bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5156: RhoC decreases tight junction stabilization in breast cancer cells, revealing a potential therapeutic target
Authors: Cheriyan Hannah , Wu Zhi-Fen , Yates Joel , Merajver Sofia D. 
Published: August 2018

Abstract
RhoC, a member of the Rho GTPase family, has been shown to have a role specifically in facilitating metastasis of breast cancer cells1, however no conclusive mechanism for how RhoC promotes metastasis currently exists. We demonstrate that CRISPR-Cas9 knockout of RhoC in SUM149 and MDA-MB231 cells results in increased epithelial morphology, cell-cell connections, and tight junction marker (ZO-1, occludin, PATJ/PALS1/CRB3) colocalization compared to wild type cells, as studied via immunofluorescent staining. These results suggest that high expression of RhoC in cancer cells destabilizes tight junctions, which could allow for the increased metastatic potential observed with RhoC-overexpressing cells in prior studies2-4. Ongoing work investigates whether these morphological and molecular changes are observed in other breast cancer cell lines, such as SUM190 and VARI-068, a low-passage PDX-derived triple negative breast cancer cell line. Furthermore, we will determine whether RhoC knockout in these cells lines actively modulates tight junction stability as measured via transepithelial electrical resistance (TEER). Taken together, these data support an important role for RhoC in tight junction modulation, thereby revealing a potential highly specific new therapeutic target in RhoC-driven metastases control.
References
1. Hakem, A. et al. RhoC is dispensable for embryogenesis and tumor initiation but essential for metastasis. Genes Dev. 19, 1974-1979 (2005).
2. van Golen, K. L., Wu, Z. F., Qiao, X. T., Bao, L. W. & Merajver, S. D. RhoC GTPase, a novel transforming oncogene for human mammary epithelial cells that partially recapitulates the inflammatory breast cancer phenotype. Cancer Res. 60, 5832-8 (2000).
3. van Golen, K. L., Wu, Z. F., Qiao, X. T., Bao, L. & Merajver, S. D. RhoC GTPase overexpression modulates induction of angiogenic factors in breast cells. Neoplasia 2, 418-25 (2000).
4. Wu, M., Wu, Z. F., Rosenthal, D. T., Rhee, E. M. & Merajver, S. D. Characterization of the roles of RHOC and RHOA GTPases in invasion, motility, and matrix adhesion in inflammatory and aggressive breast cancers. Cancer 116, 2768-2782 (2010).
Citation Format: Hannah Cheriyan, Zhi-Fen Wu, Joel Yates, Sofia D. Merajver. RhoC decreases tight junction stabilization in breast cancer cells, revealing a potential therapeutic target [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 5156.
Abstract 5164: Mechanosensor MYL9 regulates cancer cell malignancy in gastrointestinal tumors
Authors: Konno Masamitsu , Yanagisawa Kiminori , Matsushita Katsunori , Asai Ayumu , Koseki Jun , Matsusaki Michiya , Deguchi Shinji , Doki Yuichiro , Mori Masaki , Ishii Hideshi 
Published: August 2018

Abstract
Although the stiffness of tissues likely is involved in the malignant behavior of tumors, it remains to be clarified which molecules control the nature, how it is involved in the invasiveness of tumors, or whether any marker is available for the prediction of cancer patient prognosis. In the present study, we studied the role of Myl9, a non-muscle-type, myosin light chain by the experiment in vitro, and assessed the usefulness in the stratification of patients in vivo as the precision medicine. Given that Myl9 is involved in the contraction of cell skeleton, cell hardness, and alterations of cell morphology in various tissues, we first examined whether the expression of the Myl9 is associated with the clinical status of tumors by immunohistochemistry. The results of 45 cases with colon cancer and pancreatic cancer indicated that the increased expression of Myl9 is associated significantly with a reduced provability of overall survival and disease-free survival of those cancers. Moreover, we noted the differential expression of Myl9 in epithelial cancer cells and mesenchymal fibroblasts, i.e., the accumulation of Myl9 staining in cell nuclei of fibroblasts. The experiment of 3-dimensional culture with cancer cells and fibroblasts confirmed the results. Furthermore, we investigated whether Myl9 overexpression is involved in the biologic behavior of gastrointestinal cancer cells. The results showed that the rentiviral-mediated overexpression of Myl9 resulted in an increase of cell proliferation and invasion as well as tumorigenicity in mice. The present study indicates that Myl9 protein can play a fundamental role in the malignant behaviors of gastrointestinal cancer cells.
Citation Format: Masamitsu Konno, Kiminori Yanagisawa, Katsunori Matsushita, Ayumu Asai, Jun Koseki, Michiya Matsusaki, Shinji Deguchi, Yuichiro Doki, Masaki Mori, Hideshi Ishii. Mechanosensor MYL9 regulates cancer cell malignancy in gastrointestinal tumors [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 5164.
Abstract 5170: The effect of vitamin K on aggressiveness, lipid metabolism and gene expression in triple-negative breast cancer cells
Authors: Kokabee Leila , Beaudin Sarah , Welsh JoEllen 
Published: August 2018

Abstract
Vitamin K is an essential cofactor in the γ-carboxylation of glutamate (Glu) to γ-carboxyglutamate (Gla), a post-translational modification mediated by the gamma-glutamyl carboxylase enzyme (GGCX) and vitamin K oxidoreductases (VKORC1 or VCORC1L1). All three of these genes are expressed in most breast cancer cell lines and The Cancer Genome Atlas (TCGA) data indicate that upregulation of these genes is associated with reduced survival in breast cancer patients. Since vitamin K is not typically present in cell culture media, γ-carboxylation is limited in vitro despite the presence of GGCX and VKORs. To determine the functional significance of the vitamin K pathway in breast cancer cells, we assessed the effect of vitamin K treatment on cell survival, migration and mammosphere formation. SUM159 and Hs578T cells were maintained in vitamin K1 (5 ug/mL) for at least 3 passages for all assays. Cells passaged with vitamin K produced γ-carboxylated proteins and exhibited increased cell survival after 72h, increased mammosphere formation after 8 days and increased cell migration after 48h compared to cells passaged in ethanol vehicle. Inhibition of VKORC1 with 2 uM warfarin decreased the production of γ-carboxylated proteins and reduced the effects of vitamin K on cell survival and migration but not on mammosphere formation. Microarray profiling and pathway analysis indicated upregulation of several known oncogenic pathways, including PI3K-AKT, VEGF-VEGFR, RANK-RANKL and JAK/STAT in vitamin K-treated SUM159 and Hs578T cells. Thus, γ-carboxylation of proteins driven by vitamin K might indirectly activate these pathways to enhance cell survival, cell migration and mammosphere formation. Additional pathways upregulated in vitamin K-treated cells included adipogenesis, fatty acid biosynthesis and lipid metabolism. We therefore examined lipid droplet formation in cells exposed to vitamin K using Bodipy assays. We detected an increase in lipid droplet formation after 72 hours of vitamin K treatment, which was inhibited by warfarin. Since aggressive breast cancers are often associated with accumulation of cytoplasmic lipid droplets, further studies on the mechanism by which vitamin K increases lipid accumulation are warranted. In conclusion, our data indicate that vitamin K enhances γ-carboxylation, breast cancer aggressiveness and lipid metabolism, suggesting that blocking the vitamin K pathway might be of therapeutic value for patients whose tumors express GGCX and VKORs.
Citation Format: Leila Kokabee, Sarah Beaudin, JoEllen Welsh. The effect of vitamin K on aggressiveness, lipid metabolism and gene expression in triple-negative breast 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 5170.
Abstract 5177: Metastasis: Leveraging transcriptomics to identify potential therapeutics
Authors: Ung Matthew , Funt Jason M. , Lysaght Andrew C. , Zhang Jenny , Escalante-Chong Renan , Koytiger Gregory , Kolitz Sarah , Kusko Rebecca , Zeskind Benjamin , Fowler Kevin D. 
Published: August 2018

Abstract
Five-year survival rates for patients whose cancer has metastasized are dramatically lower than for those with localized disease. However, few therapeutic development efforts specifically focus on inhibiting the process of metastasis.
Here we perform a systematic pan-cancer analysis of RNA-seq data from The Cancer Genome Atlas to identify targetable gene expression changes in primary tumors that confer metastatic potential. In 4,844 patients comprising 13 different cancer types, we construct a gene expression signature that captures consistent pan-cancer alterations that differ between primary tumors associated with node-negative disease with no recorded distant metastasis (N0 not M1) and primary tumors from patients with node-positive disease (N1, N2, or N3). In order to identify potential therapeutic targets, we query this metastasis signature against drug-induced transcriptomic signatures from CMAP and LINCS. Among the top candidates is Chembl410456, which modulates the TGFβ and Wnt/β-catenin signaling pathways, among others. Genes upregulated in metastatic primary tumors and downregulated by Chembl410456 include SERPINE1 and TWIST1. We observe that low SERPINE1 and TWIST1 expression also associate with improved prognosis in multiple cancers, including colon adenocarcinoma (p=0.007 and p=0.014, respectively). Other top-ranked drugs also target the TGFβ signaling pathway. To summarize, we leverage public RNA-seq data to identify a pan-cancer metastatic signature that can be interrogated to systematically identify potential therapeutics that target metastasis. Building on this, we pinpoint antimetastasis drug candidates including several with mechanisms of action involving modulation of TGFβ and/or Wnt/β-catenin pathways.
Citation Format: Matthew Ung, Jason M. Funt, Andrew C. Lysaght, Jenny Zhang, Renan Escalante-Chong, Gregory Koytiger, Sarah Kolitz, Rebecca Kusko, Benjamin Zeskind, Kevin D. Fowler. Metastasis: Leveraging transcriptomics to identify potential therapeutics [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 5177.
Abstract 5179: Uncovering the signaling landscape controlling breast cancer cell migration identifies splicing PRPF4B as a metastasis driver
Authors: Water Bob van de , Danen Erik , Koedoot Esmee , Rogkoti Maria , Fokkelman Michiel , Devedec Sylvia Le , Martens John , Stoilov Peter , Foekens John 
Published: August 2018

Abstract
Metastasis is the major cause of death in cancer patients and migration of cancer cells from the primary tumor to distant sites is the prerequisite of metastasis formation. Here we applied an imaging-based RNAi phenotypic cell migration screen uing two highly migratory basal breast cancer cell lines (Hs578T and MDA-MB-231 to provide a repository of candidate metastasis drug targets. We screened ~4,200 individual target genes covering most cell signaling components. We discovered 133 and 113 migratory modulators of Hs578T and MDA-MB-231, respectively, of which 45 genes were common denominators of cell migration. Interaction networks of candidate migratory modulators were in common with networks of different clinical breast cancer prognostic and metastasis classifiers. The splicing factors PRPF4B and BUD31 and the transcription factor BPTF were amplified in human primary breast tumors and the expression was associated with metastasis-free survival. Depletion of PRPF4B, BUD31 and BPTF caused primarily downregulation of genes involved in focal adhesion and ECM-interaction pathways. PRPF4B was essential for TNBC cell migration and critical for breast cancer metastasis formation in vivo, making PRPF4B a candidate for further drug development. Our systematic phenotypic screen provides an important repository for signaling determinants that functionally drive cancer cell migration.
Citation Format: Bob van de Water, Erik Danen, Esmee Koedoot, Maria Rogkoti, Michiel Fokkelman, Sylvia Le Devedec, John Martens, Peter Stoilov, John Foekens. Uncovering the signaling landscape controlling breast cancer cell migration identifies splicing PRPF4B as a metastasis driver [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 5179.
Abstract 5188: MDM2 and MDMX promote p53-independent initiation of circulating tumor cells from triple negative breast cancers
Authors: Gao Chong , Xiao Gu , Piersiga Ale , Gou Jiangtao , Ogunwobi Olorunseun , Bargonetti Jill 
Published: August 2018

Abstract
Many human breast cancers overexpress the E3 ubiquitin ligase MDM2 and its homolog MDMX. The Cancer Genome Atlas (TCGA) shows that high MDM2 and MDMX expression correlates with both estrogen receptor positive (ER+) and triple negative breast cancer (TNBC) subtypes. Overexpression of MDM2 and MDMX can drive breast cancer progression through molecular pathways that do not require wild-type p53 degradation. ER positive breast cancers often express wild-type p53 but 80% of TNBC express mutant p53 (mtp53). It has not been determined how MDM2 and MDMX expression in different sub-types of breast cancer, with mutant p53, influence proliferation and metastasis. We compared MDM2 and MDMX-dependent biological outcomes for mtp53 expressing ER+ T47D (mtp53 L194F) and TNBC cell type MDA-MB-231 (mtp53 R280K). In the ER+ cells no change was detected for real time cell migration or E-cadherin protein levels when MDM2 was knocked down by stable shRNA expression but cell proliferation was reduced. In support of this in vitro data, these MDM2 knockdown cells tested in an orthotopic xenograft model in NSG mice demonstrated no MDM2-mediated change in primary tumor invasiveness but did show a reduced tumor volume. Importantly, in the TNBC MDA-MB-231 cells, knocking down either MDM2 or MDMX did not reduce cell proliferation in vitro but greatly blocked cell migration. In support of this in vitro data, these MDA-MB-231 cells with either MDM2 or MDMX knockdown in NSG mice did not show a reduced primary tumor volume but showed a significant inhibition of the production of circulating tumor cells (CTCs). Our data shows that in different subtypes of breast cancers, MDM2 and MDMX can promote alternative p53-independent biological outcomes. The estrogen-activated MDM2 strongly promotes cell proliferation while MDM2 and MDMX overexpression in TNBC promotes circulating tumor cells and potentially metastasis. Understanding the alternative p53-independent roles of MDM2 and MDMX in ER+ and triple negative breast cancers will provide insight for the development of MDMs-targeted theranostics.
Citation Format: Chong Gao, Gu Xiao, Ale Piersiga, Jiangtao Gou, Olorunseun Ogunwobi, Jill Bargonetti. MDM2 and MDMX promote p53-independent initiation of circulating tumor cells from triple negative breast cancers [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 5188.
Abstract 5199: Microenvironmental control of breast cancer subtype elicited by paracrine platelet-derived growth factor-CC signaling
Authors: Bocci Matteo , Bartoschek Michael , Roswall Pernilla , Pietras Kristian 
Published: August 2018

Abstract
Aim: Breast tumors of the basal-like, hormone receptor-negative subtype, remain an unmet clinical challenge, as patients exhibit the highest recurrence rate, the shortest time to recurrence and the worst overall survival rate due to a paucity of therapeutic targets. Also, as tumors develop, the surrounding microenvironment co-evolves into an activated state through continuous paracrine communication, creating a dynamic signaling circuitry that promotes malignant progression. Thus, novel treatment approaches that also include the targeting of stromal components are urgently required.
Methods: We used experimental mouse models of cancer (transgenic, syngeneic, human cell lines and patient-derived xenografts) for knockout studies and preclinical trials, immunostaining, in vitro assays, western-blot, gene expression analysis and RNA-sequencing, patient cohorts, data mining and bioinformatics analysis.
Results and discussion: We identified a paracrine crosstalk between cancer cells expressing platelet-derived growth factor (PDGF)-CC and cancer-associated fibroblasts (CAFs) expressing the cognate receptors in human basal-like mammary carcinomas. In translational efforts, we found that PDGF-CC expression in the malignant epithelium of breast cancer was associated with a hormone receptor-negative state. We then generated a mouse model to characterise the functional role of PDGF-CC in tumorigenesis. Moreover, we identified and validated factors secreted by CAFs to maintain a basal-like phenotype, e.g. Stanniocalcin (STC)-1, Hepatocyte Growth Factor (HGF) and Insulin Growth Factor Binding Protein (IGFBP)-3. Genetic or pharmacological targeting of PDGF-CC in experimental mouse models resulted in the transcriptional reprogramming and conversion of basal-like breast tumors into a luminal-like state characterised by Estrogen Receptor (ER)-α positivity, that conferred sensitivity to endocrine therapy in previously intrinsically unresponsive tumors.
Conclusions: Specification of the basal-like subtype of breast cancer is under microenvironmental control and therapeutically actionable in order to achieve sensitivity to endocrine therapy. Our work sheds light on previously unknown mechanisms that define breast cancer subtype and has significant therapeutic implications for patients with basal-like breast tumors.
Citation Format: Matteo Bocci, Michael Bartoschek, Pernilla Roswall, Kristian Pietras. Microenvironmental control of breast cancer subtype elicited by paracrine platelet-derived growth factor-CC signaling [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 5199.
Abstract 52: A genomic editing approach for purification of viable quiescent cancer cells
Authors: La Ting , Farrelly Margaret , Yan Xu Guang , Yari Hamed , Zhang Yuanyuan , Feng Yuchen , Tabatabaeehatambakhsh Seyed Hessamedin , Liu Tao , Jin Lei , Zhang Xu Dong 
Published: August 2018

Abstract
Recurrent and metastatic cancer lesions often undergo a period of dormancy, which is closely associated with cancer cell quiescence, a state whereby cancer cells exit the cell cycle and are reversibly arrested in G0 phase. Quiescent cancer cells are inherently resistant to cell death and refractory to therapeutic drugs. However, the mechanisms responsible for the resistance remain largely undefined. This is closely associated with the lack of understanding of their biological properties as a consequence of technical hurdles in the isolation and analysis of viable quiescent cells. Nevertheless, it is known that quiescent cells are characteristically negative for the proliferation marker Ki67 and express high levels of the cyclin-dependent kinase (CDK) inhibitor p27. In this study, we developed a CRISPR/Cas9-based system to fuse a green fluorescent protein (EGFP) gene with endogenous CDKN1B, the gene encoding p27, and a red fluorescent protein (mCherry) gene with endogenous MKI67, the gene encoding Ki67 in the genome of human melanoma cells. By using this system, we have successfully isolated viable p27high/Ki67- melanoma cells using fluorescence-activated cell sorting (FACS). The quiescent state of these cells was confirmed by dual nucleic acid staining (DNA with Hoechst-33342, and RNA with Pyronin Y). This approach provides a unique research tool for further characterization of the biological properties of quiescent cells. Comparative RNA-seq and proteomics analyses are being currently carried out to reveal molecular networks involved in regulating cell quiescence, and to identify molecular targets for overcoming resistance of quiescent melanoma cells to treatment.
Citation Format: Ting La, Margaret Farrelly, Xu Guang Yan, Hamed Yari, Yuanyuan Zhang, Yuchen Feng, Seyed Hessamedin Tabatabaeehatambakhsh, Tao Liu, Lei Jin, Xu Dong Zhang. A genomic editing approach for purification of viable quiescent 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 52.
Abstract 5214: Tumor cell-derived complement components C1r and C1s promote growth of cutaneous squamous cell carcinoma
Authors: Riihilä Pilvi , Nissinen Liisa , Farshchian Mehdi , Kallajoki Markku , Kivisaari Atte , Meri Seppo , Grénman Reidar , Peltonen Sirkku , Peltonen Juha , Kähäri Veli-Matti 
Published: August 2018

Abstract
The incidence of cutaneous squamous cell carcinoma (cSCC) is increasing worldwide. We have analyzed the role of complement components C1r and C1s in the progression of cSCC. Analysis of cSCC cell lines (n=8) and normal human epidermal keratinocytes (NHEK)(n=11) with oligonucleotide arrays, RNA-seq and quantitative RT-PCR revealed significantly elevated C1r and C1s mRNA levels in cSCC cell lines. Western blot analysis showed increased production of C1r and C1s in cSCC cells compared to NHEKs. The mRNA levels for C1r and C1s were markedly elevated in cSCC tumors (n=6) compared to normal skin (n=11) in vivo. Immunohistochemical analysis for C1r and C1s revealed strong tumor cell specific expression of C1r and C1s in invasive sporadic cSCCs (n=164) and recessive dystrophic epidermolysis bullosa-associated cSCCs (n=16), whereas the expression of both was clearly lower in cSCC in situ (n=63), in premalignant epidermal lesions (actinic keratoses, n=61), and in normal skin (n=9). Knockdown of C1r and C1s expression with two specific siRNAs inhibited proliferation of cSCC cells and growth of human cSCC xenograft tumors in vivo. These results provide evidence for the role of tumor cell-derived C1r and C1s in the progression of cSCC independently of C1q and identify these serine proteinases as putative therapeutic targets in unresectable and metastatic cSCC.
Citation Format: Pilvi Riihilä, Liisa Nissinen, Mehdi Farshchian, Markku Kallajoki, Atte Kivisaari, Seppo Meri, Reidar Grénman, Sirkku Peltonen, Juha Peltonen, Veli-Matti Kähäri. Tumor cell-derived complement components C1r and C1s promote growth of cutaneous squamous cell carcinoma [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 5214.