 2018. A total of 2743 patients with NSCLC were screened by using next-generation sequencing (NGS)-based 381 genes panel assay for detecting TP53 fusions. Results: Of this entire cohort, just four (0.15%) patients were identified with a TP53 fusion, including DNAH2-TP53 (1), TP53-MPDU1 (1), TP53-FXR2 (1), TP53-VEZF1 (1). Of the TP53 fusion NSCLC patients, 50.00% were detected in female patients. Biopsies were obtained from primary lung tumor (25.00%) and metastatic sites (75.00%). Overall TMB in the TP53 fusion was high, which had more than 20 mut/Mb. Of the TP53 fusion NSCLC, two cases (50.00%) featured EGFR SV alterations. Conclusions: The frequency of TP53 fusions in Chinese populations with NSCLC is extremely rare (0.17%). TP53 fusions may reduce responsiveness to TKIs and worsen prognosis in EGFR-mutated NSCLC patients, mainly those carrying exon 19 deletions or exon 21 L858R.

APC fusions analysis in Chinese non-small cell lung cancer (NSCLC) using comprehensive NGS panel: A multicenter study.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. e21086-e21086
Author(s):  
Liping Wang ◽  
Chunwei Xu ◽  
Wen xian Wang ◽  
You-cai Zhu ◽  
Wu Zhuang ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Next Generation Sequencing ◽  
Multicenter Study ◽  
Next Generation ◽  
Entire Cohort ◽  
Detection Techniques ◽  
Tki Treatment ◽  
Nsclc Patients ◽  
Next Generation Sequencing Ngs ◽  
Generation Sequencing

e21086 Background: Currently, with the advances in detection techniques, such as next generation sequencing (NGS), more and more rare or atypical APC fusions had been identified. Such as APC-MCC, the importance of Wnt signaling in the pathogenesis of lung cancer and the efficacy of EGFR-TKI treatment had been demonstrated. Methods: A multicenter study in China was initiated from February 2014, and NSCLC patients have been enrolled as of December 2018. To determine the frequency of the APC fusions in NSCLC and other tumors, we analyzed data from 2743 clinical NSCLC cases, each of which had results from next-generation sequencing (NGS)-based 381 genes panel assay, analogous to the index patient. Results: Of this entire cohort, just two (0.07%) patients were identified with a APC fusion, including APC-MCC (1), APC-MRPL13 (1). The genes most frequently co-altered in patients with APC fusions were TP53 (100.00%), CCND1 (50.00%), ESR1 (50.00%), TNN (50.00%), MAP2K2 (50.00%) and POM121L12 (50.00%). Overall TMB in the APC fusions was low (median 2.16 mut/Mb). For treatments, both patients chose chemotherapy based on pemetrexed for first line, and a case example of advanced APC fusion NSCLC patients responding to anlotinib was actively being sought thru our database. Conclusions: The frequency of APC fusions in Chinese populations with NSCLC is extremely rare (0.08%). NGS can identify novel fusions and increase the list of actionable variants for patients. More patients can benefit from targeted treatment. In addition, for short-or long-term responses to TKI treatment, we can use the NGS assay to explore differential gene expression in the future.

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