NSCLC Lung Research Articles

Molecularly targeted gemcitabine-loaded nanoparticulate system towards the treatment of EGFR overexpressing lung cancer

Molecularly targeted therapy emerged as a novel therapeutic strategy in the treatment of multiple cancers. In the present study, we have developed gemcitabine (GEM)-loaded cetuximab (CET) surface modified poly(lactic) acid (PLA) nanoparticles (NP) (CET-GEM/PLA NP) to target to epidermal growth factor receptor (EGFR) overexpressing non-small cell lung cancer (A549) cells. The resultant CET-GEM/PLA NP showed a very uniform particle size of∼120nm and spherical morphology. It exhibited a pH-dependent controlled release pattern. A sustained release of drug in the physiological conditions and faster release in tumor pH will greatly improve the chemotherapeutic efficiency of therapeutic system. Higher or enhanced cellular uptake of CET-GEM/PLA NP in A549 cancer cells clearly indicates the EGFR-mediated receptor based active targeting. Nearly, a two-fold increase in fluorescent intensity was observed for CET-GEM/PLA NP comparing to that of non-targeted NP in the cancer cells. EGFR-mediated internalization of the targeted NP was further confirmed by the confocal microscopy. MTT assay clearly showed the enhanced cell killing effect of CET-conjugated NP due to the selective delivery of GEM to the EGFR over expressing cancer cells. Finally, comparing to the non-targeted NP, CET-GEM/PLA NP showed greater level of cell apoptosis (early and late apoptosis∼40%). Our results showed that antibody conjugation on the surface of NP could be a potential treatment strategy for EGFR over expressing cancer cells. This suggests that CET-GEM/PLA NP could be potentially used for the treatment of NSCLC (lung cancers).

Induction of HO-1 and CO Synthesis Radiosensitizes Tumor Epithelial Cells With CO Acting as a NO Mimetic

alterations in NFE2L2 and KEAP1 confer poorer overall survival (P Z 0.02). Conclusions: The integrative, high-throughput methods shown here for large-scale profiling of radiation survival and genomic features of solidtumor derived cell lines should facilitate tumor radiogenomics and the discovery of radiation sensitizers and protective agents. The potential clinical translation of these findings includes the development of NFE2L2 molecular diagnostics and the prospective validation of the first radiotherapeutic resistance biomarker in lung NSCLC. Author Disclosure:M. Abazeed: None. C. Xu: None. D. Adams: None. P. Tamayo: None. J. Loeffler: None. J. Suh: None. M. Meyerson: G. Consultant; MM is a consultant to Foundation Medicine. R. Ownership Other; MM is an equity holder in Foundation Medicine. K. Wong: None. P. Hammerman: G. Consultant; P.S.H. reports consulting fees from ARIAD.

Assessing Superoxide Levels in Human NSCLC and Normal Lung Epithelium

Assessing Superoxide Levels in Human NSCLC and Normal Lung Epithelium

A Novel Gene Expression Signature that Robustly Predicts the Outcome of Patients Diagnosed with Stage 1 Nsclc

ABSTRACT Aim: Although the management of metastatic lung adenocarcinoma has been profoundly modified by the identification of actionable molecular aberrations, the decision making for early-stage NSCLC still relies on the tumor stage (TNM) only. Stage 1 patients are considered of good prognosis and do not receive adjuvant therapy. However a significant proportion of those patients recur within 5 years after tumor resection. The identification of those high-risk patients through high performing molecular signature would enable to treat them preventively in order to avoid or reduce the rate of recurrence and premature death. Methods: Datasets were selected from public repositories (Director's challenge (DC) n = 402, Rousseaux n = 200, Hou n = 90, Zhu n = 143; Raponi: n = 120; Bhattacharjee; n = 125) upon the following criteria: NSCLC, stage IA-IB, no adjuvant therapy, R0 tumor resection, min 3-year follow-up. Gene expression data from various data sets were normalized using RMA and rescaled. To generate robust predictors of 3-year overall survival, an iterative feature selection framework based on predictive multivariate rules was applied on DC and on a pool of data sets composed of Hou, Zhu and Raponi (Pool) (Development sets). Only highly predictive rules in both datasets were then included in a subsequent predictive model and finally assessed for external validation in Bhattacharjee and Rousseaux (Validation sets). The predictive performances of the models were evaluated by ROC-AUC and by Kaplan Meier curves. Results: Our best model was robust across three independent datasets (all AUC > 0.50, p Conclusions: We developed a robust and highly performing gene expression predictive signature of 3-year overall survival for stage I NSCLC regardless of histology. These promising results have the potential to change the decision making at bedside for early stage lung NSCLC patients by allowing high-risk patients to receive adjuvant therapy and prevent future recurrence and death. Disclosure: All authors have declared no conflicts of interest.

Prognostic and therapeutic relevance of FLIP and procaspase-8 overexpression in non-small cell lung cancer

Non-small cell lung carcinoma remains by far the leading cause of cancer-related deaths worldwide. Overexpression of FLIP, which blocks the extrinsic apoptotic pathway by inhibiting caspase-8 activation, has been identified in various cancers. We investigated FLIP and procaspase-8 expression in NSCLC and the effect of HDAC inhibitors on FLIP expression, activation of caspase-8 and drug resistance in NSCLC and normal lung cell line models. Immunohistochemical analysis of cytoplasmic and nuclear FLIP and procaspase-8 protein expression was carried out using a novel digital pathology approach. Both FLIP and procaspase-8 were found to be significantly overexpressed in tumours, and importantly, high cytoplasmic expression of FLIP significantly correlated with shorter overall survival. Treatment with HDAC inhibitors targeting HDAC1-3 downregulated FLIP expression predominantly via post-transcriptional mechanisms, and this resulted in death receptor- and caspase-8-dependent apoptosis in NSCLC cells, but not normal lung cells. In addition, HDAC inhibitors synergized with TRAIL and cisplatin in NSCLC cells in a FLIP- and caspase-8-dependent manner. Thus, FLIP and procaspase-8 are overexpressed in NSCLC, and high cytoplasmic FLIP expression is indicative of poor prognosis. Targeting high FLIP expression using HDAC1–3 selective inhibitors such as entinostat to exploit high procaspase-8 expression in NSCLC has promising therapeutic potential, particularly when used in combination with TRAIL receptor-targeted agents.

Abstract C129: TAS-116, an orally available HSP90α and β selective inhibitor, exhibits synergistic effects in combination with taxanes in EGFR-tyrosine kinase inhibitor-resistant human NSCLC xenograft models through the downregulation of key molecules involved in drug resistance of cancer chemotherapy.

Abstract Background: In advanced NSCLC, EGFR-tyrosine kinase inhibitors (EGFR-TKI) have shown limited effect on lung squamous cell carcinoma (LSCC) or NSCLC harboring either a mutation in the K-RAS oncogene or lacking an activated mutation in EGFR. Though several cytotoxic drugs have been used in these EGFR-TKI-resistant settings, clinical outcomes have been generally poor. It has been shown that several antiapoptotic molecules regulated by PI3K/AKT and MAPK signaling, are involved in drug resistance to cancer chemotherapy. One of the possible approaches to overcome such drug resistance is via simultaneous inhibition of multiple signaling pathways. We have previously reported on an orally available highly selective HSP90α and β inhibitor, TAS-116. Because TAS-116 leads to downregulation of multiple molecules involved in various signaling pathways through HSP90 inhibition, we investigated the potential utility of TAS-116 on LSCC and K-RAS mutated NSCLC in combination with taxanes. Materials and Methods: In vitro drug combination effects were evaluated with calculation of combination index. The amount and the phosphorylation status of cellular proteins were determined by western blot with appropriate antibodies. Antitumor activities of TAS-116 alone and in combination with taxanes were evaluated in human NSCLC xenograft models. The inhibition and induction study of cytochrome P450 (CYP) by TAS-116 were evaluated using human liver microsomes and human cryopreserved hepatocytes, respectively. Results: TAS-116 showed synergistic effects in combination with docetaxel (DTX) and paclitaxel (PTX) against LSCC cell lines. TAS-116 enhanced taxane-mediated apoptosis. To clarify the mode of action in these combinations, we measured the amount and the phosphorylation status of apoptosis-related proteins. TAS-116 decreased phosphorylation of BAD and YBX1, through the downregulation of upstream kinases, AKT and RAF1. TAS-116 synergistically inhibited tumor growth in both human LSCC and K-RAS mutated NSCLC xenograft models in combination with PTX or DTX. With regard to drug-drug interaction potential at effective concentrations, TAS-116 did not inhibit or induce CYP3A4 which is important in taxane metabolism. Conclusion: TAS-116 in combination with taxanes enhanced apoptosis induction and exhibited synergistic antitumor activity against LSCC and K-RAS mutated NSCLC xenograft models through downregulation of key factors involved in drug resistance in cancer chemotherapy. The combination of TAS-116 and taxanes could be a promising approach to treat EGFR-TKI-resistant NSCLC including LSCC. Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C129. Citation Format: Hiromi Muraoka, Akira Kanoh, Akihiro Hashimoto, Kenjiro Ito, Takamasa Suzuki, Kouichi Takahashi, Chihoko Yoshimura, Makoto Kitade, Kazuhiko Yonekura, Shuichi Ohkubo, Teruhiro Utsugi. TAS-116, an orally available HSP90α and β selective inhibitor, exhibits synergistic effects in combination with taxanes in EGFR-tyrosine kinase inhibitor-resistant human NSCLC xenograft models through the downregulation of key molecules involved in drug resistance of cancer chemotherapy. [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 C129.

A 4D-optimization concept for scanned ion beam therapy

Background and purposeScanned carbon beam therapy offers advantageous dose distributions and an increased biological effect. Treating moving targets is complex due to sensitivity to range changes and interplay. We propose a 4D treatment planning concept that considers motion during particle number optimization. Material and methodsThe target was subdivided into sectors, one for each motion phase of a 4D-CT. Each sector was non-rigidly transformed to its motion phase and there targeted by a dedicated raster field (RST). Therefore, the resulting 4D-RST compensated target motion and range changes. A 4D treatment control system (TCS) was needed for synchronized delivery to the measured patient motion. 4D-optimized plans were simulated for 9 NSCLC lung cancer patients and compared to static irradiation at end-exhale. A prototype TCS was implemented and successfully tested in a film experiment. ResultsThe 4D-optimized treatment plan resulted in only slightly lower dose coverage of the target compared to static optimization, with V95% of 97.9% (median, range 96.5–99.4%) vs. 99.3% (98.5–99.8%), with negligible overdose. The conformity number was comparable at 88.2% (85.1–92.5%) vs. 85.2% (79.9–91.2%) for 4D and static, respectively. ConclusionWe implemented and tested a 4D treatment plan optimization method resulting in highly conformal dose delivery.

Lung stereotactic body radiation therapy (SBRT) delivered over 4 or 11 days: A comparison of acute toxicity and quality of life

PurposeThe optimal duration over which lung SBRT should be delivered is unknown. We conducted a randomized pilot study in patients treated with four fractions of lung SBRT delivered over 4 or over 11days. MethodsPatients with a peripheral solitary lung tumor (NSCLC or pulmonary metastasis) ⩽5cm were eligible. For NSCLC lung tumors ⩽3cm, a dose of 48Gy in 4 fractions was used, otherwise 52Gy in 4 fractions was delivered. Patients were randomized to receive treatment over 4 consecutive days or over 11days. The primary end-point was acute grade ⩾2 toxicity. Secondary end-points included quality of life (QOL) assessed using the EORTC QLQ-C30 and QLQ-LC13 questionnaires. ResultsFifty four patients were enrolled. More patients in the 11day group had respiratory symptoms at baseline. 55.6% patients treated over 4days and 33.3% of patients treated over 11days experienced acute grade ⩾2 toxicity (p=0.085). Dyspnea, fatigue and coughing domains were worse in the 11day group at baseline. At 1 and 4months, more patients in the 4day group experienced a clinically meaningful worsening in the dyspnea QOL domain compared to the 11day group (44.5% vs 15.4%, p=0.02; 38.5% vs 12.0%, p=0.03, respectively). However, raw QOL scores were not different at these time-points between treatment groups. ConclusionsGrade 2 or higher acute toxicity was more common in the 4day group, approaching statistical significance. More patients treated on 4 consecutive days reported a clinically meaningful increase in dyspnea, although interpretation of these results is challenging due to baseline imbalance between treatment groups. Larger studies are required to validate these results.

Crude extracts of Antrodia cinnamomea inhibit invasion and migration of Human Hepatocellular Carcinoma Cells

A native fungus native to Taiwan, Antrodia cinnamomea is believed to be effective in preventing many diseases including apoptosis of different kinds of cancer cells like NSCLC lung cancer, breast cancer, and leukemia cancer. In this study we used the ethanol extract of Antrodia cinnamomea (EEAC) to evaluate its effects on anti‐proliferative and anti‐metastatic activity of human hepatocellular liver cancer cells HepG2 and Hep3B cells. The underlying mechanism was also conducted by Western blotting analysis. The results of MTT assay revealed non‐cytotoxic concentrations Antrodia cinnamomea dose and time‐dependently inhibited the migration of HepG2 and Hep3B cells. By gelatin zymography assay, EEAC effectively inhibited MMP‐9 and MMP‐2 activities. By Western blotting analysis, treatment with EEAC decreased phosphatidylinositol‐3‐kinase (PI3K)/AKT, MMP‐2 and MMP‐9 expressions in HepG2 and Hep3B cells. EEAC also exerted an inhibitory effect on the phosphorylation of extracellular signal‐regulated kinases 1 and 2 (ERK1/2) and inhibition of activation of nuclear factor kappa B (NF‐κB), c‐Fos, and c‐Jun. According to the above data, the growth inhibition of human hepatocellular liver cancer cells by EEAC was mediated by induction of migration and cell proliferation related proteins. In conclude, Antrodia cinnamomea may be a potential candidate for treating those patients with liver cancer metastasis.

Abstract LB-85: Ectopic expression of eukaryotic translation elongation factor eEF1A2 is a hallmark for human lung cancer and is associated with poor prognosis

Abstract Background: The elongation factor eEF1A is one of the most abundantly expressed cellular proteins. It exists in two forms, eEF1A2, expressed in neurons and striated muscle, and eEF1A1, expressed in all other cell types. No normal adult cell expresses both forms. We investigated their role in lung cancer. Methods: We studied over 500 NSCLC and SCLC lung cancers and over 170 cell lines, as well as corresponding non- malignant lung tissues and immortalized respiratory epithelial lines. Methods included microarray, qPCR, immunostaining and western blots for eEF1A1 and eEF1A2, genomic sequencing and SNP analyses. Results: All normal and malignant cells highly expressed eEF1A1. EEF1A2 was expressed in 70-80% of all forms of lung cancers and 95% of cell lines, while all non-malignant tissues were negative. Expression of eEF1A2 in neuroendocrine tumors was particular high. The ectopic expression of eEF1A2 was not the consequence of mutations or copy number gains. Use of demethylating agents and HDAC inhibitors resulted in expression of eEF1A2 in respiratory epithelial cells, indicating that epigenetic mechanisms prevented expression in non-malignant cells. Transient knockdown of the eEF1A2 expression inhibited proliferation and colony formation of lung cancer cell lines. Furthermore, the expression of eEF1A2 in resected NSCLC cancers was associated with poor survival. Conclusions: Highly specific ectopic expression of eEF1A2 gene is frequent in all types of lung cancer, plays an important role in tumor pathogenesis and is a potential diagnostic and prognostic factor. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-85. doi:1538-7445.AM2012-LB-85

Abstract B7: microRNA regulation of cell viability and drug sensitivity in lung cancer

Abstract Introduction: Lung cancer is the leading cause of cancer-related deaths, with the majority of deaths due to failed therapy from tumor drug resistance. Third-generation chemotherapeutic agents represent the standard first-line treatment for advanced small cell (SCLC) and non-small cell (NSCLC) lung cancer patients. Response rates are poor (20–40%) with a median survival of 8–10 months. Methods: In an unbiased and comprehensive approach, we have combined a high-throughput screening platform with a library of chemically synthesized microRNA mimics and inhibitors. We have used this platform to identify mimics and inhibitors that reduce cell viability in general, and those that specifically sensitize cells to taxanes. Results: We have identified several miRNAs for which over-expression or inhibition has a dramatic and selective effect on cell viability or drug response. We have demonstrated that miR-337-3p mimic sensitizes NSCLC cells to taxanes. By combining in vitro and in silico approaches, we identified STAT3 and RAP1A as direct targets that mediate the effect of miR-337-3p by enhancing taxane-induced arrest in the G2 phase of the cell cycle. We have also identified an inhibitor of miR-139-5p as a potent and selective regulator of SCLC cell viability. Inhibiting miR-139-5p decreases SCLC cell viability by over 80%, but has a minimal cytotoxic effect on that of NSCLCs or immortalized human bronchial epithelial cells. We are currently investigating the targets of miR-139-5p that mediate its effect on SCLC cell viability. Conclusion: We have identified miR-337-3p as a regulator of taxane sensitivity in NSCLC, and miR-139-5p as a modulator of viability in SCLC. Increasing levels of miR-337-3p and inhibiting miR-139-5p may therefore provide novel therapeutic tools for the treatment of NSCLC and SCLC lung tumors, respectively. Citation Format: Liqin Du, Christopher DeSevo, Borkowski Robert, Baker Michael, Adi F. Gazdar, John D. Minna, Alexander Pertsemlidis. microRNA regulation of cell viability and drug sensitivity in lung cancer [abstract]. In: Proceedings of the AACR Special Conference on Noncoding RNAs and Cancer; 2012 Jan 8-11; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(2 Suppl):Abstract nr B7.

IGDB.NSCLC: integrated genomic database of non-small cell lung cancer

Lung cancer is the most common cause of cancer-related mortality with more than 1.4 million deaths per year worldwide. To search for significant somatic alterations in lung cancer, we analyzed, integrated and manually curated various data sets and literatures to present an integrated genomic database of non-small cell lung cancer (IGDB.NSCLC, http://igdb.nsclc.ibms.sinica.edu.tw). We collected data sets derived from hundreds of human NSCLC (lung adenocarcinomas and/or squamous cell carcinomas) to illustrate genomic alterations [chromosomal regions with copy number alterations (CNAs), gain/loss and loss of heterozygosity], aberrant expressed genes and microRNAs, somatic mutations and experimental evidence and clinical information of alterations retrieved from literatures. IGDB.NSCLC provides user friendly interfaces and searching functions to display multiple layers of evidence especially emphasizing on concordant alterations of CNAs with co-localized altered gene expression, aberrant microRNAs expression, somatic mutations or genes with associated clinicopathological features. These significant concordant alterations in NSCLC are graphically or tabularly presented to facilitate and prioritize as the putative cancer targets for pathological and mechanistic studies of lung tumorigenesis and for developing new strategies in clinical interventions.

SU-E-T-545: Assessing the Impact of Proton Range Uncertainties on NSCLC Lung Patients Treated with Proton Beam-Based SBRT

Purpose: Proton therapy has been proposed as an alternative to photon beam therapy in stereotactic body radiation therapy(SBRT) of early‐stage NSCLCcancer. We investigated in the present study how proton range uncertainties can influence protonSBRTtreatment quality. Methods: Ten medically inoperable patients with stage I NSCLC were included in the current study. The treatment strategy for the SBRTtumors at MGH is to use either i)proton passive scattering or ii)photon 3DCRT for tumors with motion amplitude of less 5mm. The SBRTprotontreatment planning uses either 2 or 3 co‐planar beams. While, SBRTphotontreatment planning uses 2 or 3 non‐coplanar arcs, with a total of 8–10 beams. In the case of the proton beams an additional 3.5%+2mm is added to account for range uncertainties in the proximal and distal ends of the spread‐out‐Bragg‐peak, SOBP. Dosimetric comparisons were performed by defining a high‐ and low‐dose region representing volumes receiving more and less than 50% of the prescription dose, respectively. Results: In high‐dose regions, the volume receiving &#8805;95% of the prescription dose was larger for proton than for photonSBRT, i.e., 46.5 cc versus 33.5 cc (p=0.009), respectively. The conformity indices were 2.46 and 1.56, respectively. For tumors in close proximity to the chest‐wall, the volume of the chest‐wall receiving more‐than‐30Gy (V30) was 7 cc larger for protons than for photons (p=0.06), respectively. In low‐dose regions, the lung V5 and maximum esophagus dose were smaller for protons than for photons (p=0.019 and p<0.001, respectively). Conclusions: Protons generate larger high‐dose regions than photons because of range uncertainties. This can result in nearby healthy organs (such as chest‐wall) receiving close to treatmentdose. Future proton studies should focus on approaches to reduce dose range uncertainties and identify clinical subgroups of patients who will most benefit from the lack of exit dose.

The Rab27A effector MYRIP as a regulator of survival in non-small cell lung cancer cells.

e13537 Background: Personalising therapy for non-small cell lung cancer has been validated as an effective treatment paradigm. Somatic gene alterations confer sensitivity to inhibition of growth survival pathways irrespective of underlying resistance to chemotherapeutic agents, and enables efficient activation of BCL-2 family-dependent pro-apoptotic signalling. In platinum resistant cancers this signalling is blocked, however the intrinsic pathway remains sensitive to exogenous BH3 death signals. Methods: Microarray profiling coupled to focused RNAi screening was used to reveal those resistance genes involved in derepression of proapoptotic BCL-2 family proteins. Analysis of the effect of MYRIP expression on survival was generated from studying 6 NSCLC datasets from Gene Expression Omnibus which were paired with survival time and status information. The toxic effect of MYRIP depletion on NSCLC and non-cancerous lung cells was determined by cell viability and clonogenic assays. Apoptotic induction was examined by caspase and PARP cleavage experiments, and fractionation of mitochondrial proteins. FACS analysis was used to examine effects on cell cycle. Results: MYRIP/Slac2c, a gene hitherto associated with vesicle trafficking but not apoptosis, was overexpressed in platinum resistant NSCLC cells. In particular, cisplatin resistant cells expressed a heavier protein isoform of MYRIP which appeared to be associated with the mitochondria. Silencing of MYRIP induced BIM and BAX/BAK-dependent mitochondrial apoptosis which was selective for NSCLC, but not non-cancerous lung cells. In addition, loss of MYRIP induced S-phase arrest. Conclusions: Overexpression of MYRIP is an independent, poor prognostic factor suggesting that targeting these genes may be therapeutically relevant. In summary, MYRIP exhibits a previously unknown survival function associated with suppression of pro-apoptotic BCL-2 family proteins and maintenance of cancer cell proliferation and survival.

Neuroendocrine differentiation as an indicator of chemosensitivity and prognosis in nonsmall cell lung cancer

Context: Nonsmall cell lung cancers with neuroendocrine differentiation (NSCLC-ND) may demonstrate biologic behavior intermediate between NSCLC and small cell lung cancer (SCLC) with impact on prognosis.Methods: We analyzed 116 consecutive patients with Stage III and IV NSCLC who were diagnosed and treated between 2001 and 2006. Using immuno-histochemical staining for neuron-specific enolase (NSE), chromogranin A (ChrA), and synaptophysin (Syn), 29 (25%) NSCLC-ND were identified.Results: Expression of NSE was present in 22.4%, ChrA in 15.5% and Syn in 14.8% of patients with NSCLC. Therapeutic response was significantly better in the NSCLC-ND group and specimens with > 30% neuroendocrine (NE)-differentiated tumor cells showed favourable therapeutic response (P < 0.05). Multivariate binary logistic regression showed that percentage of NE positive tumor cells was a significant independent prognostic factor associated with a favourable outcome. Receiver operating characteristic (ROC) curves and areas under ROC curves confirmed that percentage of NE-differentiated tumor cells could be useful prediction factor of therapeutic response. Moreover, according to percentage of NE-differentiated tumor cells, optimal cutoffs and related sensitivities and specificities were determined for each markers.Conclusion: Advanced-stage NSCLC with NE tumor cells are clinically less aggressive tumors. Percentage of NE-differentiated tumor cells identifies patients with favourable therapy response to paclitaxel-cisplatin

Comprehensive CADM1 promoter methylation analysis in NSCLC and normal lung specimens

Methylation-mediated silencing of the tumour suppressor CADM1 has been functionally linked to lung cancer development. We aimed to determine whether CADM1 promoter methylation is a candidate early detection marker for lung cancer. To this end frozen tissue samples of 36 non-small cell lung cancers, 26 corresponding tumour distant normal tissue samples as well as 6 samples of normal lung from non-lung cancer patients were tested for DNA methylation at three different regions within the CADM1 promoter (M1, M5 and M9) using methylation specific PCR followed by methylation specific reverse line blot analysis.Sixty-four percentage of tumour samples tested positive at the M1 region, 47% at M5 and 74% at the M9 region, compared with 65% (M1), 23% (M5) and 46% (M9) of paired normal tissue samples. Methylation of each of these promoter regions was also detected in the majority of non-lung cancer control samples. Dense methylation, defined as methylation at ≥2 promoter regions, was detected in 66% of tumour samples compared with 38% of paired normal tissues and 67% of non-lung cancer control samples. Within the small subgroup of female patients dense methylation was found in all tumour samples but only 22% of paired normal samples. Neither methylation of individual sites nor dense methylation was correlated with disease free survival. In conclusion, CADM1 promoter methylation is a frequent event in NSCLC as well as normal lung, both of lung cancer and non-lung cancer patients. Hence, CADM1 methylation analysis is unlikely to have diagnostic value for the early detection of lung cancer in an unselected population. However, a diagnostic value for selected subjects, such as females, cannot be excluded.

MicroRNA expression distinguishes SCLC from NSCLC lung tumor cells and suggests a possible pathological relationship between SCLCs and NSCLCs

MicroRNA expression distinguishes SCLC from NSCLC lung tumor cells and suggests a possible pathological relationship between SCLCs and NSCLCs

83 poster: Stage IIIB or IV NSCLC Lung Cancer on CT, What is the Point of F-18 FDG PET/CT?

83 poster: Stage IIIB or IV NSCLC Lung Cancer on CT, What is the Point of F-18 FDG PET/CT?

Abstract A25: Establishment and characterization of primary human Biomerk Tumorgraft™ Models: Application to oncology drug development

Abstract One of the key challenges facing oncology drug development is the high attrition rates of compounds that enter the drug development pipeline, where very few achieve successful approval and marketing. Champions Biotechnology, in an effort to enhance the value of preclinical compounds and accelerate oncology drug development, has developed a novel preclinical platform derived from Biomerk Tumorgraft™ models; an innovative approach that utilizes the implantation of primary human tumors in immune-deficient mice in a manner that preserves the biological properties of the original human tumor. Biomerk Tumorgrafts™ differ from traditional xenograft models in that they are not passaged in tissue culture, and are instead exclusively passaged in vivo. Additionally, the Biomerk Tumorgraft™ models: (a) maintain the fundamental genotypic features of the original cancer, (b) represent the genetic heterogeneity of the cancer, and (c) do not change over several passages. Champions has established a bank of over 200 Tumorgraft models encompassing all major solid tumor types, all of which have been extensively characterized. Here we describe a panel of NSCLC lung, colon, and pancreatic tumorgraft models including gene expression profiling, mutational analyses and responses to standard of care agents. Of the NSCLC lung models, 6/8 demonstrated resistance to Erlotinib with 2/8 demonstrating an intermediate response, 2/6 also possessed a mutant K-ras or a mutant EGFR. Similarly, 3/6 K-ras mutant colon Tumorgraft models demonstrated sensitivity to irinotecan. In the case of the pancreatic models, 7/8 showed resistance to Erlotinib with the remaining model showing only a modest response. Through Champions' Personalized Oncology Services, we have also shown a high correlation between the Tumorgraft response and the clinical response. Together, these results demonstrate that each model differs with respect to its genetic composition and response to standard agents, and thus represents the efficacy of a compound on an individual patient. Given the diversity of models, the maintenance of the genomic and phenotypic characteristics of the original patient tumor, and the correlation between Tumorgraft and clinical responses, Champions Biomerk Tumorgraft™ models represent a novel preclinical in vivo platform capable of optimizing drug regimens for cancer patients and predicting the clinical effectiveness of preclinical drug candidates. Overall, the application of Biomerk Tumorgraft™ models has the potential to accelerate and enhance oncology drug development. Citation Information: Mol Cancer Ther 2009;8(12 Suppl):A25.

Expression of KAI1 gene in non-small cell lung cancer and its clinical significance.

Objective To investigate the expression of KAI1,as a suppressor of metastasis,in the tissues of NSCLC and its relationship with the clinicopathological indexes.Methods The expressions of KAI1/CD82 protein were detected with Western blot in 48 cases of NSCLC from patients with lung cancer and benign lung tissue of patients with benign lung disease undergoing surgery in our hospital.The RT-PCR method was also used to assess KAI1 mRNA in the same tissue samples,and the results were statistically analyzed.Results The positive rates of KAI1mRNA in NSCLC and benign lung tissues were 52%,90% respectively;the positive rates of KAI1/CD82 in NSCLC and benign lung tissues were 48%,85% respectively(P0.01).The expression level of KAI1mRNA and KAI1/CD82 protein in lung cancer were lower than those in benign lung tissue(P0.01).The expression level of KAI1/CD82 were correlated with tumor stage,grade of tumor differentiation and metastasis status(P0.05),among which the metastasis status in particular(P0.01).The expression of KAI1/CD82 had relations with its mRNA(P0.01).Conclusion The down-regulation of KAI1 may be correlated with carcinogenesis,development and metastasis in NSCLC.Its reduction may be mainly at transcriptional level,and it could be used as a marker for predicting the potential metastasis and prognosis of patients with NSCLC.