bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2897: Oncogenic Kras activation in gastric adenocarcinoma promotes cancer stem cell phenotypes including metastasis & chemotherapy resistance
Authors: Yoon Changhwan , Chang Kevin K. , Till Jacob , Ryeom Sandra W. , Yoon Sam S. 
Published: July 2017

Abstract
INTRODUCTION:
The Cancer Genome Atlas found that the receptor tyrosine kinase (RTK)-Ras signaling pathway is altered in half of gastric adenocarcinomas (GAs). We added oncogenic Kras to the Atp4b-Cre;Cdh1fl/fl;Trp53fl/fl mouse model of GA and found accelerated tumorigenesis and metastasis. We also found Kras activity to be higher in gastric cancer stem-like cells (CSCs). Thus we hypothesized that Kras activty is critical for maintenance of gastric CSCs and promotes CSC phenotypes.
METHODS:
Human and murine GA cell lines were examined. The effect of Kras pathway inhibition was examined in GA spheroid cells and monolayer cells in various in vitro assays. Kras activity and Kras pathway inhibition was examined in CSCs, primary tumors and metastases in our GA mouse model. The combination of chemotherapy and Kras pathway inhibition was tested in human GA xenografts.
RESULTS:
Kras inhibition with shRNA or MEK inhibitor PD0325901 decreased the ability of GA cells to form spheroids and deceased expression of the stem cell transcription factor, Sox2. CD44(+) gastric CSCs had 65.8-75.4% higher migration, 64.2-79.5% higher invasion, and 63.1-70.2% more anchorage-independent growth compared to unselected cells. These properties could all blocked by 71.1-82.7% with Kras shRNA or PD0325901. In our GA mouse model, PD0325901 starting at 6 weeks of life increased median survival from 76 days to 95 days. Primary tumors and metastases from these mice with and without treatment with MEK inhibition are being analyzed for level of CD44 CSCs, Kras activity, and extent of metastases. Gastric CSCs were resistant to 5-fluorouracil and cisplatin chemotherapy, and this chemotherapy resistance could be reversed in human GA xenografts transducing cells with Kras shRNA.
CONCLUSION:
Kras is more active in gastric CSCs than non-CSCs and promotes malignant phenotypes including metastasis and chemotherapy resistance. Kras pathway inhibition can block these CSC phenotypes, and thus may prove useful in various combination therapies.
Note: This abstract was not presented at the meeting.
Citation Format: Changhwan Yoon, Kevin K. Chang, Jacob Till, Sandra W. Ryeom, Sam S. Yoon. Oncogenic Kras activation in gastric adenocarcinoma promotes cancer stem cell phenotypes including metastasis & chemotherapy resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2897. doi:10.1158/1538-7445.AM2017-2897
Abstract 2902: Epithelial-type CD133+ stem-like lung cancer cells emerge higher drug resistance through MDFIC-mediated Wnt/β-catenin signaling pathway
Authors: Chen Chao-Ju , Yang Chih-Jen , Huang Ming-Shyang , Liu Yu-Peng 
Published: July 2017

Abstract
Background: The epithelial-to-mesenchymal transition (EMT) has been described to promote drug resistance and cancer stem cell (CSC) properties. Many studies demonstrated that a major proportion of circulating tumor cell (CTC) exhibits EMT and CSC characteristics. A recent finding revealed the presence of epithelial type (EpCAM+) CTC was associated with poor outcome, whereas the mesenchymal type (EpCAM-) CTC were not. We therefore hypothesize that the epithelial-type CSC may exhibit higher drug resistant ability.
Methods: We isolated epithelial type (E+) and mesenchymal type (E-) CD133+ cells and CD133- cells from PC14 lung cancer cell line by fluorescence assisted cell sorting. We used western blot, QPCR, immunofluorescence, sphere formation assay and tumor xenograft assay to characterize the 4 subpopulations. The drug resistant ability was determined by cell viability assay and in vivo drug response assays. The drug resistant signature was identified by comparing the gene expression profile from four subpopulations and Gene Expression Omnibus (GEO) database. Knockdown and overexpression of MDFIC in PC14 subpopulations were established by using lentivirus vectors. Immunoprecipitation and subcellular fractionation were performed for drug resistant mechanism investigation.
Results: The epithelial type PC14 CD133+ cells (E+/CD133+ subpopulation) exhibited higher sphere formation ability and was more resistant to the treatment of chemotherapy agents compared to the mesenchymal type CD133+ cells (E-CD133+ subpopulation) in vitro and in vivo. Gene expression profiling showed 86 genes were bioinformatically predicted as drug resistant signature and were correlated with the disease free survival of the patients with lung cancer. Among these genes, the mRNA level of 20 genes were significantly related to the patient’s prognosis in the GSE31210 dataset. Human I-mfa domain-containing protein (MDFIC) was highly expressed in E+/CD133+ subpopulation. Knockdown and overexpression of MDFIC modulates drug resistance ability in cancer cells. MDFIC increased the level of free β-catenin through binding and stabilizing the axin-GSK3-β-β-catenin destruction complex and increased the transcriptional activity of Wnt/β-catenin signaling.
Conclusion: The epithelial-type CD133+ stem-like lung cancer cells are more resistant to the chemotherapy through MDFIC-mediated Wnt/β-catenin signaling activation.
Citation Format: Chao-Ju Chen, Chih-Jen Yang, Ming-Shyang Huang, Yu-Peng Liu. Epithelial-type CD133+ stem-like lung cancer cells emerge higher drug resistance through MDFIC-mediated Wnt/β-catenin signaling pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2902. doi:10.1158/1538-7445.AM2017-2902
Abstract 2919: Quantitative mapping of epidermal growth factor receptor activation in single breast cancer cells treated with targeted therapies
Authors: Le Phuong , Kilian Kristopher , Smith Andrew 
Published: July 2017

Abstract
Twenty percent of breast cancers are triple-negative, lacking estrogen receptor, progesterone receptor, and HER2. There is currently no specific FDA-approved targeted therapy for triple-negative breast cancer patients. Approximately half of triple-negative breast cancers overexpress epidermal growth factor receptor (EGFR), however phase II trials have only shown a 5% response rate with targeted inhibition. A quantitative analysis of EGFR signaling pathway dynamics of single cells at the single molecule level has the potential to shed light on the mechanism of lack of response to targeted therapy, as well as the cell-to-cell heterogeneity of response. Here, we combine single-molecule 3D imaging of EGFR using fluorescent quantum dots (QDs) with cell geometry normalization by micro-patterning to quantitatively map EGFR endocytosis in single cancer cells. Triple negative MDA-MB-231 cells conformed to defined geometries (rectangular with different aspect ratios) were grown on soft polyacrylamide gels after micro-contact printing islands of fibronectin. Two color QD conjugates of EGFR and its ligand, EGF, are used to track ligand binding and receptor internalization upon activation in the absence and presence of different families and dosages of pharmaceutical EGFR inhibitors. We demonstrated that microcontact-printing normalizes cell geometry while maintaining EGFR localization so that EGFR signaling response can be quantitatively assessed on a single-cell basis and that QD conjugates can specifically label EGFR and EGF. Quantitative single-molecule imaging at single-cell level revealed high level of heterogeneity in the amount and spatial localization of EGF binding and EGFR endocytosis among the cell population in the presence and absence of EGFR inhibitor, gefitinib. Using this platform, we will investigate if spatial organization of EGFR clusters within individual cells contribute to single-cell heterogeneous response to EGFR inhibitors. We anticipate quantitative mapping of EGFR activity with drug treatment will help to elucidate why EGFR drug treatments fail and provide a means to develop combination therapy to address heterogeneity in triple negative-breast cancer.
Citation Format: Phuong Le, Kristopher Kilian, Andrew Smith. Quantitative mapping of epidermal growth factor receptor activation in single breast cancer cells treated with targeted therapies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2919. doi:10.1158/1538-7445.AM2017-2919
Abstract 292: Synthesis and characterization of the novel benzylindazole-based BUB1 kinase inhibitor BAY 1816032 with potent anti-tumor activity
Authors: Hitchcock Marion , Mengel Anne , Nising Carl 
Published: July 2017

Abstract
BUB1 (budding uninhibited by benzimidazoles 1) is a serine/threonine protein kinase. The protein is bound to kinetochores and plays a key role in the establishment of the mitotic spindle checkpoint and chromosome congression prior to anaphase.
Inhibition of BUB1 kinase represents a novel approach for cancer treatment: whereas cell cycle arrest is the predominant mode of action of a number of antimitotic cancer drugs (e.g. taxanes and vinca alkaloids), BUB1 inhibition results in aneuploidy and cell death by driving cells through mitosis irrespective of misattached chromosomes.
Here, we disclose for the first time the structure and functional characterization of a novel, first-in-class Bub1 kinase inhibitor. Medicinal chemistry efforts resulted in BAY 1816032 featuring high potency, long target residence time and good oral bioavailability. BAY 1816032 is highly selective for BUB1 displaying single digit nanomolar biochemical potency and double-digit nanomolar cellular potency (H2A induced HeLa-cells). Synergistic effects can be observed when BUB1 inhibitor BAY 1816032 is combined with low doses of paclitaxel affecting chromosome segregation and cell proliferation. X-ray data of benzylindoles allowed a better understanding of the binding mode. Further data on structure-activity relationships including pharmacokinetic characterization, drug metabolism and the synthesis of BAY 1816032 and analogues will be presented.
These results validate the innovative concept of interference with mitotic checkpoints and justify clinical proof of concept studies evaluating BUB1 inhibitor BAY 1816032 in combination with taxanes in order to enhance their efficacy and to potentially overcome resistance.
Citation Format: Marion Hitchcock, Anne Mengel, Carl Nising. Synthesis and characterization of the novel benzylindazole-based BUB1 kinase inhibitor BAY 1816032 with potent anti-tumor activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 292. doi:10.1158/1538-7445.AM2017-292
Abstract 2925: Distribution of copy number alterations defines clonal populations involved in colorectal cancer evolution
Authors: Quintanilla Isabel , Asensio Elena , Pellisé Maria , Castells Antoni , Cuatrecasas Miriam , Camps Jordi 
Published: July 2017

Abstract
Colorectal cancer progresses in a multi-step manner with adenoma being the most well-known precursor lesion. Malignant polyp, an adenoma that contains a focus of adenocarcinoma, is a suitable model to study the colorectal tumor evolution. Although the mutations that lead adenoma to evolve into carcinoma have been previously described, the copy number changes involved in this malignant transformation have not been fully explored. To understand how these genomic alterations contribute to carcinogenesis, we used sequential fluorescence in situ hybridization using probes for the oncogenes EGFR, MYC, CDX2, and ZNF217 and the tumor suppressor genes SMAD7 and APC in order to analyze the copy number changes in 23 cases of malignant polyps and 10 cases of low grade dysplasia (LGD) adenomas based on single cell analyses. We found the levels of genomic heterogeneity increased from LGD adenomas to malignant polyps, with the adenoma component having a lower degree of chromosomal instability than the adenocarcinoma component. Despite the intercellular heterogeneity, we observed different patterns of evolution. The gain of ZNF217 was the earliest event as it was found to be in some cases of the LGD adenoma, and was also important in the neoplastic transformation of malignant polyps. Other notable genomic imbalances we observed during the malignant transformation were the gain of CDX2 and the loss of SMAD7. Interestingly, in a significant proportion of cases we observed the gain of all loci analyzed to be the decisive step in the transition from adenoma to carcinoma, suggesting a whole genome duplication event. Lastly, in half of the malignant polyps studied the main clone in the carcinoma component was already present in the adjacent adenoma component, although in some cases at a low frequency only detected by single cell analysis. Our data suggest that copy number changes are early events in colorectal carcinogenesis that can determine the evolution from adenoma to carcinoma.
Citation Format: Isabel Quintanilla, Elena Asensio, Maria Pellisé, Antoni Castells, Miriam Cuatrecasas, Jordi Camps. Distribution of copy number alterations defines clonal populations involved in colorectal cancer evolution [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2925. doi:10.1158/1538-7445.AM2017-2925
Abstract 2926: Histologically defined intratumoral sequencing uncovers evolutionary cues into conserved molecular events driving gliomagenesis
Authors: Prabhu Antony , Kesarwani Pravin , Kant Shiva , Graham Stewart , Chinnaiyan Prakash 
Published: July 2017

Abstract
Glioblastoma (GBM) represents an archetypal example of a heterogeneous malignancy, harboring regions of invasion, necrosis, and vascularization. To begin to understand the diverse molecular consequences of this complex tumor ecology, we analyzed RNA-seq data generated from commonly identified intratumoral structures in GBM that were isolated and enriched using laser capture micro-dissection. Structures included the infiltrative, leading edge, central tumor core, and cellular regions in areas of necrosis. From this RNA-seq database consisting of 119 structural regions from 37 individual tumors generated from ~12,000 hematoxylin and eosin histologic images, we validated significant intra-tumoral heterogeneity in GBM, with cells derived from the infiltrating leading edge almost exclusively harboring the proneural molecular subtype, while cells in regions of necrosis displaying the mesenchymal subtype. In addition, we made the striking observation that when evaluating the tumors transcriptional profiles in the context of their derived structural regions, there was a relative small amount of inter-tumoral heterogeneity in GBM, with significant clustering of transcriptional profiles of specific structural regions from different tumors. To begin to understand the specific molecular events driving regional growth and their contributory role in gliomagenesis, we analyzed the transcriptional programs in the context of evolutionary progression. This level of analysis identified numerous pathways associated with neuronal receptor signaling, autophagy, and fatty acid metabolism as early events in tumorigenesis. Traditional cancer related signaling pathways emerged as key mediators driving progression, with glioma and p53 signaling ranking highest on the list. As the tumor progressed to harbor regions of necrosis, in addition to expected changes in VEGF and tumor signaling, we uncovered global metabolic changes associated with progression, with metabolic pathways comprising 10 of the top 20 pathways unique to this region. Collectively, these findings suggest that the observed intra-tumoral heterogeneity in GBM may be a conserved, predictable consequence to its complex microenvironment and therapeutic strategies rationally designed to target these unique, unequivocally present tumor cell biomes may lead to therapeutic gains in this otherwise fatal malignancy.
Citation Format: Antony Prabhu, Pravin Kesarwani, Shiva Kant, Stewart Graham, Prakash Chinnaiyan. Histologically defined intratumoral sequencing uncovers evolutionary cues into conserved molecular events driving gliomagenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2926. doi:10.1158/1538-7445.AM2017-2926
Abstract 2930: Niche microenvironments essential for tumor heterogeneity of primary cancer cells
Authors: Fujino Shiki , Miyoshi Norikatsu , Ohue Masayuki , Takayashi Yusuke , Yasui Masayoshi , Takahashi Hidekazu , Haraguchi Naoyuki , Nishimura Jhunichi , Hata Taishi , Mizushima Tsunekazu , Doki Yuichiro , Mori Masaki 
Published: July 2017

Abstract
Primary culture of cancer cells derived from each patient’s tumor can provide important information of the “individual tumor.” It is general to use cell lines in basic research filed. However, cell lines are quite different from clinical cancers. Clinical cancer tissues are composed of not only cancer cells but also tumor microenvironments such as stromal cells and tumor vessels. The primary culture method of clinical cancer with tumor niche microenvironments has not been optimized. We have developed a simple 2D-culture method for primary colorectal cancer (CRC). We obtained 30 samples from surgically resected tumor. They were mechanically and enzymatically digested and fibrotic tissue and bacteria were excluded using customized two size filters. And we cultured the obtained cells on a matrigel-coated plate with embryonic stem (ES) cells culture medium. We named these cultured cancer cells, “isolated-tumor derived Cancer Cells (iCCs).” All iCCs grew and about 80 % of iCCs were successfully passaged. Twenty-three iCCs were transplanted into the subcutaneous layer of NOD-SCID mice, and the tumor growth and histology of iCCs were examined. The morphology was similar to each parental clinical tumor. And microarray analysis showed that RNA expression of iCCs was similar to each parental tumor. Furthermore, we examined the culture medium; our modified ES culture medium (ES-cultured iCCs) and 10% FBS medium (serum-cultured iCCs). The expression of surface markers regarding cancer stem cells such as CD44 and CD24 were different between ES-cultured iCCs and serum-cultured iCCs, and drug sensitivity of iCCs were also different. FACS analysis and immunocytochemistry revealed that iCCs contained PDGFR-positive cells. The results of multi-drug sensitivity assay were different between iCCs and cell lines especially in PDGFR inhibitor. Strong correlations were observed between the results of multi-drug sensitivity assay of iCCs and clinical outcomes of chemotherapy. We report an innovative primary culture method and the in vivo and in vitro analyses of iCCs, leading to the future application.
Citation Format: Shiki Fujino, Norikatsu Miyoshi, Masayuki Ohue, Yusuke Takayashi, Masayoshi Yasui, Hidekazu Takahashi, Naoyuki Haraguchi, Jhunichi Nishimura, Taishi Hata, Tsunekazu Mizushima, Yuichiro Doki, Masaki Mori. Niche microenvironments essential for tumor heterogeneity of primary cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2930. doi:10.1158/1538-7445.AM2017-2930
Abstract 2931: Targeting membrane-bound carbonic anhydrases in breast cancer to intervene in the metastatic phenotype
Authors: Mboge Mam Y. , Chen Zhijuan , Mahon Brian P. , Tu Chingkkuang , Wolff Alyssa S. , Mathias John V. , Carta Fabrizio , Supuran Claudiu T. , McKenna Rob , Frost Susan C. 
Published: July 2017

Abstract
Breast cancer is the second leading cause of cancer related deaths among women in the United States. Despite the tremendous progress that has been made towards treating localized tumors, nearly 40,000 women die each year, predominantly from metastatic drug resistance. The tumor microenvironment plays a pivotal role in determining tumor growth, invasion, metastasis, and therapeutic success or failure. Therefore, a therapy targeting the tumor microenvironment is needed to sufficiently preserve the quality of life of cancer patients, by inhibiting metastasis. Elevated levels of Carbonic anhydrase IX (CAIX) expression in primary breast cancers is a marker for highly aggressive and metastatic tumors, especially of the triple negative subtype (TNBC). It is also associated with hypoxia, extracellular acidification, and poor prognosis. Low pH (values of ~6.5-6.8) is toxic to normal cells in the tumor microenvironment while enhancing cancer cell proliferation and tumor growth. Our goal was to compare the structure of a CAIX-mimic bound to ureidosulfonamide inhibitors with the biological activity of these inhibitors in triple negative and estrogen receptor positive (ER+) breast cancer cell lines. CAIX is a reversible enzyme and at low pH (high proton concentration), the enzyme will consume protons, raising pH. Our hypothesis is that CAIX inhibition, in the context of an acidic microenvironment, will dysregulate its ability to maintain the acidic pH preferred by cancer cells which favors their growth and migration. In this study, we have shown the interaction of sulfonamide-based inhibitors with a CAIX-mimic using X-ray crystallography. These structures show that the inhibitors make multiple contacts within the active site cavity. This is consistent with the inhibitor-induced decrease in CAIX activity and to some extent expression. We have also investigated the effect of CA inhibition on breast cancer cell growth, proliferation, activation of cell death pathways and migration. This reveals that, although CA inhibition with the sulfonamide-based compounds inhibits cell growth and migration, it does not activate apoptotic pathways. In total, these observations indicate that CAIX is a viable small molecular drug target for the treatment of metastatic breast cancer.
Citation Format: Mam Y. Mboge, Zhijuan Chen, Brian P. Mahon, Chingkkuang Tu, Alyssa S. Wolff, John V. Mathias, Fabrizio Carta, Claudiu T. Supuran, Rob McKenna, Susan C. Frost. Targeting membrane-bound carbonic anhydrases in breast cancer to intervene in the metastatic phenotype [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2931. doi:10.1158/1538-7445.AM2017-2931