A Noninvasive System for Monitoring Resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors with Plasma DNA Tomomi Nakamura, MD, Naoko.

Slides:

Advertisements

Similar presentations

Shyamala Maherswaran, Ph.D. et al. Sarah Gomez and Rachael Holmes Detection of Mutations in EGFR in Circulating Lung-Cancer Cells.

Advertisements

Frances A Shepherd, MD, Rafael Rosell, MD Journal of Thoracic Oncology

Prognostic impacts of EGFR mutation status and subtype in patients with surgically resected lung adenocarcinoma Kazuya Takamochi, MD, Shiaki Oh, MD,

Application of a Highly Sensitive Detection System for Epidermal Growth Factor Receptor Mutations in Plasma DNA Tomomi Nakamura, MD, Naoko Sueoka-Aragane,

Comparison of Clinical Outcomes Following Gefitinib and Erlotinib Treatment in Non– Small-Cell Lung Cancer Patients Harboring an Epidermal Growth Factor.

Phase II Study of Gefitinib, an Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor (EGFR-TKI), and Celecoxib, a Cyclooxygenase-2 (COX-2) Inhibitor,

Epidermal Growth Factor Receptor-Related Tumor Markers and Clinical Outcomes with Erlotinib in Non-small Cell Lung Cancer: An Analysis of Patients from.

Application of a Highly Sensitive Detection System for Epidermal Growth Factor Receptor Mutations in Plasma DNA Tomomi Nakamura, MD, Naoko Sueoka-Aragane,

Rapid Detection of the Epidermal Growth Factor Receptor Mutation in Non-Small-Cell Lung Cancer for Analysis of Acquired Resistance Using Molecular Beacons

EGFR Array: Uses in the Detection of Plasma EGFR Mutations in Non–Small Cell Lung Cancer Patients Irene Yam, BSc, David Chi-Leung Lam, MBBS, PhD, Kaimin.

Rapid detection of resistance in Mycobacterium tuberculosis: a review discussing molecular approaches D. García de Viedma Clinical Microbiology and.

T790M EGFR Mutation Detection in Cerebrospinal Fluid and Response to Osimertinib in a Lung Cancer Patient with Meningeal Carcinomatosis Hugo Gortais,

Lung Adenocarcinoma Harboring EGFR T790M and In Trans C797S Responds to Combination Therapy of First- and Third-Generation EGFR TKIs and Shifts Allelic.

Detection of EGFR Mutation Status in Lung Adenocarcinoma Specimens with Different Proportions of Tumor Cells Using Two Methods of Differential Sensitivity

Negative Thyroid Transcription Factor 1 Expression Defines an Unfavorable Subgroup of Lung Adenocarcinomas Yiliang Zhang, MD, Rui Wang, MD, PhD, Yuan.

The CTC-Chip: An Exciting New Tool to Detect Circulating Tumor Cells in Lung Cancer Patients Lecia V. Sequist, MD, MPH, Sunitha Nagrath, PhD, Mehmet.

Dramatic Response to Combination Erlotinib and Crizotinib in a Patient with Advanced, EGFR-Mutant Lung Cancer Harboring De Novo MET Amplification Justin.

Detection of Discrepant Driver Mutations in a Patient with Two Synchronous Primary Non–Small Cell Lung Cancers (NSCLCs) with Liquid Biopsy Sotirios Lakis,

Durable Response to Tyrosine Kinase Inhibitor Therapy in a Lung Cancer Patient Harboring Epidermal Growth Factor Receptor Tandem Kinase Domain Duplication

Acquired BRAF V600E Mutation as Resistant Mechanism after Treatment with Third- Generation EGFR Tyrosine Kinase Inhibitor Alessandra Bearz, MD, Elisa.

T790M EGFR Mutation Detection in Cerebrospinal Fluid and Response to Osimertinib in a Lung Cancer Patient with Meningeal Carcinomatosis Hugo Gortais,

Miliary Never-Smoking Adenocarcinoma of the Lung: Strong Association with Epidermal Growth Factor Receptor Exon 19 Deletion Eckart Laack, MD, Ronald.

Higher Dosage of the Epidermal Growth Factor Receptor Mutant Allele in Lung Adenocarcinoma Correlates with Younger Age, Stage IV at Presentation, and.

Long-Lasting Complete Remission with Tyrosine Kinase Inhibitor in Bronchioloalveolar Carcinoma with a so far Unknown EGFR Mutation Vienna Ludovini, PhD,

Geoffrey R. Oxnard, MD, Vincent A. Miller, MD, Mark E

Circulating Tumor DNA Identifies EGFR Coamplification as a Mechanism of Resistance to Crizotinib in a Patient with Advanced MET-Amplified Lung Adenocarcinoma

Small Cell Transformation and T790M Mutation as Coresistance Mechanisms for First- line Epidermal Growth Factor Receptor (EGFR) Tyrosine Kinase Inhibitor.

Combined Use of ALK Immunohistochemistry and FISH for Optimal Detection of ALK- Rearranged Lung Adenocarcinomas Lynette M. Sholl, MD, Stanislawa Weremowicz,

Tumor Cell Content for Selection of Molecular Techniques for T790M EGFR Mutation Detection in Non-small Cell Lung Cancer Nathalie Prim, Elisabeth Quoix,

EGFR Exon 19 Insertions Show Good Response to Gefitinib, but Short Time to Progression in Japanese Patients Jangchul Park, MD, Chiaki Kondo, MD, Junichi.

A Phase 1/2 Trial of Ruxolitinib and Erlotinib in Patients with EGFR-Mutant Lung Adenocarcinomas with Acquired Resistance to Erlotinib Helena A. Yu,

Matthew D. Hellmann, MD, Boris Reva, PhD, Helena Yu, MD, Valerie W

Detection rate for EGFR mutations in cfDNA.

Highly Sensitive Droplet Digital PCR Method for Detection of EGFR-Activating Mutations in Plasma Cell–Free DNA from Patients with Advanced Non–Small Cell.

Effusion Immunocytochemistry as an Alternative Approach for the Selection of First-Line Targeted Therapy in Advanced Lung Adenocarcinoma Tzu-Hsiu Tsai,

Meta-Analysis of First-Line Therapies in Advanced Non–Small-Cell Lung Cancer Harboring EGFR-Activating Mutations Benjamin Haaland, PhD, Pui San Tan,

Clinical courses of patients.

Strategies to Improve Outcomes of Patients with EGFR-Mutant Non–Small Cell Lung Cancer: Review of the Literature Caicun Zhou, MD, PhD, Luan Di Yao, MD

Impact on Disease-Free Survival of Adjuvant Erlotinib or Gefitinib in Patients with Resected Lung Adenocarcinomas that Harbor EGFR Mutations Yelena Y.

Detection rate of EGFR mutations in cfDNA by characteristics

Coexistence of Tyrosine Kinase Inhibitor-Sensitizing and Resistant EGFR Mutations in an Untreated Lung Adenocarcinoma Patient and Response to Erlotinib

V843I, a Lung Cancer Predisposing EGFR Mutation, Is Responsible for Resistance to EGFR Tyrosine Kinase Inhibitors Satsuki Matsushima, BSc, Kouki Ohtsuka,

Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors for Non–Small-Cell Lung Cancer Patients with Leptomeningeal Carcinomatosis Bin-Chi Liao,

Somatic Mutations of the Tyrosine Kinase Domain of Epidermal Growth Factor Receptor and Tyrosine Kinase Inhibitor Response to TKIs in Non-small Cell Lung.

Erratum Journal of Thoracic Oncology

EGFR Mutations Detected in Plasma Are Associated with Patient Outcomes in Erlotinib Plus Docetaxel-Treated Non-small Cell Lung Cancer Philip C. Mack,

Combination Treatment Using an EGFR Tyrosine Kinase Inhibitor plus Platinum-Based Chemotherapy for a Patient with Transformed Small Cell Carcinoma and.

Comparison of Clinical Outcomes Following Gefitinib and Erlotinib Treatment in Non– Small-Cell Lung Cancer Patients Harboring an Epidermal Growth Factor.

First Case of A859T Epidermal Growth Factor Receptor Mutation Responding to Erlotinib Laureline Le Maignan, BM, Delphine Mirebeau-Prunier, MD, PhD, Laurent.

Chien-Wei Chen, MD, Chin-Wei Kuo, MD Journal of Thoracic Oncology

Clinicopathologic Characteristics and Outcomes of Patients with Anaplastic Lymphoma Kinase-Positive Advanced Pulmonary Adenocarcinoma: Suggestion for.

Aglaya G. Iyevleva, MD, PhD Journal of Thoracic Oncology

A Noninvasive System for Monitoring Resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors with Plasma DNA Tomomi Nakamura, MD, Naoko.

XL647—A Multitargeted Tyrosine Kinase Inhibitor: Results of a Phase II Study in Subjects with Non-small Cell Lung Cancer Who Have Progressed after Responding.

EGFR delE709_T710insD: A Rare but Potentially EGFR Inhibitor Responsive Mutation in Non–Small-Cell Lung Cancer Allison Ackerman, MD, Michael A. Goldstein,

Daniel B. Costa, MD, PhD, Susumu Kobayashi, MD, PhD

Rapid Polymerase Chain Reaction-Based Detection of Epidermal Growth Factor Receptor Gene Mutations in Lung Adenocarcinomas Qiulu Pan, William Pao, Marc.

Chemotherapy Response in East Asian Non-small Cell Lung Cancer Patients Harboring Wild-Type or Activating Mutation of Epidermal Growth Factor Receptors

Driven by Mutations: The Predictive Value of Mutation Subtype in EGFR-Mutated Non– Small Cell Lung Cancer Emily Castellanos, MD, Emily Feld, MD, Leora.

Rapid Detection of Hotspot Mutations in Epidermal Growth Factor Receptor by Polymerase Chain Reaction Facilitates the Management of Non-small Cell Lung.

Highly Sensitive Detection of EGFR T790M Mutation in Pre-TKI Specimens of EGFR- Mutated NSCLC: In Cis, In Trans, or a Different Clone? Alvaro Leone, BSc.D

Daniel B. Costa, MD, PhD, Susan E. Jorge, PhD, Jason P

Activation of MET by Gene Amplification or by Splice Mutations Deleting the Juxtamembrane Domain in Primary Resected Lung Cancers Ryoichi Onozato, MD,

EGFR Mutations in Squamous Cell Lung Cancer in Never-Smokers

MET Exon 14 Alterations and New Resistance Mutations to Tyrosine Kinase Inhibitors: Risk of Inadequate Detection with Current Amplicon-Based NGS Panels

Good Response to Gefitinib in Lung Adenocarcinoma Harboring Coexisting EML4-ALK Fusion Gene and EGFR Mutation Yao-Wen Kuo, MD, Shang-Gin Wu, MD, Chao-Chi.

Waxing and Waning of MET Amplification in EGFR-Mutated NSCLC in Response to the Presence and Absence of Erlotinib Selection Pressure Joel P. Womack,

Circulating Tumor DNA Identifies EGFR Coamplification as a Mechanism of Resistance to Crizotinib in a Patient with Advanced MET-Amplified Lung Adenocarcinoma

Patterns of clinical relapse and algorithm for the therapeutic strategy when AR to EGFR TKI occurs in patients with EGFR-mutant NSCLC. *After discussion.

Ana Luque, MD, Edurne Arriola, MD, PhD Journal of Thoracic Oncology

Presentation transcript:

A Noninvasive System for Monitoring Resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors with Plasma DNA Tomomi Nakamura, MD, Naoko Sueoka-Aragane, MD, PhD, Kentaro Iwanaga, MD, PhD, Akemi Sato, PhD, Kazutoshi Komiya, MD, PhD, Tomonori Abe, MD, Norio Ureshino, MD, Shinichiro Hayashi, MD, PhD, Toshiya Hosomi, PhD, Mitsuharu Hirai, PhD, Eisaburo Sueoka, MD, PhD, Shinya Kimura, MD, PhD Journal of Thoracic Oncology Volume 6, Issue 10, Pages 1639-1648 (October 2011) DOI: 10.1097/JTO.0b013e31822956e8 Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 1 (A) Principles of the MBP-QP method. The MBP-QP method is a modified QP-system using i-densy and contains two steps, mutation-biased PCR (MBP) and a quenching probe (QP)-system. In MBP, different primer lengths for wild-type and mutant sequences, and the addition of five bases of random sequence to the reverse primers, leads to a higher efficiency of amplification of the mutant sequence relative to that of the wild type. Presence of the T790M sequence is determined by the QP-system, in which the fluorescence intensity of a TAMRA-conjugated-specific guanine quench fluorophore probe complementary to T790M is monitored. When a QProbe is hybridized to target DNA, its fluorescence is quenched by the guanine in the target, which is complementary to the modified cytosine. After PCR, the temperature is reduced to 40°C and then gradually increased. To identify the wild-type and mutant amplicons, the fluorescence intensity at the different temperatures is measured. When the QProbe dissociates from the amplicon, wild type at 59°C and mutant at 66°C, the fluorescence intensity increases. (B) The results of the MBP-QP method (left) and PCR-based direct sequencing (right) of genomic DNA isolated from H1975 cells are shown. Journal of Thoracic Oncology 2011 6, 1639-1648DOI: (10.1097/JTO.0b013e31822956e8) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 2 Detection of T790M mutation with plasma DNA using the MBP-QP method. (A) The device for the MBP-QP method. (B) The equipment used for the MBP-QP method. Four microliters of purified DNA is applied to PCR and detection tube. Reagents, primers, and the specific probes are mixed with the template DNA and reaction is started. The entire process is performed automatically. MBP-QP, mutation-biased PCR quenching probe. Journal of Thoracic Oncology 2011 6, 1639-1648DOI: (10.1097/JTO.0b013e31822956e8) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 3 Results of the MBP-QP method using plasma DNA of patients with lung adenocarcinoma with tumors that developed resistance to EGFR-TKIs. MBP-QP, mutation-biased PCR quenching probe; EGFR, epidermal growth factor receptor; TKI, tyrosine kinase inhibitor. Journal of Thoracic Oncology 2011 6, 1639-1648DOI: (10.1097/JTO.0b013e31822956e8) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 4 Representative results of serial analysis of patients 1 (A), 2 (B), 5 (C), and 3 (D) using the MBP-QP method to detect the T790M EGFR mutation in plasma DNA (plasma) or primary tumor (primary) of lung adenocarcinomas from patients whose tumors acquired resistance to EGFR-TKIs. Patient numbers are the same as in Table 4. Black and red arrows indicate the peak for T790M mutation and occurrence of PD, respectively. EGFR, epidermal growth factor receptor; TKI, tyrosine kinase inhibitor; chemo, chemotherapy; G, gefitinib, E, erlotinib, CEA, carcinoembryonic antigen; PD, progressive disease. Journal of Thoracic Oncology 2011 6, 1639-1648DOI: (10.1097/JTO.0b013e31822956e8) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 5 The results of direct sequencing using the products of the MBP-QP method. MBP-QP, mutation-biased PCR quenching probe. Journal of Thoracic Oncology 2011 6, 1639-1648DOI: (10.1097/JTO.0b013e31822956e8) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions