bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3903: Differential transcript expression in nasopharyngeal carcinoma by cDNA microarray analysis
Authors: Yip Timothy T.C. , Kwong Dora L.W. , Ngan Roger K.C. , Lim Cadmon K.P. , Cheng Wai-Wai , Ma Victor W.S. , Law Stephen C.K. , Tse Loretta
Published: August 2015

By cDNA microarray, gene expression profiling was carried out in 360 paired tumor & normal biopsies from 60 nasopharyngeal cancer (NPC) patients at 3 time points, namely, at diagnosis of NPC, on treatment with an anti-inflammatory drug, Celecoxib (Celec) and on radiotherapy (RT). We found 1 cluster of 82 transcripts with statistically significant difference in expression between tumor & normal tissues at diagnosis, a second cluster of 214 tumor versus normal transcripts on Celec treatment and finally a third cluster of 202 tumor versus normal transcripts on RT. Differential expression of selected tumor associated transcripts was confirmed by RT-PCR. These include 8 tumor up-regulated transcripts with 3 encoding for immunological/inflammatory molecules, IL8, CXCL9 & CXCL10, 1 matrix metallopeptidase, MMP12, which breaks down extracellular matrix and thus is associated with cancer metastasis, 1 osteonectin protein which is involved in cell shape changes and thus related to cancer invasion, SPARC, 1 bone morphogenic protein with a role in tissue differentiation, GREM1, 1 GTPase-activating protein for phorbol ester receptor, CHN1 and a sorting nexin family member with a role in intracellular trafficking, SNX10. Genes which were prominently down-regulated in tumor were a variety of polymorphic xenobiotic detoxification enzyme genes or genes with antioxidant function (e.g. GSTA1, GSTA2, GSTA3, CYP4B1, ALDH1A1, ADHFE1, MAOB, NQO1 & OAT). Many genes were also down-regulated by RT in tumor tissues and one notable example was TOP2A which encodes topoisomerase 2A involving in chromosome condensation & chromatid separation during DNA replication, CLDN1 which has tight junction adhesion function, ANLN which is required for cytokinesis, and STMN1 which can regulate microtubule filament. These genes had good potential in applying for clinical applications in diagnosis or treatment of NPC in the future. Further exploration of the clinical applications of these genes is underway.

Citation Format: Timothy T.C. Yip, Dora L.W. Kwong, Roger K.C. Ngan, Cadmon K.P. Lim, Wai-Wai Cheng, Victor W.S. Ma, Stephen C.K. Law, Loretta Tse. Differential transcript expression in nasopharyngeal carcinoma by cDNA microarray analysis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3903. doi:10.1158/1538-7445.AM2015-3903
Abstract 3905: DBNL is a pro-survival adaptor in ErbB2+ breast cancer
Authors: Wixted Josephine F. , Robinson Matthew K.
Published: August 2015

DBNL, also known as mAbp1, Hip-55, or SH3P7, is a fairly novel, ubiquitously expressed, multidomain adapter protein consisting of an N-terminal F-actin binding domain, an internal proline-serine-and threonine-rich domain, and a C-terminal SH3-domain. The N-terminal and C-terminal domains of DBNL are highly conserved from mammal to yeast and are implicated in a multitude of cellular processes that include, endocytosis, migration, signaling, and viability. Recently, a proteomics analysis identified DBNL as a pro-survival signaling node in various cancers. Here we demonstrate that siRNA knockdown of DBNL reduces ErbB2+ breast cancer viability by arresting cell growth at the G1/S phase and by enhancing caspase-mediated apoptosis. Furthermore, knockdown of DBNL reduces cell surface levels of ErbB2 and decreases Akt phosphorylation. Elucidating the downstream signaling mechanisms that drive DBNL-mediated pro-survival will further aid in identifying new targets to ErbB2+ cancers as well as define additional roles of DBNL in cell biology.

Citation Format: Josephine F. Wixted, Matthew K. Robinson. DBNL is a pro-survival adaptor in ErbB2+ breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3905. doi:10.1158/1538-7445.AM2015-3905
Abstract 3906: Annexin A1 overexpression preferentially predicts poor prognosis of basal-like breast cancer patients by activating m TOR-S6 signaling
Authors: Tachibana Kazunoshin , Bhardwaj Anjana , Ganesan Nivetha , Rajapakshe Kimal , Albarracin Constance T. , Gunaratn Preethi H. , Coarfa Cristian , Bedrosian Isabelle
Published: August 2015

Introduction: Annexin A1 is an anti-inflammatory protein reported to play a role in cell proliferation, apoptosis and to be deregulated in breast cancer. The role and regulation of annexin A1 in the biology and prognosis of breast cancer remain unclear because most of the studies did not consider specific differences between the different subtypes of breast cancer. Thus, in this study, we hypothesize that annexin A1 plays subtype-specific role in basal like breast cancer progression and metastasis. Methods: By mining The Cancer Genome Atlas-Breast Cancer Dataset (TCGA-BRCA) and manipulating annexin A1 levels in breast cancer cell lines, we studied the role of annexin A1 in breast cancer and underlying signaling pathways. Results: Our in-silico analysis of TCGA-BRCA dataset demonstrated that annexin A1 mRNA expression is higher in basal subtype compared to luminal and HER2 subtypes. Within the basal subtype, patients show significantly poorer survival associated with higher expression of annexin A1. In both TCGA patient samples and cell lines, annexin A1 levels were significantly higher in basal-like breast cancer tumors than luminal and Her2/neu-positive breast cancer tumors. Stable annexin A1 knockdown in TNBC cell lines suppressed the mTOR-S6 pathway. Specifically, annexin A1 knockdown inhibited phosphorylation of mTOR and S6 (18-73% inhibition in pmTOR and 50-60% inhibition in pS6). But annexin A1 knockdown had no impact on the MAPK, c-Met, and EGFR pathways. In cell migration assays, annexin A1-depleted TNBC cells showed delayed migration as compared to wild-type cells which we found to be through suppression of MMP9. Conclusions: Our data suggest that annexin A1 is prognostic only in patients with basal like breast cancer. This appears to be due in part to the role of annexin A1 in activating mTOR-pS6 pathway.

Citation Format: Kazunoshin Tachibana, Anjana Bhardwaj, Nivetha Ganesan, Kimal Rajapakshe, Constance T. Albarracin, Preethi H. Gunaratn, Cristian Coarfa, Isabelle Bedrosian. Annexin A1 overexpression preferentially predicts poor prognosis of basal-like breast cancer patients by activating m TOR-S6 signaling. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3906. doi:10.1158/1538-7445.AM2015-3906
Abstract 391: Utility of circulating tumor cell (CTC) in advanced gastric cancer
Authors: Lee Won Suk , Kim Jeong Min , Kwon Woo Sun , Lim So Jung , Kim Tae Soo , Kim Ga Yun , Choi Sung Ho , Jeon Byung Hee , Jung Min Kyu , Ahn Joong Bae , Chung Hyun Cheol , Rha Sun Young
Published: August 2015

Circulating tumor cells (CTCs), known as the “leukemic phase” of solid tumor, have been associated with poor prognosis and clinical outcome in various cancer including breast and colorectal. However, fewer studies reported characteristic and usefulness of CTC as an indicator in gastric cancer (GC). In this study, we evaluated utility of CTC in advanced GC cancer. First, we confirmed detection rate of GC cancer cell lines as a spiking test, and then defined CTC as DAPI+, EpCAM+ and CD45- cells through immunofluorescence stating. To determine a cut-off for CTC of GC, we assessed the number of CTCs in blood of 22 healthy volunteers, and observed 1 (3/22) to 2 (1/22) of CTCs. Based on these results, we determined CTC positivity of GC as more than 3 CTCs. When we evaluated CTC positive rate in Stage IV GC patients, 28 patients were

CTC positive (30.1%) with the median number of CTC 5 (range 3 to over 100). Interestingly, 3 out of 28 patients were showed more than 100 CTCs. While, in operable GC patients (Stage I to III), CTC positivity was 18.4% (7/38) and median number of CTC was 4 (range 3 to 9) in CTC positive patients. These results showed that Stage IV GC patients have high number of CTCs than operable GC patients. Tumor histology and Lauren classification did not affect to CTC positivity in Stage IV and operable GC patients. We also observed the potential use of CTC in serial follow-ups and one of the cases shows the decreasing CTCs followed by clinical response. In conclusion, this study demonstrates that detection of CTC may be useful prognostic indicator of GC cancer patients.

Citation Format: Won Suk Lee, Jeong Min Kim, Woo Sun Kwon, So Jung Lim, Tae Soo Kim, Ga Yun Kim, Sung Ho Choi, Byung Hee Jeon, Min Kyu Jung, Joong Bae Ahn, Hyun Cheol Chung, Sun Young Rha. Utility of circulating tumor cell (CTC) in advanced gastric cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 391. doi:10.1158/1538-7445.AM2015-391
Abstract 3917: PDGF induces cell growth and glycolysis in colon cancer
Authors: Moench Romana , Kannen Vinicius , Grimmig Tanja , Germer Christoph T. , Gasser Martin , Waaga-Gasser Ana Maria
Published: August 2015

The platelet derived growth factor (PDGF) plays an important role in several solid tumors. Involved in cell migration and proliferation primarily of stromal cells in cancers PDGF represents a key target in cancer therapy. The aim of this study was to analyze the specific role of PDGF on tumor cell proliferation and metabolism in colorectal cancer (CRC). The human colon cancer cell line HT-29 was cultured and stimulated time-dependently with PDGF. Additionally, inhibition of the PI3k/Akt/mTOR-pathway was performed simultaneously to PDGF stimulation. Whole cell and RNA extracts were analyzed by Western Blot and RT-q-PCR for the PI3k/Akt/mTOR-pathway and components of cellular metabolism. To investigate the effects of PDGF on proliferation MTS assays were performed. Additionally, mRNA levels of PDGF and metabolic factors in tumors from patients with CRC were analyzed by RT-qPCR. PDGF stimulation resulted in increased HT29 cell proliferation compared to untreated controls. Blocking of Akt resulted in inhibition of pS6, activation of Retinoblastoma (Rb) and reduced tumor cell growth. Additionally, under stimulation a higher glycolytic rate was observed while oxygen consumption remained unaltered. Investigated tumors from patients with CRC showed a stage-dependent increase in PDGF expression and a boosted glycolytic rate. The cytokine PDGF induced proliferation accompanied with an altered glycolysis in HT-29 colon cancer. An increased glycolysis and tumor cell proliferation support accelerated cell proliferation and tumor growth. The growth reducing effect of Akt inhibition indicates the PI3K/Akt/mTOR-pathway to play a crucial role in CRC progression and therefore could be an important target in cancer therapy.

Note: This abstract was not presented at the meeting.

Citation Format: Romana Moench, Vinicius Kannen, Tanja Grimmig, Christoph T. Germer, Martin Gasser, Ana Maria Waaga-Gasser. PDGF induces cell growth and glycolysis in colon cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3917. doi:10.1158/1538-7445.AM2015-3917
Abstract 3920: Identification of LASEP1 as a new serological and prognostic biomarker and a therapeutic target for lung cancer
Authors: Takano Atsushi , Nakamura Yusuke , Daigo Yataro
Published: August 2015

Lung cancer is the leading cause of cancer deaths in the world and the overall prognosis remains poor. The poor prognosis of lung cancer patients is largely a result of the occult metastatic dissemination. Thus, the development of early cancer detection biomarker and new treatment is required. This study aims to identify novel biomarkers or therapeutic targets for lung cancers. We used cDNA microarrays for screening genes encoding transmembrane/secretory proteins that are up-regulated in lung cancers. As a result of this process, we identified a secreted protein, LASEP1 (lung cancer-associated serum protein 1) as a candidate. Immunohistochemical staining of LASEP1 showed that LASEP1 expression was observed in 210 (56.1%) of 374 NSCLC (non-small cell lung cancer) that had undergone curative surgery. High level of LASEP1 expression was associated with poor prognosis for NSCLC patients (P<0.0001 by log-rank test). Serum LASEP1 levels were higher in NSCLC patients than in healthy volunteers. The proportion of serum LASEP1-positive cases was 127 (38.6%) of 329 NSCLCs, while 4 (3.9%) of 102 healthy volunteers were falsely diagnosed. Furthermore, reduction of LASEP1 by siRNA significantly induced the apoptosis of lung cancer cells. Furthermore, the growth activity of the LASEP1-positive cells was neutralized by addition of originally developed anti-LASEP1 monoclonal antibodies into their culture media, and the systemic administration of the anti-LASEP1 antibody to tumor-implanted mice significantly suppressed tumor growth without any adverse events. By gene expression analysis using microarray and mRNAs from lung cancer cells transfected with siRNA for LASEP1, we identified WHSC1 gene that encodes an oncogenic histone methyltransferase as one of LASEP1-target genes. These data suggest that LASEP1 is a possible diagnostic and prognostic biomarker and therapeutic target for lung cancer.

Citation Format: Atsushi Takano, Yusuke Nakamura, Yataro Daigo. Identification of LASEP1 as a new serological and prognostic biomarker and a therapeutic target for lung cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3920. doi:10.1158/1538-7445.AM2015-3920
Abstract 3926: RhoB expression is upregulated by androgen in thyroid cancer cells
Authors: Gupta Anvita , Jones Melanie , Moscatello Augustine , Shin Edward , Tiwari Raj K. , Geliebter Jan
Published: August 2015

The incidence of thyroid cancer has increased three-fold over the last thirty years and the American Cancer Society estimates that there will be 62,980 new cases and 1,890 deaths due to the disease in the United States in 2014. Papillary Thyroid Cancer (PTC) is the most common endocrine malignancy and develops in premenopausal women, at a four-fold higher incidence, than in men. However, PTC exhibits increased aggressiveness with poor prognosis in men diagnosed with the disease. These observations led us to explore the role of androgen and androgen receptor in this disease. Preliminary data from our lab indicates that androgen receptor (AR) acts as a negative regulator of growth as evidenced by a statistically significant decrease in proliferation upon DHT addition to 8505c anaplastic/PTC cells stably transfected with AR. Transcriptional profiling using RNA-Seq and gene ontology analysis, on a 48 hour DHT treated 8505c-AR transfected clone, 84E7, revealed significant changes in the expression of genes associated with proliferation (474 genes, p = 2.4E-24), cell cycle progression (129 genes, p = 6.54E-6) and migration (301 genes, p = 3.77E-30). Amongst the genes which showed the highest significant differential expression was RhoB, a Rho GTPase, with an approximately 50-fold upregulation. Rho proteins are small molecules belonging to the Ras superfamily, known to regulate cell morphology, actin/microtubule cytoskeleton reorganization along with vesicle trafficking. While RhoA and RhoC promote cancer progression and metastasis, the functions of their structural homolog, RhoB, are largely context dependent. Evidence has emerged that RhoB expression may support malignancy in certain cancers, while exhibiting a tumor-suppressor gene function in others. In our system, addition of DHT to 84E7 cells for short time periods of 1, 3 and 6 hours resulted in rapid RhoB mRNA upregulation by 2.5, 8, and 17 fold, respectively, compared to untreated cells. This quick, time-dependent increase in androgen-responsive RhoB may play an integral protective and reparative role which is possibly lost during neoplasticity of thyrocytes. Investigations into various pathways linking RhoB and effector proteins with carcinogenesis would be crucial in assessing the potential of RhoB as a bio-marker for metastatic PTC and thence opening avenues for development of viable therapies for better management of the disease.

Citation Format: Anvita Gupta, Melanie Jones, Augustine Moscatello, Edward Shin, Raj K. Tiwari, Jan Geliebter. RhoB expression is upregulated by androgen in thyroid cancer cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3926. doi:10.1158/1538-7445.AM2015-3926
Abstract 3933: The landscape and clonal architecture in lower grade glioma
Authors: Suzuki Hiromichi , Aoki Kosuke , Chiba Kenichi , Sato Yusuke , Shiozawa Yusuke , Shiraishi Yuichi , Niida Atsushi , Shimamura Teppei , Sanada Masashi , Miyano Satoru , Wakabayashi Toshihiko , Natsume Atsushi , Ogawa Seishi
Published: August 2015

Lower grade gliomas (LGGs, WHO grade II/III gliomas) account for approximately one third of all gliomas. Although LGGs are typically slowly progressive, their clinical course is invariably indolent and most patients ultimately succumb to death. In contrast to glioblastoma (GBM), our knowledge about the genetic lesions and clonal evolution in LGG is still incomplete. So, to obtain a complete registry of gene mutations involved in LGG pathogenesis and their role in clonal evolution, we performed whole exome sequencing (WES) and/or targeted sequencing of 335 LGG cases. Clonal evolution in LGG was investigated using paired primary/relapsed tumor specimens from 10 cases as well as multiple tumor specimens (median 5) from 4 cases.

Massive parallel sequencing revealed LGGs were clearly grouped into three subgroups with or without IDH1/2 mutation and 1p19q loss of heterozygous (LOH). Type I tumor with IDH1/2 mutation and 1p19q LOH had a most favorable survival and harbored mutations in TERT promoter, CIC, FUBP1 and NOTCH1. Type II tumor with IDH1/2 mutant/1p19q intact subtype represented TP53 bialleleic inactivation and/or ATRX mutations. Type III tumor with IDH1/2 intact showed GBM like mutation profile and poor prognosis.

Large scale samples allowed to obviously detect strong positive/negative correlations with each other driver genes. Extensive analysis of variant allele frequencies among co-existing mutations revealed temporal orders of gene mutations in each subtypes.

Multi regional/time-points sampling analysis suppoted mutational order and revealed regional and special heterogeneity with tumor evolution in LGGs. LGG contiguously developed and generated heterogeneity through acquiring new mutations in a complex but ordered fashion.

In conclusion, our findings delineated the landscape of gene mutations in LGG. LGG had mutually exclusive mutational patterns with hierarchical order in discrete subtypes. IDH1/2 and TERT promoter mutations and 1p19q LOH were thought to exist in the major clone and important role in tumor initiation. In contrast, common occurrence of parallel mutations found in TP53, ATRX, CIC, FUBP1 and NOTCH1 genes indicated central roles of these mutations in LGG development.

Citation Format: Hiromichi Suzuki, Kosuke Aoki, Kenichi Chiba, Yusuke Sato, Yusuke Shiozawa, Yuichi Shiraishi, Atsushi Niida, Teppei Shimamura, Masashi Sanada, Satoru Miyano, Toshihiko Wakabayashi, Atsushi Natsume, Seishi Ogawa. The landscape and clonal architecture in lower grade glioma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3933. doi:10.1158/1538-7445.AM2015-3933
Abstract 3945: PTEN loss is associated with aggressive behavior and poor prognosis in Middle Eastern Triple Negative Breast Cancer
Authors: Siraj Abdul K. , Beg Shaham , Prabhakaran Sarita , Jehan Zeenath , Ajarim Dahish , Al-Dayel Fouad , Tulbah Asma , Al-Kuraya Khawla
Published: August 2015

PTEN is a tumor suppressor that negatively regulates the PI3K-AKT signaling pathway which is involved in the pathogenesis of many different tumor types and serves as a prognostic marker in breast cancer. However, the significance of the role of PTEN in Middle Eastern ethnic breast cancer has not been explored especially with the fact that breast cancer originating from this ethnic population tend to behave more aggressively than breast cancer in the west. In order to learn more on the prevalence and clinical significance of PTEN protein expression and PTEN gene deletions, a tissue microarray containing more than 1000 primary breast cancers with clinical follow up data was used. Tissue Microarray sections were analyzed for protein expression and copy number change using immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Loss of PTEN immunostaining was observed in 77% of the cases. PTEN loss was significantly associated with large tumor size (p = 0.0030), high grade (p = 0.0281), tumor recurrence (p = 0.0333) and Triple negative breast cancers (p = 0.0086). PTEN loss in Triple negative breast cancers was significantly associated with rapid tumor cell proliferation (p = 0.0396) and poor prognosis (p = 0.0408). PTEN deletion was found only in 60 cases (6.4%) of cases. It was concluded that loss of PTEN protein expression occurs at high frequency in Middle Eastern breast cancer. PTEN inactivation may potentially lead to an aggressive behavior of tumor cells through stimulation of tumor cell proliferation. Furthermore PTEN signaling pathway might be used as potential therapeutic target in Triple negative breast cancers since loss of its expression is shown to be significantly associated with this aggressive subtype of breast cancer.

Citation Format: Abdul K. Siraj, Shaham Beg, Sarita Prabhakaran, Zeenath Jehan, Dahish Ajarim, Fouad Al-Dayel, Asma Tulbah, Khawla Al-Kuraya. PTEN loss is associated with aggressive behavior and poor prognosis in Middle Eastern Triple Negative Breast Cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3945. doi:10.1158/1538-7445.AM2015-3945
Abstract 3970: Development of novel form of mimic microRNA for lung cancer therapy
Authors: Ohno Shinichiro , Itano Karen , Harada Yuichirou , Asada Koutaro , Oikawa Keiki , Kashiwazako Mikie , Takanashi Masakatsu , Sudo Katsuko , Ikeda Norihiko , Kuroda Masahiko
Published: August 2015

Exogenously supplied tumor suppressor miR-34a induces apoptosis and cell cycle arrest in lung cancer cell lines. It is suggested that miRNA replacement therapy of miR-34a is a promising approach for the treatment of lung cancer. On the one hand, the immunological response to nucleic acid via Toll like receptors (TLRs) and RIG-I like receptors (RLRs) that restrict the practical use of nucleic acid drugs in the clinical setting. It is known that the immunological response to the nucleic acid is strongly depends the length of nucleic acid. Therefore, we have tried to develop a shorter nucleic acid drugs that can escape immune surveillance. The deletion of nucleotide with maintenance of the secondary structure of short miR-34a keeps the activity and the target specificity of miR-34a. Finally, miR-34a can be shortened from 44 to 30 nucleobase. The shortened 30 nucleotides miR-34a was named “Guide hairpin RNA (ghR) -34a” and analyzed in detail. The inhibitory activity against miR-34a targets genes such as AXL, MET and CDK6 were comparable between ghR-34a and physiological form of miR-34a at both the mRNA and protein levels. Similarly, there is no difference in the antiproliferative activity against lung cancer cell line between ghR-34a and miR-34a. On the other hand, the ghR-34a does not induce inflammatory response and unwanted gene inhibition by passenger strand. These data indicated that ghR-34a can inhibits lung cancer with low side effects. Furthermore, the shortening methods can be broadly applied to the other miRNAs. Thus, the shortening methods of miRNA developed in the study may be provide a novel, safe and effective nucleic acid drug platform and promote the clinical usefulness of nucleic acid therapy.

Note: This abstract was not presented at the meeting.

Citation Format: Shinichiro Ohno, Karen Itano, Yuichirou Harada, Koutaro Asada, Keiki Oikawa, Mikie Kashiwazako, Masakatsu Takanashi, Katsuko Sudo, Norihiko Ikeda, Masahiko Kuroda. Development of novel form of mimic microRNA for lung cancer therapy. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3970. doi:10.1158/1538-7445.AM2015-3970
Abstract 3977: Evaluation of miR10b and miR9 expression in breast cancer and correlations with distant metastases development
Authors: Barbano Raffaela , Pasculli Barbara , Copetti Massimiliano , Fontana Andrea , Coco Michelina , Vitulano Lorenzo , Poeta Maria Luana , Picardo Francesca , Marangi Gianfranco , Murgo Roberto , Ciuffreda Luigi , Valori Vanna Maria , Valori Vanna Maria , Morritti Maria , Maiello Evaristo , Fazio Vito Michele , Parrella Paola
Published: August 2015

We have recently demonstrated that the evaluation of miR10b expression relative to its normal counterparts (Relative Expression Ratio, RER) is able to predict patients risk to develop distant metastases. To confirm these results and evaluate whether other microRNAs could be predictive of outcome in breast cancer we evaluated the expression of miR10b and miR9 in paired tumors and apparently normal breast tissues 2 cm distant from tumour (NBT) on an extended cohort (n = 127) with a longer follow up (median 42 months). As control normal breast tissues from 10 healthy subjects (HBT) treated by reductive mammoplasty were analyzed. An increased expression was seen for the miR9 and miR10b in NBT samples as compared with HBT (P<0.01). Whereas the expression of each of the two miRNAs was significantly lower in tumour samples as compared to the NBT specimens with RER value of 0.15 for miR10b (P = 0.001) and 0.59 for miR9 (P = 0.05). However only higher miR10b RERs were associated with distant metastases development in multivariable analysis (HR = 1.12; P = 0.04). Our results confirm the role of miR10b as predictive biomarkers of metastases development in breast cancer. In addition our data suggest a deregulation of miRNAs expression in the normal tissue surrounding tumor as compared to normal breast of healthy individuals.

Citation Format: Raffaela Barbano, Barbara Pasculli, Massimiliano Copetti, Andrea Fontana, Michelina Coco, Lorenzo Vitulano, Maria Luana Poeta, Francesca Picardo, Gianfranco Marangi, Roberto Murgo, Luigi Ciuffreda, Vanna Maria Valori, Vanna Maria Valori, Maria Morritti, Evaristo Maiello, Vito Michele Fazio, Paola Parrella. Evaluation of miR10b and miR9 expression in breast cancer and correlations with distant metastases development. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3977. doi:10.1158/1538-7445.AM2015-3977
Abstract 4038: Role of the Rho-GTPase CDC42 in glioma migration
Authors: Okura Hidehiro , Golbourn Brian J. , Luck Amanda J. , Smith Christian A. , Rutka James T.
Published: August 2015

Introduction: Malignant astrocytoma is a highly proliferative and invasive neoplasm that infiltrates diffusely into regions of normal brain rendering total surgical extirpation impossible and effective local radiation therapy difficult. Conventional chemotherapy frequently fails because most agents administered systemically will not cross the blood-brain barrier. Despite recent reports which have shown a 2-month survival advantage when adjuvant temozolomide chemotherapy is used for glioblastoma multiforme, death ensues in most cases within 2 years of diagnosis. For truly significant advances to be made in the treatment of patients with malignant astrocytoma, we must develop a greater understanding of the basic biology of these neoplasms. PURPOSE: The aim of this study is to examine a key pathway involving the Rho-GTPase family of proteins that regulates glioma cell migration from the tumor mass into normal brain tissue. One of the Rho-GTPase family proteins, CDC42 is the major regulator of cell polarity. It plays a crucial role in controlling the direction of cell migration by determining the cell membrane localization of lamellipodia, and by moving the microtubule-organizing center and Golgi apparatus in front of the nucleus and towards the leading edge of the cell. However, there have been few reports on the role of CDC42 in human astrocytomas. METHOD: In order to examine the role of CDC42 in glioblastoma, we generated stable doxycycline (DOX) inducible cell lines expressing wild type (WT) -, constitutively active (CA) -, and dominant negative (DN) -CDC42. Cell viability, migration, and invasion assays were performed using the cell lines. We also used orthotopic xenograft mouse models and evaluated the survival by Kaplan-Meier analysis. RESULTS: Administration of DOX induced the expression of CA-CDC42 and demonstrated significantly increased migration and invasion properties compared to WT and DN-CDC42. Furthermore, CA-CDC42 bearing mice with DOX treatment had significantly decreased survival compared to WT and DN with DOX. CONCLUSION: Our results demonstrated that CA-CDC42 enhances the astrocytoma invasion and migration. Furthermore, CA-CDC42 increased the size of implanted tumor in the murine brain leading to early mortality. We concluded that CA-CDC42 plays an important role for astrocytoma malignancy.

Citation Format: Hidehiro Okura, Brian J. Golbourn, Amanda J. Luck, Christian A. Smith, James T. Rutka. Role of the Rho-GTPase CDC42 in glioma migration. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4038. doi:10.1158/1538-7445.AM2015-4038
Abstract 4048: HPV16 immunity induced by immune responses to mutations in E6 and E7 proteins
Authors: Agarwal Maria , Saint-Fleur Ashley , Fu Jie , Levitsky Hyam , Trimble Cornelia L.
Published: August 2015

Human papillomavirus (HPV) causes 30% of cancers attributable to infectious pathogens. HPV16 causes most cervical, anal, vaginal, vulvar, and oropharyngeal cancers. Although preventive vaccines targeting HPV16 and 18 are available, rates of vaccination are uneven. The incidence of HPV-associated cancers for which no screening algorithms have been validated continues to increase.

Development of squamous cervical cancer and its precursor, cervical intraepithelial neoplasia (CIN) 2/3 is associated with integration of HPV genome into the host genome, and expression of the HPV E6 and E7- gene products, which inactivate p53 and pRb respectively. Since E6 and E7 are constitutively expressed in cervical carcinomas and CIN, they present attractive immunotherapeutic targets. Development of immunotherapies has been limited, in part, by modest immunogenicity of vaccines tested to date. The purpose of this study is to augment the immunogenicity of the E6 and E7 antigens through mutational screening.

A subset of CIN2/3 do undergo complete regression. Our lab has carried out a series of prospective clinical studies, including immunotherapeutic interventions, in subjects with CIN2/3. Peripheral blood T cell responses to E6 and E7 are marginal, and do not distinguish regressors in either vaccinated or unvaccinated subjects. However, in vaccinated subjects, the lesion microenvironment is comprised of new tertiary lymphoid structures, evidence of T cell proliferation elicited by cognate antigen, and clonally expanded TCRs. Tissue T cells in vaccinated subjects access dysplastic epithelium, in contrast to T cells in persistent lesions, which were confined to the stromal compartment. These findings suggest that T cells that could mediate regression can be elicited by vaccination. However, strategies to enhance immunogenicity would likely increase therapeutic effect. Here we present an approach designed to determine the effect of mutations in E6 and E7 on antigenicity. Mutations can enhance immune responses to antigens by several mechanisms, including altering antigen processing, trafficking, glycolsylation, and affinity to major histocompatibility complexes. Using error prone PCR, we generated libraries containing random mutations in the coding sequence of E6 and E7. Clones from each library are currently being screened in vitro to identify immunogenic mutants, which could be potentially used for multi-epitope vaccines.

Citation Format: Maria Agarwal, Ashley Saint-Fleur, Jie Fu, Hyam Levitsky, Cornelia L. Trimble. HPV16 immunity induced by immune responses to mutations in E6 and E7 proteins. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4048. doi:10.1158/1538-7445.AM2015-4048
Abstract 4062: Role of CD137 signaling in immune evasion of Hodgkin and Reed-Sternberg cells
Authors: Rajendran Sakthi , Schwarz Herbert
Published: August 2015

Hodgkin lymphoma (HL), a lymphoid malignancy, is a frequently occurring cancer among adolescents and young adults. HL is marked by the presence of malignant Hodgkin and Reed-Sternberg (HRS) cells, which constitute only a small fraction of the tumor mass. HRS cells are present in a complex stroma of inflammatory leukocytes, but are often able to evade the host immune system. These cells express various receptors to recruit leukocytes and hijack them for supporting the tumor growth. Our group has reported the overexpression of CD137, a member of the tumor necrosis factor receptor (TNFR) superfamily, on the HRS cells of HL tumor samples. CD137 is a co-stimulatory molecule expressed by T cells, only upon activation. We speculated that the expression of CD137 on the HRS cells may be associated with its malignancy, as HRS cells are clonally derived from B cells which are not expressing CD137. Our study aims to analyse the functional role of CD137 expression behind the survival, progression, and cytokine release of the malignant HRS cells. We found that the induction of CD137 signaling in HRS cell lines enhanced the secretion of cytokines such as IL-6, TNF-α and IL-13. In order to investigate the effect of these cytokines on the immune microenvironment, we cultured peripheral blood mononuclear cells (PBMCs) in the supernatants derived from CD137-induced HRS cells. PBMCs, when cultured in the above mentioned conditioned supernatants, secreted lesser levels of IFN-γ (a Th1 cytokine essential for T cell and NK cell cytotoxicity), implying a suppressed state of immune cells. Further, neutralisation of IL-13 (a Th2 cytokine) in the conditioned supernatants reversed the reduction of IFN-γ observed in PBMCs. This shows that IL-13 secreted by the HRS cells upon CD137 signaling is responsible for the inhibition of IFN-γ secretion by PBMCs. To summarize, CD137 signaling in HRS cells leads to the secretion of cytokines, which drive the polarization of helper T cells towards Th2 type, thus reducing Th1 immune responses against cancers. Taken together, our findings reveal the role of CD137 signaling as a novel mechanism of immune escape in HRS cells. Hence, targeting CD137 can be a potential therapeutic option for HL patients.

Citation Format: Sakthi Rajendran, Herbert Schwarz. Role of CD137 signaling in immune evasion of Hodgkin and Reed-Sternberg cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4062. doi:10.1158/1538-7445.AM2015-4062
Abstract 4070: Prolonged silencing of diacylglycerol acyltransferase-1 induces a dedifferentiated phenotype in human liver cells
Authors: Chang Soyoung , Sung Pil Soo , Lee Jungsul , Park Junseong , Shin Eui-Cheol , Choi Chulhee
Published: August 2015

Diacylglycerol acyltransferase-1 (DGAT1), a key enzyme in triglyceride (TG) biogenesis, is highly associated with metabolic abnormalities, such as obesity and type 2 diabetes. However, the effects of DGAT1 sliencing in the human liver have not been elucidated. To investigate the effects of DGAT1 silencing in human liver cells, we compared the cellular behaviors of DGAT1-deficient Huh-7.5 cell lines with those of control Huh-7.5 cells. DGAT1-deficient cells acquired dedifferentiated and stem cell-like characteristics, such as formation of aggregates in the presence of high levels of growth factors, high proliferation rates, and loss of albumin secretion. In relation to aggregate formation, the expression level of various adhesion molecules was significantly altered in DGAT1-deficient cells. Microarray data analysis and immunostaining of patient tissue samples clearly showed decreased expression levels of DGAT1 and integrin β1 in patients who have nodular cirrhosis without fatty degeneration.

Note: This abstract was not presented at the meeting.

Citation Format: Soyoung Chang, Pil Soo Sung, Jungsul Lee, Junseong Park, Eui-Cheol Shin, Chulhee Choi. Prolonged silencing of diacylglycerol acyltransferase-1 induces a dedifferentiated phenotype in human liver cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4070. doi:10.1158/1538-7445.AM2015-4070
Abstract 4083: Deletion of the BMP receptor BMPR1a results in EMT and impairs mammary gland tumor formation and metastasis
Authors: Hover Laura D. , Pickup Michael W. , Gorska Agnieszka E. , Chytil Anna , Guo Yan , Novitskiy Sergey V. , Moses Harold L. , Owens Philip
Published: August 2015

Bone Morphogenetic Proteins (BMPs) are secreted cytokines/growth factors belonging to the Transforming Growth Factor β (TGFβ) superfamily. BMP ligands have recently been shown to be overexpressed in human breast cancers. Normal and cancerous breast display active BMP signaling as indicated by phosphorylated Smads 1, 5 and 9. We combined mice expressing the MMTV.PyVmT oncogene with mice lacking BMPR1a in mammary epithelial cells and found this deletion resulted in delayed tumor onset and extended survival significantly (p-value = <0.001). We examined the histopathology of BMPR1a knockout (cKO) tumors and found a striking loss of epithelial characteristics combined with stromal desmoplasia. Immunofluorescence staining revealed that cKO tumors co-expressed Keratin 5 and mesenchymal cell markers such as Vimentin. This indicated that epithelial-to-mesenchymal (EMT)-like transitions were occurring in cKO tumors. We performed microarray analysis on these tumors and found changes that supported EMT-like changes with increased Snail mRNA expression. We established primary tumor cell lines and found that BMPR1a cKO had slower growth in vitro and upon in vivo implantation. Additionally, cKO tumor cells had reduced migration yet not invasion in vitro. We next analyzed human databases from TCGA and survival data from microarrays to confirm BMPR1a tumor promoting functions and found high BMPR1a gene expression correlated with worse survival regardless of molecular breast cancer subtype. In conclusion, we found that loss of the BMPR1a impairs tumor formation and progression and does not exhibit tumor suppressive functions in mouse and human breast cancer.

Citation Format: Laura D. Hover, Michael W. Pickup, Agnieszka E. Gorska, Anna Chytil, Yan Guo, Sergey V. Novitskiy, Harold L. Moses, Philip Owens. Deletion of the BMP receptor BMPR1a results in EMT and impairs mammary gland tumor formation and metastasis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4083. doi:10.1158/1538-7445.AM2015-4083
Abstract 4089: CADM1 is a TWIST1 regulated suppressor of melanoma invasion
Authors: Hartsough Edward J. , Weiss Michele B. , Kugel Curtis H. , Rosenbaum Sheera R. , Aplin Andrew E.
Published: August 2015

Melanoma is the deadliest form of skin cancer; however, with early detection prior to metastatic dissemination, patients generally have a good prognosis. Recently, our lab has implicated the transcription factor TWIST1 in the progression of melanoma towards metastasis. In melanoma cells, we found that RAS-RAF-MEK-ERK (ERK1/2 pathway) signaling increases TWIST1 expression, which promotes invasive properties in the dermal microenvironment at least in part by enhancing levels of the matrix metalloproteinase, MMP1. Other TWIST1 regulated targets are poorly described. In this study, we compared expression profiling data from cells expressing shRNA against TWIST1 or cells overexpressing TWIST1 in order to determine TWIST1 regulated genes. KEGG and GO analysis revealed that TWIST1 is responsible for regulating a number of genes involved in cellular adhesion. We found that TWIST1 levels inversely correlate with levels of cell adhesion molecule 1 (CADM1) (NECL-2, IGSF4, TSLC1, SynCAM). Chromatin immunoprecipitation (ChIP) studies and promoter assays demonstrate that TWIST1 physically interacts with CADM1 promoter and this is associated with reduced CADM1 levels. Additionally, CADM1 expression is inversely associated ERK1/2 signaling and TWIST1 expression. Modulation of cellular CADM1 levels does not seem to affect proliferations rates however, overexpression of CADM1 augments cell-cell interaction, and cell aggregation. Furthermore, exogenous CADM1 inhibited serum directed migration and invasion through matrigel coated boyden chambers, while knockdown of CADM1 was associated with an enhancement in the migratory and invasion properties. Taken together, these data provide evidence that CADM1 is negatively regulated by TWIST1, and may act as a suppressor of melanoma invasion.

Citation Format: Edward J. Hartsough, Michele B. Weiss, Curtis H. Kugel, Sheera R. Rosenbaum, Andrew E. Aplin. CADM1 is a TWIST1 regulated suppressor of melanoma invasion. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4089. doi:10.1158/1538-7445.AM2015-4089
Abstract 41: The Wnt3a targetome in triple-negative breast cancer cell lines
Authors: Maubant Sylvie , Tesson Bruno , Maire Virginie , Ye Mengliang , Rigaill Guillem , Gentien David , Cruzalegui Francisco , Tucker Gordon C. , Roman-Roman Sergio , Dubois Thierry
Published: August 2015

The canonical Wnt/beta-catenin pathway has been shown to be activated in triple-negative breast cancer (TNBC). The activation of this pathway leads to the expression of specific target genes depending on the cell/tissue context. Here, we analyzed the transcriptome of two different TNBC cell lines to define a comprehensive list of Wnt target genes. The treatment of cells with Wnt3a for 6h up-regulated the expression (fold change > 1.3) of 59 genes in MDA-MB-468 cells and 241 genes in HCC38 cells. Thirty genes were common to both cell lines. Beta-catenin may also be a transcriptional repressor and we found that 18 and 166 genes were down-regulated in response to Wnt3a treatment for 6h in MDA-MB-468 and HCC38 cells, respectively, of which six were common to both cell lines. Only half of the activated and the repressed transcripts have been previously described as Wnt target genes. Therefore, our study reveals 137 novel genes that may be positively regulated by Wnt3a and 104 novel genes that may be negatively regulated by Wnt3a. These genes are involved in the Wnt pathway itself, and also in TGF-beta, p53 and Hedgehog pathways. Thorough characterization of these novel potential Wnt target genes may reveal new regulators of the canonical Wnt pathway. The comparison of our list of Wnt target genes with those published in other cellular contexts confirms the notion that Wnt target genes are tissue-, cell line- and treatment-specific. Genes up-regulated in Wnt3a-stimulated cell lines were more strongly expressed in TNBC than in luminal A breast cancer samples. These genes were also overexpressed, but to a much lesser extent, in HER2+ and luminal B tumors. We identified 72 Wnt target genes higher expressed in TNBCs (17 with a fold change >1.3) which may reflect the chronic activation of the canonical Wnt pathway that occurs in TNBC tumors.

Citation Format: Sylvie Maubant, Bruno Tesson, Virginie Maire, Mengliang Ye, Guillem Rigaill, David Gentien, Francisco Cruzalegui, Gordon C. Tucker, Sergio Roman-Roman, Thierry Dubois. The Wnt3a targetome in triple-negative breast cancer cell lines. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 41. doi:10.1158/1538-7445.AM2015-41