bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1717: Prognostic impact of cancer stem cell-related markers in non-small cell lung cancer patients treated with induction chemoradiotherapy
Authors: Shien Kazuhiko , Toyooka Shinichi , Ichimura Kouichi , Soh Junichi , Furukawa Masashi , Maki Yuho , Muraoka Takayuki , Tanaka Norimitsu , Ueno Tsuyoshi , Asano Hiroaki , Tsukuda Kazunori , Yamamoto Hiromasa , Yamane Masaomi , Oto Takahiro , Kiura Katsuyuki , Miyoshi Shinichiro
Published: August 2012

Background: Several cancer stem cell (CSC) related markers have been confirmed to be expressed in non-small cell lung cancer (NSCLC). The aim of this study was to clarify the prognostic role of CSC related markers expression in locally advanced NSCLC with induction chemoradiotherap (CRT).Methods: Fifty patients with clinically N2 or N3 NSCLC underwent induction CRT with cisplatin and docetaxel concurrently with thoracic radiation followed by surgery. Expressions of CSC related markers (CD133, ABCG2, ALDH1, and Bmi-1) were examined by immunohistochemical stain from surgical resected specimen. The prognostic factors were investigated by multivariate analysis using Cox proportional hazard model.Results: Among 50 patients, 20 patients had no pathologically residual tumor cell in the surgical resected specimen, and CSC related markers expressions were evaluated in other 30 patients. With a median follow-up duration of 71 months, the 5-year overall survival rate in the patients of CD133 or ALDH1 -positive was significantly worse than that in the patients of CD133 and ALDH1 -negative (90.0% vs. 44.9%, respectively, P = 0.042). In multivariate analysis, CD133 and ALDH1 negativity (Hazard ratio [HR] 0.16, 95% confidential interval [CI]: 0.0086-0.98, P = 0.047) and c-N2-3 single node metastasis (not multi node metastasis) (HR 0.19, 95% CI: 0.028-0.91, P = 0.03) were significant independent prognostic factors for the prolongation of survival.Discussion: The expressions of CSC related markers after CRT are significantly related to poor prognosis in NSCLC. The development of therapeutic strategy including adjuvant therapy, is a key for further improvement of prognosis in patients who underwent trimodality therapy.

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 1717. doi:1538-7445.AM2012-1717
Abstract 1718: Favorable clinical outcomes of pemetrexed treatment in anaplastic lymphoma kinase positive non-small-cell lung cancer patients were associated with low expression of thymidylate synthase in tumor
Authors: Lee Ha Yeon , Ahn Hee Kyung , Jeong Ji Yun , Kwon Mi Jung , Han Jung-Ho , Sun Jong-Mu , Ahn Jin Seok , Park Keunchil , Cha Joo-Young , Cho Yeoung-Mi , Choi Yoon-La , Ahn Myung-Ju
Published: August 2012

Introduction The purpose of this study was to investigate the clinical characteristics and treatment outcomes of patients with anaplastic lymphoma kinase (ALK)-positive non-small-cell lung cancer (NSCLC) to conventional chemotherapy in the pre-ALK inhibitor era Patients and Methods We retrospectively screened 381 consecutive NSCLC patients without known epidermal growth factor receptor (EGFR) or KRAS mutation who were diagnosed between 2007 and 2008 at a single center, and identified ALK rearrangements by fluorescence in situ hybridization Additional 44 ALK-positive patients from other period were included for the analysis of clinical outcomes Results Of the 381 tumors screened, four were excluded because the samples were unevaluable Twenty-one (5 6%) showed ALK rearrangements, with twenty adenocarcinomas and one pleomorphic carcinoma Of the entire 65 ALK-positive patients, 32 patients received pemetrexed as a second- or further-line therapy, in whom response rate was 34 4% (11/32) and median progression-free survival (PFS) was 4 0 months Among these 31 patients, 20 specimens were available for thymidylate synthase (TS) expression analysis Low expression of TS were found in 80% (16/20), with a trend toward longer PFS than TS-positive patients (median PFS 4 0 vs 1 0 months, P = 0 095) Conclusions The prevalence of ALK rearrangement was 5 6% among EGFR and/or KRAS wild-type/unknown NSCLC population Low TS protein expression might be associated with better clinical outcomes in ALK-positive NSCLC patients after pemetrexed given as a second- or further-line therapy

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 1718. doi:1538-7445.AM2012-1718
Abstract 1727: Correlation between BRCA1 and LMO4 mRNA expression levels in Erlotinib-treated non small cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) mutations
Authors: Costa Carlota , Giménez-Capitán Ana , Benlloch Susana , Tarón Miquel , Jimeno José , Viteri Santiago , Gascó Amaya , Camps Carlos , Carcereny Enric , Massuti Bartomeu , Rosell Rafael
Published: August 2012

Background Advanced NSCLC p harboring EGFR mutations show impressive progression-free survival (PFS) when treated with erlotinib. However, presence of EGFR mutations only imperfectly predicts outcome. We demonstrated that co-existence of the EGFR T790M pretreatment mutation was detected in one-third of p and, in conjunction with elevated BRCA1 mRNA levels, dramatically influenced PFS to erlotinib. Though this model is a breakthrough in outcome of EGFR-mutated NSCLC p treated with TKIs, we choose to analyze the impact of CTIP and LMO4 gene expression levels on p outcome and their correlation with BRCA1. CtIP binds to BRCA1 and LMO4 forming a complex. LMO4 is a negative regulator of BRCA1 function in sporadic breast cancers. Methods mRNA expression of LMO4 and CTIP was examined by RT-PCR in pretreatment tumor biopsies of 72 NSCLC p with EGFR mutations treated with erlotinib. Expression levels were correlated with outcome to erlotinib and BRCA1 expression levels. Results BRCA1 significantly correlated with CtIP (r=0.31;P=0.01) and LMO4 (r=0.32;P=0.02). There was no correlation between CtIP and LMO4 (r=0.09;P=0.49).PFS was not reached for p with high levels of LMO4 vs 13 months for p with low levels (P=0.006). Overall survival (OS) was not reached for p with high levels of LMO4 vs 29 months for p with low levels (P=0.10). Expression levels of CtIP did not correlate with PFS and OS. PFS was not reached for p with low levels of BRCA1 and high levels of LMO4 vs 19 months for p with low levels of both genes (P=0.04). PFS was 8 months for p with high levels of BRCA1 and low levels of LMO4 vs 18 months for p with high levels of both genes (P=0.03). Conclusions BRCA1 and LMO4 mRNA levels examined by RT-PCR could predict PFS to erlotinib in p with EGFR mutations and could be useful for development of new therapeutic strategies.

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 1727. doi:1538-7445.AM2012-1727
Abstract 1750: A phase 3 trial of weekly farletuzumab combined with carboplatin and taxane in subjects with platinum-sensitive ovarian cancer in first relapse
Authors: Weil Susan C. , Jenkins Michele , Hoffman Kimberly
Published: August 2012

Background: Ovarian cancer is the leading cause of death among women with gynecologic malignancies in North America. Despite advances in treatment, only 15%-30% of patients survive long term, underscoring the need for more effective regimens. Folate receptor ≤ (FRα) is highly expressed on all epithelial ovarian cancer (EOC) cells but largely absent from normal tissue. This study examines the efficacy and safety of farletuzumab, a monoclonal antibody that binds FRα, when combined with standard chemotherapy, in platinum-sensitive subjects with ovarian cancer after first relapse. Methods: This phase 3, randomized, double-blind, placebo-controlled trial is ongoing at 375 sites globally. Eligible subjects are β18 years of age, with a histologically confirmed diagnosis of non-mucinous EOC, including primary peritoneal and fallopian tube malignancies. Subjects must have undergone debulking surgery and achieved an objective response lasting 6-24 months following first-line platinum and taxane chemotherapy, followed by relapse confirmed by computed tomography or magnetic resonance imaging. Approximately 1080 subjects will be randomized to receive weekly intravenous farletuzumab (1.25 mg/kg or 2.5 mg/kg) or placebo, plus standard carboplatin (area under the curve [AUC] 5-6) and taxane (paclitaxel at 175 mg/m2 or docetaxel at 75 mg/m2) every 3 weeks for 6 cycles. Maintenance treatment follows, with weekly single-agent farletuzumab or placebo administered until disease progression. The primary end point is progression-free survival (PFS) based on Response Evaluation Criteria in Solid Tumors (RECIST). Secondary outcomes are overall survival, CA-125-defined PFS (based on Rustin criteria) and serologic response, best objective response and time to response, duration of first vs current remission (by RECIST), subject-reported quality of life, and resource utilization (hospitalizations, unscheduled office visits, and admissions to hospice or nursing home). Molecular markers are being sought that may be predictive of response to farletuzumab. Pharmacokinetic analysis is performed to assess interactions between agents. Safety will be evaluated by review of physical assessments, laboratory data, and adverse events; review is overseen by an independent data monitoring committee. As of November 7, 2011, 957 (of 1080 planned) subjects have enrolled in the study. The last subject will likely be enrolled by March 2012.

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 1750. doi:1538-7445.AM2012-1750
Abstract 1758: A podophyllotoxin derivative, Adva-27a, is more potent at killing multi-drug resistant breast cancer cells than etoposide
Authors: Young Rebecca S. , Liao Si , Slilaty Steve N. , Gal Susannah
Published: August 2012

A novel non-ester GEM-difluorinated C-glycoside derivative of podophyllotoxin, Adva-27a, had been designed and synthesized. The growth inhibitory properties of this drug compared to the currently used Topoisomerase II inhibitor etoposide was evaluated in MCF-7 breast cancer cells and in a multi-drug resistant (MDR) variant of this cell line. These cells were cultured in the presence of the drugs at different concentrations (0.1-100μM) for various times (1-9 days) and analyzed for viability using a mitochondrial metabolic activity dye, Alamar blue versus solvent (DMSO) treated cells. The MCF-7 cells were sensitive to Adva-27a and etoposide, resulting in an IC50 of less than 10μM for etoposide and 20μM for Adva-27a, while the IC50 for the MDR MCF-7 cells was 25μM with Adva-27a, but greater than 100μM for etoposide. The timing for killing by Adva-27a of the MDR MCF-7 cells appeared significantly different than for the MCF-7 parent cells. In this case, 100μM drug resulted in less than 50% of viability at day 3 for the MDR MCF-7 cells, while the same concentration of drug required 5-6 days to cause the same level of killing of the parent cells. To determine the mechanism of action of the drug, we analyzed the Adva-27a treated cells by flow cytometry using propidium iodide staining. We found that after 24 hours with 30μM of the drug, both MCF-7 and the MDR MCF-7 cells were accumulating in the G2/M phase of the cell cycle. Both cells are similarly killed after 7 days by this concentration of the drug. Etoposide is known to cause accumulation of cells at a similar phase, implying the Adva-27a has a similar drug target. Thus, the mechanism for the difference in sensitivity of the MDR MCF-7 cells for Adva-27a relative to etoposide is not yet understood, but will be explored in the future.

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 1758. doi:1538-7445.AM2012-1758
Abstract 1766: Persistence of drug-induced DNA interstrand cross-links distinguishes bendamustine from conventional DNA cross-linking agents
Authors: Bagnobianchi Alessia , Spanswick Victoria J. , Bingham John P. , Kiakos Konstantinos , Suarez-Henriques Paula , Smith Dean , Yong Kwee , Hochhauser Daniel , Hartley John A.
Published: August 2012

Bendamustine has demonstrated substantial clinical efficacy in the treatment of hematologic malignancies and continues to distinguish itself from other alkylating agents. The mechanistic and clinical differences associated with bendamustine may be directly related to its unique structural features, although the precise mechanism of action is still poorly understood. We have undertaken a detailed study of the drug-DNA interactions of bendamustine. It alkylates DNA primarily at guanine-N7 positions with a sequence selectivity similar to other nitrogen mustards. It produces DNA interstrand cross-links (ICLs) in both solid tumor and hematological tumor cells at doses that cause growth inhibition. ICLs peak at 8h following a 1h treatment and persist over 48h in tumor cells including those that are proficient at repairing (unhooking) ICLs formed by melphalan or cisplatin. This persistence of ICLs is also observed in multiple myeloma cells from patients who have relapsed on melphalan therapy and which efficiently repair melphalan-induced ICLs compared to cells from treatment naive patients. The peak γ-H2AX response follows the ICL peak and also persists beyond 48h. Cells defective in ICL unhooking proteins ERCC1 and XPF, or defective in homologous recombination repair, show increased sensitivity to bendamustine, but at a level less than observed for other nitrogen mustards or cisplatin. Real time PCR profiling showed that at equivalent peak levels of DNA ICLs bendamustine induced fewer changes in DNA damage signaling and DNA repair gene expression compared to either cisplatin or melphalan. These data suggest that the critical DNA damage produced by bendamustine induces a different signaling and repair response to conventional cross-linking agents, including other nitrogen mustards, which may be an important contribution to its clinical efficacy.

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 1766. doi:1538-7445.AM2012-1766
Abstract 1769: Significant in vivo activity of an APE1/Ref-1 redox inhibitor, E3330, alone and in combination with Bevacizumab in a glioblastoma mouse model analyzed by a whole slide digital imaging system and quantitative immunohistochemistry
Authors: Kumar Krishan , Jackson Jessica L. , Kelley Mark R. , Ivan Mircea , Sandsuky George
Published: August 2012

Human apurinic endonuclease/redox factor 1 (APE1/Ref-1) mediates the repair of baseless sites in DNA caused by alkylation and oxidative DNA damage as well as regulates the function of various transcription factors (TF) under redox signaling control (p53, NFkB, HIF-1, AP-1, and others). High levels of APE1 expression have been reported in numerous cancers (glioblastoma, ovarian, pancreatic, prostate, and others) such that APE1 is an emerging target for a variety of novel anticancer drugs. E3330 targets the redox signaling function of APE1 and its activity was studied in vivo using a glioblastoma xenograft mouse model. This preclinical xenograft model evaluated the use of dual anti-angiogenic agents, Bevacizumab (Bev) and E3330, in glioblastoma implanted Nude mice. Four mice groups were used to determine drug effects: E3330, Bev, a combination of E3330 and Bev, and an untreated vehicle control group. Immunohistochemistry (IHC) was used to predict the effectiveness of treatment for glioblastoma based on caspase-3, Ki67, and CD31 biomarker expression. IHC slides were quantified using both a traditional pathology hand count and an image system analysis. The positive pixel data closely mirrored the hand count for all three biomarkers. At the combined dosage, hand count data demonstrated that CD31 levels decreased significantly from the vehicle control group. Ki67 decreased slightly, and caspase-3 was not increased in any of the drug treatment groups. While each drug independently had anti-angiogenic effects, the combined drug group, E3330 and Bev, had anti-angiogenic effects that were greatly enhanced. These results support previous studies demonstrating the anti-angiogenic activity of E3330 and its enhancement of Bevacizumab.

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 1769. doi:1538-7445.AM2012-1769
Abstract 1776: Antitumor activity of Chk1 inhibitor LY2606368 as a single agent in SW1990 human pancreas orthotopic tumor model
Authors: Wu Wenjuan , Bi Chen , Bence Aimee K. , Um Suzane L. , Yan Betty , Starling James J. , Marshall Mark S. , Beckmann Richard P.
Published: August 2012

ChK1 is a key kinase in the DNA damage response signaling network, including cell cycle checkpoints, DNA repair, apoptosis and transcription, thus emerging as an attractive target in anti-cancer therapy. LY2606368, a potent and selective ATP competitive inhibitor of the Chk1 protein kinase, is currently in clinical development. LY2606368 has been reported to inhibit Chk1 auto-phosphorylation activated by DNA damaging agents, and induce phosphorylation of H2AX, a DNA damage maker in multiple cancer cell lines in vitro. In addition, LY2606368 has demonstrated potent single agent activity and potentiates the anti-tumor activity of DNA damaging agents in vivo. Pancreatic cancer is one of the least curable cancers, with an approximate 5% overall 5-year survival for all patients. In the current study, we evaluated the efficacy of LY2606368 (as its methanesulfonate salt, hydrate; LY2606368.MeSO3H.H2O = LY2940930) in pancreatic cancer cell lines in vitro and xenograft tumors (including subcutaneous and orthotopic models) in vivo. LY2606368 alone significantly inhibited the cell proliferation in a variety of pancreatic cell lines (SW1990, SU86.86, Bx-PC3, AsPC-1, CFPAC-1, Capan-2, HPAF-II) with SW1990 being the most sensitive (IC50=1.5 nM). In SW1990 subcutaneous xenograft model, LY2606368 showed substantial dose-dependent inhibition of tumor growth. In SW1990 pancreas orthotopic model, which represents the local and metastatic growth pattern seen in pancreas cancer patients, LY2606368 treatment resulted in over 92% inhibition of primary tumor growth as well as 100% inhibition of metastasis to lymph node, spleen and intestine. The anti-tumor effect of LY2606368 treatment was further demonstrated in comparing with gemcitabine (the standard of care for pancreas cancer patient) in SW1990 orthotopic model. The anti-tumor and/or anti-metastasis mechanism of LY2606368 was also investigated, including phosphorylation of H2AX, cell proliferation, cell survival, and soft agar anchorage-independent growth. Together, our findings demonstrate the therapeutic potential of LY2606368 as a single agent in the treatment of pancreatic cancer patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1776. doi:1538-7445.AM2012-1776
Abstract 1796: SAR302503: A Jak2 inhibitor with antitumor activity in solid tumor models
Authors: Greco Rita , Hurley Raelene , Sun Fangxian , Yang Lu , Yu Qunyan , Williams Juliet , Dorsch Marion , Cohen Pamela , Adrián Francisco
Published: August 2012

Numerous studies have recognized the critical role of STAT3 in malignant transformation and tumor progression. Constitutive STAT3 activation is frequently found in cancer cell lines and tumor samples and it is usually linked to the presence of IL-6. Autocrine or paracrine IL-6 loops have been described to provide tumor cells with the ability to proliferate, survive, migrate and metastasize. At the molecular level, IL-6 binding to its receptor results in activation of Jak/STAT3 signaling and other signaling cascades, namely PI3K/Akt, MEK/ERK1-2, with a well established role in cancer. We have evaluated the ability of SAR302503, a selective Jak2 inhibitor entering a PhIII clinical trial in myelofibrosis patients, to block these pathways in a panel of ∼20 tumor cell lines representing different cancer types (prostate, breast, lung, colorectal, pancreas, hepatocellular, etc). A 45 minutes treatment with different concentrations of SAR302503 (0.1-10 µM) was sufficient to block both basal or IL-6 induced STAT3 phosphorylation in a dose dependent manner. Compound concentrations equal or greater than 0.1 µM were able to reduce the phosphorylation levels of STAT3 by an extent greater than 50% in all the cell lines included in the study. The impact of SAR302503 in tumor cell proliferation and survival was evaluated using different assays including clonogenic assays. Complete inhibition of colony formation was achieved at concentrations of SAR302503 below 1 µM in most of the cell lines. The antitumor activity of SAR302503 was evaluated in mice xenotransplanted subcutaneously with DU145 human prostate cancer cells. Oral administration of SAR302503 for 10 days resulted in significant dose dependent tumor growth inhibition, near to stasis at the highest dose (T/C=19% at 100 mg/kg, bid). In summary, we demonstrate that SAR302503 negatively impacts the proliferation and survival of different solid tumor cells and our data supports a role for a selective Jak2 inhibition in treating solid tumors with activated Jak/STAT signaling.

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 1796. doi:1538-7445.AM2012-1796
Abstract 1797: Preclinical evaluation of HM-018, a potent and selective JAK inhibitor in the treatment of myloproliferative disorders
Authors: Ren Yongxin , Deng Wei , Gu Weihua , Shen Wuzhong , Yu Youjun , Yang Xiaoning , Zhang Zhixiang , Fan Shiming , Sai Yang , Wang Jian , Yan James , Li Jia , Zhao Helen , Qing Weiguo , Su Weiguo
Published: August 2012

JAK2 kinase mutation V617F is prevalent in myeloproliferative diseases (MPD), including polycythemia vera (PV, 81-99%), essential thrombocytosis (ET, 41-72%) and primary myelofibrosis (PMF, 39-57%). This point mutation constitutively activates the JAK kinase and leads to oncogenic potential of host cells, and thus making JAK2 a promising molecular target for MPD therapy. HM-018 is a small molecule inhibitor against JAK kinase and the compound's preclinical anti-MPD effects from signal transduction to biological consequences were investigated. HM-018 was found to inhibit JAK kinase 1, 2, 3 and TYK with IC50 of 0.010, 0.006, 0.040 and 0.047 μM, respectively. The compound demonstrated >100 folds selectivity against a panel of 63 kinases. In accordance with enzymatic activity, HM-018 suppressed ligand dependent or constitutive JAK activation in multiple cell lines as evidenced by the decrease of STAT3/5 phosphorylation. As a result, JAK dependent cell proliferation was significantly inhibited by HM-018. EPO-mediated mouse PV model was utilized to evaluate the in vivo efficacy of the compound. HM-018 could shrink enlarged mouse spleen, a typical symptom of PV, in a dose dependent manner accompanied with decreased STAT5 phosphorylation both in animal spleen and in bone marrow after oral dosing for 7 days. To better mimic MPD development in a more clinically-relevant manner, JAK2-V617F-tranfected 32D cells were injected into mice intravenously, and it was observed that oral treatment of HM-018 not only prolonged the animal's life span, but also reduced MPD-related symptoms, such as spleen weight increase and organ invasion by malignant cells. Meanwhile, HM-018 exhibited a favorable pharmacokinetic profile and acceptable safety window in rats. Based on the preclinical data, HM-018 demonstrated anti-MPD potency both in vitro and in vivo, and the studies have provided rationale to further develop this compound as possible MPD therapeutics.

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 1797. doi:1538-7445.AM2012-1797
Abstract 2416: Neuropilin-2 is upregulated during EMT in lung cancer
Authors: Nasarre Patrick , Roche Joelle , Potiron Vincent A. , Nair-Menon Joyce , Gemmill Robert M. , Drabkin Harry A.
Published: August 2012

Neuropilins (NRP1 and NRP2) are high affinity receptors for the class-3 semaphorins, cell guidance molecules involved in tissue development, immune responses, angiogenesis and cancer. Semaphorins affect cell protrusion, spreading and adhesion, and can trigger migratory responses often in a repulsive manner depending on the cellular context. Interestingly, neuropilins are also receptors for galectin-1 and growth factors including VEGF, PlGF, EGF and TGFβ. We previously cloned the SEMA3F gene from chromosome 3p21.3, which undergoes homozygous deletion and frequent loss of heterozygosity in lung cancer. We found that SEMA3F levels were inversely correlated with tumor aggressiveness in human lung cancer and confirmed its tumor suppressor activity in experimental xenograft models. We subsequently discovered that SEMA3F is directly downregulated by ZEB1, a transcription factor involved in the epithelial to mesenchymal transition (EMT). In the present study, we asked whether SEMA3F specific receptor, NRP2, was also regulated during the EMT process. TGFβ, a physiologic EMT inducer, stimulated NRP2 expression in two NSCLC cell lines. Forced expression of ZEB1, but not Snail, also induced NRP2. Conversely, ZEB1 and Snail inhibition blocked NRP2 upregulation by TGFβ. Importantly, inhibiting NRP2 or ZEB1 expression reduced TGFβ-induced migration in an equivalent manner. In lung cancer tissue microarrays, NRP1 and NRP2 were preferentially expressed in the tumor compartment, whereas the EMT-related transcription factors, ZEB1 and Snail, were predominantly expressed in the stroma. Of note, NRP2 was also expressed in stromal cells and was significantly associated with both ZEB1 and a higher (worse) tumor grade. Since NRP2 is the highest affinity receptor for SEMA3F, our results suggest that loss of SEMA3F coupled with increased NRP2 would facilitate the binding of growth factors to NRP2 to further promote EMT and metastasis. Therefore, targeting NRP2 could be an important therapeutic approach against EMT in lung cancer.

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 2416. doi:1538-7445.AM2012-2416
Abstract 2423: Acute in vitro exposure to cigarette smoke particulate matter does not induce epithelial-mesenchymal transition in lung epithelial cells
Authors: Breheny Damien M.
Published: August 2012

The epithelial-mesenchymal transition (EMT) program was first identified from studies on embryogenesis and organ development, but has since been found to play vital roles in tissue remodelling and tumorigenesis. EMT is a biological process during which epithelial cells change their morphology to adopt fibroblast-like characteristics. During EMT, cell adhesion is lost while cells become more motile. A number of distinct molecular events are required for a cell to undergo EMT including transcription factor activation, cytoskeletal rearrangement, changes in cell adhesion molecule expression and increased production of extracellular matrix-degrading enzymes. EMT has recently been linked to chronic inflammation and cigarette smoke exposure in the lung and is believed to be involved in lung carcinogenesis. As the specific genes and proteins involved in EMT have been well defined, they can serve as markers to demonstrate the passage of cells through EMT in the context of cigarette smoking. The purpose of this study was to develop an in vitro model of EMT using immortalised human bronchial epithelial (BEAS-2B) cells. EMT was induced in BEAS-2B cells using a 72 hour treatment with 5 ng/mL TGF-β1. Expression of the following EMT markers were assessed at the transcript level using reverse transcription real time-polymerase chain reaction (RT-PCR): alpha smooth muscle actin (α-SMA), collagen 1, fibronectin, vimentin, e-cadherin, slug and snail. Protein levels of α-SMA, fibronectin, β-catenin, e-cadherin and zonular occludin-1 were evaluated using immunocytochemistry techniques. This model was then used to determine the effects of acute (72 hour) cigarette smoke particulate matter treatment on EMT in BEAS-2B cells. Cells were treated with particulate matter alone, and in combination with TGF-β1. Particulate matter did not induce EMT directly, nor did it influence EMT induced by TGF-β1 under the treatment conditions used in this study, based on marker expression. It is possible that a more prolonged exposure to particulate matter is required for EMT induction in BEAS-2B cells.

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 2423. doi:1538-7445.AM2012-2423
Abstract 2436: [18F]FDG accumulation in tumor microenvironment -Microautoradiographic study combined with immunohistochemistry in mice-
Authors: Sugita Sachi , Yamato Masanori , Kataoka Yosky , Watanabe Yasuyoshi
Published: August 2012

[Purpose] Tumor microenvironment is characterized by the conditions including hypoxia, low nutrition, stromal cell infiltration, and acidosis due to incomplete blood vessel network. Tumor microenvironment has been recognized as a major factor influencing not only the response to anti-cancer therapies but also malignant progression and metastasis. Non-invasive imaging of such a microenvironment by positron emission tomography (PET) will provide more therapeutic merits. Accumulation of FDG, 2-deoxy-2-[F-18]fluoro-D-glucose, a most-often used PET probe, in tumor has been reported to correlate with the metastasis and malignancy. However, the FDG uptake profile in the microenvironment has remained unknown. In this study, we simultaneously visualized the intratumoral FDG distribution and the microenvironment by using microautoradiography in microscopic spatial resolution combined with immunohistochemistry. [Methods] Human epidermoid carcinoma cell line A431 was transplanted into the back of female SCID mice, and FDG was injected into the mice 2 weeks after the transplantation. The tumors were removed from mice 45 minutes after the FDG administration and the frozen sections were obtained. Overall image of the intratumoral FDG distribution was obtained by conventional autoradiography. FDG distribution and microenvironment in microscopic levels were visualized by microautoradiography followed by immunohistochemical detection of hypoxic condition and cell-specific markers. [Result & Conclusion] Conventional autoradiography and microautoradiography showed the similar pattern in FDG distribution in the tumor. Furthermore, microautoradiographic study with immunohistochemistry revealed that grains at high-density were observed in pimonidazol (hypoxia marker)-accumulated areas showing infiltration of many S100A4-immunopositive fibroblasts. These results suggested that FDG accumulation was prominent in stromal cells involved in hypoxic condition in tumor microenvironment.

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 2436. doi:1538-7445.AM2012-2436
Abstract 2437: Fluorescent imaging in animal model of human tongue squamous cell carcinoma in vivo by Nimotuzumab conjugated near-infrared quantum dots and small molecule fluorescent dye
Authors: Huang Xuelei , Ren Yipeng , Xi Qing , Zhang Lei , Fu Chongjian , Bu Rongfa
Published: August 2012

Objectives:To explore near-infrared fluorescent imaging in vivo using quantum dots Qdot800 and Alexa680 dye labeling Nimotuzumab for human tongue squamous cell carcinoma diagnosis, location. Methods: Near-infrared fluorescent quantum dots Qdot800 and Alexa680 dye were respectively conjugated with Nimotuzumab. Subsequently, the probes were used to detect fixed or live human tongue squamous cell carcinoma cell line Tca8113. After Tca8113 subcutaneous xenografts grown in, the nude mice were injected intravenously with one of the conjugates followed by fluorescent imaging using small animal imaging system. The images of targeting tumor in vivo were required and analyzed. Results: Near-infrared fluorescent quantum dots Qdot800 and Alexa680 dye were successfully conjugated with Nimotuzumab. The conjugates were able to specifically and efficiently recognize fixed and live Tca8113 with intense fluorescence. Although Nimotuzumab labeled with Qdot800 could not be detected targeting the subcutaneous Tca8113 xenografts grown in vivo, Nimotuzumab labeled with Alexa680 performed very well with bright fluorescence in contrast to surrounding normal tissue. Conclusions: The fluorescent conjugates probes Nimotuzumab conjugated with Near-infrared fluorescent Qdot800 and Alexa680 dye were able to specifically target human tongue squamous cell carcinoma cell in vitro. Alexa680 labeling with Nimotuzumab was more suitable than Qdot800 to perform fluorescent imaging in vivo for the study of human tongue squamous cell carcinoma. [Key words] tongue squamous cell carcinoma, fluorescent imaging, fluorescent probe, Nimotuzumab

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 2437. doi:1538-7445.AM2012-2437
Abstract 2438: Bevacizumab treatment hampers tumor uptake of antibodies in human tumor bearing mice
Authors: Arjaans Marlous , Munnink Thijs H. Oude , Oosting Sjoukje F. , van Scheltinga Anton G. T. Terwisscha , Timmer-Bosscha Hetty , de Hooge Marjolijn N. Lub- , Schröder Carolina P. , de Vries Elisabeth G. E.
Published: August 2012

Background: Extensive angiogenesis, defective vascular architecture, impaired lymphatic drainage and increased vascular permeability are characteristics unique for solid tumors. These characteristics result in enhanced tumor permeability and retention (EPR) effect. The EPR effect has been recognized as an important factor for intratumoral delivery of macromolecular drugs, like antibodies. Rapid changes in tumor vasculature and permeability due to antiangiogenic treatment could possibly restrain the EPR effect and thereby lead to a decreased uptake of antibodies. This would have implications for combining antiangiogenic therapy with monoclonal antibodies. We therefore investigated the effect of bevacizumab treatment on the tumor uptake of antibodies by means of immunoPET imaging. Methods: The effect of bevacizumab treatment on the antibody tumor uptake was studied with in vivo PET imaging of 89Zr-trastuzumab, 89Zr-bevacizumab and 89Zr-IgG in a SKOV-3 human xenograft tumor model. PET images were made before and during bevacizumab (5mg/kg; 2qw) or placebo (normal saline) treatment. All animals also received trastuzumab, bevacizumab or IgG labeled with the near-infrared fluorescence dye IRDye 800CW as a co-injection. Immunohistochemistry was performed to determine the mean vascular density (MVD). Immunofluorescence was used to visualize trastuzumab-800CW, bevacizumab-800CW and IgG-800CW and to determine vessel normalization by vessel pericyte coverage. Results: Bevacizumab treatment decreased 89Zr-trastuzumab, 89Zr-bevacizumab and 89Zr-IgG tumor uptake with 41 ± 5% (P<0.0001), 43 ± 4% (P<0.0001) and 28 ± 6% (P=0.0004) respectively. Placebo treatment did not affect tumor uptake of 89Zr-IgG. Immunofluorescence showed less accumulation of IgG-800CW in bevacizumab treated tumor sections compared to placebo treated tumor sections. In all groups, bevacizumab treatment decreased the MVD (P<0.0003). Remaining vessels in bevacizumab treated tumor sections showed an increase in the amount and degree of vessel pericyte coverage compared to placebo. Conclusion: Bevacizumab treatment reduces tumor uptake of trastuzumab, bevacizumab and IgG possibly by restraining the EPR effect through vessel normalization. This deserves attention in the design of clinical trials, when combining antiangiogenic treatment with antibodies.

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 2438. doi:1538-7445.AM2012-2438
Abstract 2447: Measurement of tumor vascular integrity changes in an orthotopic glioma model in rats induced by antiangiogenic treatment using DCE-MRI
Authors: Provent Peggy , Boisferon Marc Hillairet de , Koob Zina , Bichat Francis , Tizon Xavier
Published: August 2012

Background Glioblastoma is the most aggressive subtype of brain tumors. Antiangiogenic compounds are being tested in clinical trials, for this pathology, mostly in combination with other molecules. The availability of an imaging biomarker of efficacy could help optimize the dosing and scheduling of these combinations. The aim of this study was to validate DCE-MRI, using two contrast agents of different molecular weight, as an imaging biomarker of the efficacy of sorafenib (Nexavar®, BAY 43-9006) in an orthotopic glioma model in Nude rats. Methods U-87 MG human glioma cells were inoculated by stereotactic injection in the right caudate nucleus of 24 Nude rats. All rats were first imaged 10 days after cell injection to measure their baseline tumor volume (TV) using T2-weighted MRI (Bruker Pharmascan, 4.7T). Based on their TV, 20 rats were selected and imaged at D-1 (14 days after cell injection). The rats were then randomized into 2 groups of 8 rats according to their tumor vascular status following a full DCE-MRI protocol using either Gd-DTPA (Magnevist®) or P846 (two parameters: Ktrans from pharmaco-kinetic modeling of the contrast agent uptake curve, and iAUC60, the initial area under the gadolinium concentration time curve after 60 seconds). P846 is a medium-molecular weight, gadolinium-based contrast agent provided by Dr P. Robert (Guerbet). In both subgroups of 8 rats, each animal received a daily per os administration of either sorafenib (100 mg/kg/adm) or its vehicle. All 16 rats were then imaged at D1, D3 and D7. At each time point, voxelwise maps of DCE-MRI parameters were derived from imaging data and their distribution analyzed within regions of interest within the tumor and the contralateral lesion-free tissue. Results Sorafenib was well tolerated and it increased the median survival of tumor-bearing rats by 20%. At D7, despite no difference in tumor growth kinetics, DCE-MRI parameters (Ktrans and iUAC60) showed significant differences between sorafenib-treated and control groups using either Magnevist® or P846. Visual assessment showed that the heterogeneity of contrast agent uptake was reduced by antiangiogenic treatment with sorafenib. The quantification of this phenomenon by statistical analysis of the parameter distributions within the ROIs is pending. Conclusions In conclusion, we have shown that DCE-MRI, with either Magnevist® or P846 contrast agents, is a suitable non-invasive imaging technique to show changes of tumor vasculature integrity induced by antiangiogenic treatment such as sorafenib in orthotopically-implanted glioma in Nude rats.

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 2447. doi:1538-7445.AM2012-2447
Abstract 245: ST6Gal-I sialylation of the Fas (CD95) receptor provides protection against Fas mediated apoptosis
Authors: Swindall Amanda F. , Liu Zhongyu , Bellis Susan L.
Published: August 2012

The golgi glycosyltransferase, ST6Gal-I, adds the negatively-charged sugar, α2-6 sialic acid to membrane and secreted proteins. This enzyme has been shown to be upregulated in numerous human cancers including colon cancer. However, the function of ST6Gal-I in tumor progression remains unclear due to a dearth of knowledge regarding enzyme substrates, and the effect of sialylation on target function. Previously, we have shown ST6Gal-I to be upregulated by oncogenic-ras which leads to increased invasion in colon carcinoma cells. Recently a role for ST6Gal-I in the protection of tumors against apoptosis is emerging, as we have shown sialylation to block galectin-mediated apoptosis by inhibiting the binding of galectin-3. Here we identify the Fas (CD95) receptor as a target for ST6Gal-I sialylation, and show that Fas sialylation prohibits apoptotic signaling in two independent cell models. Specifically, in an shRNA knockdown of ST6Gal-I in cells with high endogenous ST6Gal-I (HD3), and in a forced expression of ST6Gal-I in cells with no endogenous sialyltransferases (SW48), we show the presence of ST6Gal-I impedes Fas apoptotic signaling. Upon treatment with both the activating antibody, CH11 and the endogenous ligand, FasL, cells expressing ST6Gal-I had decreased levels of cleaved caspases 3 and 8, as shown by western blot and immunofluorescent staining. However, there was no difference in DR4/DR5-mediated apoptosis induced by TRAIL, suggesting a specific effect of ST6Gal-I on Fas-mediated apoptosis. Additionally, we have shown ST6Gal-I sialylation of Fas decreases the binding of FADD to Fas cytoplasmic tails limiting DISC formation, and prohibits receptor internalization after activation, effectively blocking Fas apoptotic signaling. We have also shown Fas-mediated apoptosis driven by the agonistic antibody, Jo2, to be inhibited in primary macrophages from transgenic mice genetically engineered by our lab to over-express ST6Gal-I. Furthermore, we have shown ST6Gal-I to be upregulated in 7/9 human colon tumor samples, and despite Fas downregulation, the expressed Fas is hypersialylated in several human tumor tissues. In summary, these data identify ST6Gal-I sialylation of the Fas receptor as a novel mechanism in the self-protection of tumors against apoptosis, and further identify ST6Gal-I as an important regulator of apoptotic signaling.

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 245. doi:1538-7445.AM2012-245
Abstract 2454: In vivo binding of chTNT-1/B antibodies (Cotara) to DNA/histone complexes in tumors using near-infrared optical imaging
Authors: Gong Jian , Sevilla Roy , Nguyen Vince , Fisher Seth , Alldredge Jessica , Kinjo Jessie , Archer Rich , Peacock Matthew , Hughes Christopher , Chang Connie , Freimark Bruce
Published: August 2012

Tumors contain both microscopic and macroscopic areas of necrotic cells. These have abnormal cell membrane permeability, which allows passage of large macromolecules such as antibodies into the cell. 131I-chTNT-1/B antibody (Cotara) is a novel, single administration agent designed to target tumor necrosis that has completed a Phase II clinical trial for recurrent glioblastoma multiforme (GBM), showing 9.3 month median overall survival for patients. Since 131I-chTNT-1/B antibody is a high-molecular-mass protein, it cannot easily pass from the systemic circulation through the blood-brain barrier (BBB) and thus can only be delivered locally by an intratumoral catheter. We have developed a human ME-180 cervical carcinoma xenograft tumor model in nude mice to characterize the targeting of chTNT-1/B to necrotic areas using near infrared (NIR) optical imaging. The use of NIR optical imaging is a powerful translational tool to monitor in vivo antibody targeting. chTNT-1/B was labeled with a NIR fluorescent dye and kinetics of antibody retention in tumors was measured by NIR optical imaging following intratumoral injection of the NIR-chTNT-1/B. NIR-chTNT-1/B was retained in ME180 tumors for a longer duration compared to NIR dye alone. Labeling of chTNT-1/B with a low dye/protein ratio was necessary to demonstrate maximum specific targeting to tumor necrosis. Evaluation of in vivo antibody targeting to tumors using NIR imaging may facilitate improvements for treatment of patients with malignant glioma with Cotara.

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 2454. doi:1538-7445.AM2012-2454
Abstract 2558: VEGF-dependent suppression of prostaglandin transporter (PGT) expression in Kras mutant human bronchial epithelial cells
Authors: Lee Gina , Gardner Brian K. , Hazra Saswati , Kachroo Puja , Lee Mi-Heon , Krysan Kostyantyn , Walser Tonya C. , Larsen Jill E. , Girard Luc , Wistuba Ignacio I. , Minna John D. , Dubinett Steven M.
Published: August 2012

Prostaglandin transporters are expressed in cyclooxygenase-positive cells and provide the dominant mechanisms for transport of prostaglandins (PG) across the cell membrane. Specifically, the prostaglandin transporter PGT, expressed in many cell types including lung epithelia, is responsible for the uptake of prostaglandin E2 (PGE2) for intracellular metabolism and degradation by 15-prostaglandin dehydrogenase (15-PGDH). The regulation of prostaglandin transporters has not been fully explored in lung cancer. We hypothesize that loss of PGT expression is important in lung carcinogenesis and its loss may be enhanced in the presence of common mutations in premalignant bronchial epithelial cells and non-small cell lung cancer (NSCLC). Our preliminary studies show that PGT expression (protein and mRNA) and 15-PGDH are significantly downregulated in Kras mutant human bronchial epithelial cells (HBEC) and in NSCLC. It is well known that Kras mutant cancers are associated with increased production of VEGF, an important factor in tumor angiogenesis. In HBEC, we found significantly higher VEGF levels in Kras mutants compared to vector controls (2242 versus 628 pg/mg, p = 0.04). We proposed that VEGF modulates PGT expression and tested our hypothesis by treating HBEC and NSCLC with human recombinant VEGF. PGT expression is downregulated with exogenous VEGF treatment and neutralizing VEGF production (neutralizing VEGF antibody) prevented the loss of PGT expression. The MEK1/2 inhibitor PD98059 upregulated PGT expression in HBEC Kras. Our results suggest Kras mutation is associated with loss of PGT expression and may be dependent on VEGF production. Modulation of PGT expression may be important in lung carcinogenesis.

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 2558. doi:1538-7445.AM2012-2558