bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 367: The connection of vitamin D3 induced cytotoxicity with the vitamin D3 receptor
Authors: Hoshi Takaya , Abe Yuta , Sugiyama Ikumi , Sadzuka Yasuyuki 
Published: August 2019

Abstract
[Purpose] This study aims to evaluate the possible antitumor effect of vitamin D. Vitamin D is a group of lipophilic compounds comprising of vitamin D2 and vitamin D3. Vitamin D3 has been shown to play an important role in humans. It has been previously reported that the onset risk of malignant melanoma and leukemia can be decreased by maintaining high vitamin D3 levels in the body. In addition, the cytotoxic effect of vitamin D on breast cancer cells has also been shown. In this study, the relationship between the cytotoxicity of vitamin D3 and vitamin D3 receptor (VDR) expression on melanoma and leukemia cells was examined.
[Method] B16F10 melanoma (B16F10) and P388 leukemia (P388) cells were used in this study. Western blot analysis was performed to determine VDR expression in each cell type. To evaluate cytotoxicity, B16F10 or P388 cells were seeded at 1×105 cells/mL, and calcitriol as vitamin D3 (range 9 ng/mL-90 μg/mL) was added at once. After two days, cytotoxicity was evaluated by colorimetric analysis using Cell Counting Kit-8. Additionally, the cytotoxicity of P388 cells was determined following the combined treatment of vitamin D3 and retinoic acid.
[Results and Discussion] Western blot analysis revealed that both B16F10 and P388 cell types, expressed VDR with a higher expression in P388 cells. In P388 cells, there was a significant reduction in cell viability following vitamin D3 treatment, which decreased to 10% at 90 μg/mL. In contrast, the viability of B16F10 cells following vitamin D3 treatment only slightly decreased. Interestingly, these results suggest that the cytotoxicity of vitamin D3 correlates with VDR expression. VDR reportedly forms a heterodimer with the retinoid X receptor. However, the presence of retinoic acid did not alter vitamin D3 cytotoxicity. Therefore, it was considered that cytotoxicity induced by vitamin D3 was not connected with the formation of the VDR-retinoid X receptor heterodimer. In conclusion, there was a higher expression of VDR in P388 cells compared with B16F10 cells. This higher expression correlated with a higher level of vitamin D3 cytotoxicity, suggests an antitumor effect of vitamin D3. This study proposes the possibility of using vitamin D3 as a novel treatment for leukemia and other cancers.
Citation Format: Takaya Hoshi, Yuta Abe, Ikumi Sugiyama, Yasuyuki Sadzuka. The connection of vitamin D3 induced cytotoxicity with the vitamin D3 receptor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 367.
Abstract 3677: Downregulation of FOXO3a by DNMT1 promotes breast cancer stem cell properties and tumorigenesis
Authors: He Zhimin , Liu Hao , Song Ying , Zheng Guopei 
Published: August 2019

Abstract
Breast cancer stem cells (BCSCs) are tumor initiating cells that can self-renew and are highly tumorigenic and chemoresistant. Recent evidence indicated that FOXO3a plays an important role in regulating CSCs properties. However, the biologic function and detailed molecular mechanism of FOXO3a in BCSCs remains largely elusive. Here, we demonstrated that FOXO3a was hypermethylated and downregulated in breast cancer cells and tissues. Overexpression of FOXO3a potently attenuated the BCSCs properties in vitro and inhibited tumorigenesis in vivo, whereas knockdown of FOXO3a increased these properties. Further mechanistic investigation revealed that FOXO3a was functionally linked to the inhibition of the FOXM1/SOX2 pathway. Moreover, we found that SOX2 directly transactivated DNMT1 gene expression and then feedback inhibited FOXO3a expression. Clinically, we observed a significant inverse correlation between FOXO3a and FOXM1 or SOX2 expression levels. Loss of FOXO3a expression also predicted poor prognosis in breast cancer. In addition, we demonstrated that inhibition of DNMT activity suppressed tumor growth via regulating of FOXO3a/FOXM1/SOX2 signaling in breast cancer. Our findings suggest an important role of DNMT1/FOXO3a/FOXM1/SOX2 pathway in regulating BCSCs properties, identifying potential therapeutic targets for breast cancer.
Keywords: breast cancer, cancer stem cell, FOXO3a, DNMT
Note: This abstract was not presented at the meeting.
Citation Format: Zhimin He, Hao Liu, Ying Song, Guopei Zheng. Downregulation of FOXO3a by DNMT1 promotes breast cancer stem cell properties and tumorigenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3677.
Abstract 368: Next generation DNA aptamers with prolonged stability and antitumor activity
Authors: AMERO Paola , Rodriguez-Aguayo Cristian , Chaudhari Rajan R. , Zhang Shuxing , Sood Anil K. , Lopez-Berestein Gabriel 
Published: August 2019

Abstract
Aberrations in Gas6/AXL signaling is associated with many human diseases including ovarian cancer, where in patients expressing high levels of AXL have shorter overall survival than patients expressing low levels. Aptamers are short synthetic oligonucleotides (DNA or RNA) that can bind with highest specificity to various other macromolecules such as peptides, proteins and carbohydrates, based on their tertiary structures. Aptamers are similar in their functionality to antibodies in their specificity;
easier to manufacture than antibodies; have minimal side effects; and thus hold a high promise as therapeutics. The typical issues with the use of aptamers as therapeutic agents are their selectivity, stability, and bioavailability. We present the development of the next generation DNA aptamers, targeting p-AXL specifically to as a therapeutic in ovarian cancer (OC) by chemical modifications to their primary sequence. Improved bioavailability and stability, based on the comparative analysis the library of 17 DNA aptamers that differed in the position and number of fluoro- and thio- modifications on their nucleotide sequence, were used as the criteria for the selection and characterization of the best aptamer candidates to target p-AXL. Two of the best candidates, GLB-G25 and GLB-A04 thus chosen with the highest bioavailability and stability, when tested in ovarian cancer cell lines, decreased the migration and invasion of cells in vitro. Treatment of ovarian cancer orthotopic murine model animals with modified DNA aptamer candidates chosen above significantly reduced tumor growth and the number of metastases. The candidates, GLB-G25 and GLB-A04 show highest potential to be developed into therapeutics to target p-AXL that is elevated in several cancers including ovarian cancer.
Note: This abstract was not presented at the meeting.
Citation Format: Paola AMERO, Cristian Rodriguez-Aguayo, Rajan R. Chaudhari, Shuxing Zhang, Anil K. Sood, Gabriel Lopez-Berestein. Next generation DNA aptamers with prolonged stability and antitumor activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 368.
Abstract 3685: HOTAIR regulation and functionality in ovarian cancer stem cells
Authors: Wang Weini , Fang Fang , Ozes Ali , Nephew Kenneth P. 
Published: August 2019

Abstract
Ovarian cancer (OC) is the fifth leading cause of cancer-related death among American women. Persistence of OC stem cells (OCSCs) is believed to contribute to resistance to platinum-based chemotherapy and disease relapse. We have previously shown that epigenetic changes in OCSCs play a role in post-therapy OCSC persistence and demonstrated that is it possible to target OCSC using epigenetic therapies. HOXC transcript antisense RNA (HOTAIR) has been shown to be associated with chemoresistance and to be overexpressed in many types of cancers, including high-grade serous OC (HGSOC). HOTAIR interacts with Polycomb Repressive Complex 2 (PRC2) and due to its histone methyltransferases activity plays a key role in chromatin remodeling. Because HOTAIR is a known epigenetic regulator of differentiation and developmental genes in OC and other cancers, we hypothesize that HOTAIR is a key epigenetic regulator in OCSCs and therapeutically targeting HOTAIR in OCSCs will prevent tumor relapse. The goal of this study is to understand the role of HOTAIR in regulating OCSCs and further dissect the mechanism of action of HOTAIR in OCSCs. Aldehyde dehydrogenase (ALDH) activity and FACS was used to separate OCSCs from non-OCSCs in a panel of HGSOC cell lines (OVCAR3, CAOV3, OVCAR5, Kuramochi, COV362). Quantitative RT-PCR analysis revealed that HOTAIR was overexpressed in OCSC cells compared to non-OCSC cells. Knockout of HOTAIR using CRISPR-Cas9 system significantly decreased OCSC population and stemness-related phenotypes, including spheroid formation and colony formation ability. Overexpression of HOTAIR in HGSOC cells significantly increased these stem-like characteristics. Furthermore, targeting HOTAIR using peptide nucleic acid (PNA), which blocks interaction with Enhancer of Zeste Homologue 2 (EZH2), significantly decreased the OCSC population, indicating HOTAIR functions through EZH2 in regulating OCSCs. Combining the HOTAIR-targeting PNA with other epigenetic inhibitors, including the hypomethylating agent guadecitabine and the EZH2 inhibitor GSK503, significantly decreased proliferation and colony formation ability of HGSOC cells. We suggest that a better understanding of HOTAIR will facilitate identifying epigenomic alterations and chromatin landscape that contributes to OCSC phenotypes. Targeting HOTAIR in combination with epigenetic therapies may represent a therapeutic strategy to prevent tumor relapse.
Citation Format: Weini Wang, Fang Fang, Ali Ozes, Kenneth P. Nephew. HOTAIR regulation and functionality in ovarian cancer stem cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3685.
Abstract 3687: Colorectal cancer stem cell expressing POU5F1 promotes liver metastasis
Authors: Miyoshi Norikatsu , Fujino Shiki , Sasaki Masaru , Saso Kazuhiro , Takahashi Hidekazu , Haraguchi Naotsugu , Hata Taishi , Matsuda Chu , Mizushima Tsunekazu , Mori Masaki , Doki Yuichiro 
Published: August 2019

Abstract
Background: The gene POU5F1, encoding the POU domain, class 5, is a transcription factor and the expression is found in embryonic stem cells. POU5F1 gene controls self-renewal and differentiation, and regulates pluripotency and proliferation in embryonic stem cells. However, the role of POU5F1 in metastatic CRC (mCRC) is not well known. We focused on the relationship between POU5F1 gene expression and liver metastasis of CRC and clarify the role and abilities of POU5F1-positive CRC cells.
Methods: The study included 158 patients with CRC who underwent surgery from 2009 to 2011. The correlations between the POU5F1 gene expression and the clinical parameters were assessed, and liver-metastasis-free survival (LMFS) was evaluated in these patients. POU5F1-EGFP-positive cells were established by the construct with lentiviral vector, and we examine their sub-population and ability. The capacity to form liver metastasis in vivo was examined using CRC cell lines and primary cultured CRC cells.
Results: In the survival analysis, LMFS was significantly poor in the high POU5F1 expression group compared to the low-expression group (P=0.008). Multivariate analyses showed that POU5F1 expression (P=0.015) and TNM stage (p&lt;0.001) significantly correlated with LMFS. POU5F1-EGFP-positive cells highly expressed stem-cell-associated markers and they had self-renewal and differentiation abilities. POU5F1-high cells actively formed liver metastasis.
Conclusion: The POU5F1expression correlated with liver metastasis in CRC patients. POU5F1-positive cells have the self-renewal and differentiation abilities as cancer stem cells. POU5F1 contributes to the ability forming liver metastasis in CRC.
Citation Format: Norikatsu Miyoshi, Shiki Fujino, Masaru Sasaki, Kazuhiro Saso, Hidekazu Takahashi, Naotsugu Haraguchi, Taishi Hata, Chu Matsuda, Tsunekazu Mizushima, Masaki Mori, Yuichiro Doki. Colorectal cancer stem cell expressing POU5F1 promotes liver metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3687.
Abstract 3692: The effect of endothelin 1 on cancer stem cell development
Authors: Seno Akimasa , Seno Masaharu 
Published: August 2019

Abstract
Although we have been demonstrating the induction of cancer stem cells from ESCs or human/mouse iPSCs reprogrammed from normal fibroblasts using cancer cell conditioned media, the exact inducing factor is still unknown. Previously, we compared the gene expression profiles among hiPS/ESC, cancer stem like cells induced from hiPSC with cancer cell conditioned media, cancer stem cells obtained from cancer tissue or cell culture, and cancer cells. In this study, the expression of endothelin 1 was found commonly upregulated in different cancer stem cell groups. To investigate the roles of endothelin 1 in the development of cancer stem cells, the significance of endothelin 1 signaling including endothelin 1 in the cancer cell conditioned media and the expression of endothelin 1 receptors in iPSCs as well as in the developed cancer stem cells were quantitatively and qualitatively evaluated.
Citation Format: Akimasa Seno, Masaharu Seno. The effect of endothelin 1 on cancer stem cell development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3692.
Abstract 3697: Optimization of human cancer cell xenografts into zebrafish larvae for high-throughput drug screening
Authors: Dockins Stephen R. , Blackburn Jessica S. , Haney Meghan G. , Wilson Bradley 
Published: August 2019

Abstract
The use of zebrafish in cancer xenograft models has grown rapidly in recent years. This is primarily due to the fact that this model takes advantage of the ease of in vivo imaging and the high-throughput screening capabilities that zebrafish have to offer. However, researchers have yet to come to a consensus on standardized procedure to utilize zebrafish for xenograft of human cells. This study aims to optimize a zebrafish xenografting protocol for various human cancers with the intention of performing high-throughput drug screening.
We tested the survival of 3 different zebrafish strains when incubated at 28°C, 34°C and 37°C. There was no difference in survival between the AB, SAT, and Casper strains of zebrafish at any temperature. Human leukemia, breast, lung, colon, and brain cancer cell lines were fluorescently labelled with Vybrant DiI cell staining dye and injected at a concentration of 100, 250, 500, or 1000 cells per 2nL droplet volume into dechorionated 2-day-old zebrafish embryos. The injection sites used were the brain, caudal vein, duct of Cuvier (DC), eye, pericardium, perivitelline space (PVS), and yolk sac. As expected, injections became more difficult and time consuming when using higher cell counts as the injector needle clogged more often. Similarly, injection sites that required more precision like the caudal vein or eye took the longest to inject successfully and had lower engraftment rates than sites like the yolk or DC. We determined that using 500 cells per injection was optimal as engraftment level was comparable to 1,000 cell injections but had improved survival and reduced injection time. Optimal injection site varied between cell lines, but overall, injections into the duct of Cuvier resulted in consistently high survival and engraftment rates with one of the lowest injection times.
We are in the process of determining the extent to which the injection site affects chemotherapy response on human cells implanted into zebrafish. We are also preforming a high-throughput drug screen on human leukemia cells implanted into zebrafish to provide proof-of-principle that these methods are useful in identifying novel anti-cancer compounds. In total, this work will establish standard operating procedures for use of xenograft in zebrafish, providing new opportunities in personalized medicine and drug discovery.
Citation Format: Stephen R. Dockins, Jessica S. Blackburn, Meghan G. Haney, Bradley Wilson. Optimization of human cancer cell xenografts into zebrafish larvae for high-throughput drug screening [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3697.
Abstract 3703: Development of oncopig urothelial carcinoma cell lines
Authors: Qazi Aisha , Thomas Faith M. , Chaki Sulalita , Robertson Noah , Patel Shovik , Schachtschneider Kyle M. , Schook Lawrence B. 
Published: August 2019

Abstract
Urothelial carcinoma (UC), more commonly known as bladder cancer, affects approximately 81,000 people a year in the United States and is the fourth most common type of cancer among men. Age also plays a tremendous factor, as 90% of people affected by bladder cancer are older than 55, and if the cancer has metastasized the 5-year survival rate is 35%. This requires the advancement of early detection and treatment methods for bladder cancer. The development of clinically relevant systems to serve as a bridge between preclinical murine studies and human clinical practice is of vital importance. The Oncopig Cancer Model (OCM) is a novel transgenic swine platform that recapitulates human cancer through development of site/cell specific tumors after Cre recombinase induced expression of heterozygous KRASG12D and TP53R167H transgene. In this study, we standardized a protocol for the development and growth of Oncopig UC cell lines that faithfully recapitulates in vitro and in vivo features of human bladder cancer.
The focus of this preliminary research was to successfully create OCM UC cell lines that mimic features of human UC. OCM urothelial cell lines (n=7) were isolated from whole bladders collected from male Oncopigs at euthanasia. OCM urothelial cell lines were exposed to Cre recombinase 48 hours post isolation to induce expression of KRASG12D and TP53R167H transgenes, resulting in development of OCM UC cell lines. Differences in migration rates were observed between OCM urothelial and UC cell lines, confirming cellular invasiveness. RT-PCR was performed after several passages to confirm KRASG12D and TP53R167H gene expression, which was observed in OCM UC but not urothelial cell lines. Uroplakin II and pan-Cytokeratin (PCK-26) are markers for UC and urothelial cells, respectively, and are used to diagnose UC clinically. OCM UC cells stained positively for Uroplakin II and PCK-26, confirming their identity as malignant UC cells. Future work is needed to confirm OCM and human UC display similar histological and molecular features, and to develop strategies for in vivo OCM UC tumor development.
Citation Format: Aisha Qazi, Faith M. Thomas, Sulalita Chaki, Noah Robertson, Shovik Patel, Kyle M. Schachtschneider, Lawrence B. Schook. Development of oncopig urothelial carcinoma cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3703.
Abstract 3704: CRISPR screening in a xenograft model for in vivo drug moa analysis
Authors: Melton Michele , Bast Athena , Leeson-Payne Alastair , Blanck Maximilian , Lawo Steffen , Stacy Deeds , Cross Benedict C.S. 
Published: August 2019

Abstract
Functional genomic screening with CRISPR has provided a powerful and precise new way to interrogate the phenotypic consequences of gene manipulation in high-throughput, unbiased analyses. Rapid development of pooled lentivirus and deep-sequencing-led approaches have allowed us and others to exploit this technology in target ID, target validation, drug MOA analysis and patient stratification.Until now, these exquisitely powerful screening technologies have been deployed predominantly in simple in vitro analyses. Whilst these in vitro approaches allow for high quality hit discovery and at whole-genome level scale, they are not able to adequately capture the impact of tumour microenvironment and heterogeneity on the genetic perturbations under study. To address this, we present here the development and validation of a platform for pooled CRISPR knock-out screening in xenograft models of tumour growth.
A critical step in appropriate study design is the evaluation of the cell uptake following implantation. Since this directly contributes to the tumour growth and cell number, understanding cell survival dynamics post-implantation is necessary to determine the appropriate experimental scope. We used a barcoding approach coupled to next generation sequencing to determine the response and contribution of individual A375 melanoma cells to final tumour composition and used this to calibrate the experimental design for robust pooled-based CRISPR screening analysis.
Following the determination of experimental conditions, a proof-of-concept drug mechanism of action analysis was conducted using a custom designed cancer genome CRISPR knock-out library. A375 cells were first infected with the CRISPR library and selected ex vivo ahead of implantation and treatment of established tumour-bearing nude mice with a defined therapeutic regime of the BRAFV600E selective inhibitor, Vemurafenib. Tumours were harvested and the identification of genes which selectively modulate the tumour response to drug were identified by next generation sequencing using our optimised screening analysis platform.
Our results provide a robust experimental validation and define a novel platform which will allow the rapid and definitive in vivo evaluation of both drug and gene interactions using powerful CRISPR-based screening tools.
Citation Format: Michele Melton, Athena Bast, Alastair Leeson-Payne, Maximilian Blanck, Steffen Lawo, Deeds Stacy, Benedict C.S. Cross. CRISPR screening in a xenograft model for in vivo drug moa analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3704.