Detection of Therapeutically Targetable Driver and Resistance Mutations in Lung Cancer Patients by Next-Generation Sequencing of Cell-Free Circulating Tumor DNA

May combine two claims.

ctDNA NGS detected 50 driver mutations and 12 resistance mutations.

“ctDNA NGS detected 50 driver and 12 resistance mutations, and mutations in 22 additional genes for which experimental therapies, including clinical trials, are available.”

May combine two claims.

EGFR mutations were the most common variants detected, making up 20% of all ctDNA variants.

“with EGFR variants being most common”

• Also stated in: § Therapeutically targetable driver and resistance mutations detected in ctDNA · plain

  “Variants were detected in 45 genes with EGFR mutations the most prevalent at 20% of total variants (Figure 2, Supplemental Table 3).”

May combine two claims.

Actionable EGFR mutations were found in 24 tissue samples and 19 ctDNA samples, with 79% concordance.

“Actionable EGFR mutations were detected in 24 tissue and 19 ctDNA samples, yielding concordance of 79%”

• Also stated in: § Comparison of tissue and plasma results · plain

  “Among the 50 patients with matched tDNA and ctDNA, 24 therapeutically targetable driver EGFR mutations were detected in tDNA and 19 in ctDNA samples, yielding an overall concordance of 79%.”

May combine two claims.

ctDNA NGS was completed for all 102 consecutive patients, whereas tissue sequencing succeeded for only 50 patients, or 49%.

“While ctDNA NGS was completed for 102 consecutive patients, tissue sequencing was only successful for 50 patients (49%).”

• Also stated in: § Patient characteristics and study design · plain

  “While ctDNA analysis was successfully completed for all 112 samples obtained from 102 consecutively enrolled patients, tissue NGS could only be completed for 50 (49.0%) patients.”

May combine two claims.

Ten ERBB2 mutations were detected, including 5 exon 20 insertions.

“Ten ERBB2 mutations were detected, including 5 exon 20 insertions shown to confer sensitivity to ERBB2-targeted therapies (36).”

May combine two claims.

60% of variants detected in ctDNA had an allelic fraction below 4.0%, which is below the tDNA sequencing platform threshold.

“60% of variants detected in ctDNA were at an AF less than 4.0%, which is below the threshold at which calls can be made for the tDNA sequencing platform (detailed AF results for 50 patients with matched ctDNA and tissue shown in Supplemental Table 6).”

May combine two claims.

Patient #23 had one of the lowest total cfDNA yields (14.5 ng versus median 32.1 ng; range 6-390 ng), and the LOD for EGFR variants was set at 0.2% rather than 0.1%.

“the total cfDNA yield was one of the lowest among our patients (14.5 ng versus a median of 32.1 ng (range 6–390 ng); Supplemental Figure 1), and the level of detection (LOD) for EGFR variants was therefore set at 0.2% rather than 0.1%.”

Probably not a substantial claim: may not pass S3 (load-bearing).

The most common reason for ordering the ctDNA test was initial detection of targetable mutations, for 52 patients.

“the most common reason for ordering the ctDNA test was for initial detection of targetable mutations for 52 patients who either had no previous therapy (n=27), chemotherapy (n=18), or chemotherapy and immunotherapy (n=7).”

May combine two claims.

Digital ctDNA sequencing was performed on 112 samples from 102 total patients, including 100 with advanced NSCLC and two with cancer of unknown primary.

“Digital ctDNA sequencing was performed for 112 samples obtained from 102 total patients, including 100 patients with advanced NSCLC plus two patients (patients 37 and 51) with cancer of unknown primary who were treated at the University of Pennsylvania Abramson Cancer Center between February 2015 and March 2016 (Table 1).”

May combine two claims.

Sufficient cfDNA was obtained and sequenced, but no somatic variants and thus no ctDNA were detected for 16 patients.

“Sufficient input cfDNA was obtained and sequenced but no somatic variants, and thus no ctDNA, detected for 16 patients.”

May combine two claims.

When only variants covered by both panels were considered, 55 variants were detected by tDNA and 67 by ctDNA, with 41 detected by both.

“When only the variants covered by both panels were considered, the difference narrowed to 55 and 67 variants respectively, while 41 mutations were detected by both (Figure 3A).”

May combine two claims.

Concordance between ctDNA and tDNA was higher for metastatic tissue (79%) than primary tumor tissue (51%), but the difference was not statistically significant, p=0.063.

“While overall concordance between ctDNA and tDNA was higher when tDNA was obtained for metastatic (79%) versus primary tumor tissue (51%), this difference was not statistically significant (p=0.063; Figure 3C).”

May combine two claims.

The mean number of variants detected per patient was 2.8 by ctDNA and 1.5 by tDNA.

“the mean number of variants detected per patient by ctDNA was 2.8, compared to 1.5 using tDNA.”

Probably not a substantial claim: may not pass S3 (load-bearing).

For 12 patients, tissue DNA was insufficient for the full 47-gene panel, so a smaller 20-gene Penn Precision Panel was used.

“For the remaining 12 patients, the DNA extracted from submitted tissue was of insufficient quantity for the full 47-gene panel; these samples were assessed using the Penn Precision Panel for mutations in a smaller panel of 20 commonly mutated genes (Supplemental Table 2).”

May combine two claims.

Other reasons for ordering ctDNA testing were detection of resistance mutations to targeted therapy (36 patients), identification of actionable mutations in progressive disease (12 patients), and tracking a tDNA mutation to monitor response (2 patients).

“Other reasons for ordering the ctDNA test included: detection of resistance mutations to targeted therapy (36 patients), identification of actionable mutations in patients with progressive disease (12 patients), and tracking a mutation identified in tDNA to monitor response to therapy (2 patients).”

May combine two claims.

Most patients were women (68%), had adenocarcinoma (81%), and had stage IV disease (96%).

“Most were women (68%) with adenocarcinoma (81%) and stage IV disease (96%).”

May combine two claims.

Among 8 patients with discordant EGFR T790M calls, patient #23 had tDNA and ctDNA testing 18 days apart, with EGFR L858R detected in both but T790M detected at lower frequency in tDNA and not in ctDNA.

“one patient (#23) had undergone tDNA and ctDNA testing 18 days apart, with the driver EGFR L858R mutation detected in both samples but T790M detected at a lower frequency in the tDNA but not the ctDNA.”

May combine two claims.

Eleven KRAS mutations were detected, including G12C, G12D, and G12H variants.

“Eleven KRAS mutations were detected, including the G12C, G12D, and G12H variants shown to be associated with primary resistance to EGFR inhibitors in lung cancer (38).”

May combine two claims.

For patient #30, T790M was detected in tissue but not plasma, 7 months elapsed between tests, and the patient received afatinib during that time.

“For patient #30, for whom EGFR T790M was detected in the tissue but not the plasma, 7 months had elapsed between the two tests and the patient had received afatinib during that time.”

May combine two claims.

In patients #2 and #24, no EGFR T790M mutation was detected in ctDNA at either of two timepoints, so a third-line TKI was not prescribed.

“However, no EGFR T790M mutation was detected for either timepoint or patient, so a third-line TKI was not prescribed.”

May combine two claims.

In patient #82, serial ctDNA detected emergence of EGFR T790M, and changes in EGFR G719A and T790M allele fractions correlated with imaging response and progression.

“ctDNA analysis performed at that time confirmed EGFR G719A (24.1% AF), and also detected the emergence of a T790M mutation (0.5% AF).”

• Also stated in: § Real-time serial molecular monitoring of patients on therapy · plain

  “Afatanib was initiated and repeat ctDNA testing 2 months later showed a decrease in the AF of EGFR G719A to 0.4% and EGFR T790M to 0.4%, which correlated with imaging response.”

• Also stated in: § Real-time serial molecular monitoring of patients on therapy · boosted

  “ctDNA analysis two months later showed a further increase in the AF of EGFR G719A to 23.7% and EGFR T790M to 18.1% with clear progression on PET CT, and the patient was placed on osimertinib based on the EGFR T790M mutation detected only in blood.”

May combine two claims. The quote only partly states this claim.

In patient #15, serial ctDNA identified EGFR L858R, TP53 D281N, and emergence of EGFR T790M; rising allele fractions correlated with progression, and after osimertinib all three variants became undetectable.

“ctDNA sequencing performed 7 days later identified EGFR L858R (1.4% AF), TP53 D281N (1.9% AF), and the emergence of EGFR T790M (0.2% AF).”

• Also stated in: § Real-time serial molecular monitoring of patients on therapy · plain

  “However, simultaneous ctDNA analysis noted a rising AF for all three variants: EGFR L858R 17.9%, EGFR T790M 5.7%, and TP53 D281N 31.4%.”

• Also stated in: § Real-time serial molecular monitoring of patients on therapy · plain

  “A two-month follow up CT scan exhibited an interval response to therapy, and a simultaneous ctDNA test revealed that all 3 variants were now undetectable in the patient’s plasma.”

May combine two claims.

The minimum input DNA of at least 5 ng was successfully extracted from all samples and libraries were prepared.

“The minimum input DNA (≥ 5 ng) was successfully extracted from all samples and libraries prepared.”

May combine two claims.

For 6 patients with T790M detected in plasma but not tumor, the ctDNA allelic fraction was well below the 4.0% tissue NGS LOD, and 5 had received EGFR-targeting therapy between tests.

“For the 6 patients for whom EGFR T790M was detected in the plasma but not the tumor, the ctDNA allelic fraction was well below the 4.0% level of detection for the tissue NGS panel, and 5 of the patients had received an EGFR-targeting therapy between tests, thus providing a possible explanation for the emergence of the T790M resistance mutation.”

May combine two claims.

Patient #15 had T790M detected in plasma but not tDNA, received no EGFR-targeted therapy during the 7 days between tests, and imaging showed multiple metastatic sites.

“The sixth patient, #15, in whom the variant was detected in plasma but not tDNA, received no EGFR-targeted therapy during the 7 days that elapsed between ctDNA and tDNA testing. Imaging in this patient showed multiple metastatic sites.”

May combine two claims.

Among 98 patients with metastatic disease, median overall survival from metastatic diagnosis was 46 months for those with at least 3 plasma variants versus 62 months for those with fewer than 3 variants, but the difference was not statistically significant (p=0.09).

“For the 98 patients with metastatic disease, the median overall survival from time of metastatic diagnosis in patients with ≥3 variants detected in plasma was 46 months versus 62 months for those with <3 variants, although this result did not reach statistical significance (p=0.09; Figure 4A).”

May combine two claims.

In patient #72, ctDNA revealed an EML4-ALK fusion and a TP53 variant; after crizotinib, repeat ctDNA showed decreased EML4-ALK allele fraction and undetectable TP53, with concurrent imaging decrease.

“Upon referral to our hospital, ctDNA testing revealed an EML4-ALK fusion (0.4% AF) and a TP53 variant (0. 8 % AF), and the patient was initiated on crizotinib monotherapy with significant improvement in symptoms.”

• Also stated in: § Real-time serial molecular monitoring of patients on therapy · plain

  “Two months following therapy, a repeat ctDNA test showed a decrease in the EML4-ALK AF to 0.1%, and the TP53 mutation was undetectable.”

• Also stated in: § Real-time serial molecular monitoring of patients on therapy · plain

  “A CT scan performed the same day demonstrated a decrease in size of the primary lung lesion as well as liver metastatic disease.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

In patient #6, ctDNA performed twice for initial mutation screening detected no variants, and subsequent tissue NGS confirmed the absence of detectable somatic variants.

“Initial tissue testing yielded insufficient DNA, therefore ctDNA was performed twice for initial mutation screening but no variants were detected, and the patient was put on chemotherapy. Subsequent tissue NGS confirmed the lack of detectable somatic variants.”

May combine two claims.

ctDNA NGS can allow more accurate diagnosis, improved patient management, and serial sampling to monitor disease progression and clonal evolution.

“Therapeutically targetable driver and resistance mutations can be detected by ctDNA NGS, even when tissue is unavailable, thus allowing more accurate diagnosis, improved patient management, and serial sampling to monitor disease progression and clonal evolution.”

Classified as probably not a substantial claim.

NCCN guidelines recommend repeat tissue biopsies to identify resistance mechanisms in patients whose cancers progress on first-line targeted therapeutics.

“the NCCN guidelines now recommend repeat tissue biopsies to identify resistance mechanisms in patients whose cancers progress on first-line targeted therapeutics (14).”

• cites [14] Ettinger, 2015, Non-small cell lung cancer, version 6.2015, J Natl Compr Canc Netw sourceNot checked yet

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Classified as probably not a substantial claim.

Multiple molecular resistance mechanisms to ALK-TKIs have been identified.

“Multiple molecular resistance mechanisms to ALK-TKIs have also been identified (9).”

• cites [9] Awad, 2014, ALK inhibitors in non-small cell lung cancer: crizotinib and beyond, Clin Adv Hematol OncolNot checked yet

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May combine two claims.

Primary and acquired resistance to EGFR-TKI therapy can involve MET and ERBB2 amplification, EGFR, KRAS, BRAF, and PIK3CA deletions or point mutations, and PTEN loss or inactivation.

“Mechanisms of primary and acquired resistance to EGFR-TKI therapy have been identified including amplifications in MET and ERBB2, deletions or point mutations in EGFR, KRAS, BRAF, and PIK3CA, and loss or inactivation of PTEN (10, 11).”

• cites [10] Spaans, 2014, Drug resistance to molecular targeted therapy and its consequences for treatment decisions in non-small-cell lung cancer, Front Oncol sourceNot checked yet

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• cites [11] Wheeler, 2010, Understanding resistance to EGFR inhibitors-impact on future treatment strategies, Nat Rev Clin Oncol sourceNot checked yet

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May combine two claims.

A sensitizing EGFR mutation usually predicts a high likelihood of response to first-generation TKIs, but most patients develop resistance within 12 to 24 months.

“The presence of a sensitizing EGFR mutation usually indicates a high likelihood of response to first-generation tyrosine kinase inhibitors (TKIs), but most patients will develop resistance within 12–24 months of treatment initiation (7–9).”

• cites [7] Mok, 2009, Gefitinib or carboplatin-paclitaxel in pulmonary adenocarcinoma, N Engl J Med sourceNot checked yet

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• cites [8] Sequist, 2013, Phase III study of afatinib or cisplatin plus pemetrexed in patients with metastatic lung adenocarcinoma with EGFR mutations, J Clin Oncol sourceNot checked yet

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• cites [9] Awad, 2014, ALK inhibitors in non-small cell lung cancer: crizotinib and beyond, Clin Adv Hematol OncolNot checked yet

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Probably not a substantial claim: may not pass S3 (load-bearing).

EML4-ALK fusion confers sensitivity to the ALK TKI crizotinib.

“The EML4-ALK fusion, which confers sensitivity to the ALK TKI crizotinib (1), was detected in 2 samples.”

• cites [1] Solomon, 2014, First-line crizotinib versus chemotherapy in ALK-positive lung cancer, N Engl J Med sourceNot checked yet

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Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Libraries were prepared and sequenced on an Illumina MiSeq instrument and always included a no-template negative control.

“Libraries were prepared and sequenced on an Illumina MiSeq instrument, and always included a no template negative control.”

May combine two claims.

The ctDNA assay can detect 1 to 2 mutant fragments in 10 mL of blood with analytic specificity greater than 99.9999%.

“allowing the detection of 1–2 mutant fragments in 10 mL of blood with an analytic specificity greater than 99.9999%.”

Probably not a substantial claim: may not pass S3 (load-bearing).

The study was approved by the University of Pennsylvania Institutional Review Board.

“The study was approved by the University of Pennsylvania Internal Review Board (IRB).”

Probably not a substantial claim: may not pass S3 (load-bearing).

BRAF D594G is associated with resistance to EGFR-targeted therapy.

“In addition, the BRAF D594G mutation, which has been associated with resistance to EGFR-targeted therapy (35), was detected for one patient.”

• cites [35] Stewart, 2015, Known and putative mechanisms of resistance to EGFR targeted therapies in NSCLC patients with EGFR mutations-a review, Transl Lung Cancer ResNot checked yet

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Classified as probably not a substantial claim.

Clinically significant somatic alterations were distinguished from germline polymorphisms by referencing multiple SNP databases and COSMIC.

“Clinically significant somatic alterations were distinguished from germline polymorphisms by referencing multiple SNP databases and by their annotation in COSMIC (Catalogue of Somatic Mutations in Cancer; http://cancer.sanger.ac.uk/cosmic).”

Probably not a substantial claim: may not pass S3 (load-bearing).

Tissue samples were processed at the CAP/CLIA-certified University of Pennsylvania Center for Personalized Diagnostics clinical laboratory.

“All tissue samples were processed at the CAP/CLIA-certified University of Pennsylvania Center for Personalized Diagnostics clinical laboratory.”

May combine two claims.

Digital PCR assays for ctDNA have high sensitivity and specificity but are typically limited to hotspot mutations in a few genes and cannot interrogate the full spectrum of acquired resistance mutations.

“Digital polymerase chain reaction assays have been developed with high sensitivity and specificity for detection of variants in ctDNA (24, 27), however, these approaches are typically limited to hotspot mutations in a few genes, and cannot interrogate the full spectrum of mutations that may emerge in the setting of acquired resistance during targeted therapy (28).”

• cites [24] Oxnard, 2014, Noninvasive detection of response and resistance in EGFR-mutant lung cancer using quantitative next-generation genotyping of cell-free plasma DNA, Clin Cancer Res sourceNot checked yet

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• cites [27] Bettegowda, 2014, Detection of circulating tumor DNA in early- and late-stage human malignancies, Sci Transl Med sourceNot checked yet

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• cites [28] Drilon, 2015, Broad, hybrid capture-based next-generation sequencing identifies actionable genomic alterations in lung adenocarcinomas otherwise negative for such alterations by other genomic testing approaches, Clin Cancer Res sourceNot checked yet

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May combine two claims.

For patient #23, because L858R was detected at 0.3% AF in ctDNA and EGFR resistance mutations are typically at lower frequency in blood than the original driver mutation, T790M may have been present but below the plasma LOD.

“Since the L858R mutation was detected at 0.3% AF in the ctDNA, and EGFR resistance mutations are typically seen at a lower frequency in the blood compared to the originally detected EGFR driver mutations (24), it is possible that the T790M resistance mutation was present but below the LOD in the plasma.”

• cites [24] Oxnard, 2014, Noninvasive detection of response and resistance in EGFR-mutant lung cancer using quantitative next-generation genotyping of cell-free plasma DNA, Clin Cancer Res sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

The level of detection for the tissue NGS panel is 4.0%.

“The level of detection for the tissue NGS panel is 4.0%.”

May combine two claims.

A clinically validated bioinformatics pipeline was used for tumor DNA sequencing analysis, with reads mapped to hg19.

“A clinically validated bioinformatics pipeline was used for tumor DNA (tDNA) sequencing analysis (33), with reads mapped to the hg19 genome build.”

• cites [33] Daber, 2013, Understanding the limitations of next generation sequencing informatics, an approach to clinical pipeline validation using artificial data sets, Cancer Genet sourceNot checked yet

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Classified as probably not a substantial claim.

ctDNA sequencing was performed at a CLIA-certified, CAP-accredited laboratory.

“shipped overnight at ambient temperature to Guardant Health, Inc, a CLIA-certified, CAP-accredited laboratory facility.”

Probably not a substantial claim: may not pass S3 (load-bearing).

The average coverage depth of the ctDNA panel was 10,000X.

“The average coverage depth was 10,000X.”

Classified as probably not a substantial claim.

EGFR resistance mutations are typically seen at lower frequency in blood than the originally detected EGFR driver mutations.

“and EGFR resistance mutations are typically seen at a lower frequency in the blood compared to the originally detected EGFR driver mutations (24)”

• cites [24] Oxnard, 2014, Noninvasive detection of response and resistance in EGFR-mutant lung cancer using quantitative next-generation genotyping of cell-free plasma DNA, Clin Cancer Res sourceNot checked yet

  Not checked in the free run.

May combine two claims.

ctDNA can be readily detected in advanced NSCLC patients, and sensitive genotyping assays can detect mutations in ctDNA.

“ctDNA can be readily detected in advanced NSCLC patients, and several studies have shown that highly sensitive genotyping assays can detect mutations in ctDNA (20–26).”

• cites [20] Newman, 2014, An ultrasensitive method for quantitating circulating tumor DNA with broad patient coverage, Nat Med sourceNot checked yet

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• cites [21] Diaz, 2012, The molecular evolution of acquired resistance to targeted EGFR blockade in colorectal cancers, Nature sourceNot checked yet

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• cites [22] Dawson, 2013, Analysis of circulating tumor DNA to monitor metastatic breast cancer, N Engl J Med sourceNot checked yet

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• cites [23] Couraud, 2014, Noninvasive diagnosis of actionable mutations by deep sequencing of circulating free DNA in lung cancer from never-smokers: a proof-of-concept study from BioCAST/IFCT-1002, Clin Cancer Res sourceNot checked yet

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• cites [24] Oxnard, 2014, Noninvasive detection of response and resistance in EGFR-mutant lung cancer using quantitative next-generation genotyping of cell-free plasma DNA, Clin Cancer Res sourceNot checked yet

  Not checked in the free run.

• cites [25] Punnoose, 2012, Evaluation of circulating tumor cells and circulating tumor DNA in non-small cell lung cancer: association with clinical endpoints in a phase II clinical trial of pertuzumab and erlotinib, Clin Cancer Res sourceNot checked yet

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• cites [26] Kuang, 2009, Noninvasive detection of EGFR T790M in gefitinib or erlotinib resistant non-small cell lung cancer, Clin Cancer Res sourceNot checked yet

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Probably not a substantial claim: may not pass S3 (load-bearing).

KRAS G12C, G12D, and G12H variants are associated with primary resistance to EGFR inhibitors in lung cancer.

“Eleven KRAS mutations were detected, including the G12C, G12D, and G12H variants shown to be associated with primary resistance to EGFR inhibitors in lung cancer (38).”

• cites [38] Pao, 2005, KRAS mutations and primary resistance of lung adenocarcinomas to gefitinib or erlotinib, PLoS Med sourceNot checked yet

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May combine two claims.

Monitoring response to targeted therapy by measuring changes in the targeted mutation or identifying resistance mutations cannot be consistently achieved through repeat biopsy.

“However, monitoring response to targeted therapy by assaying changes in the frequency of the targeted mutation, or identifying resistance mutations cannot be consistently achieved through repeat biopsy.”

Classified as probably not a substantial claim.

The growing number of targeted therapies for lung cancer has created a need for companion diagnostics that detect therapeutically targetable genetic lesions in real time.

“The ever-expanding number of targeted therapies available for lung cancer patient treatment has been accompanied by a need for companion diagnostics for real-time detection of therapeutically targetable genetic lesions (44, 45).”

• cites [44] Neal, 2015, Developing biomarker-specific end points in lung cancer clinical trials, Nat Rev Clin Oncol sourceNot checked yet

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• cites [45] Carpenter, 2012, Targeting ALK in neuroblastoma—preclinical and clinical advancements, Nat Rev Clin Oncol sourceNot checked yet

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Probably not a substantial claim: may not pass S3 (load-bearing).

If T790M was present only in some metastases but not the biopsied site, tumor heterogeneity might explain the discordant result for patient #15.

“If the T790M mutation had been present only in some metastases, but not the site that was biopsied, this tumor heterogeneity might provide an explanation for the discordant result.”

Probably not a substantial claim: may not pass S4 (based).

ctDNA testing alone will be insufficient to detect histological sources of therapy resistance, such as a small cell phenotype.

“While ctDNA testing alone will be insufficient to detect histological sources of therapy resistance, such as a small cell phenotype, a liquid biopsy may complement histological analysis by providing additional tumor molecular characterization.”

Probably not a substantial claim: may not pass S3 (load-bearing).

ddPCR is typically used to detect hotspots in 3 to 5 genes.

“While the results we have reported are based on a panel of up to 70 clinically relevant genes, including 30 with full exon coverage, ddPCR is typically used to detect hotspots in 3 to 5 genes.”

Classified as probably not a substantial claim.

Molecular heterogeneity has been associated with poor patient outcome.

“Because molecular heterogeneity has also been associated with poor patient outcome (42, 43), we next sought to assess the prognostic significance of ctDNA molecular heterogeneity.”

• cites [42] Mroz, 2013, High intratumor genetic heterogeneity is related to worse outcome in patients with head and neck squamous cell carcinoma, Cancer sourceNot checked yet

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• cites [43] Mahrooghy, 2015, Pharmacokinetic tumor heterogeneity as a prognostic biomarker for classifying breast cancer recurrence risk, IEEE Trans Biomed Eng sourceNot checked yet

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May combine two claims.

The reported results are based on a panel of up to 70 clinically relevant genes, including 30 with full exon coverage.

“While the results we have reported are based on a panel of up to 70 clinically relevant genes, including 30 with full exon coverage, ddPCR is typically used to detect hotspots in 3 to 5 genes.”

Classified as probably not a substantial claim. May combine two claims.

Plasma-based ctDNA testing is non-invasive and offers the advantage of testing without the risks of invasive biopsies.

“Plasma-based ctDNA testing is a non-invasive means of patient monitoring and thus offers the advantage of testing without the risks associated with invasive biopsies.”

May combine two claims.

In NSCLC patients, liquid biopsies can identify resistance mutations that allow treatment with second- and third-line TKIs, or cytotoxic chemotherapy when no targetable mutation is identified.

“When applied to the blood of NSCLC patients, liquid biopsies can identify resistance mutations that allow for the treatment of patients with second- and third-line TKIs, or cytotoxic chemotherapy when no targetable mutation is identified.”

May combine two claims.

Possible explanations for detecting no mutations in cfDNA include the tumor not actively shedding ctDNA, disease being adequately controlled by therapy, or variants not being covered by the 70-gene panel or being below the assay's 0.01% detection level.

“No mutations were detected in the cfDNA for 16 of 102 patients, suggesting the tumor was not actively shedding ctDNA, the patient’s disease was adequately controlled by therapy, or plasma-based somatic variants were either not covered by the 70-gene panel or below the 0.01% level of detection for the assay.”

Probably not a substantial claim: may not pass S2 (checkable).

As resistance mechanisms and approved therapies increase, NGS may become more clinically useful than ddPCR for identifying actionable targets.

“As the number of molecular mechanisms of resistance and associated approved therapies increases, however, NGS may emerge as the more clinically useful assay for identification of actionable targets.”

May combine two claims.

The study demonstrates the feasibility and utility of plasma testing for a large subset of patients in whom matched tissue sequencing was not feasible.

“and demonstrate the feasibility and utility of plasma testing for a large subset of patients in whom matched tissue sequencing was not feasible.”

Probably not a substantial claim: may not pass S3 (load-bearing).

The EGFR T790M mutation would be detected by many current ddPCR platforms.

“The EGFR T790M mutation accounts for more than 50% of cases of acquired resistance to EGFR-TKI therapy and would certainly be detected by many current ddPCR platforms (35).”

• cites [35] Stewart, 2015, Known and putative mechanisms of resistance to EGFR targeted therapies in NSCLC patients with EGFR mutations-a review, Transl Lung Cancer ResNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S3 (load-bearing).

The EGFR T790M mutation accounts for more than 50% of cases of acquired resistance to EGFR-TKI therapy.

“The EGFR T790M mutation accounts for more than 50% of cases of acquired resistance to EGFR-TKI therapy and would certainly be detected by many current ddPCR platforms (35).”

• cites [35] Stewart, 2015, Known and putative mechanisms of resistance to EGFR targeted therapies in NSCLC patients with EGFR mutations-a review, Transl Lung Cancer ResNot checked yet

  Not checked in the free run.