Lung Squamous Cell Research Articles

Abstract 663: Development of a cancer-specific monoclonal antibody LpMab-2 specific for cancer-type podoplanin

Abstract Podoplanin is a platelet aggregation-inducing mucin-like sialoglycoprotein that is highly expressed in brain tumors, malignant mesotheliomas, testicular tumors, and lung, esophageal, or head and neck squamous cell carcinomas. Podoplanin has also been reported to be a marker to enrich tumor-initiating cells. We previously established a neutralizing monoclonal antibody (mAb), NZ-1, which inhibits the association between podoplanin and C-type lectin-like receptor-2 (CLEC-2) and blocks the podoplanin-induced platelet aggregation and tumor metastasis. A human chimeric mAb, NZ-8 possesses extremely high antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), and showed high anti-tumor activities in several xenograft models. However, those mAbs also react with normal cells including lung type I alveolar cells, renal podocytes, and lymphatic endothelial cells throughout the body. Recently, we established the platform to produce a cancer-specific mAb (CasMab) by combining glycobiology and cell engineering technologies. Herein, we report a novel cancer-specific anti-podoplanin mAb, LpMab-2. In Western-blot analyses, LpMab-2 did not react with sialic acid-deficient podoplanin and O-glycan deficient podoplanin, which were expressed in Lec2 and Lec8 cells, respectively. In enzyme-linked immunosorbent assay using several deletion mutants of podoplanin, the epitope of LpMab-2 was shown to be included in 55-88 amino acids of podoplanin, although almost all anti-podoplanin mAbs recognize platelet aggregation-stimulating (PLAG) domain near the N-terminus. Analyses of glycopeptides produced by Edman degradation revealed that O-glycans were attached to glycosylation sites at residues Thr65, Thr66, Thr70, Ser71, Ser74, and Thr76 around LpMab-2 epitope. LpMab-2 did not react with podoplanin-negative cell lines, indicating that not only glycans but also amino acids of podoplanin are necessary for LpMab-2 reaction. In flow cytometric and immunohistochemical analyses, LpMab-2 reacted with tumor cells, but not with lung type I alveolar cells and lymphatic endothelial cells, suggesting that LpMab-2 recognizes the cancer-type conformation of podoplanin. Furthermore, human chimeric LpMab-2 also showed cancer-specific reaction against podoplanin. Taken together, a cancer-specific mAb LpMab-2 should be useful for molecular targeting therapy against podoplanin without side effects such as respiratory or renal failures and lymphatic edema. The CasMab-producing platform could lead to establishment of the side effect-reducing mAbs. Note: This abstract was not presented at the meeting. Citation Format: Yukinari Kato, Mika K. Kaneko. Development of a cancer-specific monoclonal antibody LpMab-2 specific for cancer-type podoplanin. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 663. doi:10.1158/1538-7445.AM2014-663

Abstract 2920: Circulating inflammation markers and subsequent lung cancer risk: A discovery and replication study

Abstract Background: Few epidemiologic studies have investigated the role of inflammation in lung cancer etiology. We have carried out 2 independent studies nested within the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial to assess associations between circulating inflammation markers and lung cancer risk, after carefully controlling for smoking. In the discovery study, we showed associations between 11 circulating inflammation markers (CRP, SAA, sTNFRII, IL-1RA, IL-7, TGF-A, ENA 78/CXCL5, MIG/CXCL9, BCA-1/CXCL13, TARC/CCL17 and MDC/CCL22) and prospective lung cancer risk (Shiels, JNCI 2013). To assess the reproducibility of these associations, we have carried out a large, independent replication study. Methods: We designed a replication case-control study nested within the PLCO in 526 lung cancer cases and 625 controls, matched by gender, age, smoking (status, pack-years and time since quit), study visit and years of blood draw and exit. Serum levels of 51 inflammation markers were measured using Luminex bead-based assays. We combined the data from the discovery and replication studies (1052 cases and 1217 controls) and created study-specific marker categories. Conditional logistic regression was used to estimate odds ratios (ORs) overall and stratified by latency and histology in the pooled dataset. Results: In our prior study, 11 inflammation markers were associated with lung cancer risk with ORs ranging from 1.41 to 2.27. Of these 11 markers, C-reactive protein (CRP), serum amyloid A (SAA), soluble tumor necrosis factor receptor 2 (sTNFRII) and monokine induced by gamma interferon (MIG/CXCL9) were associated with prospective lung cancer risk in the replication study with ORs ranging from 1.70 to 2.09 (p-trend across quartiles3 years) (all p-interactions>0.2). Further, each marker was associated with both lung adenocarcinoma and squamous cell carcinoma (all p-trends<0.05). Conclusions: With results from two independent case-control studies, we have confirmed that pre-diagnostic circulating levels of two acute phase proteins (CRP and SAA), a receptor for a pro-inflammatory cytokine (sTNFRII), and a chemokine (MIG/CXCL9) are each associated with prospective lung cancer risk, after carefully controlling for cigarette smoking. These results provide evidence implicating inflammation in the development of lung cancer. Citation Format: Meredith S. Shiels, Hormuzd A. Katki, Allan Hildesheim, Ruth M. Pfeiffer, Eric A. Engels, Marcus C. Williams, Troy J. Kemp, Neil E. Caporaso, Ligia A. Pinto, Anil K. Chaturvedi. Circulating inflammation markers and subsequent lung cancer risk: A discovery and replication study. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2920. doi:10.1158/1538-7445.AM2014-2920

Abstract 5334: Integrated analysis of squamous tumors identifies novel targets and dissects gene regulation

Abstract Background: Studies by The Cancer Genome Atlas (TCGA) and others have identified regions of somatic copy number alteration (SCNA) in head and neck squamous cell carcinoma (HNSCC) and lung squamous cell carcinoma (LUSC). Both tumor types exhibit frequent SCNAs in chromosomes 3q, 9p, and 11q. Although CDKN2A is the putative target of 9p deletions, numerous targets have been proposed in the 3q amplicon, including SOX2, PIK3CA, and TP63. Because expression levels are affected by underlying genomic events, we hypothesized that an integrated analysis of multiple genomic data types would provide increased ability to identify target genes in SCNA regions when compared to methods based on copy number alone while providing insight into mechanisms regulating expression. Findings may be subsequently validated by examining multiple tumor types or performing training/testing analyses. Techniques: Gene expression (GE), DNA copy number (CN), DNA methylation (ME), and microRNA expression (miR) data were obtained from the TCGA studies of HNSCC and LUSC. Linear models were constructed for each gene to investigate the effect of changes in CN, ME, and miR on GE. Analysis of model output provided an approach to identify target genes in SCNA regions and assess the effect of genomic alterations on expression. Results: Genome-wide GE, CN, ME, and miR data were available for n = 279 (HNSCC) and n = 159 (LUSC) tumor samples. Univariate modeling detected a strong overall association between GE and CN, as measured by the coefficient of determination (model R2). Although some genes found in SCNA studies produced large model R2— e.g. SOX2— a number of genes with large model R2 are relatively unknown. Notable examples include DVL3, and SENP2, which were implicated as driver genes in the 3q amplicon by a recent study of LUSC but not identified in the TCGA report. Remarkably, the model R2 for DVL3 and SENP2 was also high in HNSCC, and such a finding in a distinct tumor type provides validation and merits additional study. Output from models additionally including ME and miR as covariates contributes insight into the diversity of regulation of gene expression, but the strength of the association between GE and CN could mask other effects. Thus we constructed linear models in which the response variable was the residuals from the GE/CN model and the covariates were ME or miR. The TCGA study of glioblastoma noted frequent methylation of MGMT. When modeling MGMT we detected a highly significant association between ME and the GE/CN residuals in both HNSCC and LUSC. These results illustrate a pronounced effect of ME on GE conditional on CN. Conclusion: Linear modeling techniques provide a flexible and powerful basis for performing integrated analysis of genomic data. Our approach produces predicted results when analyzing known cancer genes, highlights lesser known genes for future study, provides insight into gene regulation, and draws attention to genes relevant in multiple tumor types. Citation Format: Vonn Walter, Ying Du, Xiaoying Yin, Wei Sun, Matthew D. Wilkerson, Michele C. Hayward, Ashley H. Salazar, Charles M. Perou, David N. Hayes. Integrated analysis of squamous tumors identifies novel targets and dissects gene regulation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5334. doi:10.1158/1538-7445.AM2014-5334

Abstract LB-215: The TAM family of receptor tyrosine kinases play a role in acquired resistance to cetuximab

Abstract Cetuximab is an anti-epidermal growth factor receptor (EGFR) monoclonal antibody that has shown efficacy in metastatic colorectal cancer, head and neck squamous cell carcinoma (HNSCC) and non-small cell lung cancer (NSCLC). Clinical data suggests that patients whom initially respond to cetuximab eventually acquire resistance, highlighting the importance of understanding resistant mechanisms for the development of better treatment regimes for cancer patients. Recent studies have identified a role for the TAM family of receptor tyrosine kinases (Tyro, Axl, and Mer) in tumor biology, and more specifically the Axl receptor as playing a key role in promoting tumor growth and metastasis. In the current study, we utilized NSCLC and HNSCC models of acquired resistance to cetuximab and found that Axl was overexpressed, activated, and highly associated with EGFR as compared parental controls. Further, using siRNA technology and novel Axl targeting agents (antibody and tyrosine kinase inhibitor), Axl was found to regulate proliferation, EGFR activation, and MAPK signaling in cetuximab resistant clones (CtxR). Additionally, Axl mRNA expression was regulated by EGFR induced MAPK activation in resistant cells. To investigate the role of Axl in in vivo models of acquired resistance to cetuximab, de novo acquired resistance was created by prolonged treatment of cetuximab sensitive (CtxS) tumor xenografts with cetuximab. Analysis of resultant CtxR tumors demonstrated that Axl was hyperactivated and highly associated with EGFR as compared to vehicle treated tumors. In addition, CtxR tumors expressing the highest levels of phosphorylated Axl also expressed hyperactivated EGFR, supporting the role of Axl in EGFR activation. Finally, to expand these findings to other members of the TAM family, Tyro, Axl, or Mer were stably overexpressed in CtxS parental NSCLC cells and tested for cetuximab response. The results of this experimentation indicated that stable overexpression of each TAM receptor rendered CtxS cells resistant to cetuximab therapy. Collectively, the studies presented herein identify the TAM family of receptors as key players in acquired resistance to cetuximab and provide rationale for the clinical evaluation of anti-TAM receptor therapeutics in the resistant setting. Citation Format: Toni Michel Brand, Mari Iida, Kelsey L. Corrigan, Neha Luthar, Megan Hornung, Mahmoud Toulany, Parkash Gill, Ravi Salgia, Deric L. Wheeler. The TAM family of receptor tyrosine kinases play a role in acquired resistance to cetuximab. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-215. doi:10.1158/1538-7445.AM2014-LB-215

Abstract LB-99: Identification and characterization of small molecule thieno[3,2-d]pyrimidine inhibitors of Protein Kinase C iota (PKCι)

Abstract We describe a novel series of small molecule ATP-competitive inhibitors of the atypical class of Protein Kinase C (PKC). The PKC family of serine/threonine kinases are divided into four structurally and functionally distinct sub-types: classic (PKCα, PKCβ, PKCγ); novel (PKCδ, PKCε, PKCη, PKCθ); atypical (PKCι,PKCζ); and PKNs (PKN1, PKN2, PKN3). The PKCs have been defined as key regulators in a multitude of signal transduction pathways that impinge on diverse cellular processes such as proliferation, differentiation, and survival. The regulation and functions of the atypical PKCs are distinct from the classic and novel PKCs.Both atypical isoforms have been implicated in various models of cancer, and PKCι in particular has been described as a potential oncogene. Its abundance is frequently increased, primarily through PKCι gene amplification, in many epithelial tumours including: subsets of squamous non-small cell lung carcinoma, serous ovarian carcinoma, and squamous esophageal carcinoma. It has been widely demonstrated that deregulation of PKCι signalling leads to unconstrained cell growth, increased migratory and invasive behaviour, and aberrant cellular polarity, which is a hallmark of aggressive cancers. Together, these data make a strong case for the inhibition of PKCι as a novel therapeutic strategy. We have discovered a series of ATP-competitive thieno[3,2-d]pyrimidine- based PKCι inhibitors from a high throughput screen. Using structure-based design we have optimized the series and demonstrated potent and selective inhibition of PKCι in biochemical and cellular models. The biochemical, structural, and cellular characterization of these compounds will be described herein. Citation Format: Christelle Soudy, Emma Stanway, Bhavisha Patel, Caroline Barton, Michelle Barnard, Leon Pang, Paul Owen, Andrew Turnbull, Bruce A. Ruggeri, Bruce D. Dorsey, Mark A. Ator, Greg R. Ott, Mark Linch, Philippe Riou, Peter J. Parker, Sven Kjaer, Christian Dillon, Neil Q. McDonald, Jon Roffey. Identification and characterization of small molecule thieno[3,2-d]pyrimidine inhibitors of Protein Kinase C iota (PKCι). [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-99. doi:10.1158/1538-7445.AM2014-LB-99

Abstract 4728: FGFR1 gene amplification in small cell lung cancer

Abstract Background: Fibroblast growth factor receptor 1 (FGFR1) received special attention from many clinicians according to finding that FGFR1 played a critical role in many human cancers. Some clinical trials using FGFR1 targeting agents are in progress especially in breast cancer and squamous non-small cell lung cancer. Recently, FGFR1 amplifications were also found in about 5.6-6% of small cell lung cancer (SCLC) by gene copy number analysis and FISH. Methods: Tumor tissues from 113 patients diagnosed with SCLC from October 2009 to February 2013 in YUHS, Korea were collected. FGFR1 FISH assay was performed on the tissue microarray using FISH probe that hybridized to the band 8p12-8p11.23 (FISH probes were provided by Abbott Molecular, Abbott Park, IL). Results: Thirty-two patients with limited-stage disease (LD) and eighty-one patients with extensive-stage disease (ED) were enrolled. Fourteen patients were female. Among 113 tumor samples, 5 samples (4.4%) showed FGFR1 amplification. All 5 samples were obtained from the patients with ED. The patients with FGFR1 amplification more frequently had pleural metastasis at the time of diagnosis (80% vs. 26%, P=0.022). In ED, the patients with FGFR1 amplification represented shorter disease free survival from first line chemotherapy (Hazard ratio [HR], 4.469; 95% confidence interval [CI], 1.600-12.483; P=0.004) and second line chemotherapy (HR, 6.533; 95% CI, 1.700-25.108; P=0.006) than the patients without FGFR1 amplification in multivariate analysis. Median overall survival time was 8.2 months in the patients with FGFR1 amplification and 10.2 months in the patients without FGFR1 amplifications (P=0.296). Conclusion: FGFR1 amplification is a poor potential predictive biomarker of standard chemotherapy, necessitating further research in a large cohort of SCLC. The validity of targeting FGFR1 in SCLC should be confirmed in a clinical trial. Citation Format: Ji Soo Park, Jae-Seok Lee, Hyo Sup Shim, Hye Ryun Kim, Sun Min Lim, Joo Hang Kim, Byoung Chul Cho. FGFR1 gene amplification in small cell lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4728. doi:10.1158/1538-7445.AM2014-4728

Targeting C-Met Overexpression for Acquired Resistance to Egfr Tkis

ABSTRACT Aim: c-Met amplification and T790M are both recognized as the common molecular mechanisms of acquired resistance (AR) to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR TKIs) in advanced non-small-cell lung cancer (NSCLC). It is not clear that c-Met overexpression could be as the biomarker for AR. Methods: Advanced NSCLC patients with AR to EGFR TKIs were detected for c-Met overexpression by immunohistochemistry ≥50% tumor cells with moderate to high intensity staining were defined as c-Met positive. The statuses of EGFR, ALK, KRAS and ROS1 were also tested. Results: From January 2013 to April 2014, 80 advanced NSCLC patients with AR to gefitinib or erlotinib were enrolled prospectively. The frequency of c-Met overexpression was 28.8% (23/80), c-Met overexpression + T790M 12.5% (10/80), T790M 25.0% (20/80), small-cell lung cancer or squamous cell transformation 2.5% (2/80), KRAS mutation 1.3% (1/80), ROS1 fusion 1.3% (1/80) and unknown mechanism 28.8% (23/80), respectively. Fourteen c-Met overexpressed patients received gefitinib plus c-Met inhibitors. Among them 4 patients with c-Met overexpression only shown good response to crizotinib (2/3 cases achieving partial response [PR] and 1 with stable disease), another case shown PR to single agent axitinib and all got prominently clinical benefit. Among 10 with c-Met overexpression + T790M, 6 received combination of EGFR TKI and c-Met inhibitor and best response was progressive disease, even though one was treated with afatinib plus crizotinib. Conclusions: c-Met overexpression could be as a biomarker for AR. Combination of EGFR TKIs and c-Met inhibitor is a good strategy to overcome AR for c-Met overexpressed patients, but not effective in c-Met/T790M-coexisting cases, for whom further investigations are warranted. Disclosure: All authors have declared no conflicts of interest.

Multi-Arm, Nonrandomized, Open-Label Phase Ib Study to Evaluate Fp1039/Gsk3052230 with Chemotherapy in Nsclc and Mpm with Deregulated Fgf Pathway Signaling

ABSTRACT Background: FP1039/GSK3052230 (GSK230) is a soluble fusion protein that acts as a ligand trap by sequestering fibroblast growth factors (FGFs) involved in tumor growth. FGF ligand-dependent signaling plays an important role in cancer development and growth through autocrine production of FGFs from cancer cells, or paracrine production of FGFs from local stroma, or by tumor overexpression of FGF receptors (FGFRs) or amplification of FGFR genes. FGFR1 amplification has been identified in ∼15-20% of squamous non-small cell lung cancers (NSCLC) and is associated with shorter disease-free and overall survival. Efficacy was observed with GSK230 + chemotherapy in FGFR1 amplified in NSCLC xenograft models and in xenograft models of malignant pleural mesothelioma (MPM) where FGF2 mRNA levels were overexpressed. In a phase I study, no maximum tolerated dose (MTD) was identified and 20 mg/kg weekly was the maximum feasible dose (MFD) achieving the desired target concentration. Toxicities associated with small-molecule FGFR inhibition, namely hyperphosphatemia and retinal, nail, and skin changes, were not observed. Trial design: This is a multi-arm, nonrandomized, open-label phase IB study designed to evaluate the safety and preliminary efficacy of GSK230 in combination with paclitaxel + carboplatin in previously untreated FGFR1 amplified metastatic squamous NSCLC (Arm A), in combination with docetaxel in FGFR1 amplified metastatic squamous NSCLC that has progressed after 1 line of chemotherapy (Arm B), or in combination with pemetrexed + cisplatin in patients with untreated and unresectable MPM (Arm C). Approximately 70 patients will be enrolled. Each arm involves a dose escalation phase utilizing the 3 + 3 design, followed by a cohort expansion phase of at least 12 patients treated at either MTD or MFD. GSK230 will be administered as a 30-minute intravenous infusion once a week in a 21-day cycle. The starting dose is 5 mg/kg and will be dose escalated until MTD or MFD in combination with chemotherapy is reached. Endpoints include the combination dose of GSK230 with chemotherapy, safety, response rates and their duration, and translational objectives (NCT01868022). Disclosure: P. Garrido: I don't have any specific conflict interest related to this trial except from being part the clinical trial; E. Felip: Advisory boards for BI, Novartis, Roche, BMS and Lilly; F. Barlesi: Honorarium from GSK; P.K. Paik: Research funding provided by GlaxoSmithKline, BristolMyersSquibb; D. Morgensztern: Member of Advisory Board – Celgene Speaker – Boehringer Ingelheim; K. Reckamp: GSK provided research funding to institution. All other authors have declared no conflicts of interest.

Squire: a Randomized, Multicenter, Open-Label, Phase III Study of Gemcitabine-Cisplatin (Gc) Chemotherapy Plus Necitumumab (Imc-11F8/Ly3012211) Vs Gc Alone in the First-Line Treatment of Patients (Pts) with Stage Iv Squamous Non-Small Cell Lung Cancer (Sq-Nsclc): Update on Key Subgroups

ABSTRACT Aim: EGFR is detectable in most pts with sq-NSCLC tumors. Efficacy and safety of necitumumab (N), a human IgG1 anti-EGFR monoclonal antibody that inhibits ligand-binding and receptor activation, were evaluated in pts with advanced sq-NSCLC tumors. Methods: Pts with stage IV sq-NSCLC were randomized 1:1 to GC (G = 1250 mg/m2 iv, days 1 and 8; C = 75 mg/m2 iv, day 1) plus N (800 mg iv, days 1 and 8) (GC + N arm), or GC alone (GC arm) every 21 days for up to 6 cycles. GC + N pts with no progression continued on N alone until progressive disease or intolerable toxicity. The primary endpoint was overall survival (OS). Progression-free survival and safety were secondary endpoints. Subgroup analyses, evaluating efficacy by age, Eastern Cooperative Oncology Group (ECOG) performance status (PS), gender, and race, were also performed. Planned sample size was 1080 pts, with 90% power and a 2-sided alpha level of 0.05. ClinicalTrials.gov identifier: NCT00981058. Results: 1093 pts were randomized. Baseline characteristics, exposure to chemotherapy, and post-progression anticancer therapy were similar between GC + N and GC arms. The table shows key OS results. The addition of N to GC statistically significantly improved OS (HR = 0.84, p = 0.012), and the safety profile of GC + N was acceptable across subgroups. OS Results N mOS, GC + N* N mOS, GC* HR† ITT population 545 11.5 548 9.9 0.84 Subgroups Age, y 520 11.5 529 9.9 0.84 ≥75 25 10.3 19 7.4 0.98 ECOG PS 0 164 13.8 180 12.9 0.82 1 332 10.7 320 9.2 0.85 2 49 9.5 47 6.9 0.78 Gender Female 95 13.0 90 11.4 0.88 Male 450 11.1 458 9.7 0.84 Race Caucasian 457 11.4 456 9.7 0.86 Non-Caucasian 88 12.6 92 11.8 0.78 mOS = median overall survival. *Data = median (months) estimated by the Kaplan-Meier method. †HR Conclusions: This study met its primary endpoint. GC + N demonstrated improved OS, with an acceptable safety profile, across all subgroups. Disclosure: M.A. Socinski: Steering Committee; L. Paz-Ares: I have provided scientific advice asn received honararium from Lilly; M. Reck: Member of advisory boards (compensated): Lilly, Hoffmann-La Roche, BMS, AstraZeneca, Pfizer, Boehringer Ingelheim, Novartis; K. Park: Advisory Board; Eli Lilly; F.R. Hirsch: Research funding (through University of Colorado) from ImClone - Lilly. Participated in steering committee for the study and advisory borad for Lilly – ImClone; H. Depenbrock: Lilly Employee S. Nanda: I am a lilly employee and I have stock ownership; N. Chouaki: I am a Lilly Employee, I have stock ownership; N. Thatcher: Honoraria :Advsiory Board and Speaker Bureau Member Lilly Imclone. All other authors have declared no conflicts of interest.

Identification of featured biomarkers in different types of lung cancer with DNA microarray.

Lung cancer is a worldwide leading cause of cancer-related death. The aim of this study was to identify target genes and specific biomarkers for identification and treatment of different types of lung cancer with DNA microarray. Gene expression profile GSE6044 and miRNA microarray profile GSE17681 were downloaded from Gene Expression Omnibus database. The differentially expressed genes (DEGs) and miRNAs were screened with multtest package in R language. Then, functional enrichment analysis of identified DEGs was performed. Furthermore, the verified target genes based on screened miRNAs were selected from miRTarBase and miRecords databases. Then miRNA-target gene regulation network was constructed. APOE, CDC6 and ATP2B1were involved in most of the functions obtained for adenocarcinomas, small cell lung cancer and squamous cell carcinomas, respectively. The target DEGs of differentially expressed hsa-miR-29a included FGG in adenocarcinoma, RAN and COL4A1 in small cell lung cancer, GLUL in squamous cell carcinoma. The target DEGs of has-miR-7 were SNCA and SLC7A5 in adenocarcinoma and small cell lung cancer, respectively. ICAM1 and KIT were the target DEGs of hsa-miR-222 in adenocarcinoma and squamous cell carcinoma. The miRNAs and their differentially expressed target genes have the potential to be used in clinic for diagnosis and treatment of different kinds of lung cancer in the future.

Upregulation of annexin A5 affects the biological behaviors of lung squamous carcinoma cells in vitro

Annexin A5 is a Ca2+-dependent phospholipid-binding protein and protein kinase C inhibitory protein. It has a potential role in cellular signal transduction, inflammation, growth and differentiation. In this study, we evaluated the expression of this protein in lung tumor tissues and subsequently established a NCI-H520 cell line that stably expresses the wild-type ANXA5 gene to determine the effects of annexin A5 upregulation on the cell morphology, proliferation and metastasis potential in vitro. The effects of annexin A5 on NCI-H520 cells were tested by crystal violet staining, CCK-8 assay, scratch wound assay, and Transwell assay. The expressions of Akt, PCNA, vimentin, and E-cadherin were examined by Western blot assay. In this study, we demonstrated that annexin A5 is expressed at lower levels in tumor tissues compared with normal tissues. Additionally, the upregulation of this protein may inhibit the proliferation, migration, and invasion abilities of NCI-H520 cells in vitro. The transfected cells were arrested in the G1/S phase of the cell cycle, and the expression levels of Akt, PCNA and Vimentin were downregulated, while E-cadherin was upregulated.

P210 Cyclin-dependent kinase 4 overexpression confers an independent prognosticator of nasopharyngeal carcinoma

Background Data mining on a public domain demonstrated that cyclin dependent kinase 4 (CDK4) was highly expressed in nasopharyngeal carcinoma (NPC). Associated with cyclin-D, CDK4 phosphorylates and inactivates retinoblastoma (Rb) protein family members and mediates progression through G1 to S-phase in the cell cycle. Overexpression of CDK4 has been identified in oral squamous cell carcinoma, lung and breast cancers, sarcomas, and melanomas. However, the expression of CDK4 has never been reported in NPC. This study investigated the expression status, correlation with clinicopathological features, and prognostic implications of CDK4 in a well-defined cohort of NPC. Methods We retrospectively assessed CDK4 immunoexpression in biopsies of 124 consecutive NPC patients who did not have initial distant metastasis and who received treatment according to consistent guidelines. The results were correlated with clinicopathological features, local recurrence-free survival (LRFS), distant metastasis-free survival (DMeFS), and disease-specific survival (DSS). Findings High levels of CDK4 protein were positively correlated with T3,4 status (p = 0.024), N2,3 status (p Interpretation CDK4 overexpression represents a potential prognostic biomarker in NPC and may confer tumour aggressiveness through cell cycle dysregulation.

MP49-07 THE ANDROGEN RECEPTOR IS A DRIVER OF DNA BREAKPOINTS AND FUSION EVENTS IN PROSTATE CANCER

You have accessJournal of UrologyProstate Cancer: Basic Research IV1 Apr 2014MP49-07 THE ANDROGEN RECEPTOR IS A DRIVER OF DNA BREAKPOINTS AND FUSION EVENTS IN PROSTATE CANCER Matthew Hong, Geoff Macintyre, Niall Corcoran, Clare Sloggett, Haroon Naeem, Marek Cmero, John Pedersen, Izhak Haviv, Andrew Ryan, Pramit Phal, Anthony Costello, and Christopher Hovens Matthew HongMatthew Hong More articles by this author , Geoff MacintyreGeoff Macintyre More articles by this author , Niall CorcoranNiall Corcoran More articles by this author , Clare SloggettClare Sloggett More articles by this author , Haroon NaeemHaroon Naeem More articles by this author , Marek CmeroMarek Cmero More articles by this author , John PedersenJohn Pedersen More articles by this author , Izhak HavivIzhak Haviv More articles by this author , Andrew RyanAndrew Ryan More articles by this author , Pramit PhalPramit Phal More articles by this author , Anthony CostelloAnthony Costello More articles by this author , and Christopher HovensChristopher Hovens More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.1108AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail Introduction and Objectives The TMPRSS2-ERG fusion is the most well-known fusion event in prostate cancer. Since specific fusion genes are associated with specific tumour types, it is likely that tissue-specific mechanisms underlie the formation of structural rearrangements in cancer genomes. Recent studies have suggested that androgen receptor (AR) may play a role in the formation of TMPRSS2-ERG fusions, bringing the two loci in close proximity in the nucleus and facilitating DNA strand break and repair along with AR associated enzymes. To explore this mechanism more comprehensively, we performed whole genome sequencing on 14 prostate cancer samples from 7 patients. Methods Fresh frozen prostate cancer samples were taken from patients undergoing radical prostatectomy or biopsy. Tissue samples were cryosectioned and tumour content confirmed by a pathologist prior to mechanical homogenisation and simultaneous DNA and RNA extraction (Allprep Micro Kit, Qiagen). Whole genome sequencing was performed to 40x coverage on the Illumina HiSeq2000 platform. Results Across the 14 cancer genomes we identified a base-pair resolution high confidence set of 2,921 structural variations (some recurrent) at an average of 209 per sample and found that a large proportion of these breakpoints were in close proximity to curated androgen receptor binding sites from high-confidence ChIP-Seq data. Furthermore, when we examined breakpoints in genome datasets from 11 other cancers from the TCGA and ICGC projects, we identified a similar association with androgen (and estrogen) receptor binding sites specifically in hormone-dependent tumour types including prostate, breast, squamous cell lung, and ovarian cancers, but no such correlation in glioblastoma, renal, lung, colon or lymphoma genome datasets. Conclusions These data suggest that the androgen receptor drives genome wide breakpoints and novel fusion events in prostate cancer and potentially serves to explain why androgen exposure is necessary for prostate cancer development. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e504 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Matthew Hong More articles by this author Geoff Macintyre More articles by this author Niall Corcoran More articles by this author Clare Sloggett More articles by this author Haroon Naeem More articles by this author Marek Cmero More articles by this author John Pedersen More articles by this author Izhak Haviv More articles by this author Andrew Ryan More articles by this author Pramit Phal More articles by this author Anthony Costello More articles by this author Christopher Hovens More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...

Targeted inhibition of cell-surface serine protease Hepsin blocks prostate cancer bone metastasis

The development of effective therapies inhibiting prostate cancer progression and metastasis may substantially impact prostate cancer mortality and potentially reduce the rates of invasive treatments by enhancing the safety of active surveillance strategies. Hepsin (HPN) is a cell surface serine protease amplified in a subset of human sarcomas (7.2%), as well as in ovarian (10%), lung adeno (5.4%), lung squamous cell (4.5%), adenoid cystic (5%), breast (2.6%), uterine (1.7%) and colon (1.4%) carcinomas. While HPN is not amplified in prostate cancer, it is one of the most prominently overexpressed genes in the majority of human prostate tumors and genetic experiments in mice indicate that Hepsin promotes prostate cancer metastasis, particularly metastasis to the bone marrow. We report here the development, analysis and animal trial of the small-molecule Hepsin inhibitor HepIn-13. Long-term exposure to HepIn-13 inhibited bone, liver and lung metastasis in a murine model of metastatic prostate cancer. These findings indicate that inhibition of Hepsin with small-molecule compounds could provide an effective tool for attenuation of prostate cancer progression and metastasis.

Fibroblast growth factor receptor-1 as a potential therapeutic target in sinonasal cancer

Despite multimodal treatment, sinonasal malignancies have an unfavorable prognosis. The purpose of this study was to elucidate if these tumors harbor amplifications of the fibroblast growth factor receptor 1 (FGFR1) gene, which has recently been identified as a potential therapeutic target in squamous cell lung cancer. One hundred twelve primary tumors (including squamous cell carcinoma [SCC], carcinoma associated with an inverted papilloma, sinonasal undifferentiated carcinoma [SNUC], adenocarcinoma, adenoid cystic carcinoma [ACC], esthesioneuroblastoma, and 9 corresponding lymph node metastases) were assessed by fluorescence in situ hybridization (FISH) for FGFR1 copy number status. Human papillomavirus (HPV) status was assessed by p16 immunohistochemical as a surrogate marker. FGFR1 amplification was found in subsets of sinonasal SCCs (20%), carcinomas associated with an inverted papilloma (33%), and SNUCs (5%). In all cases, metastatic tumor samples shared the same FGFR1 amplification status as the corresponding primary tumor tissue. None of the FGFR1-amplified tumors expressed p16. FGFR1 amplification represents a potential molecular target in a subset of patients with sinonasal cancer. © 2014 Wiley Periodicals, Inc. Head Neck 36: 1253-1257, 2014.

32 Review of pemetrexed with platinum chemotherapy for locally advanced or metastatic non squamous non-small cell lung cancer (NSCLC) in Northern Ireland

32 Review of pemetrexed with platinum chemotherapy for locally advanced or metastatic non squamous non-small cell lung cancer (NSCLC) in Northern Ireland

34 Pemetrexed and carboplatin (PC) doublet in advanced non squamous non-small cell lung cancer. Real life experience from a cancer unit

34 Pemetrexed and carboplatin (PC) doublet in advanced non squamous non-small cell lung cancer. Real life experience from a cancer unit

A Trial on the Overexpression of Epidermal Growth Factor Receptor and Proliferating Cell Nuclear Antigen in NSCLC of Similar Differentiation

Objective: To explore the expression of EGFR and PCNA in lung squamous cell carcinomas and lung adenocarcinomas, and the relationships between their overexpression and cancer tissue typing or lymphatic metastasis. Method: Detect the expression of EGFR and PCNA in 44 cases of similarly differentiated cancer tissue and 16 cases of normal lung tissue by immunohistochemistry. Result: The expression of EGFR and PCNA in lung cancer tissue was significantly higher than that in normal lung tissue. Meanwhile, no significant difference existed between expression of PCNA and EGFR in lung squamous carcinomas and that in lung adenocarcinomas (P>0.05), nor between lymph node metastatic tissue and negative lymph node metastatic tissue (P>0.05). Conclusion: EGFR and PCNA were involved in the onset and development of lung cancer, and had some correlation with lung cancer. No significant correlations were observed between EGFR or PCNA overexpression and lymph node metastasis.

Abstract ED01-01: Somatic alterations in human cancer genomes.

Abstract The advent of high-throughput genome sequence analysis has enabled systematic discovery of somatic genome alterations through genome, exome, and transcriptome sequencing of paired human cancer and germline nucleic acids. These discoveries have led to the identification of somatic genome alterations in new pathways including epigenetic regulators, RNA splicing factors, and immune response regulators, in addition to previously known signal transduction, cell cycle, and transcriptional pathways. I will discuss alterations in these pathways by copy number disruption, mutation, and re-arrangement, especially as seen through the lens of recent and ongoing large-scale studies of lung adenocarcinoma and squamous cell lung carcinoma genomes, from The Cancer Genome Atlas and beyond– and will discuss the implications for the understanding of cancer pathogenesis and for the development of targeted and immunomodulatory therapies. Citation Information: Mol Cancer Ther 2013;12(11 Suppl):ED01-01. Citation Format: Matthew L. Meyerson. Somatic alterations in human cancer genomes. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr ED01-01.

Abstract CN05-02: Targeting the Fanconi anemia/BRCA pathway.

Abstract Cancer cells have genomic instability resulting from acquired defects in DNA repair. One DNA repair pathway, the Fanconi Anemia/BRCA homologous recombination pathway (Kennedy and D'Andrea, Genes & Development 19:2925, 2005), is defective in many human cancers, including breast, ovarian, pancreatic, and lung neoplasms. Disruption of the FA/BRCA Pathway results in the characteristic chromosome instability and radiation/crosslinker hypersensitivity of these tumors. In general, loss of one DNA repair pathway often leads to hyperdependence on another pathway for tumor cell survival. This hyperdependence offers a unique opportunity for the development of anticancer therapeutics. For instance, FA/BRCA pathway deficient tumors are hyperdependent on base excision repair (BER) and, accordingly, these tumors are hypersensitive to single agent treatment with PARP1 inhibitors which block BER. Our laboratory efforts are focused on profiling the FA/BRCA pathway and the other five major DNA repair pathways in tumor cells (Kennedy and D'Andrea, JCO 24: 3799, 2006). Each DNA repair pathway has a characteristic protein biomarker and repairs a specific type of DNA lesion. By profiling these pathways in primary tumor samples with activation-specific antibodies to DNA repair proteins, we are developing methods to: 1) predict the sensitivity of tumors to conventional chemotherapy and radiation (Personalized Medicine); 2) to subset tumors for their sensitivity to novel classes of DNA repair inhibitors (i.e., PARP1, Chk1, and ATM inhibitors); and 3) to screen for new small molecule inhibitors of other DNA repair pathways. This combination of novel DNA repair inhibitors, conventional DNA damaging agents, and DNA repair biomarkers offers new opportunities for developing more effective anticancer therapy. The FA/BRCA pathway was elucidated through the systematic study of the rare inherited chromosome breakage disorder, Fanconi Anemia (Moldovan and D'Andrea, Ann Rev Genet 43:223, 2009). FA is an autosomal recessive disease characterized by bone marrow failure, congenital malformations, and cellular sensitivity to cisplatin, mitomycin C, and other DNA interstrand crosslinking agents. Patients with FA develop hematopoietic malignancies and squamous cell carcinomas. Based on somatic cell fusion studies, there are sixteen FA complementation groups (A, B, C, D1, D2, E, F, G, I, J, L, M, N, O, P, Q), and the corresponding gene for each of these complementation groups has been identified. Interestingly, the sixteen FA proteins cooperate in a common cellular pathway in normal human cells, referred to as the Fanconi Anemia/BRCA pathway. Eight of the FA proteins (A, B, C, E, F, G, L, M) are assembled in a core complex (the FA core complex) which is an active ubiquitin E3 ligase. In response to DNA damage, the FA core complex modifies (monoubiquitinates) the downstream FANCD2 protein. Monoubiquitinated FANCD2 translocates to nuclear foci where it interacts with the FANCD1/BRCA2 protein and participates in the process of homologous recombination DNA repair. Additional FA proteins (namely, FANCJ/BRIP1 and FANCN/PALB2) function downstream of FANCD2 monoubiquitination. At least three of the FA genes (FANCD1, FANCJ, and FANCN) are inherited breast cancer susceptibility genes. Disruption of any step in the FA/BRCA pathway results in the common clinical and cellular phenotype of FA patients. Human tumor cells, derived from cancer patients from the general (non-FA) population, often exhibit genomic instability. Genomic instability of tumors has important clinical consequences. On the one hand, genomic instability gives the tumor the ability to break and fuse chromosomes, inactivate tumor suppressor genes, form novel oncogene fusions, and amplify drug resistance genes. Thus, the tumor with genomic instability may become more malignant and drug resistant over time. On the other hand, in order to achieve a state of genomic instability, a tumor cell must inactivate one of its major DNA repair pathways. This inactivation appears to account, at least in part, for the selective hypersensitivity of cancer cells to the cytotoxic effects of conventional anticancer radiation and chemotherapy. Recent studies indicate that some human tumors inactivate the FA/BRCA pathway. Pathway inactivation may result from somatic mutation of genes in the FA/BRCA pathway or by epigenetic silencing. For instance, methylation of one of the FA genes, FANCF, has been implicated as a mechanism for genomic instability in a wide variety of cancers, including ovarian, breast, lung, cervical, and head and neck squamous cell carcinomas. Somatic inactivation of the FA/BRCA pathway appears to account for the genomic instability and the cisplatin hypersensitivity of many of these cancers. In my presentation, I will review the features of the FA/BRCA pathway in human cells, and how knowledge of this pathway in cancers can lead to the prediction of drug response. Citation Information: Mol Cancer Ther 2013;12(11 Suppl):CN05-02. Citation Format: Alan D. D'Andrea. Targeting the Fanconi anemia/BRCA pathway. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr CN05-02.