Journal: Clinical Cancer Research

Article Title: Optimized EGFR Blockade Strategies in EGFR Addicted Gastroesophageal Adenocarcinomas

doi: 10.1158/1078-0432.ccr-20-0121

Figure Lengend Snippet: Figure 1. EGFR CNG. The graphs illustrate the percentage of tumors displaying EGFR CNG in four different cohorts. Real-time PCR analysis of IRCC gastric/gastroesophageal junction (GEJ) adenocarcinomas displaying EGFR gain (4–8 copies or >8 copies)or heterogeneity (significantly different EGFR CNG in diverse analyzed samples from the same tumor, with one tumor sample displaying >8 copies and one tumor sample having≤8 copies).CGP of FMI gastricand esophageal/gastroesophageal junction cases, FISH analysis of COG esophageal/gastroesophageal junction cases, and SISH analysis of INT gastric/gastroesophageal junction adenocarcinomas.

Article Snippet: Primary antibodies, anti-EGFR (1005: sc-03) and anti-Actin, were from Santa Cruz Biotechnology, and antibodies against phosphorylated EGFR (Tyr 845), ERK (Thr202/Tyr204), phosphorylated AKT (Ser473) (Clone D9E), total AKT, and ERK were from Cell Signaling Technology.

Techniques: Real-time Polymerase Chain Reaction

Journal: Clinical Cancer Research

Article Title: Optimized EGFR Blockade Strategies in EGFR Addicted Gastroesophageal Adenocarcinomas

doi: 10.1158/1078-0432.ccr-20-0121

Figure Lengend Snippet: Figure 2. Survival analysis of patients with EGFR CNG. A, The graphs show the cumulative survival (cum survival) of patients of the COG (left) and INT (right) cohorts related to EGFR CNG. B, The graphs show the OS (left) and the disease-free survival (right) of patients of the gastroesophageal TCGA dataset, related to EGFR CNG.

Article Snippet: Primary antibodies, anti-EGFR (1005: sc-03) and anti-Actin, were from Santa Cruz Biotechnology, and antibodies against phosphorylated EGFR (Tyr 845), ERK (Thr202/Tyr204), phosphorylated AKT (Ser473) (Clone D9E), total AKT, and ERK were from Cell Signaling Technology.

Techniques:

Journal: Clinical Cancer Research

Article Title: Optimized EGFR Blockade Strategies in EGFR Addicted Gastroesophageal Adenocarcinomas

doi: 10.1158/1078-0432.ccr-20-0121

Figure Lengend Snippet: Figure 3. Clinical history of patients treated with EGFR-targeted drugs. Summarized clinical course of INT patients with EGFR CNG. Red-lined boxes indicate periods of administration of the indicated therapeutic agents. Blue vertical lines indicate timing of tumor specimen acquisition from surgical procedures or biopsies, as well as dates of tumor assessment by CT scan. PD and SD according to RECIST 1.1. 5FU, 5-fluorouracil; CCDP, cisplatin, vinorelbine, ifosfamide, and epirubicin; EOX, epirubicin, oxaliplatin, and capecitabine; FOLFIRI, folinic acid, 5-fluorouracil, and irinotecan; OGD, esophago-gastro-duodenoscopy; TCF, docetaxel, carboplatin, and 5-fluorouracil; XELOX, capecitabine and oxaliplatin.

Article Snippet: Primary antibodies, anti-EGFR (1005: sc-03) and anti-Actin, were from Santa Cruz Biotechnology, and antibodies against phosphorylated EGFR (Tyr 845), ERK (Thr202/Tyr204), phosphorylated AKT (Ser473) (Clone D9E), total AKT, and ERK were from Cell Signaling Technology.

Techniques: Computed Tomography

Journal: Clinical Cancer Research

Article Title: Optimized EGFR Blockade Strategies in EGFR Addicted Gastroesophageal Adenocarcinomas

doi: 10.1158/1078-0432.ccr-20-0121

Figure Lengend Snippet: Figure 4. RTK/KRAS pathway activation in EGFR-amplified cases. A, The scatter plot shows a significant inverse correlation between the extent of RTKs coamplification (HER2, HER3, HER4, MET, FGFR1, FGFR2, FGFR3, FGFR4, IGF1R, PDGFR2, VEGFR1, VEGR2, and VEGFR3 ≥4 gene copies) and OS in 12 EGFR FISH–positive gastroesophageal adenocar- cinomas treated with gefitinib in the COG trial. Red dots indicate cases with high polysomy and blue dots represent cases with EGFR amplification. B, The graph shows the cooccurrence of EGFR amplification and genomic events affecting the RTK/KRAS pathway in EGFR-amplified gastro- esophageal adenocarcinoma tumors in the FMI dataset.

Article Snippet: Primary antibodies, anti-EGFR (1005: sc-03) and anti-Actin, were from Santa Cruz Biotechnology, and antibodies against phosphorylated EGFR (Tyr 845), ERK (Thr202/Tyr204), phosphorylated AKT (Ser473) (Clone D9E), total AKT, and ERK were from Cell Signaling Technology.

Techniques: Activation Assay

Journal: Clinical Cancer Research

Article Title: Optimized EGFR Blockade Strategies in EGFR Addicted Gastroesophageal Adenocarcinomas

doi: 10.1158/1078-0432.ccr-20-0121

Figure Lengend Snippet: Figure 5. Dual EGFR blockade is the most effective treatment in EGFR-amplified PDXs. Tumor growth curves in mice cohorts derived from GTR0060 (A), GTR0110 (C), and GTR0511 (D) patients treated with the EGFR inhibitors, cetuximab (CETUX), erlotinib (ERL), and lapatinib (LAP), alone or in combination, as indicated. The red lines indicate the day when treatment was started. The response in mice has been evaluated using RECIST 1.1–like criteria, that is, PD: ≥35% increase from baseline; PR: ≥ 50% reduction from baseline; and SD: intermediate variations from baseline. B, Spaghetti plot illustrating drug response in the xenotrial performed on the cohort of mice derived from GTR0060 PDX. Individual lines represent, for each mouse, the percentage variation in tumor burden, from start of treatment (day 0). Blue lines, cetuximab þ lapatinib–treated mice and red lines, cetuximab þ erlotinib–treated mice. Dashed line indicates treatment stop. E, Western blot analysis of the activation state of EGFR and its downstream targets (AKT, MAPK, and S6) in GTR0060 tumor–derived cells treated with the indicated drugs/drug combinations. Actin was used as loading control. Statistical significance is indicated (, P <0.01; , P < 0.001).

Article Snippet: Primary antibodies, anti-EGFR (1005: sc-03) and anti-Actin, were from Santa Cruz Biotechnology, and antibodies against phosphorylated EGFR (Tyr 845), ERK (Thr202/Tyr204), phosphorylated AKT (Ser473) (Clone D9E), total AKT, and ERK were from Cell Signaling Technology.

Techniques: Derivative Assay, Western Blot, Activation Assay, Control

Journal: Clinical Cancer Research

Article Title: Optimized EGFR Blockade Strategies in EGFR Addicted Gastroesophageal Adenocarcinomas

doi: 10.1158/1078-0432.ccr-20-0121

Figure Lengend Snippet: Figure 6. TSC2 inactivation is a mechanism of resistance to EGFR-targeted therapies. A, Tumor growth curves in the mice cohorts derived from GTR0078 and treated with the EGFR inhibitors, cetuximab (CETUX), erlotinib (ERL), and lapatinib (LAP), alone or in combination, as indicated. The red line indicates the day when treatment was started. B, The table shows the two TSC2 mutations identified in GTR0078 PDX. C, Tumor growth curves in the mice cohorts derived from GTR0078 and treated with erlotinib or the combination erlotinib þ everolimus (ERLþ EVEROL). The red line indicates the day when treatment was started. D, Western blot analysis of the activation state of EGFR and its downstream targets (AKT, MAPK, and S6) in GTR0078 tumor–derived cells treated with the indicated drugs/drug combinations. Actin was used as loading control. Statistical significance is indicated (, P < 0.001).

Article Snippet: Primary antibodies, anti-EGFR (1005: sc-03) and anti-Actin, were from Santa Cruz Biotechnology, and antibodies against phosphorylated EGFR (Tyr 845), ERK (Thr202/Tyr204), phosphorylated AKT (Ser473) (Clone D9E), total AKT, and ERK were from Cell Signaling Technology.

Techniques: Derivative Assay, Western Blot, Activation Assay, Control

Journal: Archives of Pathology & Laboratory Medicine

Article Title: Application of Immunohistochemistry in the Diagnosis of Pulmonary and Pleural Neoplasms

doi: 10.5858/arpa.2016-0550-ra

Figure Lengend Snippet: Figure 3. Moderately differentiated adenocarcinoma of the lung. The biopsy shows a moderately differentiated adenocarcinoma with an acinar pattern (hematoxylin-eosin, A). The tumor cells express the EGFR mutant protein, E746-A750 (B) (hematoxylin-eosin, original magnification 3200 [A]; original magnification 3200 [B]).

Article Snippet: The 2 most common EGFR mutations include inframe deletions in exon 19 (E746_A750del) and a point mutation at codon 858 in exon 21 (L858R), together representing 85% to 90% of EGFR mutations in NSCLC patients.41,42 Mutation-specific antibodies for these 2 genetic mutations were developed43 in 2009, and numerous studies have since demonstrated that IHC may be used as a reliable prescreening test for detecting EGFR mutations in NSCLC.44–47 A moderately differentiated adenocarcinoma of the lung shows an acinar pattern of growth (Figure 3, A) and contains an EGFR mutant with E746-A750 (Figure 3, B) detected by using rabbit monoclonal antibody against EGFR E746-A750 (clone 6B6; Cell Signaling Technology, Danvers, Massachusetts); a poorly differentiated adenocarcinoma of the lung exhibits a solid pattern of growth (Figure 4, A) and harbors an EGFR mutant with L835R (Figure 4, B) detected by using rabbit monoclonal antibodies against EGFR L835R (clone 43B2; Cell Signaling Technology).

Techniques: Mutagenesis