bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1404: A red-shifted fluorescent substrate for aldehyde dehydrogenase, AldeRed 588-A, for labeling viable ALDH-positive cells
Authors: Chu Vi , Taganov Konstantin , Ramesh D , Asbrock Nick , Pomper Martin
Published: August 2015

Normal and cancer stem cells can be isolated based upon the enzymatic activity of aldehyde dehydrogenase I (ALDH1), a detoxifying enzyme responsible for oxidation of hazardous aldehyde byproducts. ALDH1 has been used to isolate cancer stem cells of various human malignancies including bladder, breast, cervical, colon, head and neck, liver, lung, pancreas, prostate and ovary. Currently, the ALDEFLUORTM assay which uses a fluorescent substrate is the only commercially available reagent for ALDH detection. The substrate used in this assay primarily emits in the green region of the electromagnetic spectrum (512nm). For researchers with valuable cell and transgenic animal models in which the target gene of interest has been tagged with eGFP, ALDEFLUOR therefore cannot be used. Selection of cells positive for aldehyde dehydrogenase (ALDH) activity from a green fluorescent background is thus difficult with existing reagents. We now describe a red-shifted fluorescent substrate for ALDH, AldeRed 588-A, that provides additional flexibility for utilizing ALDH as a marker for stem cell and cancer stem cell isolation. The activity of AldRed 588-A was compared with the ALDEFLUOR reagent and demonstrated similar ability to fractionate ALDHpos cells in a number of cell lines tested.

Citation Format: Vi Chu, Konstantin Taganov, D Ramesh, Nick Asbrock, Martin Pomper. A red-shifted fluorescent substrate for aldehyde dehydrogenase, AldeRed 588-A, for labeling viable ALDH-positive 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 1404. doi:10.1158/1538-7445.AM2015-1404
Abstract 1414: Molecular and functional characterization of ovarian cancer stem cells
Authors: Lupia Michela , Bertalot Giovanni , Colombo Nicoletta , Confalonieri Stefano , Di Fiore Pier Paolo , Cavallaro Ugo
Published: August 2015

Epithelial Ovarian Cancer (OC) is one of the leading causes of death for women, and no significant therapeutic progress has been made in the last decades.

Recent data suggest that one of the mechanisms accounting for resistant and/or relapsing disease is a subpopulation of cells in human tumors with stem-like characteristics (cancer stem cells, CSCs). CSCs are defined as a small subpopulation of cells within the tumor bulk that possess the capacity, on one hand, to self-renew and, on the other hand, to give rise to all heterogeneous cancer cell lineages that compose the tumor of origin. The CSC hypothesis provides an attractive cellular mechanism to explain the therapeutic refractoriness, dormant behavior, and relapse of the disease.

Our study aims at assessing ovarian cancer stem cells (OCSC) as causal players in OC etiology and progression and at defining their molecular and functional profile. Specifically, we are pursuing the following objectives through the accomplishment of these milestones: 1) collection of normal and pathological samples; 2) identification of OCSC based on functional properties; 3) comparison of gene expression profiles between cancer stem cells and their normal counterpart; 4) characterization of novel genes/pathways involved in OCSC function (clonogenicity, tumorigenicity, quiescence, chemoresistance, etc.).

This workflow is being applied to a series of fresh surgical samples of OC as well as to normal ovarian surface epithelium (OSE) and fallopian tube epithelium (FTE), namely the tissues of origin of OC. Thus, we obtained a collection of primary cells that recapitulate many traits of the original tissue both in vitro and in vivo. The gene expression profiles of sphere-forming SC has been compared to that of adherent, parental cells, aimed at identifying stemness-associated genes. This screening has been extended to OC, OSE, and FTE, resulting in a set of genes that are differentially expressed in OCSC as compared to their normal counterparts.

A subset of such genes has been selected for the subsequent validation, both as OCSC biomarkers that could have clinical applications and as drivers in the biological and pathogenic function of OCSC.

Our study might set the stage for innovative therapeutic approaches aimed at the selective elimination of OCSC, thus preventing tumor recurrence and chemoresistance, namely the two main causes of ovarian cancer lethality.

Citation Format: Michela Lupia, Giovanni Bertalot, Nicoletta Colombo, Stefano Confalonieri, Pier Paolo Di Fiore, Ugo Cavallaro. Molecular and functional characterization of ovarian cancer stem 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 1414. doi:10.1158/1538-7445.AM2015-1414
Abstract 1426: Lung epithelial cells induce both phenotype alteration and senescence in breast cancer cells
Authors: Furukawa Masashi , Wells Alan , Wheeler Sarah , Clark Amanda M.
Published: August 2015

Purpose: Breast cancer is the most common cancer in women. The lung is one of the most common sites of breast cancer metastasis. While metastatic seeding is often accompanied by a dormancy-promoting mesenchymal to epithelial reverting transitions (MErT), we aimed to determine whether lung epithelial cells can impart this phenotype on aggressive breast cancer cells.

Methods: Co-culture experiments of normal lung epithelial cell lines (SAEC, NHBE or BEAS-2B) and breast cancer cell lines (MCF-7 or MDA-MB-231) were conducted. Flow cytometry analysis, immunofluorescence staining for E-cadherin or Ki-67 and senescence associated beta-galactosidase assays assessed breast cancer cell outgrowth and phenotype.

Results: Co-culture of the breast cancer cells had different effects on breast cancer cells. The epithelial MCF-7 cells were increased in number but still clustered even if in a slight more mesenchymal spindle morphology. On the other hand, the mesenchymal MDA-MB-231 cells survived but did not progressively grow out in co-culture. These aggressive carcinoma cells underwent an epithelial shift as indicated by cuboidal morphology and increased E-cadherin. Disruption of E-cadherin express in MDA-MB-231 using shRNA prevented this phenotypic reversion in coculture. In addition, we had noted enlarged MDA-MB-231 cells in the co-cultures with SAEC and NHBE but not BEAS-2B cells as suggestion of senescence. A subset of these cells stained with the senescence associated marker β-galactosidase (SA-β-gal). Lung cells limited cancer cell growth kinetics as noted by both (1) some of the cells becoming larger and positive for senescence markers/negative for proliferation marker Ki-67, and (2) Ki-67 positive cells significantly decreasing in MDA-MB-231 and MCF-7 cells after co-culture.

Conclusions: Our data indicate that normal lung epithelial cells can drive an epithelial phenotype and suppress the growth kinetics of breast cancer cells by changing their phenotypes.Similar to the hepatocytes, these normal cells may promote the survival, seeding and even dormancy of aggressive mesenchymal carcinoma cells, with phenotypes similar to MDA-MB-231 cells, but also provide progressive signaling when stressed accounting for the spindle morphology and slightly increased growth rates of the epithelial carcinoma cells, such as those MCF-7 represents. In this latter manner, insults the small airway may provoke an outgrowth from dormant breast cancer cells.

Citation Format: Masashi Furukawa, Alan Wells, Sarah Wheeler, Amanda M. Clark. Lung epithelial cells induce both phenotype alteration and senescence in breast 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 1426. doi:10.1158/1538-7445.AM2015-1426
Abstract 1427: Embigin is overexpressed in pancreatic ductal adenocarcinoma and regulates cell motility through epithelial to mesenchymal transition via the TGF-β pathway
Authors: Jung Dawoon E. , Kim Jeong Mi , Kim Chanyang , Song Si Young
Published: August 2015

Embigin is a member of the immunoglobulin superfamily and encodes a transmembrane glycoprotein. Strong expression of Embigin has been observed in the mouse endoderm during early postimplantation embryogenesis and in the gut and visceral endoderm at the somite stage. There have been reports of Embigin involvement in neuromuscular junction formation and plasticity; however, the molecular functions of Embigin in other organs are unknown. Pancreatic cancer is an extremely aggressive malignancy, and it is necessary to understand the biology of pancreatic cancer to better detect and treat this disease. Our aim was to investigate the possible role of Embigin in pancreatic cancer. In pancreatic ductal adenocarcinoma tissues, Embigin expression was higher than that in normal pancreatic tissues. Immunohistochemical analysis revealed expression of Embigin in pancreatic cancer cells, as well as expression of monocarboxylate transporter 2 (MCT2) in cancer tissues. To gain further insight, we transfected BxPC-3 and HPAC pancreatic cancer cells with siRNA or shRNA targeting Embigin and observed reductions in cell proliferation, migration, invasion, wound healing, and reduced levels of matrix metalloproteinases-2 and -9. Silencing of Embigin increased intracellular L-lactate concentration by 1.5-fold and decreased MCT2 levels at the plasma membrane. Furthermore, Embigin silencing led to a reduced expression of PI3K, GSK3-β, and Snail/Slug. Upon treating BxPC-3 cells with transforming growth factor-β (TGF-β), we observed elevated expression of Snail/Slug, Embigin, and Vimentin; meanwhile, when treating cells with SB-216763, a GSK3-β inhibitor, we noted decreases in GSK3-β, Snail/Slug, and Embigin expression, suggesting that the TGF-β signaling cascade, comprising PI3K, GSK3-β, Snail/Slug, and Embigin signals, mediates epithelial to mesenchymal transition (EMT) in pancreatic cancer cells. These findings indicate the involvement of Embigin in EMT in pancreatic cancer progression and suggest Embigin as a putative target for the detection and/or treatment of pancreatic cancer.

Citation Format: Dawoon E. Jung, Jeong Mi Kim, Chanyang Kim, Si Young Song. Embigin is overexpressed in pancreatic ductal adenocarcinoma and regulates cell motility through epithelial to mesenchymal transition via the TGF-β pathway. [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 1427. doi:10.1158/1538-7445.AM2015-1427
Abstract 1430: Transcriptional regulatory loops among SNAI1, TWIST1, ZEB1, and ZEB2 defines the epithelial-mesenchymal transition (EMT) spectrum in epithelial ovarian cancer (EOC)
Authors: Tan Ming , Chuang Vin Yee , Tan Tuan Zea , Thiery Jean Paul , Huang Ruby Yun-Ju
Published: August 2015

Epithelial-mesenchymal transition (EMT) refers to a sequential phenotypic transformation process from epithelial to mesenchymal states, which is well defined in embryonic development. During cancer progression, cancer cells undergo EMT progressively loss epithelial characteristics and gain mesenchymal properties and are correlated with acquisition of increased capacity of metastasis and resistance to chemo- and immunotherapy. Although EMT is intensively studied in various types of cancers, its role in epithelial ovarian cancer (EOC), the most lethal gynaecological malignancy in the world, has yet to be elucidated. Previously, our group proposed the concept of an EMT Spectrum which encompasses the hybrid “intermediate” phenotype as well as the bona-fide epithelial and mesenchymal phenotypes. EOC cells harboring intermediate phenotypes showed distinct functional differences in in vitro aggressiveness. We hypothesize that EMT transcriptional drivers, SNAI1, TWIST1, ZEB1, and ZEB2, contribute to different biological consequences and show different hierarchy to induce step-wise phenotypic changes along the EMT Spectrum. EMT models were established by overexpressing classic EMT drivers in selected EOC cell lines with different EM phenotypes at different positions of EMT spectrum to test the intrinsic transcriptional regulation networks. Quantitative PCR (QPCR) was performed to examine the cross regulation among the EMT transcriptional drivers and their downstream targets. There was no cross regulation detected in the most epithelial cell line, PEO1, while a simple, repressive regulatory net was found in the OVCA420 cell line. The cell line with an intermediate phenotype, OVCA429, showed a complex double positive feedback loop among the EMT drivers. The divergence in the transcriptional regulatory loops correlated with the differences of in vitro EMT functions. These results indicate that there is a dynamic EMT transcriptional regulatory network to further define the EMT spectrum. In conclusion, this study demonstrates that cancer cells with an intermediate phenotype are more easily to be pushed along the EMT spectrum due to their ability to initiate a complex transcriptional feedback regulatory loop.

Citation Format: Ming Tan, Vin Yee Chuang, Tuan Zea Tan, Jean Paul Thiery, Ruby Yun-Ju Huang. Transcriptional regulatory loops among SNAI1, TWIST1, ZEB1, and ZEB2 defines the epithelial-mesenchymal transition (EMT) spectrum in epithelial ovarian cancer (EOC). [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 1430. doi:10.1158/1538-7445.AM2015-1430
Abstract 1443: BMP-2 induces motility and invasiveness of colon cancer through upregulation of cancer stem cell
Authors: Kim Bo Ram , Kim Jung Lim , Park Seong Hye , Jeoung Yoon A , Lee Sun Il , Min Byoung-Wook , Oh Sang Cheul
Published: August 2015

The majority of deaths in cancer patients are will present with metastasis and relapses. One of the key steps in metastasis is the migration, and Cancer stem cells(CSCs) activate migration. So, targeting of CSC is an important strategy for cancers. Bone morphogenetic protein(BMP) signaling, member of the TGF-β superfamily, plays an important role in development of bone and in regulation of stem cell properties. In Previous study, Bone morphogenetic protein(BMP) signaling has been associated with various types of cancers. BMPs contributes through autocrine and paracrine mechanisms to tumor development, invasion and metastasis. However, their roles and mechanisms are unclear. In this study, we investigated the function of BMP-2 and relevance BMP-2 and CSCs in Colon cancer cells.

First, We compared the endogenous protein level of parental and spheroid cells by western blotting. The spheroid cells showed higher level of EMT(Epithelial mesenchymal transition) markers N-Cadherin and Snail, CSC markers (EpCAM and CD133), and p Smad1/5 known BMP-2 downstream than the corresponding parental cells. Then expression of EpCAM and CD133 was confirmed by flow cytometry. We found that BMP-2 plays a key role in Cancer stem cell. To determine function of BMP-2 in Colon cancer cells, we stimulated the colon cancer cells with recombinant human BMP-2. Stimulation of BMP-2 increased EMT, CSC markers(EpCAM and CD133). In addition, BMP-2 enhanced cell motility, and invasiveness by wound healing and matrigel invasion assay. These results indicated BMP-2 signaling pathway enhance metastasis through increase of CSC portion. Blockade of BMP-2 pathway by Noggin(BMP2 antagonist) or Smad1/5 siRNA inhibited BMP2-induced EMT marker, MMP2, and CSC portion. The morphologic changes though the EMT process were monitored by immunofluorescence staining and Western blot for EMT markers. And Blockade of BMP-2 pathway inhibited cell motility and invasiveness.

Our findings suggest that BMP-2 signaling regulates metastatic potential through regulation of CSCs in colon cancer cell.

Citation Format: Bo Ram Kim, Jung Lim Kim, Seong Hye Park, Yoon A Jeoung, Sun Il Lee, Byoung-Wook Min, Sang Cheul Oh. BMP-2 induces motility and invasiveness of colon cancer through upregulation of cancer stem cell. [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 1443. doi:10.1158/1538-7445.AM2015-1443
Abstract 1444: Docosahexaenoic acid reduces cancer cell migration may link with actin binding proteins and miRNA 17-92 cluster expressions changes
Authors: Ali Mehboob , Heyob Kathryn M. , Rogers Lynette K.
Published: August 2015

Metastasis and progression, reflected by higher migration and longer cell survival, are major contributing factors in making cancers the leading cause of death worldwide. Cancer cells exhibit higher migration rates and greater cell viability facilitated by actin binding protein-mediated cytoskeletal remodeling and secondary messengers (cAMP and/or cGMP) signaling. Studies have shown that docosahexaenoic acid (DHA), a poly unsaturated fatty acid, inhibits cancer cell metastatic phenotype. Moreover, miRNA-17∼92 cluster has been reported to be highly expressed in metastasis. Therefore, we put forward a hypothesis that DHA will induce changes in actin binding protein dynamics and miRNA-17∼92 cluster expression to inhibit cancer cell migration and viability. Non-cance (MLE12) and cancer (A549) cells were treated with 8-Br-cAMP and/or DHA for 30 min, 3h, 6h and 24h. Actin binding proteins (profilin, cofilin, vimentin and gelsolin) and miRNA-17∼92 cluster expression were analyzed by western blot and qRT PCR respectively. Cell migration was estimated by wound assay and transwell apparatus. F-actin content and actin binding proteins were measured using confocal microscopy at the leading edges of wound. Proliferation and cell viability were estimated by flow cytometer, IF using ki-67, and cleaved-caspase-3 antibodies. F-actin content, cell migration and proliferation were increased while cell viability was decreased by cAMP. DHA treatment reverses the increase in actin content and cell migration in cancer cells but not in non-cancer cells. These findings correlate with changes in actin binding protein content and miRNA 17∼92 cluster expression. Thus, DHA specifically inhibits cancer cell migration, proliferation, and viability and this effect correlates with decreases in actin binding protein levels and miRNA-17∼92 cluster expression. In conclusion, DHA supplementation suppresses cellular changes related to metastatic potential in cancer cells and could provide therapeutic potential against cancer cell metastasis. Moreover, actin binding proteins and miRNA-17∼92 cluster expression could serve as innovative therapeutic targets and biomarkers for cancer progression.

Citation Format: Mehboob Ali, Kathryn M. Heyob, Lynette K. Rogers. Docosahexaenoic acid reduces cancer cell migration may link with actin binding proteins and miRNA 17-92 cluster expressions changes. [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 1444. doi:10.1158/1538-7445.AM2015-1444
Abstract 1453: Methods for developing patient derived xenografts in athymic nude mice
Authors: Marlow Laura A. , Miller James L. , Edenfield Brandy , Mathias Adam C. , Dawson Louis K. , Durham William F. , Mullin Robert J. , Small Daniel L. , Synnott Aidan J. , Wu Kevin , Copland John A.
Published: August 2015

Patient derived xenografts (PDX) mouse models for many types of cancer have been developed recapitulating therapeutic responses in the patient for which the PDX model was derived. Essentially, live surgically resected or biopsy tumor tissues are directly implanted into immune incompetent mice to develop in vivo models to test therapies, follow tumor progression (gene mutations that occur with patient tumor progression do so in mice) or to better understand drug resistance. However, the use of PDX models also has its limitations and challenges. Our laboratory has attempted to develop PDX models in athymic nude mice for numerous cancer histotypes such as bladder, brain, breast, colon, kidney, lung, melanoma, ovarian, pancreas, sarcoma and thyroid. While melanoma and colon cancers have very high tumor take rates (73 - 100%), the other cancer histotypes do not (14 - 50%). We share our experience for each cancer histotype and the techniques used to enhance the success rates in athymic nude mice. We also address the other challenges that include expansion of tumors after cryopreservation and conversion of human tumors to mouse tumors and spontaneous lymphomas. We provide methods to test for human versus mouse content that include QPCR and immunohistochemistry

Citation Format: Laura A. Marlow, James L. Miller, Brandy Edenfield, Adam C. Mathias, Louis K. Dawson, William F. Durham, Robert J. Mullin, Daniel L. Small, Aidan J. Synnott, Kevin Wu, John A. Copland. Methods for developing patient derived xenografts in athymic nude mice. [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 1453. doi:10.1158/1538-7445.AM2015-1453
Abstract 1456: Efficacy of photo-immunotherapy in combination with surgical resection on a pancreatic cancer patient derived orthotopic xenograft (PDOX)
Authors: Hiroshima Yukihiko , Maawy Ali , Murakami Takashi , Magalhães Nzola De , Kawaguchi Daisuke , Matsuo Kenichi , Tanaka Kuniya , Bouvet Michael , Hoffman Robert M. , Endo Itaru
Published: August 2015

Complete tumor resection improves overall survival of pancreatic cancer patients. We have previously showed that UVC irradiation in combination with fluorescence-guided surgery (FGS) completely cured metastatic pancreatic cancer by eradicating minimal residual cancer after FGS. Recently, photoimmunotherapy (PIT), that uses a target-specific photosensitizer based on a near-infrared (NIR) phthalocyanine dye, IR700, demonstrated in vivo tumor shrinkage after irradiation with NIR light without any side effects. In this study, we used a clinically applicable model of pancreatic cancer to examine the use of PIT to eradicate minimal residual cancer of a patient derived orthotopic xenograft (PDOX). We found that anti-CEA-IR700 could label and illuminate PDOX strongly enough for PIT in both subcutaneous and orthotopic models. Furthermore, we succeeded in detecting fluorescence from the minimal residual cancer of the PDOX. The local recurrence rate of BLS only and BLS + PIT group was 85.7% and 28.6%, respectively. BLS + PIT tended to reduce the recurrence rate compared to BLS only, and there was marginally significant difference (p = 0.051). The average recurrent tumor weight of BLS only and BLS + PIT group was 1149.0 ± 794.6 mg and 210.8 ± 336.9 mg, respectively. The average recurrent tumor weight of BLS + PIT group was significantly less than BLS only (p = 0.015). In conclusion, anti-CEA-IR700 is able to label and illuminate the PDOX strongly enough for PIT. PIT reduces recurrence by eradicating microscopic disease undetectable with BLS, and might be applicable to pancreatic cancer patients.

Citation Format: Yukihiko Hiroshima, Ali Maawy, Takashi Murakami, Nzola De Magalhães, Daisuke Kawaguchi, Kenichi Matsuo, Kuniya Tanaka, Michael Bouvet, Robert M. Hoffman, Itaru Endo. Efficacy of photo-immunotherapy in combination with surgical resection on a pancreatic cancer patient derived orthotopic xenograft (PDOX). [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 1456. doi:10.1158/1538-7445.AM2015-1456
Abstract 1458: Characterization of novel thyroid PDX models and their response to combination therapies
Authors: Marlow Laura A. , Mathias Adam C. , Dawson Louis K. , Durham William F. , Meshaw Kenneth A. , Mullin Robert J. , Small Daniel L. , Synnott Aidan J. , Milosevic Dragana , Netzel Brian C. , Grebe Stefan K. , Wu Kevin , Smallridge Robert C. , Copland John A.
Published: August 2015

Patient derived xenografts (PDX) mouse models for many types of cancer have demonstrated therapeutic responses similar to those seen in patients. Thus, resected patient tumor tissue directly implanted into immune compromised mice followed by therapy and tumor growth analysis is thought to be the closest preclinical model to predict patient therapeutic responses. To date, few models are available for the different histotypes of thyroid cancer derived from follicular thyrocytes; these include papillary, follicular, Hurthle cell, squamous and anaplastic thyroid carcinoma. We have developed eight PDX models in athymic nude mice representing all of these subtypes. The models have been extensively characterized for mutational status (i.e. BRAF, telomerase, RAS, PTEN, TP53, RB, etc) as well as validation by short tandem repeat (STR) analysis to match that of the originating patient tumor tissue. Squamous and anaplastic thyroid cancers are rare tumor types with no FDA approved therapies. Each model demonstrated their own unique responses to radiation, cytotoxic or molecular targeted therapies such as doxorubicin, sorafenib (tyrosine kinase inhibitor), cisplatin, paclitaxel, sunitinib (tyrosine kinase inhibitor), erlotinib (EGFR inhibitor), trametinib (MEK inhibitor) and vemurafenib (BRAF inhibitor). We expect that these models may provide useful in vivo models for thyroid cancer research as well as models for therapeutic guidance based upon histotype, mutational status and response to therapies.

Citation Format: Laura A. Marlow, Adam C. Mathias, Louis K. Dawson, William F. Durham, Kenneth A. Meshaw, Robert J. Mullin, Daniel L. Small, Aidan J. Synnott, Dragana Milosevic, Brian C. Netzel, Stefan K. Grebe, Kevin Wu, Robert C. Smallridge, John A. Copland. Characterization of novel thyroid PDX models and their response to combination therapies. [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 1458. doi:10.1158/1538-7445.AM2015-1458
Abstract 146: The long noncoding RNA MALAT1 promotes hypoxia-driven angiogenesis by upregulating pro-angiogenic gene expression in neuroblastoma cells
Authors: Tee Andrew E. , Liu Peiyan , Maag Jesper , Song Renhua , Li Jinyan , Cheung Belamy B. , Haber Michelle , Norris Murray D. , Marshall Glenn M. , Dinger Marcel , Liu Tao
Published: August 2015

Neuroblastoma is the most common solid tumour during early childhood, and accounts for 15% of all childhood cancer death. One of the key features of neuroblastoma is aggressive progression due to tumour-driven angiogenesis. However, the mechanism through which neuroblastoma cells drive angiogenesis is unclear. Here we showed that the long noncoding RNA MALAT1 was over-expressed in human neuroblastoma cells under hypoxic condition, the condition which triggers pro-angiogenic switch and angiogenesis. In vitro angiogenesis assays demonstrated that conditioned medium from neuroblastoma cells transfected with MALAT1 siRNAs, compared with conditioned medium from neuroblastoma cells transfected with control siRNAs, induced considerably less endothelial cell migration, invasion and vascular sprouting under hypoxic condition. Affymetrix microarray differential gene expression study showed that one of the genes most significantly down-regulated by MALAT1 siRNAs in human neuroblastoma cells under hypoxic condition, was fibroblast growth factor (FGF). RT-PCR and immunoblot analyses confirmed that MALAT1 siRNAs reduced FGF mRNA and protein expression, and in vitro angiogenesis assays demonstrated that forced over-expression of FGF in neuroblastoma cells blocked MALAT1 siRNA-mediated reduction in endothelial cell migration, invasion and vascular sprouting. Taken together, our data suggest that over-expression of MALAT1 in human neuroblastoma cells plays an important role hypoxia-driven angiogenesis by up-regulating FGF mRNA expression.

Citation Format: Andrew E. Tee, Peiyan Liu, Jesper Maag, Renhua Song, Jinyan Li, Belamy B. Cheung, Michelle Haber, Murray D. Norris, Glenn M. Marshall, Marcel Dinger, Tao Liu. The long noncoding RNA MALAT1 promotes hypoxia-driven angiogenesis by upregulating pro-angiogenic gene expression in neuroblastoma 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 146. doi:10.1158/1538-7445.AM2015-146
Abstract 1462: Characterization of patient-derived bladder cancer xenografts: role of xCT in response to cisplatin
Authors: Elbanna May F. , Ciamporcero Eric , Ramakrishnan Swathi , Adelaiye Remi , Shen Li , Orillion Ashley , Ku Sheng-Yu , Chintala Sreenivasulu , Pili Roberto
Published: August 2015

Background: A major challenge in cancer drug development has been largely attributed to the inability of cell lines to recapitulate the heterogeneity of human tumors. Patient derived xenografts (PDX) represent a major advance as they are more representative of the clinical setting. However, appropriate characterization of PDXs is necessary to guide biomarker driven drug discovery research. Methods: H&E staining was used to compare the original patient tumors with PDX developed from cystectomy specimens to validate morphological similarity. Immunohistochemical (IHC) staining for cytokeratins (CK) 5/6 & 20 and EMT markers E cadherin and vimentin were used to identify whether these tumors represent a basal or a luminal subtype. In addition, Western blot and RT-PCR analysis were performed to assess markers of epithelial-to-mesenchymal transition (EMT). Therapeutic response to cisplatin was evaluated both in vitro and in vivo. Furthermore, expression of xCT (cystine transporter), one of the known cisplatin resistance markers, was determined by RT-PCR, Western blot and IHC in PDXs and cells. Results: Morphologically, we observed that our PDX models (RP-B-01 and RP-B-02) did not drift from the original patient tumors and they still retained their characteristic stromal features. Based on cytokeratin (CK) staining, we found both models to be CK 5/6 positive and CK 20 negative, suggesting a basal subtype of bladder cancer. Both PDXs, similarly to the original tumors, were E cadherin positive. However, cells derived from these tumors and cultured in vitro lost E cadherin and acquired vimentin expression, suggesting an EMT phenotype. When cells from PDX were transplanted back into mice they re-expressed E cadherin and lost vimentin expression, suggesting a reversion to MET. Despite no difference in cytokeratins and EMT markers between the two models, both PDX and cells isolated from PDXs had differential expression of xCT and possessed differential response to cisplatin treatment. RP-B01 expressed higher levels of xCT and was less responsive to cisplatin as compared to RP-B-02 which expressed lower levels of xCT. Conclusion: Differential expression of xCT in bladder cancer PDXs and cells correlated with cisplatin response, suggesting that pharmacological targeting of xCT in combination therapies will enhance cisplatin efficacy in bladder cancer.

Citation Format: May F. Elbanna, Eric Ciamporcero, Swathi Ramakrishnan, Remi Adelaiye, Li Shen, Ashley Orillion, Sheng-Yu Ku, Sreenivasulu Chintala, Roberto Pili. Characterization of patient-derived bladder cancer xenografts: role of xCT in response to cisplatin. [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 1462. doi:10.1158/1538-7445.AM2015-1462
Abstract 148: Transcriptome sequencing reveals PCAT5 - new ERG-regulated non-coding transcript in prostate cancer
Authors: Ylipää Antti , Kivinummi Kati , Annala Matti , Kohvakka Annika , Latonen Leena , Scaravilli Mauro , Kartasalo Kimmo , Leppänen Simo-Pekka , Karakurt Serdar , Seppälä Janne , Yli-Harja Olli , Tammela Teuvo L.J. , Zhang Wei , Visakorpi Tapio , Nykter Matti
Published: August 2015

Background: Prostate cancer (PC) is the second most common cancer among men. Most PC-related deaths are due to invasive tumors that are treated with therapies inhibiting androgen production or androgen receptor (AR) activity. After an initial response, tumors invariably progress to castration resistant prostate cancers (CRPCs) for which no effective cure exists.

Methods and Results: We used transcriptome sequencing to study fresh-frozen tissue specimens from 12 benign prostatic hyperplasias (BPHs), 28 PCs, and 13 CRPCs. Reference-based transcriptome assembly uncovered 145 previously unannotated intergenic PC and CRPC associated long non-coding transcripts (lncRNAs) or isoforms. One third of the transcripts were CRPC-specific. We showed that one of the novel transcripts, Prostate Cancer Associated Transcript 5 (PCAT5), expressed in half of the tumors, was likely regulated by ERG, the key transcription factor in ∼50% of prostate cancers. Genome-wide expression analysis of a PCAT5-positive prostate cancer cell line after PCAT5 knockdown suggested significant alterations in proliferation pathways. In vitro validation of the pathway alterations revealed concordantly dramatic effects in phenotype: stalling of cell growth, migration, invasion, and colony forming potential, and increase in the rate of apoptosis.

Conclusions: We identified the key differences between PC and CRPC in transcriptome level, and validated the oncogenic potential of a novel lncRNA in ERG-positive prostate cancers, PCAT5. Our study presents a number of putative lncRNA biomarkers for CRPC, and opportunities for therapeutic intervention.

Citation Format: Antti Ylipää, Kati Kivinummi, Matti Annala, Annika Kohvakka, Leena Latonen, Mauro Scaravilli, Kimmo Kartasalo, Simo-Pekka Leppänen, Serdar Karakurt, Janne Seppälä, Olli Yli-Harja, Teuvo L.J. Tammela, Wei Zhang, Tapio Visakorpi, Matti Nykter. Transcriptome sequencing reveals PCAT5 - new ERG-regulated non-coding transcript in prostate 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 148. doi:10.1158/1538-7445.AM2015-148
Abstract 1499: SUPR peptides as novel targeted molecular imaging agents for Her2
Authors: Kelderhouse Lindsay , Hardy Amanda , Takahashi Terry T. , Ornelas Argentina , Gammon Seth , Howell Shannon , Yang Peiying , Wang Pin , Roberts Richard W. , Fiacco Steve , Millward Steven W.
Published: August 2015

Ideal molecular imaging agents would combine the affinity and selectivity of monoclonal antibodies with the rapid clearance and pharmacokinetics of small molecules Here we describe the use of directed evolution to design Scanning Unnatural Protease Resistant (SUPR) peptides as novel molecular scaffolds for in vivo optical and PET imaging of the Her2 receptor. SUPR peptides were obtained through selection against Her2-expressing breast cancer cells in culture using focused biological display peptide libraries pre-selected for protease resistance. SUPR peptide libraries were programmed with unnatural amino acids for enhanced stability and chemically cyclized after translation to enhance affinity. Peptides isolated from selection selectively bind the Her2 receptor in vitro with low nanomolar affinity and do not compete with Trastuzumab or Pertuzumab. Cy5-labeled SUPR peptides show rapid and Her2-specific tumor uptake in vivo and minimal retention in non-tumor tissues after 24 hours of washout. SUPR peptides can be efficiently labeled with 18F via the copper-catalyzed azide-alkyne cycloaddtion and the resulting radiotracers used for nuclear imaging of Her2-postive tumors by PET/CT. Unlike antibodies and their derivatives, SUPR peptides are synthesized chemically and show rapid clearance from systemic circulation. SUPR peptides also show dramatically enhanced stability relative to linear peptides obtained from phage display or combinatorial chemistry while still retaining affinities compatible with molecular imaging applications. While SUPR peptides are selected from biological display libraries, they share similar chemical composition and structure with peptides derived from non-ribosomal peptide synthesis. We believe that this combination of affinity, stability, evolvability, and synthetic accessibility make SUPR peptide technology a general approach for generating highly potent compounds for targeted molecular imaging of cancer biomarkers.

Citation Format: Lindsay Kelderhouse, Amanda Hardy, Terry T. Takahashi, Argentina Ornelas, Seth Gammon, Shannon Howell, Peiying Yang, Pin Wang, Richard W. Roberts, Steve Fiacco, Steven W. Millward. SUPR peptides as novel targeted molecular imaging agents for Her2. [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 1499. doi:10.1158/1538-7445.AM2015-1499
Abstract 1501: Evaluation of tumor vascularity in colorectal cancer using contrast enhanced ultrasonograpy in comparison with microvessel density
Authors: Okawa Takaomi , Nogami Hiromi , Takaoka Munenori
Published: August 2015

Background : The relationship between tumor vascularity and malignant potential in colorectal cancer a(CRC) is reported by many authors. Therefore, to assess the tumor vascularity before surgical treatment is thought to be one of very important factors in CRC patients. Contrast enhanced ultrasonography (CEUS) can visualize the tumor vascularity vividly. We aim to determine the reliability of CEUS for the assessment of vascularity of CRC and to analyze the relationship between tumor vascularity and clinicopathological characteristics. In addition, to find the clinical application, we assessed the efficacy of chemotherapy in unresectable advanced CRC patients by CEUS.

Methods : 56 patients undergoing surgical resection of CRC were studied. An ultrasound system (GE health care, LOGIC 7) is used and visualized the colo-rectal tumor using a 4.5MHZ probe. The 2nd generated ultrasonic contrast medium (Belfulbuttan, Sonazoid) is injected intravenously. Approximately 20 s after injection, microbubbles were obtained to enhance the B-mode image. Using the tumor image showing the strongest vascularity, region of interest were determined to measure mean echo intensity in the tumor. Their images were analyzed with NIH image program (developed by the US National Institutes of health) and calculated tumor vascularity as Vascular Index (VI). The clinicopathological features are based on the TNM classification from UICC. Using surgical specimens, tumor vascularity was assessed by immunohistochemistry as microvessel density in order to do statistical analysis for the correlation between VI, clinicopathologic features and microvessel density.

Results : CEUS could visualize the tumor vascularity in all patients. In advanced CRC patients, we observed that the tumor invaded into other organ, such as prostate gland, small intestine and so on. The VI correlated significantly with depth of tumor invasion, liver metastasis and microvessel density. Furthermore, in patients with abundant vascularity in tumor before treatment, chemotherapy led to the reduction of VI prior to the radiographically visible difference in tumor size.

Conclusion : CEUS can evaluate the tumor vascularity non-invasively and is very useful for the evaluation of their pathological diagnosis and the efficacy of chemotherapy in CRC.

Citation Format: Takaomi Okawa, Hiromi Nogami, Munenori Takaoka. Evaluation of tumor vascularity in colorectal cancer using contrast enhanced ultrasonograpy in comparison with microvessel density. [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 1501. doi:10.1158/1538-7445.AM2015-1501
Abstract 151: Transcriptome-wide expression profiling of long noncoding and small nucleolar RNAs in head and neck squamous cell carcinomas identifies novel transcripts associated with survival
Authors: Honda Thomas K. , Zou Angela , Yu Vicky , Zheng Hao , Kuo Selena , Saad Maarouf , Xuan Yinan , Singh Pranav , Wang-Rodriguez Jessica , Ongkeko Weg M.
Published: August 2015

Head and neck squamous cell carcinoma (HNSCC) persists as one of the most common and deadly malignancies, with early detection and effective treatment still posing formidable challenges. To expand our currently sparse knowledge of the epigenetic alterations involved in this disease and identify potential biomarkers and therapeutic targets, we globally profiled the dysregulation of small nucleolar (snoRNA) and long non-coding RNAs (lncRNA) in head and neck tumors. Using next-generation RNA-sequencing data from 40 pairs of tumor and matched normal tissues, we profiled lncRNA expression by mapping each RNA-seq library to a reference annotation containing 32,108 lncRNAs. Using edgeR, we found 19,681 lncRNA transcripts significantly differentially expressed by a fold change magnitude ≥ 4 with a false discovery rate <10-4. We selected the 10 most dysregulated lncRNAs and further confirmed the dysregulation of these lncRNAs in-vitro by performing quantitative reverse transcriptase PCR (RT-qPCR) on two normal oral epithelial cell lines and five HNSCC cell lines. Using Kaplan-Meier analysis, we demonstrated a correlation between high expression of several lncRNAs, including JPH1-7, upregulated by 40-fold in HNSCC, with overall poor survival in a cohort of over 400 HNSCC patients (HR = 1.62, p < 0.01). Meanwhile, RNA-sequencing analysis of 31 tumor-normal pairs yielded 33 significantly dysregulated snoRNA. We identified one snoRNA, SNORD116-20, downregulated by 4-fold in HNSCC, whose expression levels correlated significantly with overall patient survival (HR = 1.64, p < 0.05), suggesting its functional significance and clinical relevance in head and neck cancer pathogenesis. Similarly, in in-vitro experiments, we found reduced expression of this snoRNA in HNSCC cell lines compared to normal oral epithelial cell lines. The precise molecular roles of these transcripts identified warrants further characterization, but our data suggest that they are likely to play substantial roles in head and neck cancer pathogenesis and are significantly associated with patient survival.

Note: This abstract was not presented at the meeting.

Citation Format: Thomas K. Honda, Angela Zou, Vicky Yu, Hao Zheng, Selena Kuo, Maarouf Saad, Yinan Xuan, Pranav Singh, Jessica Wang-Rodriguez, Weg M. Ongkeko. Transcriptome-wide expression profiling of long noncoding and small nucleolar RNAs in head and neck squamous cell carcinomas identifies novel transcripts associated with survival. [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 151. doi:10.1158/1538-7445.AM2015-151
Abstract 1526: HOXA5 inhibits cell plasticity and stemness in breast cancer cells by reinforcing epithelial traits
Authors: Teo Wei Wen , Merino Vanessa , Cho Sean , Korangath Preethi , Neumann Neil M. , Ewald Andrew J. , Sukumar Saraswati
Published: August 2015

Members of the Retinoic acid receptor (RAR) and HOX gene families are known to be associated with cellular differentiation. Nonetheless, how HOX genes drive differentiation in mammalian cells is poorly understood. HOX genes are transcription factors that orchestrate body segmentation during embryogenesis. But the continued expression of several members in adulthood indicates the necessity of their biological functions in specifying cell fate in adulthood. The mammary gland is an organ that continues to go through multiple cycles of development during life. Here, loss of specific HOX genes could be detrimental to normal development.

We have previously shown that expression of HOXA5, a downstream target of the retinoic acid signaling pathway, is often lost in breast cancer. Loss of HOXA5 expression correlates with increasing pathological grades of breast cancer. Expression array analysis of HOXA5-depleted MCF10A (immortalized line of human breast epithelium) revealed its potential role in regulating several traits of the epithelial lineage, including expression of E-cadherin and CD24. Depleting HOXA5 in MCF10A enhanced cell motility, invasiveness and increased stem-cell population (CD24lo/CD44hi). Using an inducible HOXA5-depletion system, differentiation of the stem cells in MCF10A was shown to be mediated by HOXA5. Conversely, ectopic expression of HOXA5 in SUM149 breast cancer cells significantly reduced plasticity and stem cell population, confirmed by in vivo limiting dilution assays in mice. MCF10A-KRAS-HOXA5 depleted cells formed invasive structures in 3D matrigel cultures, and formed larger tumors, implanted orthotopically in mice. MCF10A-KRAS-tumors were histologically dedifferentiated and lost expression of luminal cell markers such as E-cadherin and cytokeratin 18. Collectively, our findings demonstrate the role of HOXA5 in retinoid-induced cell differentiation by regulating cell fate specific surface adhesion molecules.

Citation Format: Wei Wen Teo, Vanessa Merino, Sean Cho, Preethi Korangath, Neil M. Neumann, Andrew J. Ewald, Saraswati Sukumar. HOXA5 inhibits cell plasticity and stemness in breast cancer cells by reinforcing epithelial traits. [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 1526. doi:10.1158/1538-7445.AM2015-1526