bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 673: The role of MYPT1/2, ASPP2 and MYH9 in invasive lobular carcinoma
Authors: Schipper Koen , de Ruiter Julian , Kas Sjors , Schut Eva , Koudijs Marco , Addams David , Jonkers Jos 
Published: July 2016

Abstract
Invasive lobular carcinoma (ILC) accounts for 10-15% of breast cancer cases in women. One of the characteristics of this type of cancer is the loss of intercellular adhesion which is facilitated by inactivating mutations in E-cadherin. However the loss of E-cadherin alone is not enough to induce ILC. An in vivo Sleeping Beauty insertional mutagenesis screen was performed to identify genes that together with E-cadherin loss contribute to ILC development in mice. In around 85% of the tumors that were analyzed during this screen one of four hits were observed, namely MYPT1, MYPT2, ASPP2 and MYH9. Interestingly these four hits appear to be mutually exclusive indicating a shared mechanism of action.
To investigate how these genes might affect tumorigenesis we first looked at the location of the transposon insertions. This revealed that for MYPT1, MYPT2 and ASPP2 the transposons appeared to localize to a specific region in the gene indicating that these insertions could result in a truncation variant. In the case of MYH9 no clear localization of insertions was found which in combination with the recently published data indicates a loss of function. Northern blot analysis revealed the presence of truncation variants for MYPT1/2 and ASPP2 which for MYPT1 were confirmed to also result in truncated protein variants.
In order to identify the tumorigenic potential of truncated MYPT1 and ASPP2 we overexpressed them in spontaneously immortalized mammary epithelial cell lines HC11 and NMuMG. We are using these cell lines to analyse differences in cell proliferation, anchorage independent growth, sensitivity to apoptosis in vitro and tumor formation in vivo. To further analyse the hits in an E-cadherin negative setting we made use of primary mammary epithelial cells (MECs) isolated from genetically engineered mice with mammary gland specific E-cadherin loss. These MECs normally do not grow past several passages in vitro. However, these MECs when we overexpress truncated MYPT1 or ASPP2, or use shRNA mediated knockdown of MYH9 can be passaged and grown in vitro consistently.
Finally we are generating conditional mouse models with mammary gland specific E-cadherin loss in combination of expression of the truncated variants of MYPT1 or ASPP2, or loss of MYH9. We will to monitor whether these mice are prone to developing ILC.
Citation Format: Koen Schipper, Julian de Ruiter, Sjors Kas, Eva Schut, Marco Koudijs, David Addams, Jos Jonkers. The role of MYPT1/2, ASPP2 and MYH9 in invasive lobular carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 673.
Abstract 68: Neuroblastoma drug response profiles are associated with gene expression profiles
Authors: Bond Jeffrey P. , VanSickle Elizabeth , Dexhemier Thomas S. , Neubig Richard R. , Sholler Giselle L. 
Published: July 2016

Abstract
Background: High risk neuroblastoma patients have a poor response to initial chemotherapy, with 20% unable to achieve a complete response. Genomic studies have identified 6 targeted agents whose pathways have been identified in some high risk neuroblastoma tumors. The low frequency of established driver mutations, along with the variety of molecular mechanisms for cancer pathway dysregulation, suggest development of gene expression-based predictive biomarkers in addition to sequence-based biomarkers. Our platform for developing gene expression-based biomarkers includes primary neuroblastoma cell lines, high throughput quantification of cell survival, exome sequencing, and genome-wide expression profiling.
Methods: Cell survival was quantified for each of thirty primary neuroblastoma cell lines in the presence of each of six drugs (bortezomib, crizotinib, dasatinib, lapatinib, sorafenib, and vorinostat) using high throughput cell survival measurements. Titration curves were based on treatment with vehicle as well as sixteen concentrations of each drug (replicated within and between plates). Summary descriptions of the response of each cell line to each drug were obtained using the four parameter logistic model as well as the area under the curve. Exome sequences were obtained using Illumina HiSeq technology. Genome-wide expression profiles were obtained using Affymetrix GeneChip technology. Inference was based on parametric univariate and non-parametric multivariate tests.
Results: For three of the drugs (bortezomib, dasatinib, and vorinostat) the variation in response between cell lines was statistically significant (p &lt; 0.05; F ≥3) and assay results were reproducible in the context of the differences between cell lines (for the other three drugs the estimated standard deviation of log10 EC50 ≤ 0.2). Genome-wide expression profiles were associated with variation in drug response between cell lines, for example, high PDGFRB expression indicates sensitivity to dasatinib (p &lt; 0.001).
Conclusion: High throughput cell survival measurements provided for identification of predictive biomarkers of neuroblastoma drug sensitivity. These biomarkers are currently being studied in the PEDS-PLAN (Pediatric Precision Laboratory Advanced Neuroblastoma Therapy) clinical trial.
Citation Format: Jeffrey P. Bond, Elizabeth VanSickle, Thomas S. Dexhemier, Richard R. Neubig, Giselle L. Sholler. Neuroblastoma drug response profiles are associated with gene expression profiles. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 68.
Abstract 681: Using a genome-editing approach for the stepwise establishment of zebrafish models of pediatric high-grade gliomas and MPNSTs
Authors: Oppel Felix , He Shuning , Tao Ting , Zimmerman Mark W. , Durbin Adam D. , Weichert Nina , Ki Dong H. , Look A Thomas 
Published: July 2016

Abstract
By high-throughput sequencing of cancer patient's genomes, driver mutations have recently been described in high-grade pediatric glioma, an aggressive type of brain tumor in children. The most prominent recurrent mutations were found in four genes, namely NF1, TP53, ATRX and H3F3A. NF1 and TP53 are tumor suppressor genes and their loss of function is characteristic for various human malignancies. However, the mechanisms through which precise missense mutations in H3F3A and loss-of-function of ATRX influence tumor biology remain largely unknown. In our project, we employ the genome-editing technology CRISPR-Cas9 to develop a zebrafish model of pediatric high-grade glioma, in order to examine the impact of these mutations on tumor onset and progression. We previously created a zebrafish line that is deficient for tp53 (tp53-/-) and has a loss-of-function mutation in 3 of 4 alleles of nf1 (nf1a+/-, nf1b-/-), which is duplicated in zebrafish. These fish develop high-grade gliomas with low penetrance and malignant peripheral nerve sheath tumors (MPNSTs) with high penetrance. By CRISPR-Cas9 we further incorporated early frameshift mutations of atrx in the background of tp53-/-, nf1a+/-, nf1b-/-. All analyzed fish injected with Cas9-mRNA and CRISPR guide-RNAs (gRNAs) targeting atrx coding sequences showed germline transmission of mutant alleles, of which one third were frameshift mutations. Primary CRISPR-Cas9 atrx-gRNA injected tp53-/-, nf1a+/-, nf1b-/- fertilized eggs developed as mosaics and exhibited increased tumor penetrance and faster onset of MPNSTs and these tumors harbored atrx mutations in 8/10 analyzed samples. This indicates that atrx loss-of-function mutations cooperate with loss of tp53 and nf1 tumor suppressors. Gliomas were not observed in mosaic atrx mutated fish, and these will be reanalyzed in stable atrx mutant lines. Next, we will add previously described missense mutations of h3f3a. To unravel the synergy between oncogenic mutations in atrx and h3f3a in tumor progression, mutant stable lines for both genes will be investigated independently and in a combined manner. The zebrafish model of pediatric high-grade gliomas and MPNSTs that we are developing will be useful to dissect the mechanisms underlying cooperation among driver mutations and for small molecule screens to identify specific inhibitors of cell growth and survival in these malignancies.
Citation Format: Felix Oppel, Shuning He, Ting Tao, Mark W. Zimmerman, Adam D. Durbin, Nina Weichert, Dong H. Ki, A Thomas Look. Using a genome-editing approach for the stepwise establishment of zebrafish models of pediatric high-grade gliomas and MPNSTs. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 681.
Abstract 686: MNK1 promotes the progression of breast ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC)
Authors: Guo Qianyu , del Rincon Sonia V. , Goncalves Christophe , Miller Wilson H. 
Published: July 2016

Abstract
The mechanism by which breast ductal carcinoma in situ (DCIS) progresses to invasive ductal carcinoma (IDC) is poorly described. Previous studies have revealed that DCIS and IDC share similar genomes and transcriptomes. We hypothesize that one mechanism contributing to the DCIS to IDC conversion involves aberrations in mRNA translation. The Mnk/eIF4E axis has a critical role in promoting the translation of tumor promoting and pro-invasive mRNAs, thus we propose to study whether aberrant activation of this axis promotes the transition of DCIS to IDC. Our model system involved overexpressing constitutively active Mnk1 (caMNK1) in MCF10 variants: (1) MCF10A immortalized mammary epithelial cells and (2) MCFDCIS.com ductal carcinoma in situ cells (termed DCIS cells). Our in vitro results suggest that caMNK1 enables DCIS cells to gain invasive properties. First, overexpression of caMnk1 increased colony formation, larger acinar size in 3D culture, and promoted the migration and invasion of MCF10A and MCFDCIS cells. Second, SEL201, a novel Mnk1/2 inhibitor, can inhibit proliferation, colony formation, 3D acini formation, and the migration and invasion capacity of DCIS-pBABE and -caMNK1 expressing cells. Tumor xenografting in athymic nude mice revealed that caMNK1 facilitates the conversion of DCIS to IDC as the DCIS-caMNK1 expressing tumors progress to IDC, while DCIS-pBABE derived tumors retain DCIS-like structures. Breast tumor samples from patients with DCIS, IDC or mixed DCIS-IDC will also be obtained to examine the levels of phospho-Mnk1 and phospho-eIF4E.
Citation Format: Qianyu Guo, Sonia V. del Rincon, Christophe Goncalves, Wilson H. Miller. MNK1 promotes the progression of breast ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC). [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 686.
Abstract 693: Oncogenic and osteolytic function of histone demethylase NO66 in castration resistant prostate cancer
Authors: Bagheri-Yarmand Rozita , Navone Nora , Wan Xinhai , Logothetis Christopher , Huard Johnny , Sinha Krishna 
Published: July 2016

Abstract
Androgen ablation by pharmacological use or castration is a standard therapy for treating prostate cancer (PCa), however, at the later stage PCa becomes castration-resistant and is metastasized primarily to bones. Epigenetic changes that cause dysregulated gene expression during progression and metastasis of androgen-independent prostate cancer are not clearly understood. We previously reported that the jumonji C domain-containing NO66 is a histone demethylase specific for H3K4me3 and H3K36me3, which negatively regulates osteoblast differentiation and bone formation. By analyzing a multi-cancer tissue microarray, the serendipitous observation was made that NO66 is overexpressed in a variety of cancers. Our objective was to examine NO66's role in the pathogenesis of prostate cancer and bone metastasis.
Our findings indicate that NO66 knockdown resulted in significant decreases in the proliferation and invasiveness of prostate cancer cells (PC3 and DU145) while overexpressing NO66 promoted their proliferation and invasion. Furthermore, NO66 depletion resulted in a two- to three fold decrease in colony formation and anchorage-independent growth of PCa cells suggesting an oncogenic function of NO66. RNAseq analysis comparing between control and NO66-depleted DU-145 cells revealed a repertoire of differentially expressed genes. NO66 up-regulates genes with proliferative activity such as chemokine CXCL5, growth factor IGFBP5, IL6, WNT and MAPK4 pathway, which are known to drive PCa proliferation. Furthermore, ChIP sequencing for NO66 and H3K9AC in NO66-overexpressing PC3 cells revealed that interaction sites of NO66 overlap with peaks of H3K9AC in majority of the genes. These data clearly indicate that NO66 activates a cohort of genes which are involved in tumor progression and metastasis. Strikingly, implanting NO66-overexpressing prostate cancer cells into the femur of SCID male mice caused massive femoral bone loss after three weeks, which suggests that NO66 causes osteolytic bone lesions similar to the clinico-pathological features of advanced PCa. Finally, analysis of tumor xenografts derived from patients by immunohistochemistry showed high expression of NO66 in high grade advanced PCa compared to low grade and hyperplasia suggesting that NO66 may be used as a biomarker to distinguish aggressive prostate cancer from indolent stage. Our data suggest that NO66 plays an important role in cell proliferation, cell invasion and bone metastasis of prostate cancer.
Citation Format: Rozita Bagheri-Yarmand, Nora Navone, Xinhai Wan, Christopher Logothetis, Johnny Huard, Krishna Sinha. Oncogenic and osteolytic function of histone demethylase NO66 in castration resistant prostate cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 693.
Abstract 695: SHP-2-upregulated ZEB1 is important for PDGFRá-driven glioma epithelial-mesenchymal transition and invasion in mice and humans
Authors: Zhang Lei , Zhang Weiwei , Li Yanxin , Li Zuoqing , Wang Yinfang , Song Lina , Lv Deguan , Nakano Ichiro , Hu Bo , Cheng Shi-Yuan , Feng Haizhong 
Published: July 2016

Abstract
A critical clinical challenge in glioblastoma therapy is robust tumor growth and invasion driven by aberrant activation of oncogenic tyrosine receptor kinases (RTKs) including PDGFRá. Here, we report that a SHP-2-upregulated epithelial-mesenchymal transition (EMT)-inducer, ZEB1 is important for PDGFRá-driven glioma EMT, invasion and glioma stem cell renewal in mice and humans. ZEB1 and activated PDGFRá were co-expressed in invasive regions of mouse glioma xenografts and human clinical glioma specimens. Glioma patients with high levels of both p-PDGFRá and ZEB1 had significantly shorter overall survival compared with those with low expression of p-PDGFRá and ZEB1. Knockdown of ZEB1 inhibited PDGF-A/PDGFRá-stimulated glioma EMT, tumor growth, invasion and glioma stem cell renewal. PDGFRá mutant deficient of SHP-2 binding (PDGFRá-F720) or PI3K binding (PDGFRá-F731/42), knockdown of SHP-2 or treatments of pharmacological inhibitor for PDGFRá-signaling effectors attenuated PDGF-A/PDGFRá-stimulated ZEB1 expression, cell migration and glioma stem cell proliferation. Importantly, SHP-2 acts together with PI3K/Akt to regulate a ZEB1-miR-200 feedback loop in PDGFRá-driven gliomas. Together, our findings uncover a new pathway in which ZEB1 functions as a key regulator for PDGFRá-driven glioma EMT, invasion and glioma stem cell renewal, suggesting that ZEB1 as a potential therapeutic target for human gliomas with high PDGFRá activation.
Citation Format: Lei Zhang, Weiwei Zhang, Yanxin Li, Zuoqing Li, Yinfang Wang, Lina Song, Deguan Lv, Ichiro Nakano, Bo Hu, Shi-Yuan Cheng, Haizhong Feng. SHP-2-upregulated ZEB1 is important for PDGFRá-driven glioma epithelial-mesenchymal transition and invasion in mice and humans. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 695.
Abstract 719: ICAM3 and CCL16, inflammation genes screened by high-throughput siRNA library, govern cancer stemness in malignant tumors
Authors: Shen Wenzhi , Xie Junling , Du Renle , Jiang Shan , Luo Xiaohe , He Huiwen , Xiang Rong , Luo Yunping 
Published: July 2016

Abstract
Cancer stem cells (CSCs) have the ability of both self-renewal and differentiation, were thought to be the main root of tumorigenesis, metastasis, recurrence and drug resistance. Current studies have focused on the maintenance of cancer stem cell abilities. Moreover, existing studies have shown that tumor associated inflammation microenvironment (TIM) play a pivotal role in different stages of tumor development. The linkage between inflammation and tumor stemness has been established. However, the evidence of TIM function on maintaining and nourishing CSCs properties remains insufficient. Here, we performed a high throughput siRNA interference platform to systemically screen out the inflammatory genes that regulate tumor stemness via OCT4 promoter linked reporter system in HMLE-Snail cells. 10 candidates were screened out which knockdown could both down-regulate OCT4 expression and decrease ALDH+ subpopulation. Furthermore, we validated the function of genes ICAM3 and CCL16 and found that knockdown ICAM3 or CCL16 inhibit cell migration, sphere formation in 231, A549 and HepG2 cell lines. We also found ICAM3 or CCL16 deficiency decrease side population and reduce chemo-resistance. In addition, mice bearing ICAM3 or CCL16 knockdown breast cancer cells develop smaller tumors and less lung metastases verse controls. Taken together, our study demonstrated that the inflammation related genes ICAM3 and CCL16 govern cancer cell migration and stemness in malignant tumors in vitro and in vivo, and that will provide new possible targets to CSCs for cancer therapy.
Citation Format: Wenzhi Shen, Junling Xie, Renle Du, Shan Jiang, Xiaohe Luo, Huiwen He, Rong Xiang, Yunping Luo. ICAM3 and CCL16, inflammation genes screened by high-throughput siRNA library, govern cancer stemness in malignant tumors. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 719.
Abstract 72: Therapeutic compound discovery targeting a recurrent glioblastoma (GBM) phenotype using LINCS compounds via QUADrATiC analyses
Authors: Al Rashid Shahnaz T. , O’Reilly Paul , Dunne Philip D. , Alderdice Matthew , Flannery Thomas J. , Prise Kevin M. , Zhang Shu-Dong , McArt Darragh G. 
Published: July 2016

Abstract
Glioblastoma (GBM) is the most prevalent and aggressive of malignant primary brain tumours. Despite multi-modal therapy, including surgical resection, radiotherapy and chemotherapy, GBM continues to be uniformly lethal, with a median survival of 12-18 months from the time of diagnosis. The resistance to therapy and ultimate recurrence is thought to originate from a sub-population of cells residing in the initial tumour, identified as glioma-initiating or glioma-stem cells (GSC). These GSC have been shown to be intrinsically more invasive, with enhanced repair and drug-efflux pathways mediating resistance to chemo-radiation therapies. The purpose of this study was to use and validate a novel connectivity mapping procedure to identify candidate compounds targeting therapy-resistant and recurrent GBM. This procedure was developed by O’Reilly and colleagues (“QUADrATiC: A Scalable Approach to Repurposing FDA-approved therapeutics”, under review in Bioinformatics), and in this study, was performed on publicly available NCBI GEO microarray datasets.
Based on the cancer stem cell theory whereby the resistant GSCs repopulate and are enriched in recurrent tumours, primary and recurrent GBM gene expression profiles were compared and the connections between the identified gene lists and drug responses were mapped. To validate that this connectivity mapping procedure is reliable at identifying effective treatments to target therapeutic resistance and recurrence, we tested one of the top ten compounds identified, rosiglitazone (rosi), a ligand of the peroxisome proliferator activated receptor gamma (PPARγ). The efficacy of rosi alone and in combination with the current standard of care therapies, temozolomide and radiation, at killing GSC was assessed in vitro using the clonogenic survival assay. Our data indicated that rosi significantly reduced survival of both non-stem glioma cells and GSC, alone and in combination with radiation and temozolomide. The p53 pathway was identified as a pathway by which rosi may induce permanent cell cycle arrest and cell death. Further work is ongoing to fully elucidate the downstream mechanisms of cell death. This work is proof-of-principle that the current connectivity mapping procedure can be instrumental in identifying more targeted therapeutic interventions for GBM resistance and prevent recurrence.
Citation Format: Shahnaz T. Al Rashid, Paul O’Reilly, Philip D. Dunne, Matthew Alderdice, Thomas J. Flannery, Kevin M. Prise, Shu-Dong Zhang, Darragh G. McArt. Therapeutic compound discovery targeting a recurrent glioblastoma (GBM) phenotype using LINCS compounds via QUADrATiC analyses. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 72.