bluuebunny/crossref_metadata_2025 huggingface dataset:
Weakly Compact Uniform Attractor for the Nonautonomous Long-Short Wave Equations
Authors: Cui Hongyong , Xin Jie , Li Anran
Published: March 2013

Solutions and weakly compact uniform attractor for the nonautonomous long-short wave equations with translation compact forces were studied in a bounded domain. We first established the existence and the uniqueness of the solution to the system by using Galerkin method and then obtained the uniform absorbing set and the weakly compact uniform attractor of the problem by applying techniques of constructing skew product flow in the extended phase space.
John Nyman and the Economics of Health Care Moral Hazard
Authors: Kelman Sander , Woodward Albert
Published: March 2013

In 2003, John Nyman published The Theory of Demand for Health Insurance. His principal contributions are (1) to replace the previously unexamined axiom of risk avoidance with the axiom of welfare maximization; (2) to uncover a misinterpretation in the literature on moral hazard, namely, the insurance payoff as a price reduction, rather than as an income transfer. The immediate consequence of these reformulations is to recognize insurance-induced health care utilization as resulting in an increase in social welfare. Despite its evident validity and enormous implications, Nyman’s work has received very little attention or recognition in the health economics literature.
Robust On-Demand Multipath Routing with Dynamic Path Upgrade for Delay-Sensitive Data over Ad Hoc Networks
Authors: Kumar Sunil , Khimsara Siddharth , Kambhatla Kashyap , Girivanesh Kalyani , Matyjas John D. , Medley Michael
Published: March 2013

Node mobility in mobile ad hoc networks (MANETs) causes frequent route breakages and intermittent link stability. In this paper, we introduce a robust routing scheme, known as ad hoc on-demand multipath distance vector with dynamic path update (AOMDV-DPU), for delay-sensitive data transmission over MANET. The proposed scheme improves the AOMDV scheme by incorporating the following features: (i) a routing metric based on the combination of minimum hops and received signal strength indicator (RSSI) for discovery of reliable routes; (ii) alocal path updatemechanism which strengthens the route, reduces the route breakage frequency, and increases the route longevity; (iii) akeep alivemechanism for secondary route maintenance which enables smooth switching between routes and reduces the route discovery frequency; (iv) apacket salvagingscheme to improve packet delivery in the event of a route breakage; and (v) low HELLO packet overhead. The simulations are carried out in ns-2 for varying node speeds, number of sources, and traffic load conditions. Our AOMDV-DPU scheme achieves significantly higher throughput, lower delay, routing overhead, and route discovery frequency and latency compared to AOMDV. For H.264 compressed video traffic, AOMDV-DPU scheme achieves 3 dB or higher PSNR gain over AOMDV at both low and high node speeds.
Applications of Littlewood-Paley Theory forB˙σ-Morrey Spaces to the Boundedness of Integral Operators
Authors: Komori-Furuya Yasuo , Matsuoka Katsuo , Nakai Eiichi , Sawano Yoshihiro
Published: March 2013

The boundedness of the various operators onB˙σ-Morrey spaces is considered in the framework of the Littlewood-Paley decompositions. First, the Littlewood-Paley characterization ofB˙σ-Morrey-Campanato spaces is established. As an application, the boundedness of Riesz potential operators is revisted. Also, a characterization ofB˙σ-Lipschitz spaces is obtained: and, as an application, the boundedness of Riesz potential operators onB˙σ-Lipschitz spaces is discussed.
Optimal Power Flow Using Adaptive Fuzzy Logic Controllers
Authors: Abusorrah Abdullah M.
Published: March 2013

This paper presents an approach for optimum reactive power dispatch through the power network with flexible AC transmission systems (FACTSs) devices, using adaptive fuzzy logic controller (AFLC) driven by adaptive fuzzy sets (AFSs). The membership functions of AFLC are optimized based on 2nd-order fuzzy set specifications. The operation of FACTS devices (particularly, static VAR compensator (SVC)) and the setting of their control parameters (QSVC) are optimized dynamically based on the proposed AFLC to enhance the power system stability in addition to their main function of power flow control. The proposed AFLC is compared with a static fuzzy logic controller (SFLC), driven by a fixed fuzzy set (FFS). Simulation studies were carried out and validated on the standard IEEE 30-bus test system.
Abstract A31: Predictive and prognostic value of KRAS mutations in advanced non-small cell lung cancer (NSCLC) patients treated with chemotherapy
Authors: Mellema Wouter W. , Dingemans Anne-Marie C. , Smit Egbert F. , Derks Jules , Heideman Daniëlle A.M. , van Suylen Robertjan , Thunnissen Frederik B.
Published: March 2013

Introduction: K-Ras mutation is thought to be related with poor outcome of NSCLC patients with an advanced stage of disease. The role of K-Ras mutation as a predictor of response for these patients treated only with chemotherapy is poorly understood.

Patients and methods: From a retrospective database from 2 university hospitals all patients with advanced stage, nonsquamous, NSCLC treated with palliative platinum containing chemotherapy were selected. Patients were included when archive tumour tissue was available for DNA extraction. High resolution melting followed by PCR sequencing was performed for KRAS exon1 and 2. Response to treatment was assessed by RECIST. The databases were matched by treatment.

Results: A total of 179 patients were included in this study. All patients were treated with chemotherapy and were no candidate for treatment with curative intent. There were 59 patients with a K-Ras mutation (32.9%). There was no significant difference in basic characteristics between the group with a K-Ras mutation and the group with K-Ras wild-type. Mean age 59 years (range 32–83 years), 108 males and 71 females. ECOG performance score 0/1/2/3 73/77/13/2. Thirty patients with stage IIIb disease, 149 patients with stage IV disease. Progression as best response to treatment was reported in 21/59 patients with a K-ras mutation (35.6%) and 34/120 patients (28.3%) with a K-Ras wild type, this was not significant (p=0.611). The median progression free survival (PFS) in the group with a K-Ras mutation was 4.1 months (95% CI 3.12–5.0 months) compared to 5.4 months (95% CI 4.4–6.4 months) in patients with K-Ras wild type (p=0.202). The overall survival (OS) in the group with a K-Ras mutation, was 8.3 months (95% CI 5.2–11.4 months) compared to 12.5 months (95% CI 10.2–14–7 months) in patients with K-Ras wild type (p=0.037).

Conclusion: In our series K-Ras mutation was not predictive for response to platinum containing chemotherapy in patients with advanced NSCLC. K-Ras mutational status was prognostic for overall survival.
Abstract A5: Characterizing and targeting B cells in lung cancer patients
Authors: Bruno Tullia C. , Kern Jeffrey , Slansky Jill E.
Published: March 2013

Lung cancer is the leading cause of cancer death in men and women, both in the United States and the world. The current 5-year survival rate for lung cancer in the United States for all races is a discouraging 16% and has not changed dramatically during the past 30 years. The biggest reason for the discouraging survival statistics is that the majority of lung cancer subjects present with late-stage disease that is not curable by current therapies i.e. chemotherapy and radiation. However, in the last several years novel therapies have emerged to make lung cancer therapy better tolerated and more effective. As tumor immunologists, we are particularly interested in developing an immunotherapy that will help to engage the immune system in targeting and destroying tumor cells. In a comprehensive survey of the immune infiltrate in non-small cell lung cancer (NSCLC) patients, we discovered that the frequency of the B cell infiltrate in the tumor versus the normal tumor-adjacent tissue was significantly increased in comparison to other immune subsets, specifically, CD4, CD8, and regulatory T cells as well as NK and NKT cells. Further preliminary studies on the characterization of these B cells suggest that they have proliferated and acquired a memory phenotype. We will continue to characterize the B cells in the tumor microenvironment to determine if they are activated as well as functional. If the B cells in the tumor microenvironment are activated, we will test if the cells are responsive to antigens currently in clinical trial like MUC-1 as well as new antigens in the cancer-testis classification like XAGE-1b. We will test if the antigens are stimulating the B cells through the BCR and if these antigens will be presented to CD4 T cells in vitro, which would lead to the stimulation of the CD4 T cells. If the B cells are not activated, we will then query whether or not the B cells could have a suppressive phenotype, which will be determined by assaying the types of cytokines produced by the B cells using a cytokine array. As we continue to further characterize the B cells, we will also continue to monitor the various immune subsets in the three subtypes of NSCLC: adenocarcinoma, squamous cell carcinoma and large cell carcinoma. This immunophenotyping of the immune subsets will aid us in the identification of the appropriate immune cell populations to target for immunotherapy in the three subtypes of NSCLC. Ultimately, we aim to identify a novel target for an immunotherapy clinical trial in lung cancer patients.
Abstract B12: Predictibility of response to anti-EGFR treatment with respect to assessment of receptor status in different geographical population with lung cancer (NSCLC)
Authors: K V Veerendra Kumar
Published: March 2013

Lung cancer affecting populations in different geographical areas varies with respect to incidence, smoking history, histology and response to treatment with monoclonal antibodies inhibiting EGFR.

Assessment of EGFR status in lung cancer varies with respect to methodology, type of antibody etc, resulting in wide variation in reported incidence of positivity. In addition EGFR receptor is a polyclonal antigen (eg: EGFR1, EGFR2) and standardization of antigen methodology of assessment and interpretation of the test results is not yet uniformly accepted worldwide. Response to treatment with monoclonal antibodies to EGFR in lung cancer patients vis a vis receptor status is varied in different geographical areas. Study in Erlotinib has shown response in selected sample of patients in western population whereas there are no well established clinical trials in Asian population establishing relevance of receptor status with respect to clinical response to EFGR antibodies.

It is well known fact from different clinical trials that Asian especially Southeast Asian populations with lung cancer respond differently in comparison to European population (cacuasions). But receptor status and response to treatment not known.

Hence it is opinioned that Asian and especially Southeast Asians population's treatment with monoclonal antibodies should be given a clinical trial irrespective of receptor status or unknown receptor status. However clinical trials with respect to response v/s receptor status should be conducted. Till then these populations should not be denied treatment with monoclonal antibodies in lung cancer (NSCLC).
Abstract B19: Fetal lung adenocarcinoma: Characterization of primary cell culture and in vitro drug response
Authors: Menna Cecilia , Ibrahim Mohsen , Peruzzi Daniela , Pacini Luca , Ciancamerla Michela , Calogero Antonella , Rendina Erino Angelo
Published: March 2013

Purpose: Fetal lung adenocarcinoma (FLAC) is an extremely rare subtype of pulmonary adenocarcinoma estimated to be the 0.5% of all lung cancers. Therefore a standardized model of FLAC primary cell culture and evidence-based treatment guidelines are not available. The aim of this study is to characterize FLAC primary cell culture, to evaluate the biological features of this uncommon tumor and the in vitro drug response.

Methods: FLAC specimen was obtained from a 59-years-old female patient, cultured in vitro with DMEM-F12 medium supplemented with 5% FBS and maintained in culture for more than 6 passages. Primary cells from a lung adenocarcinoma specimen were as well cultured and used as control for all the in vitro experiments. Immunohistochemistry for β-catenin, TTF-1, Ki67, cKit, MDM2, p53, S-100, EGFR, E-cadherin, cytokeratin 7 and 8/18 was performed on paraffin-fixed specimen. Immunofluorescence on cells in culture was assessed for MDM2, p53, CD133. Specimen and cells in culture were analyzed by real time PCR for oncogenic mutations (KRAS, EGFR) and for a stem cells gene panel. In vitro tumorigenicity was tested by plating efficency and soft agar colony formation assays. Cell proliferation was determined and a cell viability test was performed after treatment of FLAC cells with tyrosine kinase inhibitors (erlotinib and gefitinib), temozolomide (TMZ), cisplatin, gemcitabine and cysplatin+gemcitabine by MTS assay.

Results: A primary cell culture of human FLAC was characterized and standardized. FLAC histotype was confirmed by immunohistochemistry. Immunofluorescence showed a positivity for MDM2, but not for p53 and CD133. By molecular analysis, DNA from both specimen and cells in culture was found to harbor epidermal growth factor receptor (EGFR) kinase domain non-sense mutation in exon 20 and no mutations in the K-ras gene. Primary FLAC cells showed a colony formation activity and the soft agar assay revealed an anchorage-indipendent growth. At 48 hours FLAC cells proliferation was inhibited of 56% and 41% by erlotinib and gefitinib respectively, with an IC50 value of 10 μM, compared with the control proliferation (lung adenocarcinoma cells) inhibited of 25% and 60% by erlotinib and gefitinib respectively (p=0,17; p=0,005). However, FLAC cell viability at 48 hours after being treated with cisplatin 10 μM was 3%, compared with the control viability that was 16% (p=0,007).

Conclusion: This well-characterized FLAC primary cell culture provide a unique model for future studies in lung cancer biology and drug response. Although chemosensitivity to cisplatin was determined, erlotinib and gefitinib were effective against FLAC cells and appear to represent a novel targeting approach for the treatment of this rare tumor.
Abstract B36: Micronome-wide explorations of NSCLC: microRNA-seq discovery
Authors: Gombar Saurabh , Shi Miao , Locker Joseph , Zhu Changcheng , Keller Steven , Suh Yousin , Spivack Simon
Published: March 2013

Background: Recent studies have revealed that microRNAs play important regulatory roles in carcinogenesis. Some key miRNAs may regulate lung carcinogenesis and be useful for lung cancer early detection. For discovery, we used high-throughput next-gen sequencing technology to investigate miRNA expression profiles in a study of 25 lung adeno- and squamous cell carcinomas, paired with far-adjacent noncancerous lung tissues from the same donors.

Method: Total RNA was isolated by Trizol from 14 pairs of lung adenocarcinomas, 11 squamous cell cancers, and paired noncancerous lung tissues, from the same patients. Small RNA library of each total RNA was constructed with Illumina's Small RNA Sample Prep Kit, followed by high-throughput sequencing using the Illumina IIa platform. In parallel, transcriptome microarrays (Affymetrix GA 1.0) were run on these same paired samples. Statistical differences in microRNA expression between groups were analyzed by GenePattern software, logistic regression, and other methods.

Results: For adeno, 128 micros were differentially expressed and 63 were two-fold differentially expressed between the tumors and non-tumor tissue; for squamous, the numbers of altered microRNAs were 42 overall, and 25 >two-fold. Among adenocarcinoma, the top 10 upregulated were miRs-147b, 577, 877, 556, 449c, and others; and downregulated were miRs 516b, 1251, 486, 139, 516a, and others. Among squamous cell carcinomas, the top up-regulated miRs were 708, 1259, 494, 200a, 216b, and others; and down-regulated miRs were 184, 1251, 144, 516a, 584, and others. MiRs 1259, 135b, 200a, 21, 494, 708 were upregulated in both tumor histologies. When the highest expressed microRNAs were correlated with the most altered mRNA transcripts, putative mRNA targets were more often anti-correlated to the individual microRNAs (Chi Square, p = 4.6 × 10-6), and this anticorrelation generally held true for the top differentially expressed microRNAs and their putative mRNA targets, and within histology strata.

Conclusion: The lung cancer microRNA complement differs from non-tumor tissue, adeno- differs from squamous, and many of the most dysregulated miRs appear to be anticorrelated with putative mRNA targets. Which of the putative mRNA targets are actual biological targets awaits experimental functional work. Once constructed, the combined microRNA:mRNA signature of lung cancer is likely to be more informative for biologic potential than either signature alone.

[Funding source: NCI 1RC1 CA145422-01; 1K24-CA139054-01]
Abstract B43: SNP Q787Q of EGFR gene and efficacy of EGFR-TKI in patients with non-small cell lung cancer
Authors: Young-Chul Kim , Kyu-Sik Kim , In-Jae Oh , Hee-Jung Ban , Kook-Joo Na , Sung-Ja Ahn , Sang-Yun Song , Song Choi , Yoo-Duk Choi
Published: March 2013

Background: Many activating mutations in epidermal growth factor receptor (EGFR) gene have been correlated with sensitivity to EGFR tyrosine kinase inhibitors (TKIs). Single nucleotide polymorphism (SNP) in exon 20 of EGFR gene (2361G>A transition) does not alter the amino acids of glutamine at codon 787 (Q787Q). We reviewed the relationship between this SNP and EGFR-TKI sensitivity.

Patients and Methods: Twenty-five patients (male 19, female 6) with only SNP Q787Q in EGFR exon 20 were analyzed. Tissue or cytologic specimens were used for analysis of mutations in exon 18 to 21 of EGFR gene by direct sequencing. Treatments with EGFR-TKIs were performed in 16 patients and response evaluations were eligible in 15 cases.

Results: In 15 response evaluable patients, there were 2 partial responses (PR) and 5 stable diseases (SD) [response rate: 13.3%, disease control rate: 46.7%]. Median time to progression (TTP) was 392 days (range: 105∼511). Five patients showed longer than 3 months of TTP (PR 2 and SD 3, adenocarcinoma 4 and squamous cell carcinoma 1, gefitinib 4 and erlotinib 1).

Conclusion: Thirteen percent of NSCLC with 2361G>A transition without other activating mutations responded to EGFR-TKIs.
Abstract B45: Upfront genomic testing in non-small cell lung cancer (NSCLC) patients: Preliminary result of the MSN study
Authors: Planchard David , Lacroix Ludovic , Le Chevalier Thierry , Soria Jean-Charles , Besse Benjamin , Celebic Aljosa , Ngocamus Maud , Auger Nathalie , Lueza Beranger , Saulnier Patrick , Rahal Arslane , Kalai Khadja , Dorfmuller Peter
Published: March 2013

Background: Recent advances in lung cancer have identified potential driver mutations that may be targeted. To identify new predictors of response as well as novel targets for therapy, we have initiated a comprehensive large-scale sequencing analysis of genes potentially mutated in NSCLC.

Methods: Genomic DNA was extracted prospectively from untreated advanced NSCLCs. All tumors were obtained on IRB-approved protocols and after patients' consent (MSN trial “Melanoma – Small-cell lung cancer – Non-small cell lung cancer”). Pathology specimens were macrodissected in order to obtain more than 30% of tumor cells and, after DNA extraction, 96 selected exons from 30 genes were analyzed by Sanger sequencing. ALK rearrangements were detected by fluorescence in-situ hybridization. All results were discussed monthly in a molecular thoracic multidisciplinary staff. Second or third lines of treatment were adapted to mutation profiling.

Results: Thus far (between 09/01/2010 and 06/01/2011), 82 tumors have been analyzed for AKT, ALK, APC, BRAF, CTNNB1, EGFR, ERBB2, FBXW7, FGFR2-3, GNAQ, GNAS, HRAS, KIT, KRAS, MAP2K1-2, MET, NOTCH1, NRAS, PDGFRA, PI3KCA, PTEN, RET, STK11, TP53, and VHL mutations. The median age was 60 years (range 26–76), 29 (35%) were female, 58 (70%) had adenocarcinoma, 72 (67%) were former/current smokers. Nine patients had incomplete genomic analysis due mostly to insufficient tumor cells in the specimen or poor quality DNA. Median tumor cells ratio was 50%. Mutations were identified in 43/82 (52%) patients (EGFR: 9; KRAS: 10; STK 11: 8; BRAF: 4; MET: 2; PTEN: 2; NRAS: 1; ERBB2: 1; PIK3CA: 1…) of whom 11 had concurrent mutations. EGFR and KRAS mutations were mutually exclusive. No mutations were identified among 5 never smokers analyzed. The median time to complete testing for this initial phase was 30 days. Half of patients with genomic alterations were treated with molecularly targeted therapy based on their genetic alteration.

Conclusions: Mutational profiling of NSCLC is feasible, can distinguish relevant molecular subsets of lung cancer, and may present an impact on treatment at our cancer institute. Furthers sequencing are in progress and updated results will be presented in January 2012.
Abstract IA12: Redox pathways as platforms for drug development
Authors: Tew Kenneth D.
Published: March 2013

While oxygen based metabolism is the most evolved and efficient method for producing energy, the generation of reactive oxygen electrophiles creates physiological stress. Sulfur has variable valencies, and as a consequence its nucleophilicity allows it to react with electrophilic species and exist in a variety of oxidation states. Lung cancer cells, in common with most cancers have aberrant redox homeostasis, a property that has provided a platform for drug development for both therapeutic and prevention approaches. There are >200,000 cysteines encoded by the human genome and the most common non-protein form of sulfur in cells is glutathione (GSH). Glutathione S-Transferase P (GSTP) is one member of the family of GST's that contribute to cellular thiol and redox balance. GSTP is characteristically expressed at high levels in some tumors (including lung) and in drug resistant cancer cells. Targeting of GSTP with small molecules has identified a specific inhibitor (Telintra) that has myeloproliferative properties and may be useful in combination drug regimens. There are also a number of preclinical and clinical (e.g. Telcyta) GSTP activated pro-drug candidates that are at various stages of testing. A recently completed Phase III trial in NSCLC used NOV-002 (a glutathione disulfide mimetic) to alter the toxicity profiles of standard anticancer drugs by modulating intra- and extracellular redox balance. Changes in cellular redox homeostasis play an essential role in numerous signaling cascades, including those associated with proliferation, inflammatory responses, apoptosis, and senescence. Moreover, GSTP has been implicated in catalyzing S-glutathionylation (a post-translational modification adding glutathione to cysteine residues) of a number of proteins that fall into functional clusters important to these signaling events. Recent studies suggest that S-glutathionylated proteins may serve as clinical biomarkers of drug response. Finally, many chemoprevention drugs are designed to stimulate the production of protective Phase II detoxification enzyme systems in target normal tissues. Many of these agents are sulfur based and most contain electrophilic centers that show selectivity for the Nrf2 transcription factor and genes that contain antioxidant response elements (ARE). Chronic, low dose exposure to these agents may have short-term prevention potential, but the down sides of long term impact of low levels of electrophilic stress remains to be answered.
Abstract IA17: TSLP-driven inflammation fosters development of epithelial tumors
Authors: Palucka Anna Karolina , Xu Kangling , Pedroza Alexander Gonzalez , Wu Te-Chia , Zurawski Sandra , Marches Florentina , Zurawski Gerard , Liu Yong-Jun , Banchereau Jacques
Published: March 2013

Solid tumors are often associated with aseptic inflammation. There are two types of inflammation that have opposing effects on tumors, chronic inflammation that promotes cancer cell survival, and metastasis, and acute inflammation which triggers cancer cell destruction. Chronic inflammation is often linked with the presence of type 2-polarized macrophages (M2), which are induced by Th2 cytokines, IL-4 and IL-13.

Our recent studies have demonstrated the presence in breast cancer tumors of inflammatory Th2 cells, which produce IL-13, IL-4, and TNF. These CD4+ T cells appear to play a key role in the disease as they accelerate breast tumor development in a xenograft model through the production of IL-13. Breast tumors appear to play a critical role in conditioning the infiltrating myeloid DCs (mDCs) to induce such inflammatory Th2 cells. Our most recent results suggest that thymic stromal lymphopoetin (TSLP) secreted by cancer cells plays a role in mDCs conditioning. Breast cancer cell lines and primary tumors from patients show TSLP protein expression. TSLP-neutralizing antibodies block the upregulation of OX40L by mDCs exposed to tumor supernatant and consequently block mDCs capacity to generate inflammatory Th2 cells in vitro. The TSLP production is mediated by PAR2-signaling in cancer cells. PAR2 is expressed by all 4 breast cancer cell lines, and TSLP production can be induced upon PAR2-agonist peptide treatment. Similar observations were made by us and others in pancreatic cancer as well as lung cancer. Thus, TSLP could serve as therapeutic target in tumors of epithelial origin.
Abstract IA6: Gene fusions in lung cancer
Authors: Beer David G.
Published: March 2013

Gene fusions represent one type of genetic alteration recently detected in lung cancer that results from chromosomal rearrangements. A fusion joining the echinoderm microtubule-associated protein-like 4 (EML4) and anaplastic lymphoma kinase (ALK) genes was detected as a transforming gene in a small subset of lung adenocarcinomas. Importantly, patients with these fusions show responsiveness to ALK kinase inhibitors. Additional studies have identified multiple EML4-ALK gene fusion variants and the histopathological characteristics of the lung tumors identified. Various methods utilized to detect lung gene fusions have included retrovirus-mediated expression screening with 3T3 cell transformation assays, FISH, mRNA/protein expression, RACE-coupled PCR sequencing, massively parallel paired-end and transcriptome sequencing. Additional novel gene fusion events in lung cancer have recently been identified using these and newly defined approaches in small percentages of primary lung tumors. Potential mechanisms underlying the production and triggers for gene fusion events will be discussed.
Abstract IA8: Role of biomarkers in the early detection of lung cancer
Authors: Massion Pierre P.
Published: March 2013

While lung cancer remains the leading cause of cancer-related death in the world with a 16% overall survival rate, we now know that early detection by low dose chest CT saves lives. The implementation of screening programs represents a huge challenge. Research is needed to address who will benefit from screening, for how long, and with what frequency we should screen. Using biomarkers to select the most at-risk population, to detect the disease while measurable and yet not clinically apparent, and to predict survival, has been the goal of many investigations. Recent advances in molecular strategies and analytical platforms including genomics, epigenomics, proteomics, and metabolomics, have identified increasing numbers of potential biomarkers. These molecular approaches are applied to a variety of samples collected by non invasive methods including blood, urine, exhaled breath condensate, bronchial specimens, saliva, and sputum. None have yet moved to the clinical setting. In this presentation, we will define clinical contexts where diagnostic biomarkers may have utility in the management of lung cancer, we will review the state of development of candidate diagnostic biomarkers, and finally, we will discuss critical aspects of study design in biomarkers research.
Abstract PR5: A phase II study of sorafenib in patients with stage IV non-small cell lung cancer (NSCLC) with a K-Ras mutation
Authors: Mellema Wouter W. , Dingemans Anne-Marie C. , Groen Harry J.M. , van Wijk Atie , Burgers Sjaak , Kunst Peter W.A. , Thunnissen Frederik B. , Heideman Daniëlle A.M. , Smit Egbert F.
Published: March 2013

Background: In a pilot study (1) we found sorafenib to display clinical activity against patients with K-Ras positive NSCLC, sufficient for formal phase II testing. Methods: Patients with K-Ras mutated NSCLC that progressed after at least 1 platinum containing regimen with adequate organ reserve, ECOG 0–2, who provided written informed consent according to local IRB regulations were eligible. A tumor biopsy confirming the presence of a K-Ras mutation was mandatory. Treatment consisted of sorafenib 400 mg BID until disease progression or unacceptable toxicity. Dose reductions and delays were specified per protocol in the face of CTC toxicities grade 3 and 4. Primary endpoint: Rate of No Progression (NPR) at 6 weeks. Secondary endpoints: duration of response, progression free survival (PFS), overall survival and treatment related toxicities. A 2-stage design was implemented (Simon's optimal design; p0=40%, p1=60%, alpha=0.05, beta=0.20) for a total number of 48 pts. Results: 59 patients were entered between May 1st 2010 and February 18 2011. Median age was 58 (range 46–79) years, 17 Male/42 Female, ECOG PS 0/1/2 23/32/4. 57 patients started treatment. At 6 weeks 5 PR, 25 SD, 27 PD were observed; NPR 52.6%. Fifteen patients stopped treatment before 6 weeks, of which 10 patients stopped due to (clinical) progression. Median duration of treatment was 9 (range 0–62) weeks, 2 patients are still on treatment. Median duration of response was 32 (range 5–58) weeks. Median PFS was 2.3 months, median OS was 4.9 months, 14 patients still alive. Dose modifications were realized in 21 patients, of whom 4 discontinued treatment. Most reported adverse events were fatigue (6.4%), hand-foot reaction (5.7%), dyspnea (5.6%), anorexia (3.7%) diarrhea (3.6%) and cough (3.6%). Sorafenib related grade 3-4 toxicity was reported in 10 patients. Five patients with grade 3 skin toxicity, 4 patients with grade 3 gastrointestinal toxicity and 1 patient with both grade 3 metabolic abnormalities and a grade 3 pneumonitis. Conclusion: Treatment with sorafenib has relevant clinical activity in patients with K-Ras mutational status. Further randomized study with this agent is warranted.

This abstract is also presented as Poster B18.
Abstract A17: Loss of DEPTOR increases resistance to EGFR tyrosine kinase inhibitors by upregulating EGFR and MAPK signaling
Authors: Cowell Catherine F. , De Bruin Elza C. , Howell Michael , Downward Julian
Published: March 2013

Epidermal growth factor (EGFR) activating mutations occur in 10% to 40% of non-small cell lung cancer (NSCLC), and can be successfully targeted with EGFR tyrosine kinase inhibitors (EGFR-TKIs), achieving up to 80% response rate in patient populations selected for presence of activating mutations. However, almost all patients develop resistance to treatment within a year, and in at least 30% of cases the cause of resistance is unknown. To identify novel mechanisms of resistance we executed a genome-wide siRNA screen in PC9 cells, an EGFR-TKI sensitive NSCLC cell line. We completed the screen in untreated and drug treated conditions, and selected hits as those genes that had a significantly different effect on survival in drug treated conditions compared to untreated controls. RNAi-mediated knockdown of DEPTOR, a negative regulator of mTOR, increased resistance of PC9 cells to erlotinib (an EGFR-TKI). Cells depleted of DEPTOR have higher levels of mTOR signaling as seen by greater phosphorylation of AKT, and SGK1 substrate NDRG1. DEPTOR loss leads to enhanced resistance to erlotinib treatment by increasing abundance of EGFR protein, thereby augmenting MAPK signaling in basal and erlotinib treated conditions. This upregulation of EGFR appears dependent on mTOR activation, as mTOR kinase inhibitors can resensitize DEPTOR deficient cells to erlotinib treatment. Also, combination treatment of erlotinib with a MEK inhibitor restores sensitivity to erlotinib of cells in which DEPTOR expression has been reduced, indicating increased MAPK signaling contributes to resistance in these cells. mTOR activation is known to cause DEPTOR degradation, therefore our results may have implications for a wide subset of NSCLC where mTOR signaling is upregulated.
Abstract A19: A qPCR assay, OncoScore Colon, predicts resistance to cetuximab in formalin-fixed, paraffin-embedded colorectal cancer tissue independent of KRAS status
Authors: Eddy Sean , Lee Byung-In , Macarulla Teresa , Tabernero Josep , Monforte Joseph , Rhodes Daniel R. , Williams Paul , Tomilo Mark , Sadis Seth , Wyngaard Peter , Vo Lien , Oades Kahuku , Kim Hyunsoo , Wang Yipeng
Published: March 2013

Gene expression modules derived from an unsupervised analysis of 20 independent microarray datasets comprising more than 2,000 colorectal cancer patients were identified. Each module represents a set of highly co-expressed genes related to an important aspect of underlying cancer variability. Modules containing genes related to epithelial and mesenchymal biology associated with sensitivity and resistance to EGFR family targeted inhibitors (gefitinib and lapatinib), respectively. In retrospective analysis of clinical samples, the epithelial-mesenchymal axis associated with cetuximab response in two independent patient cohorts. The first study was a Phase II clinical trial (Khambata-Ford et al., J Clin Oncol, 2007) with accompanying microarray data from pre-treatment metastatic colorectal tumor biopsies. Expression of the modules was determined by normalizing and averaging co-expressed module genes. Patients with a more epithelial and less mesenchymal module expression profile were enriched for cetuximab response. An independent cohort of patients was analyzed using module scores that were generated from a qPCR gene expression module test, OncoScore™ Colon, which quantifies modules by averaging three representative module genes relative to housekeeping genes using formalin-fixed-paraffin-embedded primary tumor samples. In these patients, presence of the mesenchymal module was significantly associated with a decrease in progression free survival. Notably, the status of the mesenchymal module was independent of KRAS mutation status—as KRAS mutations occurred in both mesenchymal module-positive and -negative patients. Further clinical studies are ongoing to continue to support the development of the OncoScore™ Colon assay and to further test the predictive capacity of the module with regards to cetuximab resistance and other MAPK pathway inhibitors. This study demonstrates the value of a gene expression module-based qPCR panel for stratifying colorectal cancer patients for treatment response, and suggests that our approach may have immediate utility for cetuximab treatment response prediction.
Abstract A31: Roles of Nogo-B receptor (NgBR) in regulating Ras translocation and modulating breast cancer cell sensitivity to chemotherapeutic drug and irradiation
Authors: Zhao Baofeng , Liu Zhong , Kumar Suresh , Miao Robert Q.
Published: March 2013

Ras, a critical oncogene, is one of small GTPase family members. It is active when bound to GTP and inactive when bound to GDP. The activation process needs membrane translocation of Ras because its activators are recruited by membrane receptors, such as EGF receptors. In addition, appropriate localization of activated Ras also is critical for interacting with the downstream effectors. Therefore, regulation of Ras membrane translocation is very important for Ras function.

Nogo-B receptor (NgBR) is a type I receptor with a single transmembrane domain. Our previous work has shown that axon guidance gene family Nogo-B and its receptor (NgBR) are essential for chemotaxis and morphogenesis of endothelial cells in vitro and blood vessel formation in Zebrafish. Here, we demonstrate that NgBR can bind and recruit farnesylated Ras to plasma membrane, which results in the activation of PI3K-Akt and Raf-MEK-ERK signaling pathways in human breast cancer cells. Direct interaction between NgBR and farnesylated Ras has been demonstrated by co-immunoprecipitation of MBP-NgBR from incubation of purified recombinant farnesylated Ras and MBP-NgBR protein. Knockdown of NgBR in MDA-MB-231 human breast cancer cells remarkably reduces the amount of membrane-associated Ras. These results suggest that NgBR is a potential modulator for Ras translocation. Knockdown of NgBR expression in MDA-MB-231 breast cancer cells decreases cancer cell colony formation, and abolishes EGF-stimulated Ras activation and phosphorylation of Akt in MDA-MB-231 cells. In addition, overexpression of NgBR caused the transformation of NIH-3T3 cells via increasing Ras plasma membrane translocation and enhancing EGF-stimulated Ras signaling. Knockdown of NgBR decreases the survival and colony formation of MDA-MB-231 cells after irradiation and increases the cisplatin-induced apoptosis and Caspase-3 activity of MDA-MB-231 Cells. These results suggest that NgBR can increase MDA-MB-231 breast cancer cell sensitivity to chemotherapeutic drug and irradiation. Consequently, our preliminary results further demonstrated that inducible knockdown of NgBR in tumor xenograft of MDA-MB-231 cells in nude mice significantly retards tumor growth. Thus, these data demonstrate that NgBR is a unique receptor that recruits farnesylated Ras to the plasma membrane via its hydrophobic cytoplasmic domain and then activates both PI3K-Akt and Raf-MEK-ERK pathways in human breast cancer cells. Collectively, it is the first study to demonstrate that NgBR is a potential therapeutic target for inhibiting Ras-dependent growth of breast cancer and decreasing the sensitivity of breast cancer cells to chemotherapeutic drug and irradiation.
Abstract A32: Notch signaling regulates recurrence in a model of HER2 targeted therapy
Authors: Abravanel Daniel L. , Sterner Christopher J. , Pan Tien-chi , Belka George K. , Chodosh Lewis A.
Published: March 2013

Breast cancer is the leading cause of cancer-related deaths among women worldwide. While mortality rates have improved over the past thirty years, physicians still lack effective tools for the prevention or treatment of breast cancer recurrence. An essential first step in designing such therapies will be to elucidate the pathways that contribute to therapeutic resistance, dormancy, and recurrence.

Our laboratory has developed a series of mouse models that permit the conditional activation of oncogenes in the mammary glands of mice and can be used to recapitulate key features of breast cancer progression including dormancy and recurrence after targeted therapy. In the MMTV-rtTA;TetO-neu (MTB/TAN) model, treatment with doxycycline (dox) permits mammary specific activation of HER2/neu and drives primary tumor formation. Upon removal of dox and resultant down-regulation of HER2/neu, primary tumors regress as a consequence of oncogene addiction. However, a small population of tumor cells persists in a histologically identifiable residual lesion. After a period of cellular dormancy, residual tumor cells reenter the cell cycle in a stochastic manner and give rise to recurrent tumors, independent of HER2/neu signaling.

Recent evidence suggests that Notch signaling may play a role in breast cancer recurrence. For example, elevated Notch signaling in ductal carcinoma in situ lesions has been found to be associated with early recurrence after surgery. To build on these data, we analyzed microarray datasets from breast cancer patients and found that NOTCH1 expression levels are correlated with recurrence risk, suggesting that this pathway may play a role in human disease. Furthermore, in our mouse model system, we have found that Notch signaling is activated following HER2/neu downregulation. To assess the functional significance of this phenomenon, we performed in vitro clonogenic assays and in vivo recurrence assays. Activation of Notch signaling accelerated colony formation and recurrence, whereas Notch repression inhibited colony formation and recurrence. These data suggest that Notch signaling is both necessary and sufficient to promote recurrence following HER2/neu downregulation.

Taken together, our data support a model in which the Notch signaling pathway provides a mechanism by which HER2/neu driven tumors can escape therapy and cause recurrent disease. As gamma secretase inhibitors are currently in late-stage clinical trials for the treatment of breast cancer, work to determine whether Notch signaling contributes to the survival and recurrence of breast cancer cells could have a significant clinical impact. The observation that Notch signaling is required for recurrence would raise the possibility that these drugs could be used to target residual tumor cells and prevent breast cancer recurrence.
Abstract A42: Sensitivity and resistance to a p-AKT inhibitor in an erlotinib-sensitive model for lung cancer
Authors: El Meskini Rajaa , Martin Philip L. , Gumprecht Michelle , Kulaga Alan , Guerin Theresa , Baran Maureen , Iacovelli Anthony J. , Van Dyke Terry , Weaver Zoe
Published: March 2013

Activating mutations in EGFR, such as the L858R mutation, are effective predictors of response to EGFR tyrosine kinase inhibitor (TKI) therapy. While most patients initially respond to EGFR TKIs, the majority relapse within 6-12 months, underscoring the importance of identifying therapies that enhance and prolong EGFR TKI activity in patients. Agents targeting additional nodes in the EGFR pathway have the potential to overcome such resistance but single-agent activity must be understood first. We utilized mice engineered to develop erlotinib-sensitive lung adenocarcinomas driven by the human EGFR L858R mutation to evaluate the efficacy of a p-AKT inhibitor, MK-2206, currently in clinical trials for a variety of solid tumor malignancies. MK-2206 concentration and target activity were assessed in short-term PK/PD studies in both the lungs and blood of tumor-bearing mice. MK-2206 was then evaluated in an efficacy study using a twice-weekly dosing regimen, both as a single agent and in combination with other TKIs. MK-2206 alone resulted in tumor regression or tumor growth inhibition in ∼50% of the mice, but exhibited greater efficacy in combination with other upstream pathway inhibitors. Temporal evaluation of tumor growth inhibition indicated that MK-2206 was most effective at early time points and did not produce a sustained pathway and growth response as treatment continued. Analysis of gene expression changes in mice treated with MK-2006 alone vs. MK-2206 in combination with EGFR TKIs highlighted the pathways perturbed by both drugs.
Abstract A7: A novel class of selective estrogen receptors degraders regresses tumors in pre-clinical models of endocrine-resistant breast cancer
Authors: Darimont Beatrice , Govek Steven , Lu Nihn , Nagasawa Johnny , Prudente Rene , Qian Jing , Sensintaffar John , Shao Gang , Heyman Richard , Rix Peter , Smith Nicholas , Hager Jeffrey , Joseph James , Grillot Katherine , Aparicio Anna , Bischoff Eric , Kahraman Mehmet , Kaufman Josh , Lai Andiliy , Lee Kyoung-Jin
Published: March 2013

Eighty percent of all breast cancers express the estrogen receptor alpha (ERα) and thus are treated with antihormonal therapies that directly block ERα function (e.g., tamoxifen) or hormone synthesis (aromatase inhibitors). While these therapies are initially effective, acquired resistance invariably emerges and disease progression ensues. Importantly, the majority of these tumors continue to depend on ERα for growth and survival via both ligand-dependent and ligand-independent pathways. The emerging evidence that ERα can signal in the absence of estrogens supports the development of agents that are not only competitive ERα antagonists but also reduce steady state levels of the receptor and thus limit both ligand dependent and independent signaling.

We have identified novel, nonsteroidal ERα antagonists that induce degradation of ERα at picomolar concentrations resulting in significant reduction in steady state ERα protein levels in breast cancer cell lines. Using peptide-based conformational profiling, we show that these ligands induce estrogen receptor conformations that are distinct from both fulvestrant and tamoxifen indicating novel mechanism of action. Importantly, these compounds yield tumor regression in both tamoxifen-sensitive and -resistant models of breast cancer in vivo. Based on their unique in vitro profile, and good pharmacokinetics following oral dosing, these compounds represent a novel class of selective estrogen receptor degraders (SERDs) that hold promise as a next-generation therapy for the treatment of ER+ breast cancer as monotherapy, as well as in combination with agents that target other pathways involved in both intrinsic and acquired endocrine resistance.
Abstract A9: The cocaine and amphetamine-regulated transcript (CART) is an independent prognostic factor in lymph node-negative breast cancer and predicts outcome in tamoxifen treated patients
Authors: O'Connor Darran , Brennan Donal , Laursen Henriette , Bernards Rene , Crown John , Jirstrom Karin , Gallagher William
Published: March 2013

The successful identification and translation of informative biomarkers to aid clinical decision making is central to the effective implementation of personalized cancer therapeutic regimens. Antibody-based proteomics occupies a pivotal space in the cancer biomarker discovery and validation pipeline, facilitating the high-throughput evaluation of candidate markers. We have applied this approach to identify the cocaine- and amphetamine-regulated transcript (CART) as an independent poor prognostic marker in early stage breast cancer. The management of lymph node-negative breast cancer is the subject of much debate and two large ongoing clinical trials with particular emphasis on the requirement for adjuvant chemotherapy. Here we demonstrate, by tissue microarray and automated image analysis using a consistently applied threshold for definition of CART levels (median intensity +/− 1 standard deviation), that the neuropeptide hormone CART is expressed in primary and metastatic breast cancer and is an independent poor prognostic factor in ER-positive (p=0.0003), lymph node-negative (p=0.0025) tumors in two large breast cancer cohorts (n>1000). Kaplan-Meier analysis and the log rank test were used to illustrate differences between overall survival (OS) and recurrence free survival (RFS) according to CART protein and mRNA expression. Cox regression proportional hazards models were used to estimate the relationship between outcome and CART expression, grade, age, nodal status, ER status, PR status, Her2, Ki-67 and tumor size. We also show that CART mediates an autocrine/paracrine effect leading to an amplification of the CART signal in breast cancer cells. Furthermore, we additionally demonstrate that CART increases the transcriptional activity of ERα in a ligand-independent manner via the mitogen-activated protein kinase (MAPK) pathway and that CART-High tumors express increased levels of ER-target genes. Finally, we show that ectopic expression of CART in two independent ER+ cell lines protects against tamoxifen-mediated cell death (p=0.025) and that high CART expression predicts outcome in 3 independent tamoxifen-treated cohorts (p=0.002). We clearly demonstrate that both pre- and postmenopausal patients with CART-positive tumors have a poor prognosis and are resistant to tamoxifen and therefore should receive aggressive adjuvant chemotherapy. We believe that CART profiling will facilitate stratification of lymph node-negative breast cancer patients into high- and low-risk categories and allow the personalization of therapy.