Journal: Cell reports

Article Title: EGFR-phosphorylated GDH1 harmonizes with RSK2 to drive CREB activation and tumor metastasis in EGFR-activated lung cancer

doi: 10.1016/j.celrep.2022.111827

Figure Lengend Snippet:

Article Snippet: Plasmid: pBabe-puro-EGFR WT , Greulich et al. , Addgene plasmid: EGFR WT; Cat#11011.

Techniques: Microarray, Recombinant, Purification, Membrane, SYBR Green Assay, Viability Assay, Phospho-proteomics, Kinase Assay, Reverse Transcription, Transcription Factor Assay, Enzyme-linked Immunosorbent Assay, Multiplex Assay, Extraction, Isolation, shRNA, Sequencing, Plasmid Preparation, Software

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Acquired Resistance Mechanism of EGFR Kinase Domain Duplication to EGFR TKIs in Non–Small Cell Lung Cancer

doi: 10.4143/crt.2021.385

Figure Lengend Snippet: Clinical history and genomic features for a patient with EGFR KDD. (A) Clinical history of a patient with EGFR -KDD. Numbers beneath the line represent months after diagnosis. Patient’s computerized tomography images indicated tumor masses pre-erlotinib treatment (first), post-erlotinib with partial response (second), post-erlotinib with progressive disease (third), post-osimertinib with partial response (fourth). A patient-derived cell line SNU-4784 was established with pleural effusion upon emerging erlotinib resistance, but without the EGFR T790M mutation. (B) EGFR -KDD breakpoint in a patient cDNA from pleural effusion. (C) The EGFR T790M mutation confirmed by droplet digital polymerase chain reaction (orange dots, blank; green dots, wild type; blue dots, T790M; red dots, T790M plus WT). EGFR -KDD, epidermal growth factor receptor kinase domain duplication.

Article Snippet: The pBabe EGFR T790M was purchased from Addgene (Addgene, Watertown, MA; Addgene plasmid #32070).

Techniques: Biomarker Discovery, Tomography, Derivative Assay, Mutagenesis, Digital PCR

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Acquired Resistance Mechanism of EGFR Kinase Domain Duplication to EGFR TKIs in Non–Small Cell Lung Cancer

doi: 10.4143/crt.2021.385

Figure Lengend Snippet: Characteristics of the EGFR -KDD T790M Ba/F3 cell lines. (A) Cell viability assays in EGFR -KDD WT and EGFR -KDD T790M Ba/F3 cell lines. Cells were exposed to EGFR TKIs for 72 hours. Graphs represent mean±SD values. (B) Immunoblot assays of EGFR -KDD WT and EGFR -KDD T790M Ba/F3 cell lines. Cells were exposed to erlotinib, afatinib, or osimertinib for 4 hours. Cell viability assays and immunoblot assays were independently repeated three times. EGFR -KDD, epidermal growth factor receptor kinase domain duplication; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: The pBabe EGFR T790M was purchased from Addgene (Addgene, Watertown, MA; Addgene plasmid #32070).

Techniques: Western Blot

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Acquired Resistance Mechanism of EGFR Kinase Domain Duplication to EGFR TKIs in Non–Small Cell Lung Cancer

doi: 10.4143/crt.2021.385

Figure Lengend Snippet: Clinical history and genomic features for a patient with EGFR KDD. (A) Clinical history of a patient with EGFR -KDD. Numbers beneath the line represent months after diagnosis. Patient’s computerized tomography images indicated tumor masses pre-erlotinib treatment (first), post-erlotinib with partial response (second), post-erlotinib with progressive disease (third), post-osimertinib with partial response (fourth). A patient-derived cell line SNU-4784 was established with pleural effusion upon emerging erlotinib resistance, but without the EGFR T790M mutation. (B) EGFR -KDD breakpoint in a patient cDNA from pleural effusion. (C) The EGFR T790M mutation confirmed by droplet digital polymerase chain reaction (orange dots, blank; green dots, wild type; blue dots, T790M; red dots, T790M plus WT). EGFR -KDD, epidermal growth factor receptor kinase domain duplication.

Article Snippet: The pBabe EGFR WT was kindly provided by Matthew Meyerson (Dana-Farber Cancer Research Institute, Boston, MA; Addgene plasmids #11011).

Techniques: Biomarker Discovery, Tomography, Derivative Assay, Mutagenesis, Digital PCR

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Acquired Resistance Mechanism of EGFR Kinase Domain Duplication to EGFR TKIs in Non–Small Cell Lung Cancer

doi: 10.4143/crt.2021.385

Figure Lengend Snippet: Characteristics of the patient-derived EGFR -KDD cell line SNU-4784. (A) Cell viability assay in SNU-4784 cell line. Cells were exposed to erlotinib, afatinib, and osimertinib for 72 hours. (B) Immunoblot assay of SNU-4784 cell line. Cells were exposed to erlotinib, afatinib, or osimertinib for 4 hours. Cell viability assay and the immunoblot assay were independently repeated three times. EGFR -KDD, epidermal growth factor receptor kinase domain duplication; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: The pBabe EGFR WT was kindly provided by Matthew Meyerson (Dana-Farber Cancer Research Institute, Boston, MA; Addgene plasmids #11011).

Techniques: Derivative Assay, Viability Assay, Western Blot

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Acquired Resistance Mechanism of EGFR Kinase Domain Duplication to EGFR TKIs in Non–Small Cell Lung Cancer

doi: 10.4143/crt.2021.385

Figure Lengend Snippet: Characteristics of the EGFR -KDD T790M Ba/F3 cell lines. (A) Cell viability assays in EGFR -KDD WT and EGFR -KDD T790M Ba/F3 cell lines. Cells were exposed to EGFR TKIs for 72 hours. Graphs represent mean±SD values. (B) Immunoblot assays of EGFR -KDD WT and EGFR -KDD T790M Ba/F3 cell lines. Cells were exposed to erlotinib, afatinib, or osimertinib for 4 hours. Cell viability assays and immunoblot assays were independently repeated three times. EGFR -KDD, epidermal growth factor receptor kinase domain duplication; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: The pBabe EGFR WT was kindly provided by Matthew Meyerson (Dana-Farber Cancer Research Institute, Boston, MA; Addgene plasmids #11011).

Techniques: Western Blot

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Acquired Resistance Mechanism of EGFR Kinase Domain Duplication to EGFR TKIs in Non–Small Cell Lung Cancer

doi: 10.4143/crt.2021.385

Figure Lengend Snippet: ENU mutagenesis screening to identify the potential resistance mechanism underlying the EGFR -KDD BDT mutation. (A) Graphical scheme of ENU mutagenesis screening. EGFR -KDD BDT Ba/F3 cells were exposed to 50 μg/mL ENU and selected with 2 μM osimertinib. (B) Sanger sequencing of osimertinib-resistant EGFR -KDD BDT Ba/F3 cells. EGFR C797S mutation in kinase domain 2. EGFR -KDD, epidermal growth factor receptor kinase domain duplication; ENU, N-ethyl-N-nitrosourea.

Article Snippet: The pBabe EGFR WT was kindly provided by Matthew Meyerson (Dana-Farber Cancer Research Institute, Boston, MA; Addgene plasmids #11011).

Techniques: Mutagenesis, Sequencing

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Acquired Resistance Mechanism of EGFR Kinase Domain Duplication to EGFR TKIs in Non–Small Cell Lung Cancer

doi: 10.4143/crt.2021.385

Figure Lengend Snippet: Characteristics of the EGFR -KDD T/T+C Ba/F3 cell line. (A) Cell viability assay of the EGFR -KDD T/T+C Ba/F3 cell line. Cells were exposed to EGFR TKIs for 72 hours. (B) Immunoblot assays of the EGFR -KDD T/T+C Ba/F3 cell line. Cells were exposed to EGFR TKIs for 4 hours. (C) Cell viability assay in the EGFR -KDD T/T+C Ba/F3 cell line. Cells were exposed to EGFR TKIs and cetuximab for 72 hours. For combination treatment, 10 μg/mL cetuximab was added. (D) Growth curve of the EGFR -KDD Ba/F3 cell lines. Cells were grown in interleukin-3 free media and the Ba/F3 parental cells were used as controls. EGFR -KDD, epidermal growth factor receptor kinase domain duplication; TKI, tyrosine kinase inhibitor.

Article Snippet: The pBabe EGFR WT was kindly provided by Matthew Meyerson (Dana-Farber Cancer Research Institute, Boston, MA; Addgene plasmids #11011).

Techniques: Viability Assay, Western Blot