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Esophageal cancer incidence is increasing and has few treatment options. In studying receptor tyrosine kinases associated with esophageal cancers, we have identified EPHB4 to be robustly overexpressed in cell lines and primary tumor tissues. In total, 94 squamous cell carcinoma, 82 adenocarcinoma, 25 dysplasia, 13 Barrett esophagus, and 25 adjacent or unrelated normal esophageal tissues were evaluated by immunohistochemistry. EPHB4 expression was significantly higher in all the different histologic categories than in adjacent normal tissues. In 13 esophageal cancer cell lines, 3 of the 9 SCC cell lines and 2 of the 4 adenocarcinomas expressed very high levels of EPHB4. An increased gene copy number ranging from 4 to 20 copies was identified in a subset of the overexpressing patient samples and cell lines. We have developed a novel 4-nitroquinoline 1-oxide (4-NQO)-induced mouse model of esophageal cancer that recapitulates the EPHB4 expression in humans. A specific small-molecule inhibitor of EPHB4 decreased cell viability in a time- and dose-dependent manner in 3 of the 4 cell lines tested. The small-molecule inhibitor and an EPHB4 siRNA also decreased cell migration (12%–40% closure in treated vs. 60%–80% in untreated), with decreased phosphorylation of various tyrosyl-containing proteins, EphB4, and its downstream target p125FAK. Finally, in a xenograft tumor model, an EPHB4 inhibitor abrogated tumor growth by approximately 60% compared with untreated control. EphB4 is robustly expressed and potentially serves as a novel biomarker for targeted therapy in esophageal cancers. Cancer Res; 73(1); 184–94. ©2012 AACR.

Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKI) show antitumor activity in a subset of non–small cell lung cancer (NSCLC) patients. However, the initial tumor response is followed by recurrence. Several studies have suggested the importance of other receptor tyrosine kinases (RTK) and downstream kinases as potential targets in the treatment of NSCLC. We used the multiple-RTK inhibitor AEE788, which inhibits EGFR, vascular endothelial growth factor receptor, and human epidermal growth factor receptor 2, with and without the downstream kinase inhibitor RAD001 (an inhibitor of mammalian target of rapamycin). AEE788 inhibited cell growth more effectively than did erlotinib in three NSCLC cell lines examined (A549, H1650, and H1975). However, in the EGFR-TKI–resistant cell line H1975 harboring T790M resistance mutation, cell growth inhibition by AEE788 was only mild, and the phosphorylation of its leading targets such as EGFR and vascular endothelial growth factor receptor 2 was not inhibited. In H1975, AEE788 induced significantly greater cell growth inhibition when combined with RAD001 than when used alone. This cooperative effect was not seen with the combination of erlotinib and RAD001. We found that c-Met was highly phosphorylated in this cell line, and the phosphorylated c-Met was inhibited effectively by AEE788. Using a phospho-RTK array, the phosphorylation of c-Met and insulin-like growth factor-I receptor was inhibited by AEE788. These results suggest that upstream RTK inhibitor overcomes the acquired resistance to EGFR-TKI when combined with downstream kinase inhibitor. Thus, the combined inhibition of upstream and downstream RTKs is a promising strategy for the treatment of NSCLC. Mol Cancer Res; 8(8); 1142–51. ©2010 AACR.

We dissected the hindlimb of a female western lowland gorilla and determined the muscle dimensions (mass, fascicle length, and physiological cross-sectional area: PCSA). Comparisons of the muscle parameters of the measured gorilla with corresponding reported human data demonstrated that the triceps surae muscles were larger and had more capacity to generate force than the other muscle groups in both species, but this tendency was more prominent in the human, probably as an adaptation to strong toe-off during bipedal walking. On the other hand, PCSAs of the extrinsic pedal digital flexors and digiti minimi muscles were larger in the western lowland gorilla, suggesting that the foot, particularly the fifth toe, has a relatively high grasping capability in the lowland gorilla.

Background Alterations in the processing of the genetic information in carcinogenesis result from stable genetic mutations or epigenetic modifications. It is becoming clear that nucleosomal histones are central to proper gene expression and that aberrant DNA methylation of genes and histone methylation plays important roles in tumor progression. To date, several histone lysine methyltransferases (HKMTs) have been identified and histone lysine methylation is now considered to be a critical regulator of transcription. However, still relatively little is known about the role of HKMTs in tumorigenesis. Results We observed differential HKMT expression in a lung cancer model in which normal human bronchial epithelial (NHBE) cells expressing telomerase, SV40 large T antigen, and Ras were immortal, formed colonies in soft agar, and expressed specific HKMTs for H3 lysine 9 and 27 residues but not for H3 lysine 4 residue. Modifications in the H3 tails affect the binding of proteins to the histone tails and regulate protein function and the position of lysine methylation marks a gene to be either activated or repressed. In the present study, suppression by siRNA of HKMTs (EZH2, G9A, SETDB1 and SUV39H1) that are over-expressed in immortalized and transformed cells lead to reduced cell proliferation and much less anchorage-independent colony growth. We also found that the suppression of H3-K9, G9A and SUV39H1 induced apoptosis and the suppression of H3-K27, EZH2 caused G1 arrest. Conclusion Our results indicate the potential of these HKMTs in addition to the other targets for epigenetics such as DNMTs and HDACs to be interesting therapeutic targets.

We report a case of X-linked agammaglobulinemia who presented with bronchiectasis. The patient had suffered pneumonia about every five years since childhood until he presented to our hospital at age 34 years old. CT showed bronchiectasis predominantly in the right middle lobe, lingula, and lower lobes. Administration of antibiotics resulted in symptomatic relief. Episodes of recurrent pulmonary infection and bronchiectasis indicated congenital immunodeficiency disorder. Investigation of lymphocyte subsets and serum immunoglobulin values showed remarkable reduction of B cells, IgG 772 mg/dl, IgA 216 mg/dl, and IgM 29 mg/dl. Flow cytometric assessment combined with genetic analysis was performed, and the results showed decreased expression of monocyte Bruton's tyrosine kinase (BTK) and missense mutation of Btk gene. We diagnosed X-linked agammaglobulinemia. IgG remained above 600 mg/dl in this case, we have not administered immunoglobulin after discharge. He suffered from pneumonia in 2004 and 2006 and bronchiectasis has progressed. In this report, we present a case including CT findings over a period of 8 years.

Rationale: Recently, the importance of fibroblast growth factor (FGF) and its receptor (FGFR) signals has been emphasized in various types of cancer, including lung cancer. FGFs and FGFRs are reported to be related to drug resistance, cell proliferation, differentiation and motility. Among various FGFs, FGF9 is a developmentally important FGF associated with epithelial airway branching, and its expression is repressed in normal adult lungs. We have previously shown that FGF9 is highly expressed in NSCLC cells, and FGF9 expression was associated with poor prognosis of NSCLC patients. Furthermore, we reported that induction of FGF9 in adult lung results in the rapid formation of adenocarcinoma in specific gene engineered mouse model. However, the exact roles of FGF9 in lung epithelial cells remain elusive. Objectives: The objective of this study is to clarify the roles of FGF9 in lung epithelial cells. Methods: For in vitro experiments, a stable cell line with constitutive expression of FGF9 in MLE12 (a mouse lung alveolar type II cell line transformed by SV40 large T antigen) was established by retroviral infection. The effect of FGF9 on proliferation, colony formation capacity and downstream signaling was evaluated by MTS assay, softagar colony formation assay and Western blotting, respectively. For in vivo experiments, the cells were transplanted into immunodeficient mice subcutaneously, and the size of tumor was measured. To evaluate the efficacy of FGFR inhibitors for FGF9-driven lung cancers, AZD4547, selective FGFR inhibitor, was orally administered. For pathological characterization of the tumors, immunohistochemical staining was performed. Fifteen surgically-resected SCLC samples were obtained and the expression of FGF9 was evaluated by immunohistochemistry. Results: FGF9 has oncogenic ability in vitro and its effect may be exerted by the activation of MAPK pathway through FGFR1 and FGFR3 in MLE12 cells. Unexpectedly, pathological analysis revealed FGF9-driven tumors exhibited SCLC histology. Selective FGFR inhibitor, AZD4547 suppressed tumor growth of FGF9-driven MLE12 tumors. FGF9 is highly expressed in human SCLC samples (67%). Conclusions: These results suggest that FGF9 has roles of tumor initiation and progression in lung cancer, especially in SCLC. SCLC which highly expresses FGF9 may be a target of FGFR inhibitors. Citation Format: Kota Ishioka, Kenzo Sejima, Hiroyuki Yasuda, Junko Hamamoto, Ahmed Hegab, Tetsuo Tani, Aoi Kuroda, Daisuke Arai, Keiko Ohgino, Takashi Sato, Terai Hideki, Ichiro Kawada, Katsuhiko Naoki, Yuichiro Hayashi, Tomoko Betsuyaku. Oncogenic potential of FGF9 in lung cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3916. doi:10.1158/1538-7445.AM2015-3916

Purpose: Alectinib is a highly selective ALK inhibitor and showed a striking efficacy in non-small cell lung cancers (NSCLC) harboring EML4-ALK gene rearrangement. Recently, it was reported that alectinib had an objective response rate of 93.5% in ALK positive patients in a clinical study. However, cancer cells usually acquire resistance to molecular-targeted drugs. The mechanisms of acquired resistance to alectinib are not yet well clarified. The purpose of this study is to clarify the mechanism of acquired resistance to alectinib in ALK translocated lung cancer cells. Experimental design: We have established alectinib resistant cells (H3122-AR) from H3122, which harbors EML4-ALK rearrangement, by long term exposure to alectinib. The mechanism of acquired resistance to alectinib in H3122-AR cells was evaluated in vitro by cDNA sequencing of ALK tyrosine kinase (TK) domain, or by phospho-RTK array to estimate activation of bypass pathway. The efficacy of combination therapy targeting ALK and EGFR was evaluated in vitro and in vivo by using alectinib, afatinib, or siRNA for EGFR. For in vivo experiment, we used mouse xenograft model. Human tumor samples were evaluated by immunohistochemistry. Results: No additional mutation in ALK TK domain was found by cDNA sequencing. Phospho-RTK array revealed that the phosphorylation level of EGFR was increased in H3122-AR cells compared with H3122 parental cells, which was subsequently confirmed by western blotting. We found no additional mutation in EGFR TK domain by cDNA sequencing. Among 5 ligands for EGFR, the expression of TGFα was significantly increased in H3122-AR cells compared with H3122 parental cells. We found the phosphorylation level of EGFR was decreased in TGFα gene knockdown, therefore TGFα and EGFR contributed to the acquired resistance to alectinib. The combination therapy targeting ALK and EGFR by using alectinib, afatinib in vitro and in vivo, or siRNA for EGFR in vitro showed efficacy. We found the increased phosphorylation of EGFR after long term ALK-TKI treatment was also detected in human tumor samples. Conclusion: Activation of EGFR signaling pathway through TGFα overexpression induces acquired resistance to alectinib in ALK-translocated lung cancer cells. Citation Format: Tetsuo Tani, Hiroyuki Yasuda, Junko Hamamoto, Aoi Kuroda, Daisuke Arai, Kota Ishioka, Keiko Ohgino, Ichiro Kawada, Katsuhiko Naoki, Hayashi Yuichiro, Tomoko Betsuyaku, Kenzo Soejima. Activation of EGFR bypass signaling through TGFα overexpression induces acquired resistance to alectinib in ALK-translocated lung cancer cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 746. doi:10.1158/1538-7445.AM2015-746

Introduction: Many anti-tumor drugs have been developed, however, prognosis of NSCLC remains poor and new approaches for NSCLC treatment are expected. Lung is a unique organ whose epithelial cells are directly exposed to oxygen. Hyperoxia produces reactive oxygen species (ROS) and induces cell damage. Therefore we attempted to investigate the potential of hyperoxia as an anti-tumor treatment in NSCLC cells in this study. Methods: We used oxygen-concentration-adjustable incubators and validated effect of hyperoxia on various lung cancer cell lines in vitro. The effect of hyperoxia on cell proliferation, apoptosis and gene expression was evaluated using MTS assay, flowcytometry and cDNA microarray, respectively. Results: We found that the growth of lung cancer cell lines (A549, NCI-H1975) was inhibited by 50% of hyperoxia in a dose-dependent manner, while that of normal airway epithelial cells (NHBE, BEAS-2B) was not. Two independent pathway analysis using cDNA microarray revealed the activation of NF-kB and AMPK pathways, and the production of nitric oxide and ROS in hyperoxia (50%) treated lung cancer cell lines compared to untreated ones. The combined treatment with ABT-263, Bcl-2 inhibitor, and hyperoxia (50%) induced synergistic growth inhibition and apoptosis in NCI-H1975 and SK-MES-1 cells. Moreover, the combination of ABT-263 and activator of either ROS, NF-kB or AMPK also showed synergistic growth inhibition in those cells. Conclusions: Hyperoxia (50%) showed anti-tumor effect in NSCLC cell lines through the activation of NF-kB and AMPK pathways, and the production of ROS. The effect was augumented in combination with Bcl-2 inhibitor. Hyperoxia may be a treatment option for NSCLC patients. Citation Format: Junko Hamamoto, Hiroyuki Yasuda, Ichiro Nakachi, Michael G. Edwards, Masayoshi Miyawaki, Makoto Nishino, Aoi Kuroda, Tetsuo Tani, Daisuke Arai, Kota Ishioka, Ichiro Kawada, Katsuhiko Naoki, Tomoko Betsuyaku, Kenzo Soejima. Hyperoxia may be a treatment option for NSCLC. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3006. doi:10.1158/1538-7445.AM2015-3006

Rationale: ABT-263 (Navitoclax) is one of the BH3 mimetics targeting anti-apoptotic B-cell lymphoma-2 (Bcl-2) family proteins such as Bcl-2, Bcl-XL, and Bcl-w, thereby inducing apoptosis. It has been reported that the response to ABT-263 is associated with expressions of myeloid cell leukemia-1 (Mcl-1), an anti-apoptotic protein. Given its effectiveness as a single agent in preclinical studies, ABT-263 is currently being evaluated in clinical trials for small cell lung cancer (SCLC) and leukemia. However, the efficacy of ABT-263 in non-small cell lung cancer (NSCLC) has not been fully evaluated. We examined the effect of ABT-263 on cell proliferation of NSCLC cell lines and investigated the underlying mechanisms. Methods: The following 9 NSCLC cell lines were examined: SK-LU-1, A549, H358, Calu3, H3122, H1975, H460, H441, and BID007. The effects of ABT-263 in NSCLC cell lines were evaluated by MTS assay. Apoptosis was examined by flowcytometry using staining for annexin V and propidium iodide (PI), and also western blotting for cleaved PARP. Quantitative RT-PCR was carried out to assess the mRNA expression levels of anti-apoptotic genes and pro-apoptotic genes. Immunoprecipitation and western blotting were performed to compare the levels of anti-apoptotic and pro-apoptotic proteins between the sensitive and resistant cell lines. In addition, knockdown of Mcl-1 was performed by siRNA. Results: By screening 9 NSCLC cell lines using MTS assay, we found Calu3 and BID007were sensitive to ABT-263. We also confirmed that apoptosis was induced only in the ABT-263 sensitive lines but not in the ABT-263 resistant cell lines after ABT-263 treatment. However, the expression levels of Bcl-2 family proteins, including Mcl-1, did not differ significantly among the ABT-263 sensitive and resistant cell lines. Unlike the results in previous reports regarding SCLC, Mcl-1 was not decreased in the sensitive cell lines. The ABT-263 resistant cell lines became sensitive to ABT-263 after knockdown of Mcl-1 by siRNA, while the ABT-263 sensitive cell lines maintained the same sensitivity. Conclusion: We found that Calu3 and BID007 were sensitive to ABT-263. In the sensitive NSCLC cell lines, ABT-263 induces apoptosis irrespective of Mcl-1 expression levels. Citation Format: Aoi Kuroda, Keiko Ohgino, Hiroyuki Yasuda, Junko Hamamoto, Daisuke Arai, Kota Ishioka, Tetsuo Tani, Shigenari Nukaga, Ichiro Kawada, Katsuhiko Naoki, Kenzo Soejima, Tomoko Betsuyaku. ABT-263 is effective in a subset of non-small cell lung cancer cell lines. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4. doi:10.1158/1538-7445.AM2015-4

The gross anatomy of the mastication system of the giant anteater (Myrmecophaga tridactyla) was examined by means of three-dimensional image analysis. The anteater rotates the mandibles medially and laterally to control its tongue when it is elongated and to house it when it is relaxed. Three-dimensional CT image analysis demonstrated that the shape and size of the oral cavity changes drastically when the mandibles are rotated. The oral cavity expands bilaterally when the dorsal part of the mandibles bend medially. Macroscopic observations and muscle-weight data supported the observation that the superficial temporal and medial pterygoid muscles act as the main medial and lateral rotators of the mandible, respectively. The low height of the mandibular ramus and the incomplete zygomatic arch in this species represent adaptations for the rotational movement of the mandibles, since they both contribute to the medially oriented transmission of force from the temporal muscles and to preventing collision between the mandibles and the cranium during the rotational movement.

We re-examined tooth variation in specimens of the Siberian mole, Talpa altaica, from the collection of the Siberian Zoological Museum and discuss the mechanisms of dental evolution. The number of teeth counted in 1789 specimens ranged from 34 to 47, and supernumerary, absent, and connate teeth were observed. The most frequent tooth anomaly was an absent tooth in the premolar region (200 maxillary first premolars and 190 mandibular third premolars), which does not support Fujita and Kirino's terminal reduction hypothesis in the mandible [Fujita T, Kirino T. Ha No Kaibougaku. 21st ed. Tokyo: Kanehara Publishers Inc.; 1976 (in Japanese)]. Supernumerary teeth were found in premolar rows and in the incisor and molar regions. An maxillary fourth molar, positioned distal to the normal third molar, was thought to result from a genetically programmed atavistic event during the natal stages. Connate teeth were observed only in the premolar rows and were thought to have developed with the fusion of two independent tooth germs. Connate premolars appeared to result from an expression of an incomplete division of tooth germ at an early developmental stage or a reunion of independent tooth germs, based on the morphological similarity of the normal and supernumerary premolars. These extraordinarily frequent tooth anomalies of T. altaica are of much interest both in terms of tooth development and classification.

Ultra-low pressure RO membrane module “ES series” and high performance under operating pressure of 0.7 MPa. The ES series requires about half of the operating pressure as most commonly available aromatic polyamide composite RO membranes, so they can save energy costs and initial costs.The ES series consist of 3 types, that is, “ES10”, “ES15” and “ES20”. “ES10”, is a base grade, in which NaCl rejection is 99.5% and permeate flux is 30 m3/day at 0.7 MPa. “ES15” has higher permeate flux, and “ES20” has higher NaCl rejection than that of ES10. All types have 2 grades for their usage, that is, desalination of tap water and making of ultra pure water.

Metastasis to the tongue seldom occurs, and lingual metastasis as an initial sign of cancer occurs even less frequently. We report a case of lung cancer in which the patient's initial symptom was related to the tongue metastasis. A 63-year-old man had a submucosal tumor on the left posterolateral aspect of the tongue and a biopsy specimen of the tongue tumor showed poorly differentiated squamous cell carcinoma. A chest X-ray showed a mass in the right lung and cytological examination of the specimen obtained by bronchial brushing showed poorly differentiated squamous cell carcinoma, whose appearance was similar to that of the tongue. Based on these findings, the tongue lesion was diagnosed a metastatic tumor from the lung cancer. The patient received radiation therapy combined with systemic chemotherapy, however, he died 5 months after the diagnosis of lung cancer. An autopsy revealed a lung cancer in the right lower lobe with metastatic tumors in the tongue, right middle lobe, left upper lobe, liver, adrenal gland, pericardium, heart, and subcutaneous tissues. No other possible primary cancer that may have been the cause of the metastases was identified.

We describe a case of lung tumor in a 33-year-old woman with tongue cancer. She had noticed a painful sensation in her tongue in April 2000. The results of a physical examination showed a 22 x 11 mm ulcerated lesion on the right side of her tongue. A biopsy specimen showed a moderately differentiated squamous cell carcinoma. A preoperative chest radiograph showed no evidence of pulmonary metastasis. No neck lymphadenopathy was found. The tongue cancer was resected in September 2000, and the pathological stage was T2 N0 M0. A solitary pulmonary lesion appeared 8 months after the surgery. A Histological examination of a transbronchial lung biopsy specimen showed a moderately differentiated squamous cell carcinoma. It was difficult to distinguish histopathologically or immunohistochemically between a second primary tumor and a metastasis, but the pulmonary lesion was considered to be a second primary tumor on the basis of a single endobronchial lesion that appeared after the T2 tongue cancer with no regional recurrence or cervical lymphadenopathy. Treatment with chemotherapy and irradiation was not effective and the patient died 3 months after the diagnosis of lung cancer.

Introduction: Lung cancer can be a heterogeneous group of disorders. We have identified that the MET receptor tyrosine kinase is overexpressed, mutated, and/or amplified in various subsets of lung cancer. Several clinical trials of MET inhibitors that are underway have documented cases of acquired resistance. Based on evidence that the RON tyrosine kinase receptor (family member of MET receptor) can also be overexpressed in NSCLC, we determined the effect of sequential and concurrent inhibition of MET and RON with gene silencing. We identified that both MET and RON gene silencing led to inhibition of signal transduction, but that the combination was more effective. We evaluated the potent MET/RON dual kinase inhibitor LY2801653 in this setting and found it to be more efficacious than the MET/ALK/RON/ROS inhibitor crizotinib with a distinct pattern of changes in downstream signaling. Using the protein phosphorylation detection platform PamGene®, we found that inhibition of MET and RON was associated with decreased phosphorylation of CBL, PI3K and STAT3. A systematic analysis of the PamGene® data allowed us to define abnormalities of the signal transduction pathways. We further report that LY2801653 also has biological and tumor inhibitory effect both in vitro and in vivo. In mouse flank and orthotopic lung xenografts of NSCLC, LY2801653 decreased tumor growth, dramatically inhibiting mitotic events and angiogenesis.. Taken together, our results argued that specific targeting of the MET/RON kinases could provide robust inhibition of cell proliferation and tumor growth in multiple in vitro and in vivo models of NSCLC. These findings offer a robust preclinical proof of concept for MET/RON targeting by LY2801653 as a promising small molecule treatment modality. Methods: LY2801653 was provided by Eli Lilly and Company (Indianapolis, IN). NSCLC cell lines purchased from ATCC were utilized to conduct in vitro and in vivo studies. Cell proliferation assays were performed using MTT, standard immunoblot assays and the high-throughput protein phosphorylation detection platform PamGene® was used to demonstrate phosphorylation events. To determine drug effects in vivo, the mouse flank xenograft model and an orthotopic lung tumor model was used. In the orthotopic model, tumor progression was monitored by bioluminescence using Xenogen IVIS®. Results: We have reported that LY2801653 specifically inhibited proliferation of NSCLC cell lines at IC50 as low as 6-600 nM with inhibition of phosphorylation in downstream signaling of MET/RON. The PamGene® platform also showed a network of downstream signaling molecules affected by MET/RON inhibition, such as CBL, PI3K and STAT3. We further report that LY2801653 also has tumor growth inhibitory effects in mouse xenografts and dramatic effects on mitosis and angiogenesis. Conclusions: Specific inhibition of MET/RON by a small molecule inhibitor LY2801653 was able to abrogate cell proliferation, tumor growth, as well as affect downstream signal transduction of MET/RON in NSCLC. These results demonstrate the promising therapeutic potential of LY2801653 in treating lung cancer. Citation Format: Rifat Hasina, Ichiro Kawada, Qudsia Arif, Jeffrey Mueller, Aliya N. Husain, Everett E. Vokes, Ravi Salgia. Inhibition of MET/RON in 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 LB-239. doi:10.1158/1538-7445.AM2014-LB-239

Malignant pleural mesothelioma (MPM) is an aggressive disease with a poor prognosis. Studies have shown that both MET and its key downstream intracellular signaling partners, PI3K and mTOR, are overexpressed in MPM. Here we determined the combinatorial therapeutic efficacy of a new generation small molecule inhibitor of MET, ARQ 197, and dual PI3K/mTOR inhibitors NVP-BEZ235 and GDC-0980 in mesothelioma cell and mouse xenograft models. Cell viability results show that mesothelioma cell lines were sensitive to ARQ 197, NVP-BEZ235 and GDC-0980 inhibitors. The combined use of ARQ 197 with either NVP-BEZ235 or GDC-0980, was synergistic (CI

Despite substantial improvements in screening, diagnosis, and treatment of esophageal cancer, the prognosis of this disease remains bleak. Survival at 5 years for all esophageal cancer patients taken together, with amenable treatments and with or without surgery ranges from 5-20% (Cancer Facts and Figures. American Cancer Society 2011). Although a multidisciplinary approach to include surgery, radiation, and chemotherapy, alone or in combination, attempts to improve the survival of this aggressive disease, these statistics underscore the continued need for attention to this disease and for identification of new targets in its treatment. We have been studying the role of receptor tyrosine kinases (RTKs) in cancer, particularly the EphB4 receptor, which has become increasingly associated with the pathobiology of adult cancers over the past several years. It has been shown to be aberrantly expressed and/or to play an oncogenic role in cancers of the breast, bladder, ovary, uterus, colon, head and neck, and prostate through its effects on cellular motility, growth, and migration, as well as on tumors’ ability to induce neoangiogenesis. To date, its role and potential as a target for therapy in different cancers have not been thoroughly investigated and remain poorly understood. Targeted inactivation of EphB4 and its ligand Ephrin-B2 have demonstrated that both are essential for angiogenic remodeling and embryonic survival (Adams and Klein 2000; Kim, Hu et al. 2008). In this report, we determine that EphB4 is overexpressed, has increased gene copy number and is involved in enhanced motility and migration in esophageal cancer. Archival patient samples consisting of 93 squamous cell carcinoma, 100 adenocarcinoma and 25 adjacent normal control samples as well as four adenocarcinoma and nine squamous cell carcinoma cell lines were used for the studies. Extensive mouse modeling of esophageal cancer was also used. We have reported that there is consistently higher expression of EphB4 in both squamous and adenocarcinoma compared to adjacent normal tissue with a statistically significant correlation between EphB4 expression and higher grades of squamous cell carcinoma. In a chemically induced esophageal squamous cell carcinoma model in mice, EphB4 was found overexpressed compared to normal controls. This study identifies EphB4 to be an important pathway in esophageal neoplastic lesions. It would now be useful to bring this to clinical fruition. 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 1861. doi:1538-7445.AM2012-1861

Paxillin is a 68 kDa focal adhesion protein that provides multiple docking sites at the plasma membrane for an array of signaling molecules and helps form a structural link between the extracellular matrix and the actin cytoskeleton. It is essential in actin filament assembly and focal adhesion formation. Focal adhesion complexes are the drivers of cell spreading and migration, adhesion to the extracellular matrix and matrix remodeling by fibrillar adhesions in which paxillin plays a major role. It has been shown that the modulation of tyrosine phosphorylation of paxillin regulates both the assembly and turnover of adhesion sites and phosphorylated paxillin enhances lamellipodial protrusions whereas non-phosphorylated paxillin is essential for fibrillar adhesion formation. We have previously reported that in lung cancer, paxillin was over expressed, amplified and mutated in a significant number of patient samples and the protein upregulated in higher stages of lung cancer compared with lower stages (Jagadeeswaran et al, 2008). We recently showed paxillin gene to be amplified in some pre-neoplastic lung lesions. Among the mutations we described, A127T enhanced cell proliferation, focal adhesion formation and colocalization with Bcl-2 in lung cancer cells. We have now analyzed 50 epithelioid, 16 sarcomatoid, and one mixed malignant pleural mesothelioma tissue, and compared to 40 normal adjacent lung parenchyma (pneumocytes and endothelial cells) by immunohistochemistry. Automated cellular imaging was used to calculate average staining intensity as an integrated optical density (IOD). Whereas normal lung had IODs of 45 for paxillin, epithelioid MPM had IODs of 268 and sarcomatoid 331. This demonstrates that paxillin is highly upregulated in both epithelioid as well as sarcomatoid. To further study the effects of activating mutations of paxillin, we cloned the most commonly occurring paxillin mutations in a GFP tagged vector and transiently transfect HEK-293 cells. Utilizing live-cell imaging to systematically study the effects of wild-type paxillin versus mutants, we created a mathematical model that recapitulates the salient features of the measured dynamics, and conclude that compared to wild-type, some mutant clones confer a) enhanced focal adhesion formation, b) increased filopodia and lamellipodia formation c) increased mobility and d) increased cell displacement in transiently transfected HEK-293 cells. We believe that paxillin is an important molecule in thoracic cancer including lung and malignant mesothelioma, and its therapeutic potential needs to be explored further. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2341. doi:10.1158/1538-7445.AM2011-2341

There are a number of molecular abnormalities that can occur in normal cells to induce a malignant phenotype. Recently, the receptor tyrosine kinase anaplastic lymphoma kinase (ALK) has been shown to have gain-of-function when partnered with different proteins. As an example, on chromosome 2p, with inversion, there is translocation with generation of EML4-ALK tyrosine kinase in lung cancer. In a phase I trial, EML4-ALK patients were selected to determine the response to a potent small molecule tyrosine kinase inhibitor crizotinib (previously identified as PF02341066). Dramatic durable responses were observed with crizotinib at 250 mg twice a day (orally). Interestingly, crizotinib also has activity against MET receptor tyrosine kinase. We have previously shown that MET can be overexpressed, sometimes mutated, or sometimes amplified in lung cancer. Thus, this review will emphasize the characteristics of crizotinib, and detail the clinical experience.

7071 Background: Somatic mutations in the tyrosine kinase domain of the epidermal growth factor receptor (EGFR) gene are reportedly associated with sensitivity of non-small cell lung carcinomas (NS...

Ternary copper(I) complexes with alkynes and 1,10-phenanthroline (phen) have been prepared and characterized by their formation constants, IR spectra, 1H and 13C NMR spectra. The structures of three complexes, [Cu(phen)(HCCR)]ClO4(R = H 1, Ph 2 or CO2Et 3) have been determined using single-crystal X-ray methods. Each copper atom in all three complexes is co-ordinated to two nitrogen atoms of phen and an alkyne molecule in a trigonal-planar arrangement. The alkyne is sideways bonded onto the copper atom. The CC bond distances of 1.190(7)(1), 1.218(13)(2) and 1.193(10)A(3) are slightly longer than those of the free alkynes and indicate that π back donation from copper to the alkyne is weak. The reduction of the CC stretching frequency for metal–alkyne complexes is correlated to the acetylenic bend-back angles and the lenghthening of the CC distance of the alkyne upon co-ordination. The copper–carbon and –nitrogen distances for 1 and 2 are essentially the same as those for the corresponding olefin complexes.

A series of alumina substrates were prepared with controlled variation of normal parameters — alumina content, density, forming method, and minor constituents. Variations in dielectric constant at X-band frequencies were determined by different methods, including microwave cavities, and the results interpreted.