bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2441: Effects of hypoxia on Src family members' activity in prostate cancer cells
Authors: Dai Yao , Siemann Dietmar 
Published: August 2018

Abstract
Metastasis is the major reason of disease progression and poor prognosis in prostate cancer. Src family kinases (SFKs), including c-Src, Lyn and Fyn, are non-receptor tyrosine kinases that have been shown to play an essential role in castration resistance and metastasis in prostate cancer. Hypoxia, a typical hallmark of solid malignancies, is able to promote metastasis-associated functions. However, whether SFK members are upregulated under hypoxia is unclear. The current project will detect the impact of hypoxia on cell functions and molecular features in prostate cancer cells. For hypoxic exposure, prostate cancer cells were exposed to low oxygen tensions (1% O2) for varied durations (0, 2, 6, 24 h). To test cell migration, “wound-healing” assay was used. Cell invasion was detected by seeding cells into Matrigel-coated transwell chamber and cells on the bottom membranes were counted after treatment. Clonogenic assay was performed to test cell survival. At the molecular level, SFKs' phosphorylation in cancer cells and patient tissues was detected by Western blotting, and gene knockdown was accomplished by siRNA transfection. To detect the effects of Src inhibitors, cells were treated with saracatinib for indicated time periods. Phosphorylation of all three typical SFKs, e.g., c-Src, Lyn and Fyn, were highly expressed in prostate cancer patient tissues. At the cellular level, while short term hypoxic exposure (2-6 h) induced greater effects than prolonged hypoxia (24 h) in PC-3ML, these cellular functions were increased with 24 h hypoxic exposure in C4-2B cells. Further, hypoxia enhanced SFK phosphorylation in the pattern that was consistent with cell functions in both cell lines. Knockdown SRC, but not LYN or FYN, abolished hypoxia-induced invasion and p-SFK expression. Lastly, SFK inhibitor saracatinib showed stronger inhibition on functional behaviors facilitated under hypoxia than normal conditions. Our data show that hypoxia is able to enhance metastatic phenotypes by activating SFKs in prostate cancer cells. Interestingly, c-Src may be the most important signaling molecule for hypoxia-mediated behaviors. More importantly, SFK inhibitors are able to impair hypoxia-induced cell functions, suggesting their therapeutic potential by suppressing tumor metastasis that is driven under hypoxia in prostate cancer.
Citation Format: Yao Dai, Dietmar Siemann. Effects of hypoxia on Src family members' activity in prostate cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2441.
Abstract 2451: Long noncoding RNA OVAAL promotes melanoma cell proliferation through translational suppression of p27
Authors: ZHANG YUNA YUAN , Yan Xu Guang , Farrelly Margaret , Yari Hamed , Feng Yuchen , La Ting , Tabatabaee Hessam , Jin Lei , Zhang Xu Dong 
Published: August 2018

Abstract
Evading apoptosis and limitless replicative potential are two hallmarks of malignant cells. Here we report that melanoma cells surviving the induction of apoptosis by TNF-related apoptosis-inducing ligand (TRAIL) or the Mcl-1 inhibitor UMI-77 have even higher proliferative potential, and that this is associated with the increased expression of the long noncoding RNA (lncRNA) OVAAL which competes with p27 mRNA for binding to polypyrimidine tract binding protein 1 (PTBP1) and thus represses p27 mRNA translation. Prolonged exposure to TRAIL or Mcl-1 resulted in stable melanoma cell populations with increased proliferation activity. Comparative RNA-seq analysis identified OVAAL as the most significantly upregulated lncRNA in melanoma cells resistance to TRAIL and Mcl-1. ShRNA knockdown of OVVAL reversed the increased proliferative potential of resistant cells, and also inhibited proliferation in a subset of parental melanoma cell lines that constitutively expressed relatively high levels of OVAAL. RNA pulldown followed by Mass Spectrometry analysis revealed that among proteins that were physically associated with OVAAL was a protein called PTBP1 that bound to and enhanced the activity of the internal ribosomal entry site (IRES) in the 5' untranslated region (5'-UTR) of p27 mRNA. Indeed, OVAAL knockdown resulted in an increase in the association between PTBP1 and p27 IRES leading to upregulation of the p27 protein, whereas overexpression of OVAAL caused reduction in binding of PTBP1 to the IRES of p27. Furthermore, co-knockdown of PTBP1 abolished upregulation of p27 triggered by OVAAL knockdown. These results suggest that OVAAL competes with p27 mRNA for binding to PTBP1 and thus inhibits p27 expression and promotes cell proliferation. This was substantiated by binding assays using in vitro-synthesized biotin-labelled OVAAL, p27 IRES and purified PTBP1 in a cell-free system. Our results showed that the increased c-Myc activity was responsible for upregulation of OVAAL in melanoma cells selected for resistance to apoptosis induced by TRAIL or UMI-77. Collectively, these results identify the expression of OVAAL as an important mechanism promoting melanoma cell proliferation, with potential implications for targeting OVAAL in combination with apoptosis-inducing therapeutics in the treatment of melanoma.
Citation Format: YUNA YUAN ZHANG, Xu Guang Yan, Margaret Farrelly, Hamed Yari, Yuchen Feng, Ting La, Hessam Tabatabaee, Lei Jin, Xu Dong Zhang. Long noncoding RNA OVAAL promotes melanoma cell proliferation through translational suppression of p27 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2451.
Abstract 2456: The PIWI-interacting RNA pathway in prostate is regulated by vitamin D and altered in cancer
Authors: Baumann Bethany , Gao Shang , Lugli Giovanni , Nonn Larisa 
Published: August 2018

Abstract
Low serum vitamin D is a risk factor for aggressive prostate cancer. piRNAs and their interacting partners, PIWI-like (PIWIL) proteins, are aberrantly expressed in many cancers, but few studies have focused on piRNAs in the prostate. We used laser-capture microdissected (LCM) human prostate epithelium from frozen samples collected for a clinical trial of high dose vitamin D to examine the small non-coding RNA population by next-generation sequencing. The 8 patients for this study were selected for having either high (avg=48.7pg/ml) or low (avg=21.4pg/ml) prostatic vitamin D (1,25(OH)2D) levels. VDR ChIP-seq was performed as well, using human primary epithelial cells treated for 3hr with EtOH or 50nM vitamin D. In LCM-collected prostate epithelium unexpectedly piRNAs represented ~20% of the mapped reads. Of all the small RNA species investigated, piRNAs were the most affected by vitamin D status, increasing in abundance in tissues wish high vitamin D (p=0.036, t-test). The LCM data were compared to small-RNA sequencing data from benign prostate available from The Cancer Genome Atlas (TCGA), which represent whole prostate tissue and may contain stromal bias. The LCM-collected samples had a distinct, more consistent profile of small RNAs than TCGA samples. Two unambiguous piRNAs that were highly expressed in LCM data, but not in TCGA data, were verified as epithelial-specific in prostate by in situ hybridization. Several piRNA-biogenesis genes lie within 100kb of VDR ChIP peaks, including TDRKH, which was found to be vitamin D-regulated experimentally. The expression of piRNA-interacting PIWIL proteins was assessed by RT-qPCR, immunoblot and immunofluorescence on a tissue microarray containing paired benign and cancer tissues (N=23). Of the 4 human PIWIL proteins, PIWIL1, 2 and 4 were present in normal prostate epithelial cells by PCR. PIWIL1 protein was also detected in PrE and RWPE-1 benign cells, but PIWIL1 was not detected in the PC3 prostate cancer cell line. By immunofluorescence, PIWIL1 protein in prostate epithelium significantly decreased in prostate cancer (p=0.016, paired t-test). In conclusion, vitamin D status associated with the expression of piRNA in prostate epithelium in clinical trial samples. PIWIL1 protein levels were lower in prostate cancer, suggesting a pro-differentiation role for the PIWI pathway that may be affected in vitamin D-deficient patients.
Citation Format: Bethany Baumann, Shang Gao, Giovanni Lugli, Larisa Nonn. The PIWI-interacting RNA pathway in prostate is regulated by vitamin D and altered in cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2456.
Abstract 2477: Exosomal RNA mediates transformation of nontumorigenic human bronchial lung epithelial cells
Authors: Kitdumrongthum Sarunya , Humphrey Sean , Kubo Hana , Tian Feng , Allen Sarah , Atallah Nadia , Hasan Humna , Kasinski Andrea L. 
Published: August 2018

Abstract
Exosomes are nanovesicles with a diameter between 30-150 nm. Exosomes are produced and released by all cell types into body fluids and, thus, play a major role in cell-cell communication through the transfer of proteins, lipids, and nucleic acids into recipient cells. Uptake of bioactive cargo into recipient cells can impart physiologic and biologic changes in the recipient cell. Especially in cancer, exosome released from cancer cells can reprogram cells, driving various hallmarks of cancer, such as proliferation, migration, invasion, and angiogenesis. In this study we evaluated the effects of exosomes, isolated from non-small cell lung cancer (NSCLC) cells, on nontumorigenic human bronchial epithelial cells (HBECs). Firstly, we determined that cancer cells generate more exosomes than nontumorigenic cells and that cancer cell-derived exosomes are readily taken up by nontumorigenic cells. The nontumorigenic cells exposed to NSCLC-derived exosomes gained the ability to migration and invade. In parallel, we observed that NSCLC-derived exosomes promoted changes in epithelial-to-mesenchymal gene profiles, supporting the conclusion that HBECs were becoming transformed. Exosomes were also capable of permeablizing a monolayer of human endothelial cells, suggesting that NSCLC-derived exosomes may disrupt the vasculature enhancing metastatic capabilities. Importantly, the transformative effects of the exosomes were mediated mostly by exosomal RNA. Comparing the RNA profile from a panel of cancerous exosomes to RNAs contained inside of exosomes isolated from nontumorigenic, one long noncoding RNA (lncRNA) was highly elevated. Knockout of this lncRNA greatly diminished the capability of the cancerous exosomes to promote the observed hallmarks of cancer, suggesting that a single lncRNA is responsible for the majority of the tumorigenic capacity of the exosomes. In vivo mouse modeling and patient-based evidence are currently under way to support these cell-based findings.
Citation Format: Sarunya Kitdumrongthum, Sean Humphrey, Hana Kubo, Feng Tian, Sarah Allen, Nadia Atallah, Humna Hasan, Andrea L. Kasinski. Exosomal RNA mediates transformation of nontumorigenic human bronchial lung epithelial cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2477.
Abstract 2485: CYP24A1-induced vitamin D insufficiency promotes breast cancer growth
Authors: Osanai Makoto , Ono Yusuke , Takasawa Akira , Takasawa Kumi , Murata Masaki , Sawada Norimasa 
Published: August 2018

Abstract
Vitamin D plays a critical role in the maintenance of tissue homeostasis by regulating the expression of genes affecting cell proliferation, differentiation, and apoptosis. The vitamin D 24-hydroxylase CYP24A1 functions in vitamin D target tissues to degrade the hormonal form of vitamin D. Existing knowledge regarding dysregulated CYP24A1 expression supports its candidacy as a putative oncogene. In our presentation, we demonstrate that the suppression of constitutive CYP24A1 expression by CYP24A1-specidic shRNA conferred target cells with increased susceptibility to apoptosis and consequently inhibited anchorage-independent growth in breast carcinoma cells. In addition, suppression of vitamin D metabolism following knockdown of CYP24A1 significantly reduced tumor growth in vivo. Since these data provide substantial evidence for a pro-survival and stimulatory oncogenic effect of CYP24A1 in breast carcinoma cells, we will discuss the potential feasibility of CYP24A1-inhibitory cancer therapies.
Citation Format: Makoto Osanai, Yusuke Ono, Akira Takasawa, Kumi Takasawa, Masaki Murata, Norimasa Sawada. CYP24A1-induced vitamin D insufficiency promotes breast cancer growth [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2485.
Abstract 2507: Axl suppression inhibits cell migration, invasion and survival in breast and prostate cancer cell lines
Authors: Tanaka Mai , Dykes Samantha S. , Siemann Dietmar W. 
Published: August 2018

Abstract
Cancer is the second leading cause of death in the United States with approximately 90% of cancer-related deaths resulting from metastatic disease. In tumor cells, upregulation of certain signaling pathways promote metastatic phenotypes characterized by enhanced invasion, migration, survival, proliferation and induction of angiogenesis. One pathway of considerable interest involves the receptor tyrosine kinase Axl that is expressed in a variety of tumor types and is associated with poor prognosis and metastasis. Furthermore,
Axl's ligand, Growth arrest specific 6 (Gas6) protein, can activate a number of downstream signaling pathways that promote metastatic phenotypes. The purpose of the current study was to characterize Axl and Gas6 in a number of human tumor cell lines and to determine the effect of Axl suppression on the metastatic phenotypes, specifically cell migration, invasion and survival. A panel of human tumor cell lines, including prostate, breast, colorectal, renal, and osteosarcoma, were characterized for Axl expression by Western blot and qPCR. Gas6 expression was assessed by qPCR and Gas6 secretion was determined in conditioned media by ELISA. Axl was genetically inactivated by shRNA
in prostate (DU145) and breast (MDA-MB-231) cancer cell lines. The efficiency of knockdown was determined by Western blot. The effect of Axl knockdown on the migratory and invasive potentials of these cell lines was evaluated by transwell migration and invasion assays. A clonogenic cell survival assay was used to assess the impact of Axl knockdown on tumor cell viability. Axl-expressing tumor cells expressed and secreted its ligand Gas6, suggesting that Axl is co-expressed with Gas6, and may be mediated by autocrine regulation. Genetic inactivation of Axl (shRNA) inhibited cell migration, invasion and reduced clonogenic cell survival in both the MD-MB-231 breast and DU145 prostate cancer cell lines. These findings suggest that Axl may be a novel therapeutic target to inhibit the dissemination of tumor cells. Ongoing experiments are evaluating the impact of the Axl-targeting agent R428 on Axl signaling and metastatic phenotype.
Citation Format: Mai Tanaka, Samantha S. Dykes, Dietmar W. Siemann. Axl suppression inhibits cell migration, invasion and survival in breast and prostate cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2507.
Abstract 2550: Generation of CAR-iPS-T cells expressing CD8β
Authors: Ueda Tatsuki , Iriguchi Shoichi , Kawai Yohei , Minagawa Atsutaka , Miyoshi Hiroyuki , Terakura Seitaro , Uemura Yasushi , Woltjen Knut , Kodama Yuzo , Seno Hiroshi , Hitoshi Yasumichi , Nakatsura Tetsuya , Tamada Koji , Kaneko Shin 
Published: August 2018

Abstract
Chimeric antigen receptor (CAR) is an artificial protein that provides HLA-independent antigen specificity to T cells. CAR-T therapy has shown remarkable clinical responses especially in hematologic malignancies. But this therapy needs cell preparation for each patient. That limits its applicability. We have reported regeneration of T cells from iPSCs (Cell Stem Cell 2013). This technology provides unlimited number of T cells, so CAR-T therapy using iPSCs can broadens its applicability. Regenerated T cells previously described in some reports including ours have some different characters from peripheral T cells. In particular, they express only CD8α, and do not express CD8β. We successfully generate CAR-iPS-T cells expressing CD8β with some modification of differentiation protocols. We assayed the function of CAR-iPS-T cells in view of the difference between CD8β positive and negative iPS-T cells. We found CD8β positive CAR-iPS-T cells showed enhanced function to suppress tumor progression compared with CD8β negative CAR-iPS-T cells in subcutaneous xenograft model. In vivo kinetics study revealed that CD8β positive CAR-iPS-T cells have superior function to traffic to target expressing tumor site and enhanced sustainability in vivo compared with CD8β negative CAR-iPS-T cells. These findings indicate that CD8β positive CAR-iPS-T cells may be a potent cell source for iPSC-based cancer immunotherapy.
Citation Format: Tatsuki Ueda, Shoichi Iriguchi, Yohei Kawai, Atsutaka Minagawa, Hiroyuki Miyoshi, Seitaro Terakura, Yasushi Uemura, Knut Woltjen, Yuzo Kodama, Hiroshi Seno, Yasumichi Hitoshi, Tetsuya Nakatsura, Koji Tamada, Shin Kaneko. Generation of CAR-iPS-T cells expressing CD8β [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2550.
Abstract 2571: An integral genomic/expression analysis reveals a cooperative role of APC and TP53 mutations in identifying cetuximab-sensitive colorectal cancer
Authors: Yang Mingli , Schell Michael J. , Loboda Andrey , Nebozhyn Michael V. , Li Jiannong , Teer Jamie K. , Pledger W. Jack , Yeatman Timothy J. 
Published: August 2018

Abstract
The epidermal growth factor receptor (EGFR) is a major therapeutic target in metastatic colorectal cancer (CRC). Mutations in KRAS/NRAS have been identified as the resistance mechanisms to anti-EGFR antibody therapy (cetuximab/pantiumab). However, genomic understanding of the sensitivity to the EGFR-targeted therapy is lacking. A pre-specified, 203-gene cetuximab sensitivity (CTX-S) expression signature score was first validated on two previously published datasets: (1) 77 CTX-treated CRC tumors (progression free survival (PFS)&gt;7 days), and (2) 147 in vitro CTX-treated CRC cell lines. The CTX-S signature was found to be consistently correlated with a newly developed 24-gene APC (truncating) mutations-specific Wnt/β-catenin score on four independent CRC datasets that also include Moffitt (n=468) and TCGA (n=624) CRC datasets. This promoted us to carry out a variety of analyses that revealed a cooperative role of mutant APC + mutant TP53 in identifying cetuximab sensitive subpopulations, which was further evidenced by additional validation analysis on two CRC PDX datasets (n=52 and n=98). While debating or inconclusive, other potential predictive markers of EGFRi therapies have been reported, including BRAF(V600E), PIK3CA mutations, loss of PTEN, and overexpression of HER2 as negative indicators, as well as increased EGFR copy number, and overexpression of EGFR ligands epiregulin and amphiregulin as positive indicators. Here, for the first time, we report that the mutant genes may cooperatively act as positive indicators of EGFRi response. Our study suggests that addition of the routine sequencing of APC and TP53 to extended RAS testing may refine the patient selection strategy to improve cetuximab/pantiumab response and potentially expand the therapeutic opportunity to previously-excluded mutant KRAS/NRAS patients who have “APC + TP53” mutations.
Citation Format: Mingli Yang, Michael J. Schell, Andrey Loboda, Michael V. Nebozhyn, Jiannong Li, Jamie K. Teer, W. Jack Pledger, Timothy J. Yeatman. An integral genomic/expression analysis reveals a cooperative role of APC and TP53 mutations in identifying cetuximab-sensitive colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2571.
Abstract 258: Targeting Nanog in 3D explant models for the evaluation of cancer prevention agents
Authors: Khan Sam , Karmokar Ankur , Sidat Zahirah , Foreman Nalini , Moore David , Higgins Jennifer , Parrott Emma , Theofanous Despoina , Hobbs Dominic , Howells Lynne , Thomas Anne , Brown Karen 
Published: August 2018

Abstract
Background: The transcription factor Nanog is crucial for the self-renewal of cancer stem-like cells (CSCs). Nanog expression in colorectal cancer (CRC) tissue correlates with lymph node metastasis and poor prognosis. Since Nanog is not expressed in most tissues, including normal adult stem-cells, it represents a therapeutic target specific to cancer cells. Curcumin inhibits proliferation and expansion of CSCs derived from CRC and adenomas. In NOD/SCID mice bearing xenografts from patient-derived CRC CSCs, curcumin significantly inhibited tumour growth and improved survival. In addition, curcumin binds directly to Nanog recombinant protein (quantified using microscale thermophoresis). We have developed 3D in-vitro primary human explant models to further characterise the effects of curcumin on Nanog. In this work the hypothesis is tested that Nanog may be an early marker of response for CRC prevention agents.
Methods: Patient-derived CRC and adenoma tissue was cubed (2x2x2mm) and treated for 24 hours with curcumin. Following treatment, explant tissues were processed for analysis by immunohistochemistry (IHC) (n=6) and flow cytometry (n=17). The effect of curcumin on CSCs (defined by expression of aldehyde dehydrogenase (ALDH) or Nanog) and differentiation (via Mucin 2 expression) was analysed using IHC. Additionally, cells expressing Nanog (Nanog+) or Nanog plus proliferation marker Ki67 (Nanog+Ki67+) were assessed using flow cytometry.
Results: A range of adenoma (n=5) and Dukes stage A-C CRC (n=18) samples were studied. Following exposure to curcumin, a 30% reduction was observed in Nanog+ and Nanog+Ki67+ cells. Nanog+ cell number was decreased in a curcumin concentration-dependent fashion in 6 samples and concentration-independently in a further 8. No response was observed in 3 samples. A reduction in Nanog and ALDH with concurrent increase in differentiation was observed via IHC in one sample.
Conclusion: Our data suggest Nanog is targeted by curcumin in adenoma and CRC tissues. Nanog may serve as a biomarker in clinical trials to identify individuals most amenable to treatment with curcumin alone or in combination treatment. Crucially, this will help select those who are likely to benefit from curcumin as a cancer prevention agent. Ultimately, this concept may be applicable to the evaluation of novel CRC prevention agents.
Citation Format: Sam Khan, Ankur Karmokar, Zahirah Sidat, Nalini Foreman, David Moore, Jennifer Higgins, Emma Parrott, Despoina Theofanous, Dominic Hobbs, Lynne Howells, Anne Thomas, Karen Brown. Targeting Nanog in 3D explant models for the evaluation of cancer prevention agents [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 258.