bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4605: STK17A is a potential therapeutic target in glioblastoma
Authors: Mao Pingping , Jardine Mary P. , Rahme Gilbert J. , Yang Eric C. , Tam Janice , Kodali Anita , Biswal Bijesh , Fadul Camilo E. , Gaur Arti B. , Israel Mark A. , Pletnev Alexandre , Spinella Michael
Published: May 2015

Current targeted therapies for glioblastoma multiforme (GBM) fail to significantly improve clinical outcome. Identifying new molecular targets driving GBM tumorigenesis is imperative. Our previous study demonstrated that STK17A, a serine-threonine kinase in the death-associated protein family, is a bona fide p53 target gene. In silico analyses indicated that STK17A is highly overexpressed in GBM patients in a tumor grade-dependent manner. Furthermore, high STK17A expression correlated with poor clinical outcome and decreased survival of patients from multiple datasets. This correlation was independent of age, tumor subtype and known biomarkers such as EGFR, NF1, and IDH, suggesting STK17A may contribute to GBM development and progression.

In vitro experiments confirmed increased mRNA and protein expression of STK17A in GBM cells compared to immortalized normal human astrocytes and other cancer types. ShRNA mediated STK17A knockdown in GBM cells decreased cell survival and sensitized cells to genotoxic stress. In addition, STK17A knockdown led to reduced tumor cell proliferation and clonogenicity, suggesting a tumor-promoting role for STK17A in GBM. Interestingly, STK17A depletion resulted in a cell morphological change from a spindle-like phenotype to a phenotype with a flattened, enlarged and more rounded shape that was associated with induction of actin stress fibers. This cytoskeleton remodeling was associated with impaired cell migration and invasion. In contrast STK17A overexpressing cells displayed a pronounced needle-like elongated phenotype. In addition genome-wide expression analysis of STK17A knockdown GBM cells revealed regulation of genes involved in glycolysis including PKM2, PGAM1 and HK2, suggesting that STK17A may also promote tumor growth and survival through regulating metabolism. Small molecule inhibitors that block the kinase activity of STK17A decreased cell survival of GBM cells cultured under both serum and serum-free conditions. Further investigation is required to understand the precise role of STK17A in GBM.

Citation Format: Pingping Mao, Mary P. Jardine, Gilbert J. Rahme, Eric C. Yang, Janice Tam, Anita Kodali, Bijesh Biswal, Camilo E. Fadul, Arti B. Gaur, Mark A. Israel, Alexandre Pletnev, Michael Spinella. STK17A is a potential therapeutic target in glioblastoma. [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 4605. doi:10.1158/1538-7445.AM2014-4605
Abstract 4608: Exploring anti-oncogenic properties of riluzole in breast cancer
Authors: Bukhsh Miriam A. , Speyer Cecilia L. , Hachem Ali A. , Nassar Mahdy , Assi Ali A. , Gorski David H.
Published: May 2015

Introduction: Triple negative breast cancer (TNBC) is a subtype of breast cancer that lacks the three receptors that current therapy targets (estrogen (ER), progesterone, HER2). Because of this, it is a very aggressive type of breast cancer with a high mortality rate. In a previous study, we discovered the drug, riluzole, to be an mGluR1 inhibitor and to reduce cell proliferation and tumor growth in triple negative breast cancer (TNBC), resulting in apoptosis. This was a significant finding because there are currently no major therapies available to successfully treat TNBC. It is now being thought that riluzole also has properties that may allow it to act as a therapy against ER-positive breast cancer (ER+).

Methods: The goal of this study was to learn more about how riluzole functions by analyzing the anti-oncogenic properties of riluzole. To do this, we conducted a series of transformation assays including cell proliferation (MTT), invasion, soft agar, adhesion, and in vivo metastasis using 4T1-12B cells (a luciferase-expressing clone of the 4T1 mouse mammary tumor cell line).

Results: The results of the assays completed thus far indicate that both TNBC and ER+ cells are sensitive to riluzole, with TNBC being more sensitive. Riluzole significantly inhibits invasion of TNBC cells but not ER+ cells. These findings are consistent with the previous study's results showing that riluzole targets mGluR1 pathways, since ER+ cells contain significantly smaller amounts of mGluR1 compared to TNBC cells. Additionally, riluzole inhibits both cell proliferation and anchorage independent growth of both cell types, suggesting both mGluR1 dependent and independent pathways are involved.

Conclusion: The results of this study suggest that riluzole can be developed as an anti-breast cancer therapy for both triple negative and ER-positive breast cancer patients. Further studies silencing mGluR1 in TNBC and overexpressing mGluR1 in ER+ cell lines will help to determine any significance of mGluR1 in mediating riluzole's mechanism of action.

Citation Format: Miriam A. Bukhsh, Cecilia L. Speyer, Ali A. Hachem, Mahdy Nassar, Ali A. Assi, David H. Gorski. Exploring anti-oncogenic properties of riluzole in breast 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 4608. doi:10.1158/1538-7445.AM2014-4608
Abstract 4609: MEF2 plays a critical role in RENCA macrobead-induced tumor cell growth inhibition
Authors: Martis Prithy C. , Dudley Atira , Laramore Melissa A. , Smith Barry H. , Gazda Lawrence S.
Published: May 2015

Regulation of cell proliferation and cell death is often dependent on the coupling of extracellular signals to the activation or repression of specific intracellular signaling pathways. We have previously demonstrated the capacity of mouse renal adenocarcinoma (RENCA) cells encapsulated in agarose macrobeads to form numerous small tumor colonies within the macrobeads over a period of several months. As the encapsulated tumor colonies mature and reach a maximum size, the macrobeads themselves, or conditioned media from cultured macrobeads, markedly inhibit the in vitro and in vivo growth of epithelial-derived tumor cells outside the macrobead microenvironment. Myocyte enhancer factor 2 (MEF2) is a family of four transcription factors that have been shown to play a central role in linking signaling pathways to the genes responsible for cell division, differentiation and death. In this study, we evaluated the expression and role of MEF2 in regulating cell proliferation in mouse (RENCA) and human cell lines (J82, MCF7) in response to RENCA macrobeads. Freely growing RENCA target cells transiently transfected with a MEF2 transcriptional response element and exposed to conditioned media from variously aged macrobeads demonstrated increased response element activation in parallel with the increasing age of the macrobeads. Similarly, growth inhibition of the freely growing RENCA cells increased with older macrobeads. Suppression of the expressed MEF2 isoforms in target RENCA cells, either individually (MEF2a, MEF2b, and MEF2d) or in combination (MEF2pool) using synthetic small interfering RNA (siRNA) markedly reduced the growth inhibitory effects of RENCA macrobeads. In J82 and MCF7 cell lines, MEF2D expression was responsive to replete media from RENCA macrobeads, increasing 2.0-fold and 2.7-fold respectively. Degradation of MEF2D mRNA transcripts in the human cell lines studied resulted in abatement of the growth inhibitory effect of the RENCA macrobeads. These findings reveal an essential role for MEF2 in RENCA macrobead-induced cancer cell growth inhibition and raise interesting questions about the molecular basis of this response.

Citation Format: Prithy C. Martis, Atira Dudley, Melissa A. Laramore, Barry H. Smith, Lawrence S. Gazda. MEF2 plays a critical role in RENCA macrobead-induced tumor cell growth inhibition. [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 4609. doi:10.1158/1538-7445.AM2014-4609
Abstract 4613: Identification of tumor necrosis factor receptor II as a regulatory T cell target for cancer immunotherapy using designed ankyrin repeat protein phenotypic selections
Authors: Williams Geoff , Anderton Judith , Bedian Vahe , Ulrichsen Jane Coates , Gonzalez-Munoz Andrea , Guillard Sandrine , Harris Olivia , Hair James , Leishman Andrew , Lewis Arthur , Moissan Jacques , Minter Ralph , Mistry Bina , Parmentier Julie , Poon Edmund , Popple Amy , Rust Steve , Sandercock Alan , Stewart Ross , Valge-Archer Viia , Wilkinson Robert W.
Published: May 2015

Regulatory T cells (Treg cells) are observed within multiple tumor types and contribute to immune evasion. Effective and specific targeting of Treg cells by antibodies is hampered by the lack of a unique surface marker. With the aim of identifying such a marker, human Treg cells were isolated from peripheral blood, expanded in vitro and used as a source of antigen in phage display selections with a diverse library of designed ankyrin repeat proteins (DARPins). The resulting DARPins were screened for binding to multiple cell-types, and approximately thirty with preferential binding to Treg cells were identified. All of these DARPins were determined to bind to the third and/or fourth cysteine-rich extracellular domains of tumor necrosis factor receptor II (TNFRII, also known as TNFRSF1B or CD120b). Expression of TNFRII has previously been demonstrated on Treg cells, and TNFRII is shown here also to be expressed on CD8+ T cells following exposure to cognate antigen and on the surface of tumor-infiltrating T cells in the 4T1 mouse tumor model. Furthermore, TNFRII mRNA was detected at increased levels in tumor-infiltrating leukocytes isolated from cancer patients. Functional characterisation of the anti-TNFRII DARPins and mAbs revealed an ability to enhance T-cell activation in response to polyclonal and antigen specific stimuli, and in Treg cell suppression assays. These studies demonstrate the utility of DARPins for phenotypic selections to identify drug targets using primary human lymphocytes and highlight TNFRII as a potential therapeutic target enriched on Treg cells.

Citation Format: Geoff Williams, Judith Anderton, Vahe Bedian, Jane Coates Ulrichsen, Andrea Gonzalez-Munoz, Sandrine Guillard, Olivia Harris, James Hair, Andrew Leishman, Arthur Lewis, Jacques Moissan, Ralph Minter, Bina Mistry, Julie Parmentier, Edmund Poon, Amy Popple, Steve Rust, Alan Sandercock, Ross Stewart, Viia Valge-Archer, Robert W. Wilkinson. Identification of tumor necrosis factor receptor II as a regulatory T cell target for cancer immunotherapy using designed ankyrin repeat protein phenotypic selections. [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 4613. doi:10.1158/1538-7445.AM2014-4613
Abstract 4621: Hedgehog acyltransferase as a novel target in breast cancer
Authors: Matevossian Armine , Resh Marilyn
Published: May 2015

Hedgehog acyltransferase (Hhat) catalyzes the transfer of a palmitoyl group onto Sonic Hedgehog (Shh), a modification that is essential for the signaling activity of Shh. Recent large scale genomics data show Hhat amplifications in a variety of tumor types including breast, liver, lung and uterine cancers. Breast invasive carcinomas have the highest percentage of Hhat amplification. To determine the functional significance of Hhat expression in breast cancers, we used a panel of estrogen receptor-α (ERα) positive and negative cell lines. Here, we show that Hhat is a novel target for inhibition of ERα positive breast cancer. Depletion of Hhat with lentivirally delivered shRNAs decreased both anchorage-dependent and anchorage-independent proliferation of ERα positive, but not ERα negative, breast cancer cells. Treatment with RU-SKI 43, a small molecule inhibitor of Hhat recently developed by our group, also reduced ERα positive cell proliferation, whereas a structurally related, inactive compound had no effect. To identify signaling pathways that mediate the growth defect in response to Hhat inhibition, we used a commercially available microarray which analyzes the phosphorylation of 18 different intracellular signaling components. Phosphorylation of Akt, S6 ribosomal protein, Bad, and PRAS40 were reduced after treatment of the ERα positive cell line MCF7 with RU-SKI 43. Of note, the Smoothened inhibitor erismodegib had no effect on ERα positive cell proliferation, indicating that cell growth is regulated by non-canonical signaling pathways. Overexpression of Hhat in ERα positive cells not only rescued the growth defect in the presence of RU-SKI 43, but also resulted in increased cell proliferation in the absence of drug treatment. Together, these data suggest that Hhat plays a critical role in ERα positive breast cancer and highlight the potential promise of Hhat inhibitors for therapeutic benefit.

Citation Format: Armine Matevossian, Marilyn Resh. Hedgehog acyltransferase as a novel target in breast 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 4621. doi:10.1158/1538-7445.AM2014-4621
Abstract 4622: Discovery of a novel target for monoclonal antibody therapy of breast and ovary cancers
Authors: Grandi Alberto , Campagnoli Susanna , Parri Matteo , Camilli Elisa D. , Jin Boquan , Sarmientos Paolo , Grandi Guido , Terracciano Luigi , Viale Giuseppe , Pileri Piero , Grifantini Renata Maria
Published: May 2015

In our recent research activities, we identified a panel of novel candidate markers for prevalent cancers by a systematic immune-histochemical screening with a large collection of polyclonal antibodies (approximately 1600) raised against membrane-associated and secreted human proteins currently marginally characterized. Of utmost interest were those proteins over-expressed detected in one or more tumors showing surface exposure on cancer cells. Among them, here we describe the molecular characterization of a novel surface-associated protein (EXN36) belonging to the lectin-binding protein family for which very little is presently known. We found that the protein is mainly over-expressed in breast and ovary cancers (frequency of approximately 30-40% of tested cases, based on analysis of 50 patients) and it is involved in cell proliferation, migration and invasiveness, as demonstrated by gene silencing. With the intent of exploiting EXN36 as therapeutic target for monoclonal antibody (mAb) therapy, highly specific murine mAbs were generated and proved to recognize the protein on the surface of selected breast and ovary cell lines. Of particular interest are two mAbs, which, upon binding, are efficiently internalized by breast and ovary cancer cells. These two mAbs influence the viability of breast and ovary cells in vitro. One of them shows antibody-dependent cell-mediated cytotoxicity in vitro in breast cancer cells. An analysis of their biodistribution in normal human tissues showed that these mAbs have limited cytoplasmic reactivity in some tissues. Overall, the functional properties of the anti-EXN36 mAbs make them amenable for the generation of antibody-drug conjugates (ADCs). Moreover, they could be exploited for immune-nanoparticles targeted delivery systems. In vivo efficacy in breast and ovary xenograft cancer models are ongoing to evaluate their therapeutic activity either as naked antibodies or ADCs. Overall, data indicate that EXN36 and the specific mAbs represent innovative and promising tools for a targeted mAb therapy of breast and colon cancer, either alone or in combinatorial therapeutic strategies.

Citation Format: Alberto Grandi, Susanna Campagnoli, Matteo Parri, Elisa De Camilli, Boquan Jin, Paolo Sarmientos, Guido Grandi, Luigi Terracciano, Giuseppe Viale, Piero Pileri, Renata Maria Grifantini. Discovery of a novel target for monoclonal antibody therapy of breast and ovary cancers. [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 4622. doi:10.1158/1538-7445.AM2014-4622
Abstract 4623: HDAC6 inhibition is a novel strategy to selectively impact inflammatory breast cancer cell survival
Authors: Putcha Preeti , Yu Jiyang , Califano Andrea , Silva Jose
Published: May 2015

Inflammatory breast cancer (IBC) is a lethal and understudied variant of breast cancer, presenting with a signature clinical phenotype of rapidly metastasizing diffuse tumor emboli and rapid resistance to traditional chemotherapeutics. In an unbiased, highthroughput search for novel genes and therapeutics which specifically impact IBC cell survival, we performed a genome wide shRNA screen on two IBC cell lines and identified Histone Deacetylase 6 (HDAC6) as vital for IBC vs non-IBC breast cancer cell line survival. In IBC vs non-IBC breast cancer cell lines, individual knockdown of HDAC6 decreases cell viability and causes apoptosis via cleaved PARP expression and annexin V assays. Importantly, HDAC6 is a therapeutically viable candidate. The orally bioavailable next generation HDAC6-selective inhibitor, Rocilinostat, also preferentially impacts IBC vs non-IBC cell survival. We transitioned these findings in vivo by treating a well-established mouse xenograft model of IBC with Rocilinostat and found significant slowing of tumor growth rates. Taken together, these data represent a new and tailored treatment option for IBC.

Note: This abstract was not presented at the meeting.

Citation Format: Preeti Putcha, Jiyang Yu, Andrea Califano, Jose Silva. HDAC6 inhibition is a novel strategy to selectively impact inflammatory breast cancer cell survival. [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 4623. doi:10.1158/1538-7445.AM2014-4623
Abstract 4624: Molecular target identification of quinolinone based anticancer compounds
Authors: Rylova Gabriela , Dzubak Petr , Janostakova Anna , Frydrych Ivo , Konecny Petr , Holub Dusan , Ozdian Tomas , Dolezal Dalibor , Soural Miroslav , Hlavac Jan , Hajduch Marian
Published: May 2015

Quinolinones, compounds derived from quinolone antibiotics, are well known for their antibacterial, antiprotozoal, cytotoxic and immunosuppressive effects. Screening of collection of small molecules revealed 2-phenyl-3-hydroxy-4(1H)-quinolinones as molecules with highly selective cytotoxic activity against tumor cell lines. Moreover, in vivo activity assessment of the most active compounds against mouse tumor hollow fibre model of CCRF-CEM cells, showed anticancer activity comparable to doxorubicin. Furthermore, cell biology tests revealed that DNA/RNA synthesis was inhibited together with block of tumor cells in G1 phase within the cell cycle.

With these promising results, we decided to continue to identify molecular target and mechanism of action of these compounds and based on such understanding to further improve its biological activity and pharmacological properties.

As a first step, the biotinylated molecules of original derivatives were synthesized, which were used for immunoafinity pull down target identification. By mass spectrometry analysis we have reproducibily identified complex of proteins, which are involved in glucose metabolism, translational regulation and are building blocks of cytoskeleton. Involvement of these pathways was further supported by metabolomics, proteomics profiling and by microarray expressional analysis. Identified molecular targets were further validated by western blots, in reporter cell lines and in the cell-free system by measurement of enzymatic activity and monitoring of complex interaction of identified partners, suggesting new mechanisms of regulation. Currently, we are working on the state-of-art proof by structural analysis of target and chinolinone ligand. Acknowledgement: This work was supported by internal grant of Palacky University (LF_2013_016) and BIOMEDREG (CZ.1.05/2.1.00/ 01.0030).

Citation Format: Gabriela Rylova, Petr Dzubak, Anna Janostakova, Ivo Frydrych, Petr Konecny, Dusan Holub, Tomas Ozdian, Dalibor Dolezal, Miroslav Soural, Jan Hlavac, Marian Hajduch. Molecular target identification of quinolinone based anticancer compounds. [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 4624. doi:10.1158/1538-7445.AM2014-4624
Abstract 4650: Pharmacokinetic and pharmacodynamic study of adjuvant gemcitabine therapy of biliary tract cancer following major hepatectomy (KHBO1101)
Authors: Fujiwara Yutaka , Kobayashi Shogo , Nagano Hiroaki , Kanai Masashi , Hatano Etsuo , Toyoda Masanori , Ajiki Tetsuo , Takashima Yuki , Hamada Akinobu , Minami Hironobu , Ioka Tatsuya
Published: May 2015

Background: Gemcitabine at a standard dose of 1000 mg/m2 on Day 1, 8 and 15 every 4 weeks is not tolerated in biliary tract cancer (BTC) patients who have undergone surgical resection with major hepatectomy due to severe toxicities such as myelosuppression. Our dose-finding study of adjuvant gemcitabine therapy determined that the recommended dose is 1000 mg/m2 on Day 1 and 15 every 4 weeks. Here, we evaluated the pharmacokinetics and pharmacodynamics of gemcitabine at this recommended dose. Methods: This study evaluated BTC patients scheduled to undergo surgical resection with major hepatectomy followed by postoperative gemcitabine therapy. Pharmacokinetic evaluation of gemcitabine and 2′,2′-difluorodeoxyuridine (dFdU) was conducted at the initial administration of gemcitabine, which was given by intravenous infusion over 30 min at a dose of 800 to 1000 mg/m2. Physical examination and adverse events were assessed for two weeks. Results: Thirteen patients were enrolled from August 2011 to January 2013, with 12 completing the study. Eight patients had hilar cholangiocarcinoma, three had intrahepatic cholangiocarcinoma, and one had distal extrahepatic cholangiocarcinoma. Median interval from surgery to first administration of gemcitabine was 65.5 days (range, 43-83 days). In the twelve patients, the following disorders at all grades of severity were observed: leukopenia (83.3%), neutropenia (66.7%), and thrombocytopenia (50.0%). Further, Grade 3 of neutropenia was observed in 16.7% of these patients. The dose-normalized AUC of gemcitabine and dFdU in patients with major hepatectomy was 10.22 ± 2.54 and 94.38 ± 45.38 mg/L/hr, respectively. These values were generally similar to those of patients with pancreatic cancer who did not undergo hepatectomy in a previous study. Conclusion: This study demonstrated that major hepatectomy did not affect the pharmacokinetics of gemcitabine and dFdU, despite an increase in hematological toxicity.

Citation Format: Yutaka Fujiwara, Shogo Kobayashi, Hiroaki Nagano, Masashi Kanai, Etsuo Hatano, Masanori Toyoda, Tetsuo Ajiki, Yuki Takashima, Akinobu Hamada, Hironobu Minami, Tatsuya Ioka. Pharmacokinetic and pharmacodynamic study of adjuvant gemcitabine therapy of biliary tract cancer following major hepatectomy (KHBO1101). [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 4650. doi:10.1158/1538-7445.AM2014-4650
Abstract 4658: Use of a defined oleander extract by advanced cancer patients: Case reports on toxicity and tumor-related outcomes
Authors: Block Keith I. , Gyllenhaal Charlotte , Newman Robert A.
Published: May 2015

The present study reports cases of advanced cancer patient taking an extract of Nerium oleander now in Phase 1 clinical trials in cancer patients. A phytochemical constituent of the extract, oleandrin, has multiple anticancer activities including inhibition of NF-kappaB and induction of apoptosis. We present case reports of 9 metastatic cancer patients who received the extract on personal use exemptions. Five of the patients also received full integrative programs at an integrative medicine center. Cases included 4 colon cancer patients, 3 pancreatic cancer patients (1 neuroendocrine and 2 adenocarcinoma), 1 renal cell carcinoma and 1 lung cancer patient. For the patients at the integrative center, all of whom took the extract during periods when they were receiving chemotherapy, toxicity grading of chemotherapy side effects was performed. We analyzed the case records for potential effects of the extract on tumor regression/stability in the absence of chemotherapy, survival time, use concurrent with chemotherapy and toxicity. The renal cell patient experienced tumor regression for 12 months with the extract alone; the 3 pancreatic cancer patients and a colon cancer patient experienced disease stabilization. Survival times were extended for all patients with survival times ranging from 32 months to 11 years for these patients with metastatic disease. Chemotherapy was well tolerated by colon cancer patients with only grade 1-2 toxicity during as many as 70 cycles of chemotherapy. Mild toxicities thought to be related to the extract included lightheadedness and mild tingling and burning of the mouth with use of the sublingual preparation. Severe toxicities including one myocardial infarction and one deep-vein thrombosis were observed but not suspected to be related to the extract. Good clinical outcomes and low toxicity were thus observed for the extract in these patients.

Citation Format: Keith I. Block, Charlotte Gyllenhaal, Robert A. Newman. Use of a defined oleander extract by advanced cancer patients: Case reports on toxicity and tumor-related outcomes. [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 4658. doi:10.1158/1538-7445.AM2014-4658
Abstract 466: LMO1 is a novel oncogene in neuroendocrine lung cancer
Authors: Zhao Zhenze , Ma Xiuye , Yu Xiaojie , Hsiao Tzu-Hung , Chen Yidong , Suraokar Milind , Wistuba Ignacio , Minna John D. , Pertsemlidis Alexander , Du Liqin
Published: May 2015

Introduction. The aim of the study is to characterize the role of LMO1 in tumorigenesis of neuroendocrine lung cancer. LMO1 is a transcription factor that belongs to the LMO protein family containing the cysteine-rich LIM domains. LMO1 has been demonstrated to function as an important oncogene in the development of T-lineage leukemia and lymphoma. A recent study in neuroblastoma suggests that aberrant overexpression of LMO1 may play a critical role in the development of neuroendocrine cancers. The role of LMO1 in neuroendocrine lung cancer, however, was not investigated.

Experimental Approaches. By analyzing a large panel of cell lines including lung cancer cell lines and normal lung epithelial cells, we compared the relative expression of LMO1 in different subtypes of lung cancers, and examined the correlation of LMO1 expression levels with neuroendocrine differentiation markers. Using in vitro approaches, we investigated the function of LMO1 in regulating lung cancer cell growth. To investigate the clinical significance of LMO1 dysregulation in lung cancer patients, we analyzed the correlation of LMO1 tumor levels with lung cancer patient survival.

Results. We show that LMO1 is significantly overexpressed in small cell lung cancer (SCLC), an aggressive and major subtype of neuroendocrine lung cancers, relative to non-small cell lung cancer (NSCLC) and normal lung cells. In addition, in NSCLC cells, LMO1 mRNA levels are significantly correlated with expression of neuroendocrine differentiation markers. Functional in vitro investigations indicate that LMO1 overexpression significantly promotes growth of cultured lung cancer cells, suggesting its oncogenic function in lung cancer. More strikingly, our investigations of two independent lung cancer patient populations indicate that high tumor LMO1 mRNA level is an independent predictor of poor patient survival.

Conclusions. Altogether, our findings strongly suggest that LMO1 is a neuroendocrine-specific oncogene in lung cancer that plays an important role in determining the cancer aggressiveness. Further studies are certainly warranted to define the mechanisms underlying the oncogenic function of LMO1 in neuroendocrine lung cancer and to further evaluate the clinical significance of LMO1 as a diagnostic and prognostic marker for neuroendocrine lung cancers in prospective studies.

Citation Format: Zhenze Zhao, Xiuye Ma, Xiaojie Yu, Tzu-Hung Hsiao, Yidong Chen, Milind Suraokar, Ignacio Wistuba, John D. Minna, Alexander Pertsemlidis, Liqin Du. LMO1 is a novel oncogene in neuroendocrine 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 466. doi:10.1158/1538-7445.AM2014-466
Abstract 4663: KRAS, BRAF, and EGFR mutational analysis in ovarian, colon, and lung cancers by highly multiplex PCR/barcoded-magnetic-bead (BMB) suspension-array assays
Authors: Lei Jason , Hsu Julia , Jen Peggy , Huang Daniel , Chung Andre , Kao Lloyd , Chang Miller , Yuan Chiou-Chung , Lee Wei-Hwa , Yeh Chi-Tai , Tsao Dean
Published: May 2015

Anti-epithelial growth factor receptor (EGFR) monoclonal antibodies and small molecule EGFR inhibitors have been used for treating metastatic colorectal cancer (mCRC) and non-small cell lung cancer (NSCLC), respectively. However, since a relatively large proportion of the mCRC and NSCLC patients carry the activating KRAS/BRAF mutations that can render these EGFR-targeted therapies ineffective, the KRAS/BRAF mutation testing is now required for such prescription. In contrast, epithelial ovarian cancer (EOC) thus far has been treated by surgery, radiotherapy, and/or chemotherapy depending on the tumor stages. However, it has been shown that a high percentage of tumors of the mucinous, endometrioid, low-grade serous, and other types of EOC patients also contain KRAS or BRAF somatic mutations, suggesting that drugs targeting the mediators of the EGFR pathway may have implications on treating EOC of certain types. The conventional analysis of the KRAS, BRAF, and EGFR mutations has been done by multiple reactions with one mutation target per reaction, thus requiring a large amount of precious patient sample for complete testing. Here we present three highly multiplex molecular diagnostic assays, utilizing amplification by allele-specific PCR and automatic detection by a platform designed around the barcoded-magnetic-bead (BMB) suspension-array technology, for detection of 12 KRAS (in codons 12 and 13), six BRAF (in codon 600), and 21 EGFR (in exons 18-21) mutations in mCRC, NSCLC, and EOC patient samples. The results indicate that, albeit all the reaction components are in single wells, these highly multiplex PCR/BMB assays, by comparing with the sequencing results, not only are sensitive and specific but also can conserve precious tissue specimens, save operating time and labor, reduce turnaround time, and increase assay throughput.

Citation Format: Jason Lei, Julia Hsu, Peggy Jen, Daniel Huang, Andre Chung, Lloyd Kao, Miller Chang, Chiou-Chung Yuan, Wei-Hwa Lee, Chi-Tai Yeh, Dean Tsao. KRAS, BRAF, and EGFR mutational analysis in ovarian, colon, and lung cancers by highly multiplex PCR/barcoded-magnetic-bead (BMB) suspension-array assays. [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 4663. doi:10.1158/1538-7445.AM2014-4663
Abstract 4667: The impact of IASLC/ATS/ERS classification of lung adenocarcinoma on survival after recurrence in resected adenocarcinoma
Authors: Hung Jung-Jyh , Chou Teh-Ying , Hsu Wen-Hu , Wu Yu-Chung
Published: May 2015

Objective: Factors predicting post-recurrence survival (PRS) in lung adenocarcinoma have not been reported. This study aimed to investigate the prognostic factors of PRS, including the new classification of lung adenocarcinoma proposed by the International Association for the Study of Lung Cancer (IASLC), the American Thoracic Society (ATS), and the European Respiratory Society (ERS), in resected lung adenocarcinoma after recurrence.

Methods: The clinicopathological characteristics of 140 patients with recurrence after complete resection of lung adenocarcinoma at Taipei Veterans General Hospital were retrospectively reviewed. The prognostic factors of PRS were analyzed.

Results: The pattern of recurrence included local only in 22 (17.1%), distant only in 40 (31.0%), and both local and distant in 84 (51.9%) of patients. The 2- and 4-year PRS were 67.2% and 39.1%, respectively. The most common organ sites of metastasis were the contralateral lung (40.7%), followed by the brain (37.1%) and bone (30.7%). Univariate analysis showed that N2 status (vs. N0 and N1) (P = 0.007), stage III (vs. stage I and II) (P = 0.043), micropapillary/solid predominant pattern group (P = 0.012), liver metastasis (P = 0.033), and no treatment for recurrence (P < 0.001) were significant predictors of worse PRS. Multivariate analysis revealed that N2 status (P = 0.036), micropapillary/solid predominant pattern group (P = 0.018), and no treatment for recurrence (P < 0.001) were still significant predictors of worse PRS.

Conclusions: N2 status, micropapillary/solid predominant pattern group, and no treatment for recurrence predict worse PRS in patients with resected lung adenocarcinoma. The new classification of lung adenocarcinoma had significant impact on PRS.

Citation Format: Jung-Jyh Hung, Teh-Ying Chou, Wen-Hu Hsu, Yu-Chung Wu. The impact of IASLC/ATS/ERS classification of lung adenocarcinoma on survival after recurrence in resected adenocarcinoma. [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 4667. doi:10.1158/1538-7445.AM2014-4667
Abstract 4668: A next generation sequencing assay to detect the fusion products of ALK, ROS1 and RET with any fusion partner genes and hotspot mutations in FFPE samples
Authors: Xu Qiang , Wang Guan , Li Bo , Qian Maoxiang , Fang Douglas , Sun Lele , Chen Yundi , Sun Hongye
Published: May 2015

Targeted drugs have been widely used in the treatment of lung cancer, especially for non small cell lung cancer (NSCLC) patients. NCCN guideline lists multiple drugs which are particularly effective in the subgroup of patients with specific gene mutation variants, including the fusion transcripts of ALK, ROS1 and RET, and the mutations of EGFR, KRAS, BRAF and HER2. Multiple techniques have been developed to detect fusion transcripts in tumor samples, such as FISH, IHC and PCR assays. FISH is the most accurate method for fusion detection currently. The split probe FISH assay, which does not require the pre-knowledge of what is the fusion partner with ALK for example, can detect unknown ALK fusion products as well. However, it fails to identify which gene is fused with ALK (or other oncogene) in the chimera products, therefore is not informative for the mechanism or drug resistance study. In addition, due to the limitation on the fluoresce dye, it is quite difficult to multiplex fusion FISH probes for different genes into one assay.

We describe here a next generation sequencing assay to detect a panel of fusion transcripts and mutations in one single assay. Our single assay can detect the fusion products of ALK, ROS1 and RET with any known or unknown fusion partners, together with the hotspot mutations in EGFR, KRAS, BRAF and HER2. Our assay can identify not only the exact sequence of the unknown fusion partners, but also any mutations within the full coding regions of ALK, ROS1 and RET. This information is critical for the resistance mechanism study and the next generation inhibitor drug development, since about half of the ALK inhibitor resistance was caused by the mutations in ALK gene itself. We have identified a novel ALK fusion gene with a never-reported fusion partner in a lung cancer patient. The xenograft model derived from this patient responds to crizotinib well in the in vivo study (published on Journal of Thoracic Oncology). We have demonstrated that our assay can reliably detect both the novel fusion and the well known EML4-ALK fusion from only 1-2 5um FFPE slides of the lung tumor tissues, together with other possible fusions and mutations as well.

Note: This abstract was not presented at the meeting.

Citation Format: Qiang Xu, Guan Wang, Bo Li, Maoxiang Qian, Douglas Fang, Lele Sun, Yundi Chen, Hongye Sun. A next generation sequencing assay to detect the fusion products of ALK, ROS1 and RET with any fusion partner genes and hotspot mutations in FFPE samples. [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 4668. doi:10.1158/1538-7445.AM2014-4668
Abstract 4669: Prevalence of high risk human papillomavirus infection in different cervical cytological features among health examination of women in National Cancer Institute, Thailand
Authors: Sangrajrang Suleeporn , Laowahutanont Piyawat , Kalalak Anant , Jamsri Paphawin
Published: May 2015

High-risk (HR) human paillomavirus (HPV) testing is important in cervical cancer screening for triage colposcopy. The objective of the study was to evaluate the prevalence of HPV HR infection in different cytological features among health examination women. DNA was extracted from 2896 residual liquid-based cytology specimens collected during routine liquid-based Papanicolaou tests at National Cancer Institute. Overall, HPV HR prevalence was 9.3% including 1.6% of HPV-16 and 0.4% of HPV-18. HPV prevalent was highest in women younger than 35 years of age, decreasing in women of older age. Among women with normal cytology, HPV HR positive were found in 6.7%. In abnormal cytology, HPV HR were found in 46.7% atypical squamous cells (ASC), 54.8% were low-grade squamous intraepithelial lesions (LSIL) and 80.0% were high-grade squamous intraepithelial lesions (HSIL). HPV-16 were detected in 8.6%, 6.5% and 12.0% of ASC, LSIL and HSIL, respectively.The results of this study provide baseline information on the HPV type distribution, which may useful for clinicians to decide who should be monitored or treated more aggressively.

Note: This abstract was not presented at the meeting.

Citation Format: Suleeporn Sangrajrang, Piyawat Laowahutanont, Anant Kalalak, Paphawin Jamsri. Prevalence of high risk human papillomavirus infection in different cervical cytological features among health examination of women in National Cancer Institute, Thailand. [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 4669. doi:10.1158/1538-7445.AM2014-4669
Abstract 4673: ARID1A expression and its relation with microsatellite instability and clinicopathological characteristics in colorectal and gastric cancers of Korean patients
Authors: Lee Soo Young , Kim Duck-Woo , Lee Hye Seung , Ihn Myong Hoon , Oh Heung-Kwon , Kang Sung-Bum
Published: May 2015

AT-rich interactive domain-containing protein 1A (ARID1A) is a subunit of the SWI-SNF chromatin-remodeling complex, which is considered to perform a tumor suppressor function. Somatic mutations of ARID1A were reported to be detected in about 50% of colorectal cancer (CRC), but loss of expression seems vary according to tissue type and microsatellite instability (MSI) status. So, we designed this study to identify the frequency of loss of ARID1A expression and its clinical relevance in CRC and gastric cancer (GC) of Korean patients. Immunohistochemistry for ARID1A was performed using tissue microarray blocks containing 197 CRCs and 275 SCs, and paired normal mucosa. Data on MSI status were available in 178 of 197 CRCs and 233 of 275 GCs. Clinicopathological data of the patients were collected and analyzed. We identified 12 (12/197, 6.1%) CRCs showing loss of ARID1A expression, all of which were microsatellite stable. In 178 CRCs with identified MSI status, 3 (1.7%) MSI-H and 6 (3.4%) MSI-L tumors showed normal expression of ARID1A, contrary to MSS tumors (ARID1A loss: 10/169, 5.9%). Among GCs, 8.0% (22/275) showed loss of ARID1A expression and 6.4% (15/233) were MSI-H. Tumors with loss of ARID1A expression accounted for 13.3% (2/13) of GCs with MSI-H whereas 5.0% (11/207) of GCs with MSS. However, there was no association between ARID1A expression and MSI status in both CRCs (p=0.754) and SCs (p=0.200). Expression of ARID1A in paired normal mucosal epithelial cells was normal in all patients. Clinicopathological factors were not associated with ARID1A expression in both cancers except that CRCs with loss of ARID1A expression showed less frequent lymphatic invasion than those with normal expression (25.0% vs. 69.0%, p=0.003). There was no association between ARID1A expression and 3-year disease-free survival or 5-year overall survival in both cancers. In this study population, loss of ARID1A expression was uncommon and not relevant to MSI-H tumors. Its clinical implication for the prognosis of CRC and GC patients seems to need further studies with more accumulated data.

Citation Format: Soo Young Lee, Duck-Woo Kim, Hye Seung Lee, Myong Hoon Ihn, Heung-Kwon Oh, Sung-Bum Kang. ARID1A expression and its relation with microsatellite instability and clinicopathological characteristics in colorectal and gastric cancers of Korean patients. [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 4673. doi:10.1158/1538-7445.AM2014-4673
Abstract 4676: In-vitro adenosine triphosphate-based chemotherapy response assay (ATP-CRA) results comparison between primary and recurrent gynecologic cancer using a unique cohort
Authors: Nam Eun J. , Kim Young Tae , Kim Sang Wun , Kim Sunghoon , Yim Ga Won
Published: May 2015

Purpose: The standard chemotherapy regimen in gynecologic cancer treatment is not established, particularly in cases of recurrence. The objective of this study is to evaluate the role of in vitro adenosine triphosphate-based chemotherapy response assay (ATP-CRA) in recurrent gynecologic cancer.

Methods: We examined 20 matched primary and recurrent tumor pairs of each patient with recurrent gynecologic cancer, using ATP-CRA. Tumor samples were obtained from primary and secondary debulking or palliative surgery. Inhibition rates of the chemotherapeutic agents were compared.

Results: The diseases of patients analyzed in this study were composed of ovarian/peritoneal cancer [11 patients (55%)], uterine cancer [7 patients (35%)], and cervical cancer [3 patients (15%)]. The average of inhibition rate was 36.3% in samples obtained from primary surgery and 33.6% in samples from secondary debulking or palliative surgery (P>0.1). In the primary surgery cases, the drugs showing high inhibition rate were ifosfamide (56.4%), cisplatin (52.6%), and irinotecan (43.3%) in the following order, and in the secondary operation cases, ifosfamide (54.3%), cisplatin (44.3%), and Topotecan (39.9%). The number of cases in which the ATP-CRA result had actual influence in choosing the regimen, were 0 (0%) in primary surgery cases, and 7 (35%) in secondary surgery cases (P<0.05).

Conclusion: The drug inhibition rate did not decrease in the recurrent tumors compared to the primary tumors. The actual influence of ATP-CRA results increase significantly in the secondary surgery compared to the primary one.

Note: This abstract was not presented at the meeting.

Citation Format: Eun Ji Nam, Young Tae Kim, Sang Wun Kim, Sunghoon Kim, Ga Won Yim. In-vitro adenosine triphosphate-based chemotherapy response assay (ATP-CRA) results comparison between primary and recurrent gynecologic cancer using a unique cohort. [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 4676. doi:10.1158/1538-7445.AM2014-4676
Abstract 4681: The use of patient derived malignant effusions as in vitro models for a personalized healthcare-approach in anticancer therapy
Authors: Ruiz Christian , Vlajnic Tatjana , Arabi Leila , Wyttenbach Sina , Baschiera Betty , Rothschild Sacha , Kustermann Stefan , Zippelius Alfred , Roth Adrian , Bubendorf Lukas
Published: May 2015

Background

The use of patients' own cancer cells for in vitro selection of the most promising treatment is an attractive concept in personalized medicine. Human carcinoma cells from malignant effusions (MEs) are well suited for this purpose since they have already adapted to the liquid environment in the patient and are not enclosed by stromal cell compartments. Aim of this study was the establishment of a consistent approach for the in-vitro ex-vivo culture of MEs and the comprehensive analysis of the effect of chemotherapeutic as well as targeted drugs.

Methods

MEs from patients with lung or breast cancer were selected for this study. After morphological and molecular characterization, epithelial cells were cultured in cell culture medium supplemented with 30% of patient derived sterile filtered effusion supernatant. Growth characteristics were monitored in real-time using the xCelligence system. MEs were treated with targeted therapeutics according to their genomic profile or with chemotherapeutics based on the recommendation by the oncologist.

Results

We have developed a reliable system for the ex-vivo culture of MEs and the application of drug tests in-vitro. The use of an enrichment system for epithelial cells allowed for culturing of effusions with only moderate tumor cell content. Experiments using drugs revealed dose-dependent effects and specific growth inhibitory effects of targeted therapeutics, such as Crizotinib and Erlotinib.

Conclusions

In this study, we show that cancer cells from patient derived MEs can be cultured and treated in-vitro. Based on our developed setup, these MEs can be genomically characterized by whole genome approaches in order to identify genomic vulnerabilities and then subjected to growth tests by using the respective targeted drugs. This approach could be an important step towards personalized healthcare, at least for patients with malignant effusions.

Citation Format: Christian Ruiz, Tatjana Vlajnic, Leila Arabi, Sina Wyttenbach, Betty Baschiera, Sacha Rothschild, Stefan Kustermann, Alfred Zippelius, Adrian Roth, Lukas Bubendorf. The use of patient derived malignant effusions as in vitro models for a personalized healthcare-approach in anticancer therapy. [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 4681. doi:10.1158/1538-7445.AM2014-4681