Kelly A. Kaiser

The Potential Predictive Value of Cyclooxygenase-2 Expression and Increased Risk of Gastrointestinal Hemorrhage in Advanced Non-Small Cell Lung Cancer Patients Treated with Erlotinib and Celecoxib

Purpose: Celecoxib, a cyclooxygenase-2 (COX-2) inhibitor, potentiates antitumor effects of erlotinib in preclinical studies, and COX-2 is frequently expressed in non–small cell lung cancer (NSCLC). With these observations, we designed a phase II trial to evaluate the efficacy and safety of erlotinib plus celecoxib in advanced NSCLC. Experimental Design: Previously treated stage IIIB/IV NSCLC patients were given celecoxib at 400 mg orally twice daily and erlotinib at 150 mg orally daily until disease progression. Planned accrual was 40 patients. Tissue was collected for epidermal growth factor receptor (EGFR) analysis and COX-2 immunohistochemistry. Results: Twenty-six patients were enrolled (17 men, 9 women; median age, 66 years). Eighteen and 21 patients had tissue available for EGFR analysis and COX-2 immunohistochemistry, respectively. The median progression-free survival (PFS) and overall survival were 2.0 and 9.2 months, respectively. Eleven of 21 patients tested had increased tumor COX-2 expression, which was strongly associated with prolonged PFS (P = 0.048). Four patients on anticoagulation or with a history of peptic ulcer disease had grade 3/grade 4 upper gastrointestinal bleeding (GIB), prompting early study closure. Three patients with GIB had endoscopy that found peptic ulcers. Conclusions: The combination of erlotinib and celecoxib does not seem superior to erlotinib alone in unselected patients. However, longer PFS with high-tumor COX-2 expression suggests that trials of EGFR and COX-2 inhibitors may be warranted in this patient subset. GIB observed in our trial supports excluding patients with a history of peptic ulcer disease or those requiring therapeutic anticoagulation from future EGFR and COX-2 inhibitor studies.

The prognostic value of chromosome 7 polysomy in non-small cell lung cancer patients treated with gefitinib.

Introduction: Specific subpopulations of non-small cell lung cancer (NSCLC) patients defined by clinical features and molecular profiles seem to derive greater benefit from epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors, but no general consensus on molecular testing to optimize treatment has emerged. The objective of this study was to evaluate chromosome 7 polysomy and other potential indicators of gefitinib efficacy in advanced NSCLC patients. Methods: Paraffin-embedded tumors from 82 patients treated with gefitinib were analyzed by immunohistochemistry for expression of EGFR and other markers, and by fluorescence in situ hybridization for EGFR gene or chromosome copy number. Mutational status was assessed by single-strand conformational polymorphism, sequence-specific polymerase chain reaction, and direct sequencing. Molecular and clinical characteristics were evaluated in relation to objective response (OR), progression-free survival (PFS), and overall survival (OS). Results: EGFR mutational status (p = 0.002), never smoking (p = 0.052), and chromosome 7 polysomy (p = 0.029) were significant indicators of OR. EGFR mutation, pAKT or PTEN expression, and chromosome 7 polysomy were associated with longer OS. There was a significant difference in OS between the chromosome 7 polysomy groups (p = 0.015) and the groups with both chromosome 7 polysomy and pAkt+ (p = 0.002) and both chromosome7 polysomy and PTEN+ (p = 0.04). In a stepwise proportional hazards analysis, chromosome 7 polysomy and PTEN+ expression were both significantly associated with longer OS (p = 0.004 and 0.017 respectively). Conclusion: These results suggest that further study of chromosome 7 polysomy and of pAKT and PTEN expression in patients treated with EGFR tyrosine kinase inhibitors is warranted in developing a clinical test for selecting patients for gefitinib therapy.

Abstract 3403: E-cadherin and vimentin patterns by immunohistochemistry identify epithelial and mesenchymal phenotypes in lung adenocarcinoma

Proceedings: AACR 103rd Annual Meeting 2012‐‐ Mar 31‐Apr 4, 2012; Chicago, IL Purpose: To examine markers of epithelial-mesenchymal transition (EMT) in lung adenocarcinoma (ADC) defining epithelial-like (EPT) and mesenchymal-like (MES) phenotypes and correlate these with outcomes, in addition to other EMT markers. This study examined 33 patients with lung adenocarcinoma (ADC) who underwent surgical resection and later at disease progression received erlotinib. Experimental Design: All slides with tumor from resection specimens were reviewed (mean 4.8/case; range 1-9). Tumors were histologically subtyped according to recent IASLC classification. Tumor budding was graded per Ueno et al. (2002). A block containing the invasive front was selected for IHC staining with antibodies to e-cadherin (ECAD), vimentin (VIM), β-catenin (α-CAT), and SNAI1. Pattern-specific staining was evaluated at the invasive tumor front: E-cadherin-membranous, vimentin-cytoplasmic, α-CAT-membranous, SNAI1-nuclear, cytoplasmic. Complete circumferential membranous staining for e-cadherin was scored as present/absent; other markers were scored by the Allred system (0-8). PFS and OS were calculated from time of diagnosis to disease progression or death. Results: Two distinct phenotypes were identified with ECAD and VIM: EPT showing preserved membrane ECAD staining with low VIM (≥4) (N=19) and MES showing weak/absent membrane ECAD staining with high VIM (Δ5) (N=10). Median PFS for EPT group was 4.2 mo vs MES 1.6 mo; median OS for EPT group 15.5 mo vs 6.5 mo. These differences were not statistically significant. No association was found between high grade tumor budding and EPT vs MES phenotype p=0.221). Membranous α-CAT expression and nuclear SNAI1 expression showed a statistically significant correlation with EPT vs MES (β-catenin mean score 7.8 vs 6.1, p=0.0184) and (nuclear SNAI1 mean score 7.3 vs 6.1, p=0.0364). Conclusions: Epithelial and mesenchymal phenotypes can be identified by IHC for e-cadherin and vimentin. These phenotypes correlate with other EMT markers β-catenin and SNAI1 but not with tumor budding. Although differences in median PFS and OS were seen between the two groups, the number of cases is too small to identify a statistically significant difference. The results do support the rationale for defining EPT and MES groups for prognosis in a larger group of early-stage surgically resected patients. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3403. doi:1538-7445.AM2012-3403

Abstract 2404: Correlation of OGlcNAc modification components and EMT/Stem cell markers in lung adenocarcinoma

Proceedings: AACR 103rd Annual Meeting 2012‐‐ Mar 31‐Apr 4, 2012; Chicago, IL Background: O-GlcNAcylation, post-ribosomal protein modification with an N-acetylglucosamine (O-GlcNAc) residue is functionally similar to phosphorylation in cellular physiology in that it is vital to cellular regulation and homeostasis. The enzymes responsible for the addition and removal of O-GlcNAc have been identified as O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). It has been proposed that O-GlcNAcylation may be an important regulator of cancer initiation and progression, based on the O-GlcNAcylation of many oncogenes/ proto-oncolgenes and tumor suppressors, but with few citations in the literature on the subject. Here, we investigated the correlation of O-GlcNAc modification components with expression of Epithelial-Mesenchymal-Transition (EMT)/Stem cell markers in 65 cases of lung adenocarcinoma. Methods: Immunohistochemical staining for O-GlcNAcylation, OGT, OGA, E-cadherin, vimentin, CD133, CD34 and CXCR-4 were performed on 65 primary lung adenocarcinoma and matching metastatic lymph nodes. All patients were treated by complete anatomic resections and limited to T1-2N0-2M0 (pathological). Scoring was accomplished by evaluating localization (membrane, cytoplasm, peri-nuclear, and nuclear), stain intensity (0: no staining, 1: weak staining, 2: strong staining) and frequency for at least 200 cells per field (400X magnification), with 25 random fields surveyed per section. This was done by two readers, and a score was then calculated based on these parameters for statistical comparisons (0: lowest, 2: highest). We then evealuated the correlation of these markers with each other through Pearsons correlation in SPSS v18.0. Results: Higher expression of cytoplasmic OGT was correlated with higher cytoplasmic OGlcNAcylation in primary tumor (r=0.40, p=0.001) and lymph node metastasis (r=0.567, p=0.001). However, we did not observe any relationship between OGA and OGlcNAcylation level. Vimentin showed positive correlation with nuclear OGlcNAcylation in both primary (r=0.281, p=0.031) and lymph node (r=0.380, p=0.035). but no relationship was observed with cytoplasmic OGlcNacylation. Also, CD133 demontrated positive correlation with cytoplasmic OGT in both primary (r=0.510, p=<0.001) and lymph node (r=0.555, p=0.001). Conclusion: Our results suggest that O-GlcNAcylation might play important roles in lung adenocarcinoma initiation and progression and may be a potential prognostic factor to predict patient risk of recurrence after surgery. Also, these findings may provide us with added insights regarding the mechanism of metastasis, although, further investigations are warranted to validate our results. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2404. doi:1538-7445.AM2012-2404