Presentation is loading. Please wait.

Presentation is loading. Please wait.

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

Published byJohnathan Carter Modified over 6 years ago

Similar presentations

Presentation on theme: "A Noninvasive System for Monitoring Resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors with Plasma DNA  Tomomi Nakamura, MD, Naoko."— Presentation transcript:

1 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 (October 2011) DOI: /JTO.0b013e e8 Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

2 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, DOI: ( /JTO.0b013e e8) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

3 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, DOI: ( /JTO.0b013e e8) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

4 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, DOI: ( /JTO.0b013e e8) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

5 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, DOI: ( /JTO.0b013e e8) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

6 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, DOI: ( /JTO.0b013e e8) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

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

Similar presentations

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

Shyamala Maherswaran, Ph.D. et al. Sarah Gomez and Rachael Holmes Detection of Mutations in EGFR in Circulating Lung-Cancer Cells.
Frances A Shepherd, MD, Rafael Rosell, MD  Journal of Thoracic Oncology 

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,

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,

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.

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,

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.

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,

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 

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.

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.

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,

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.

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 

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.

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.

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.

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,

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 

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.

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

Similar presentations

Ads by Google