bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2025: CD147/EMMPRIN mediates epithelial-mesenchymal transition of head and neck squamous cell carcinoma
Authors: Suzuki Shinsuke
Published: August 2018

Epithelial-mesenchymal transition (EMT) is a key mechanism in the progression of solid tumors. This process is characterized by the development of more migratory, invasive cell behavior. Transforming growth factor-β (TGF-β) has been well characterized as a potent inducer of EMT. CD147, also known as extracellular matrix metalloproteinase inducer (EMMPRIN), a member of the immunoglobulin superfamily, is highly expressed in cancer cells and induces malignant characteristics in various tumors, including head and neck squamous cell carcinoma (HNSCC). Accumulating evidence indicates a contribution of CD147/EMMPRIN to tumorigenesis of solid tumors. However, the function of CD147/EMMPRIN in EMT and TGF-β-induced tumorigenicity in HNSCC has not yet been explored. To determine the role of CD147/EMMPRIN in EMT and TGF-β-induced tumorigenic behavior, SAS, a tongue squamous cell carcinoma cell line, was stimulated with TGF-β. After stimulation with TGF-β, we found that SAS cells acquired a spindle-shaped morphology. Immunoblotting analysis showed that the expression of vimentin was upregulated and E-cadherin was downregulated after TGF-β treatment. This result suggests that EMT was induced by TGF-β in SAS cells. In addition, expression of CD147/EMMPRIN was upregulated in SAS cells using TGF-β treatment. TGF-β stimulation enhanced the migratory ability of the cells. Furthermore, the TGF-β-induced migratory ability, and the change in cellular morphology was diminished by CD147/EMMPRIN knockdown. These results suggest that CD147 is involved in TGF-β-induced EMT and the migratory capacity of HNSCC.

Citation Format: Shinsuke Suzuki. CD147/EMMPRIN mediates epithelial-mesenchymal transition of head and neck 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 2025.
Abstract 2028: ZEB2 promotes cancer cell survival and angiogenesis by cooperating with transcription factor Sp1
Authors: Ko Dongjoon
Published: August 2018

The metastatic cascade essential for tumor progression consists of local invasion, intravasation, extravasation, micrometastasis, cancer cell survival in the circulation, and metastatic colonization. In particular, epithelial-mesenchymal transition (EMT) has been demonstrated a critical process whereby epithelial cells undergo molecular changes to acquire mesenchymal phenotypes through upregulation of EMT-inducing transcription factors. We previously showed that ZEB2, through cooperation with the transcription factor Sp1, plays an important role in the transcriptional upregulation of mesenchymal genes such as integrin α5 and vimentin during cancer cell invasion. Here, we show that ZEB2 cooperates with Sp1 for promoting not only EMT and invasion but also diverse cellular functions during metastasis. ZEB2 induces expression of Sp1-regulated genes such as VEGF, survivin, bcl-2 and cyclin D1 in an Sp1-dependent manner. Collaboration between ZEB2 and Sp1 induces cancer cell survival and proliferation, endothelial cell activation, and tumor angiogenesis. Furthermore, Sp1 stabilizes ZEB2 protein, supporting the presence of a positive feedback loop between ZEB2 and Sp1. Clinical data show that ZEB2 expression was correlated to Sp1 expression, and that those factors were a crucial for predicting survival in cancer patients. Our findings demonstrate cooperative roles of ZEB2 and Sp1 in regulating cancer cell survival and angiogenesis beyond invasion during cancer progression.

Citation Format: Dongjoon Ko. ZEB2 promotes cancer cell survival and angiogenesis by cooperating with transcription factor Sp1 [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 2028.
Abstract 203: Regenerable altruism drives chemotolerance in clonal cancer cells
Authors: Lee Kee Wah , Masroni Muhammad Sufyan , Tan Karen Meiling , Li Mo-Huang , Zhou Lihan , Tucker Steven , Ngo Lynette Su Mien , Chang Chan Fong , Bay Boon Huat , Tan Soo Yong , Hartman Mikael , Chua Huiwen , Loh Tze Ping , Putti Thomas , Leong Sai Mun , Koay Evelyn Siew-Chuan
Published: August 2018

Introduction: Cancer is thought to represent a breakdown of multicellular cooperation and reversion to Darwinian dynamics characterized principally by self-interested competition. However, there is increasing evidence that cancer cells can behave as communities, manifesting social behaviors that influence cancer progression.

Methods: Circulating tumor cells enrichment, microRNA extraction, qRT-PCR, MTS assays, RNA in situ hybridization, immunohistochemical staining, SILAC-based mass spectrometry, flow cytometry, ChIP-sequencing, and western blotting.

Results: We report here evidences of altruistic cooperation in clonal breast cancer cells. Heterogeneity in expression of a microRNA, miR-125b, leads to sectoring of clonal breast cancer cell populations into minority miR-125b-high and majority miR-125b-low subpopulations. Enhanced population-wide tolerance to taxane-based chemotherapy is mediated by the miR-125b-high minority, in part through increased secretion of extracellular public goods such as insulin-like growth factor binding protein 2 (IGFBP2) and chemokine (C-C motif) ligand 28 (CCL28). Cost-benefit analysis established this helping behavior to be altruistic, as survival benefits conferred to the miR-125b-low cells, via public goods sharing, occurred at a fitness cost to the miR-125b-high cells. Notwithstanding this fitness cost, miR-125b-high altruists regenerate readily from isolated populations of miR-125b-low defectors, via Kruppel-like factor 2-mediated epigenetic mechanism of histone deacetylation. As therapeutic proof-of-principle, we demonstrated that targeting extracellular IGFBP2 and CCL28, through ligand neutralization, markedly blunted the tolerance response of cancer cells towards taxane chemotherapy.

Conclusions: Our results demonstrate how positive social engagement such as altruism may be employed by heterogeneous clonal cancer communities to drive chemotolerance, and understanding of such social behaviors could help formulate more effective treatment strategies.

Citation Format: Kee Wah Lee, Muhammad Sufyan Masroni, Karen Meiling Tan, Mo-Huang Li, Lihan Zhou, Steven Tucker, Lynette Su Mien Ngo, Chan Fong Chang, Boon Huat Bay, Soo Yong Tan, Mikael Hartman, Huiwen Chua, Tze Ping Loh, Thomas Putti, Sai Mun Leong, Evelyn Siew-Chuan Koay. Regenerable altruism drives chemotolerance in clonal 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 203.
Abstract 2034: Traf2-and Nck-interacting kinase (TNIK) inhibitor, NCB-0846, suppresses TGF-β1-induced epithelial-mesenchymal transition in lung cancer
Authors: Sugano Teppei , Masuda Mari , Uno Yuko , Goto Naoko , Seike Masahiro , Sawa Masaaki , Gemma Akihiko , Yamada Tesshi
Published: August 2018

Epithelial mesenchymal transition (EMT) has been implicated in cancer cell migration, invasion, and metastasis. Transforming growth factor-β (TGF-β) signal was known to be an essential EMT promotor. Development of therapeutics for targeting EMT through the TGF-β signal may be beneficial for the management of cancer metastasis. Recently, we found that a Traf2- and Nck-interacting kinase (TNIK) inhibitor, named NCB-0846, was capable of attenuating tumor-initiating cells among human colorectal cancer (1, 2). The cross link between EMT and cancer stemness has been revealed in several studies. Therefore we evaluated whether this small-molecule compound could have efficacy to inhibit TGF-β-induced EMT. NCB-0846 reduced the expression of mesenchymal markers (Vimentin and N-cadherin) and upregulated the expression of epithelial marker E-cadherin in A549 and H2228 cells. NCB-0846 suppressed the phosphorylation and nuclear translocation of Smad proteins and also inhibited migration, invasion, and metastasis. NCB-0846 inhibited TGF-β1-induced EMT through the down-regulation of TGFBR1 in mRNA levels. MiR-186-5p and miR-320 family were identified as candidate miRNAs that could target TGFBR1 and inhibited TGFBR1 protein expression.NCB-0846 might be a novel therapeutics drugs that targets the invasion and metastasis through inhibiting TGF-β-induced EMT in lung cancer.

1. Masuda M, Sawa M, Yamada T. Therapeutic targets in the Wnt signaling pathway: Feasibility of targeting TNIK in colorectal cancer. Pharmacology & therapeutics. 2015;156:1-9. Epub 2015/11/07.2. Masuda M, Uno Y, Ohbayashi N, Ohata H, Mimata A, Kukimoto-Niino M, et al. TNIK inhibition abrogates colorectal cancer stemness. Nature communications. 2016;7:12586. Epub 2016/08/27.

Citation Format: Teppei Sugano, Mari Masuda, Yuko Uno, Naoko Goto, Masahiro Seike, Masaaki Sawa, Akihiko Gemma, Tesshi Yamada. Traf2-and Nck-interacting kinase (TNIK) inhibitor, NCB-0846, suppresses TGF-β1-induced epithelial-mesenchymal transition in lung 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 2034.
Abstract 2046: Structural analysis of VEGFR2 function blocking antibodies
Authors: Wary Anita
Published: August 2018

The vascular endothelial growth factor receptor-2 (VEGFR2) is a receptor tyrosine kinase that is expressed highly in endothelial cells (ECs). By binding to VEGF165 (VEGF-A) VEGFR2 signals in ECs mediate the formation of new blood vessels, a process known as angiogenesis. Growth of tumor and metastasis depends on angiogenic activities of ECs. Thus, an agent that blocks VEGFR2 signaling specifically in tumor ECs could inhibit tumor growth and metastasis. The IMC-1121B is an antiangiogenic agent that inhibits the VEGFR2 signaling. Here we modeled the structure of the 1121B Fab fragment in complex with domain-3 of VEGFR2, and the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain-3. The two Fab fragments interact at opposite ends of VEGFR2 domain-3; 1121B blocks VEGF binding, whereas 6.64 appears to prevent receptor dimerization by perturbing the domain-3:domain-4 interface. Blocking of VEGFR2 signaling activities by two different antibodies could be a powerful approach to block angiogenesis, thereby inhibiting growth of tumor and metastasis.

Key reference: Franklin MC et al., Structure 2011;19:1097-1107.

Citation Format: Anita Wary. Structural analysis of VEGFR2 function blocking antibodies [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 2046.
Abstract 2056: CPAP promotes angiogenesis via interacting with and enhancing the activity of STAT3 in HCC
Authors: Chen Ruo Yu
Published: August 2018

Hepatocellular carcinoma (HCC) is one of the malignant hypervascular tumors in which angiogenesis plays a vital role in tumor growth and metastasis. Our previous studies indicated that CPAP is overexpressed in HCC, and is positively correlated with recurrence and vascular invasion. In this study, we demonstrated that overexpressed CPAP increases tumor growth, angiogenesis, as well as metastasis ex vivo and in vivo. The expression level of CPAP has a positive correlation with activated STAT3 in HCC. Mechanistically, CPAP can act as a transcriptional co-activator of STAT3 to enhance IL-6-induced STAT3 activation by directly binding with STAT3. Overexpressed CPAP up-regulates several target gene expressions of STAT3 that are involved in angiogenesis, such as IL-8, CD44 and MCAM. The expression of CPAP mRNA is positively correlated with the levels of IL-8, CD44 and MCAM mRNAs in HCC. Importantly, IL-6/STAT3-mediated angiogenesis is significantly increased by overexpressed CPAP, and can be blocked by decreased expression of IL-8. Taken together, our results suggest that overexpressed CPAP interacts with STAT3 to enhance its activity and therefore leads to angiogenesis. Our findings shed light on the importance of CPAP to act as a potential therapeutic target in inhibiting the angiogenesis pathway.

Citation Format: Ruo Yu Chen. CPAP promotes angiogenesis via interacting with and enhancing the activity of STAT3 in HCC [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 2056.
Abstract 2060: The pseudogene FLT1P1 functions as a tumor metastasis suppressor in a preclinical model of colorectal cancer
Authors: Ye Xiangcang , Fan Fan , Bhattacharya Rajat , Boulbes Delphine R. , Wang Rui , Xia Ling , Ellis Lee M.
Published: August 2018

Chromosome 3p21.3 is one of the unstable genomic loci that are frequently altered by epigenetic modifications or interstitial deletions in human solid tumors. Several genes in 3p21.3 are known to be tumor suppressors. This locus includes a non-coding gene, fms-related tyrosine kinase 1 pseudogene 1 (FLT1P1), of which the role is largely unknown. Our previous published study (Ye et al. MCR, 2015) found that FLT1P1 was transcribed primarily as an antisense that inhibits VEGFR1 protein expression through interaction with miR-520a in human colorectal cancer (CRC) cells. In this study, we showed that the expression of FLT1P1 antisense is silenced by promoter hyper-methylation in xenograft tumors. Our data further showed that the loss of the FLT1P1 antisense not only up-regulated the expression of VEGFR1 and other genes relating to tumor angiogenesis, but also induced the EMT phenotype in tumor cells. Moreover, the FLT1P1 silenced cells demonstrated an increase in the number of metastasis to the liver following spleen injection in a mouse model. These observations suggest that the transcribed FLT1P1 antisense is a tumor metastasis suppressor. Our finding warrants further investigation of the genetic and epigenetic alterations of FLT1P1 and their impact on metastasis of CRC cells.

Citation Format: Xiangcang Ye, Fan Fan, Rajat Bhattacharya, Delphine R. Boulbes, Rui Wang, Ling Xia, Lee M. Ellis. The pseudogene FLT1P1 functions as a tumor metastasis suppressor in a preclinical model of 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 2060.
Abstract 2062: Selective gene dependencies in MYCN-amplified neuroblastoma include the core transcriptional regulatory circuitry
Authors: Durbin Adam D. , Zimmerman Mark W. , Dharia Neekesh V. , Abraham Brian J. , Abraham Brian J. , Balboni-Iniguez Amanda , Weichert-Leahey Nina , He Shuning , Krill-Burger John M. , Root David E. , Vazquez Francisca , Tsherniak Aviad , Hahn William C. , Golub Todd R. , Young Richard A. , Look A. Thomas , Stegmaier Kimberly
Published: August 2018

Childhood neuroblastomas with MYCN gene amplification form a particularly high-risk subset of this disease and are difficult to treat effectively. This has focused attention on tumor-specific gene dependencies that reflect important pathways in tumorigenesis, and thus could provide valuable targets for the development of novel therapeutics. Using genome-scale CRISPR-Cas9 approaches that allow unbiased detection of genes critically involved in tumor cell growth and survival, we identified 147 candidate genes associated with selective vulnerabilities in nine MYCN-amplified neuroblastoma cell lines, compared to findings in over 300 other human cancer cell lines representing multiple tumor cell types. We then used genome-wide ChIP-seq analysis to test the hypothesis that a small number of transcription factors - MYCN, HAND2, ISL1, PHOX2B, GATA3, and TBX2, all represented in the selective dependency group - are members of the transcriptional core regulatory circuitry (CRC) that underlies cell state in MYCN-amplified neuroblastoma. We show that these transcription factors bind as dense clusters at defined epicentres within the enhancers of their own genes, as well as those of the other CRC transcription factor genes, creating a positive feed-forward autoregulatory loop that establishes and maintains high levels of gene expression. To disable the CRC, we tested a combination of BRD4 and CDK7 inhibitors, which we postulated would act synergistically by targeting both transcriptional initiation and elongation required to synthesize regulatory transcription factors. MYCN-amplified neuroblastoma cells treated with both drugs were killed synergistically, in vitro and in vivo, and accompanied by rapid downregulation of CRC transcription factor gene expression. This study defines a set of critical dependency genes in MYCN-amplified neuroblastoma, a subset of which comprises the oncogenic transcriptional regulatory circuitry that underlies cell state and survival in this tumor.

Citation Format: Adam D. Durbin, Mark W. Zimmerman, Neekesh V. Dharia, Brian J. Abraham, Brian J. Abraham, Amanda Balboni-Iniguez, Nina Weichert-Leahey, Shuning He, John M. Krill-Burger, David E. Root, Francisca Vazquez, Aviad Tsherniak, William C. Hahn, Todd R. Golub, Richard A. Young, A. Thomas Look, Kimberly Stegmaier. Selective gene dependencies in MYCN-amplified neuroblastoma include the core transcriptional regulatory circuitry [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 2062.
Abstract 2063: SAMD9/SAMD9L mutations in familial monosomy 7
Authors: Bryant Victoria , Wong Jasmine , Schwartz Jason , Lamprecht Tamara , Ma Jing , Mullighan Charles , Loh Mignon , Shannon Kevin , Klco Jeffery
Published: August 2018

The purpose of this study was to define the mutations causing familial monosomy 7 with myelodysplastic syndrome (MDS) and/or acute myeloid leukemia (AML). We have previously reported frequent germline mutations in the genes SAMD9 and SAMD9L in pediatric MDS. We obtained samples on 32 individuals in 7 families with multiple siblings affected with MDS/AML and monosomy 7. We conducted targeted sequencing of genes frequently mutated in MDS/AML and amplicon sequencing of SAMD9 and SAMD9L. We performed cell cycle analysis assays on SAMD9/9L mutations to study their effects on cell proliferation. Our data show that 5 of the 7 families have germline pathologic mutations in SAMD9 (n=1) and SAMD9L (n=4), with significant clinical variability in the phenotypes displayed, ranging from transient monosomy 7 to overt AML with monosomy 7. Consistent with previous data, the mutant SAMD9 or SAMD9L allele was underrepresented in the bone marrow. This decrease in the mutant allele paralleled the extent of monosomy 7, suggesting that only the wild-type allele was retained in cells with monosomy 7. Functional analysis revealed that all germline SAMD9/9L mutations displayed a gain-of-function ability to suppress cell cycle progression. The patients with more aggressive disease were found to harbor somatic mutations in SETBP1, ETV6 or RUNX1. In many individuals we found additional somatic revertant mutations in cis with the germline mutation, which overcame the growth-suppressive effects of the germline mutation. Our data show that germline mutations in SAMD9/9L represent a new class of predisposition genes for familial monosomy 7 with MDS/AML and should be screened for in all children with MDS.

Citation Format: Victoria Bryant, Jasmine Wong, Jason Schwartz, Tamara Lamprecht, Jing Ma, Charles Mullighan, Mignon Loh, Kevin Shannon, Jeffery Klco. SAMD9/SAMD9L mutations in familial monosomy 7 [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 2063.
Abstract 2074: Single-cell RNA-seq reveals heterogeneity for stem cell markers, LRG5 and CD271 to predict a subpopulation in Ewing sarcoma cells with lower levels of BRAF
Authors: Khoogar Roxane , Houghton Peter
Published: August 2018

Introduction: Ewing Sarcoma (ES) is an aggressive bone and soft tissue tumor, which has a high rate of recurrence among all childhood cancers. This suggests the need for a better understanding of heterogeneous cancer populations. In many cancers, recurrence and therapy resistance has been suggested to be due to the relative state of quiescent cells. Our previous work has shown that low EWSR1-FLI1 expressing cells have a significantly lower proliferative activity, which is similar to a quiescent state. Therefore, we used cells with low EWSR1-FLI1 expression to model and predict cells with a quiescent state.Methods: Using siEWSR1 to suppress EWSR1-FLI1, we studied proliferative quiescence. The Fluidigm C1 system was used to perform SMART-Seq whole transcriptome amplification using EW8 cells. We sequenced single- cell RNA-Seq libraries prepared from 700 ES cells. We developed a computational analysis to profile single cells. Then, we used gene expression signatures derived from the quiescent population to classify parental cells with ensemble classifiers. Results: Results revealed that EW8 cells show considerable heterogeneity at the transcriptome level in control (scrambled RNAi) as well as in the quiescent populations. We identified a subpopulation of EW8 cells with a negative correlation between the expression of LGR5 and Nerve Growth Factor Receptor (NGFR/CD271). Our preliminary data from 255 single cells indicate that LGR5 (0-43.57 TPM, median 0) and CD271 (0-80.05 unique transcripts, median 6.33) were correlated (r= -0.12, P<0.0001) in EW8 cells. CD271 is a surface membrane protein, and a mesenchymal stem cell marker, that regulates stemness, and heterogeneity. A subpopulation of cells with low CD271 was distinguished and studied for its co-expression pattern with LRG5 and BRAF. Conclusions: Our results suggest that there are multiple subpopulations within EW8 parental (Control) cells. We can characterize them with proliferation, stemness, motility and dormancy-associated markers. Ongoing studies are aimed to determine, if EW8 cells having elevated LGR5 exhibit different sensitivity to cytotoxic agents. Data derived from this work are expected to provide insights into the extent to which CD271 contributes to dormancy, and ES heterogeneity. Whether CD271 regulates proliferation to promote a dormant state in this subpopulation is being studied.

Citation Format: Roxane Khoogar, Peter Houghton. Single-cell RNA-seq reveals heterogeneity for stem cell markers, LRG5 and CD271 to predict a subpopulation in Ewing sarcoma cells with lower levels of BRAF [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 2074.
Abstract 2102: UFH-001: A novel triple-negative, CAIX-positive cell line for small-molecule targeting of human breast cancer
Authors: Mboge Mam Y. , Chen Zhijuan , Ai Lingbao , Tu Chingkuang , Carta Fabrizio , Supuran Claudiu , Frost Christopher J. , Zhang Zaihui , McKenna Robert , Heldermon Coy , Frost Susan C.
Published: August 2018

Specific treatment options for patients with metastatic breast cancers, especially of the triple negative subtype (TNBC), are limited and most patients develop resistance to radiation and/or chemotherapy. Because of this, the mortality rate in women with TNBC remains high. Carbonic anhydrases (CAs), most specifically membrane-bound CAIX, has been shown to play an important role in tumor progression and as a result is validated as a therapeutic target in several aggressive cancers. In breast cancer, CAIX expression has been shown to be ubiquitous in hypoxic TNBC tissues and is considered a prognostic marker and a potential therapeutic target. Therefore targeting CAIX activity, using small molecule inhibitors, in these highly aggressive and metastatic TNBCs may serve to improve overall disease free survival and therapeutic outcome in clinics. Our goal for this study was to 1) Identify and/or develop a cell line that expresses CAIX in a hypoxia independent manner and is representative of TNBCs and 2) To target CAIX activity in this cell line and observe the effects of CAIX inhibition on its growth and metastasis. Our hypothesis is that CAIX inhibition, in the context of a hypoxic and/or acidic microenvironment, will dysregulate its ability to maintain the acidic pH preferred by cancer cells which favors their growth and migration. To achieve our aims, we newly developed and characterized an endogenous CAIX-expressing cell line named UFH-001. Our results show that CAIX expression and activity in this cell line, although endogenous under normoxic conditions, is increased more than 2 fold under hypoxic conditions. This cell line is also representative of TNBCs (ER/PR and HER2 negative) with a more epithelial-like morphology and phenotype. UFH-001 cells grow aggressively and have the ability to migrate/invade and form tumors in vivo. In these studies we have also shown how inhibition of CAIX activity, using sulfonamide-based inhibitors, affects UFH-001 growth in an apoptosis independent manner and decrease the migratory/inhibitory capacity of these cells. Taken together, our observations indicate that CAIX is a viable small molecular drug target for the treatment of patients with TNBCs.

Citation Format: Mam Y. Mboge, Zhijuan Chen, Lingbao Ai, Chingkuang Tu, Fabrizio Carta, Claudiu Supuran, Christopher J. Frost, Zaihui Zhang, Robert McKenna, Coy Heldermon, Susan C. Frost. UFH-001: A novel triple-negative, CAIX-positive cell line for small-molecule targeting of human breast 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 2102.
Abstract 2114: 3D co-culture of pancreatic tumor sphroids and stellate cells using minipillar array as a microtumor model of pancreatic ductal adenocarcinoma
Authors: Hwang Hyun Ju , Oh Min-Suk , Kim Seul-Ki , Kuh Hyo-Jeong
Published: August 2018

Purpose: Tumor microenvironmental (TME) factors play important roles in cancer progression and chemoresistance by inducing epithelial-mesenchymal transition (EMT) and related mechanisms. Pancreatic stellate cell (PSC) is a highly-enriched cell type in TME of pancreatic ductal adenocarcinoma (PDAC). Three dimensional (3D) tumor spheroids (TS) are considered as an in vivo-like tumor model demonstrating its clinical relevancy. We previously developed a minipillar array histochip for 3D culture and immunohistological analysis of TS in an increased throughput manner. Here, we present a minipillar-based co-culture of TS and PSC as a novel in vitro microtumor model of pancreatic ductal adenocarcinoma. Methods: Minipillar chips were obtained from MBD Co. (Cellvitro™ 55Cryo). 3D TS of Panc-1, human PDAC cells and PSCs (Sciencell) were cultured on minipillars and at the bottom of 96 well plates, respectively, as embedded in type I collagen. Changes in expression of extracellular matrix (ECM) and EMT-related factors in TS were assessed by confocal microscopy following cyrosection preparation. Results: Co-culture condition was optimized for the growth of TS, activation of PSC, and their interaction. Expression of EMT markers such as TGF-β1 and vimentin increased in co-cultured TS than mono-cultured TS. ECM remodeling as indicated by increased matrix deposition (collagen I, fibronectin) and fiber organization was observed in TS co-cultured with PSC compared to mono-cultured TS. Upon drug exposure (gemcitabine and paclitaxel), survival advantage was prominent in TS co-cultured with activated PSC. Conclusions: We established 3D model of PDAC by co-culturing TS and PSC using minipillar histochips. EMT-related changes including ECM remodeling and drug resistance were recapitulated in the model. Overall our results indicate usefulness of the present model not only in studying the role of TME factors and their interactions towards tumor progression but also in the evaluation of therapeutic efficacy of drug candidates and combinations.

Citation Format: Hyun Ju Hwang, Min-Suk Oh, Seul-Ki Kim, Hyo-Jeong Kuh. 3D co-culture of pancreatic tumor sphroids and stellate cells using minipillar array as a microtumor model of pancreatic ductal adenocarcinoma [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 2114.
Abstract 2119: Bone Morphogenetic Protein Receptor 1a in myeloid cells regulates mouse prostate cancer growth
Authors: Straign Desiree M. , Novitskiy Sergey V. , Owens Philip
Published: August 2018

The Bone Morphogenetic Protein (BMP) pathway is a member of the TGFβ signaling family and has mirrored the complex roles in diverse contexts during cancer development. BMP signaling is rarely mutated and can be frequently overexpressed in many human cancers. The dichotomous role of BMPs as both tumor promoters and suppressors appears to be largely context based in both the cancer cell and the surrounding microenvironment. Myeloid cells including macrophages and neutrophils have been shown to be tumor promoting when stimulated from BMP signaling. We discovered that conditional deletion of BMPR1a in myeloid cells (LysM-Cre) would affect tumor progression and found smaller tumors in a syngeneic FVBn prostate cancer model. Specific changes occurred in myeloid cells both in tumor-bearing mice and non-tumor naïve mice throughout multiple tissues. We profiled myeloid subsets both in the bone marrow, spleen and primary tumor. We found that BMPR1a loss in myeloid cells altered the differentiation and lineage capability of distinct populations by histologic, flow cytometry and high-dimensional mass cytometry analysis. These results indicate a distinct genetic requirement for BMP signaling in myeloid cells during tumor progression.

Citation Format: Desiree M. Straign, Sergey V. Novitskiy, Philip Owens. Bone Morphogenetic Protein Receptor 1a in myeloid cells regulates mouse prostate 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 2119.
Abstract 2120: Inhibition of multiple myeloma exosomes prevents bone loss and reduces tumor growth
Authors: Faict Sylvia T. , Veirman Kim De , Maes Ken , Bruyne Elke De , Heusschen Roy , Caers Jo , Vanderkerken Karin , Schots Rik , Menu Eline
Published: August 2018

Multiple myeloma, one of the most common hematological malignancies, will often manifest osteolytic lesions during disease development, throughout the whole body. MM is still considered an incurable malignancy, with a supportive micro-environment in the bone marrow. Extracellular vesicles such as exosomes are known to play an important tumor-promoting role in this micro-environment, by inducing angiogenesis and immune suppression. The aim of our study was to examine the effects of MM exosomes on osteolysis in vitro and in vivo, and to determine whether inhibition of exosome secretion can lead to a delay in tumor growth and prevent bone loss. We used the 5TGM1 model which is a syngeneic murine MM model, presenting typical MM characteristics such as osteolysis, angiogenesis and a serum M-spike. We examined the effects of conditioned medium from these MM cells and MM-derived exosomes on cell viability and proliferation of the MC3T3 pre-osteoblast cell line and on their ability to produce alkaline phosphatase (ALP). We saw a decrease in viability and ALP activity when adding exosomes or conditioned medium of the 5TGM1 myeloma cells to the osteoblasts. Moreover, RT-PCR analysis showed a decrease of various osteogenic differentiation genes when osteoblasts were exposed to 5TGM1 exosomes, demonstrating a diminished capacity of osteoblastic differentiation. We next added the exosome secretion inhibitor GW4869 to the 5TGM1 cells, thereby reducing the number of exosomes in the conditioned medium. This was confirmed through western blot of the typical exosome markers. The negative effects of the 5TGM1 conditioned medium on osteoblasts could be overturned by reducing the exosome content therein. In vivo, we injected 5TGM1 exosomes intravenously in healthy mice for three weeks and examined the effects on osteolysis by microCT and TRAP staining. We saw an increase in osteolysis, comparable to the typical osteolytic lesions in tumor-bearing mice. Furthermore, when inhibiting the exosome secretion in vivo in 5TGM1 mice, we observed a significant reduction in tumor load. Through this study we can conclude that exosomes from multiple myeloma cells can induce osteolytic lesions by inhibiting osteoblast proliferation and differentiation. These effects can be reversed by inhibiting the secretion of exosomes.

Citation Format: Sylvia T. Faict, Kim De Veirman, Ken Maes, Elke De Bruyne, Roy Heusschen, Jo Caers, Karin Vanderkerken, Rik Schots, Eline Menu. Inhibition of multiple myeloma exosomes prevents bone loss and reduces tumor 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 2120.
Abstract 2125: Cancer-associated fibroblasts-stimulated IL-11 promotes metastasis of gastric cancer cells mediated by upregulation of MUC1
Authors: Wang Xiaoxun , Che Xiaofang , Bai Ming , Fan Yibo , Liu Yunpeng , qu xiujuan
Published: August 2018

Background: Cancer-associated fibroblasts (CAFs) are major components of the tumor stroma and regulators of tumor progression. However, the molecular mechanism by which CAFs promote gastric cancer progression should be further explored.

Methods: Mouse model was used to certify that CAFs could promote peritoneal metastasis of gastric cancer cells. The expression of interleukin-11 (IL-11) was analyzed in two gastric cancer cell lines by real-time RT-PCR and ELISA. The role of IL-11 in gastric cancer cell migration and invasion explored by Transwell. Western blotting was conducted to detect the activation of STAT3 and ERK signaling pathway.

Results: In our study, we reveal that when CAFs co-cultured with gastric cancer cells secreted significant amounts of IL-11. CAFs promote the ability of migration and invasion through the production of IL-11 by stimulating JAK/STAT3 and MAPK/ERK pathways to upregulate MUC1 in gastric cancer cells. On the other hand, when IL-11 was deprived using a neutralizing antibody or inhibition of JAK/STAT3 and MAPK/ERK pathways with specific inhibitors, Stattic and PD-98059, markedly attenuated these CAFs-induced phenotypes in gastric cancer cells.

Conclusions: Taken together, these results revealed that CAFs play a significant role in the gastric cancer progression in the tumor microenvironment through IL-11-STAT3/ERK signaling by upregulating MUC1. Also, IL-11 targeted therapy can achieve an effective treatment against gastric cancer indirectly by exerting their action on stromal fibroblasts.

Citation Format: Xiaoxun Wang, Xiaofang Che, Ming Bai, Yibo Fan, Yunpeng Liu, xiujuan qu. Cancer-associated fibroblasts-stimulated IL-11 promotes metastasis of gastric cancer cells mediated by upregulation of MUC1 [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 2125.
Abstract 2130: Novel ADAM9 variant induces prostate cancer metastasis
Authors: Hsieh Chia-Ling , Sung Shian-Ying
Published: August 2018

Prostate cancer often progresses from an androgen to castration-refractory prostate cancer and metastasizes to the bone and lymph nodes. However, there is no biomarker to predict the initiation of cancer metastasis. Therefore, it is an unmet need to understand molecules which induce metastasis to replace the prediction by PSA. Previous studies by us demonstrate the elevation of ADAM9 in PCa malignant progression. However, the role of ADAM9 in the tumor microenvironment is still not fully understood. Using laser capture microdissection of prostate tumor and stromal cells, indicating the increased of ADAM9 and reactive oxygen species in tumor-associated stromal cells. These inductions correlated with the increased expression of SOD-2 in tumor-associated stromal cells. Both electron microscope analysis and conditioned medium of stromal cells demonstrated the increased of soluble ADAM9 level in the medium. ELISA analysis of patient serum confirmed the increased of ADAM9 serum levels in prostate cancer patients. PCR and sequencing analysis confirmed a novel soluble ADAM9 (sADAM9v2) with deletion of membrane domain that induces cell migration through the activation of FAK and AKT phosphorylation. Our studies demonstrated the release of sADAM9 in tumor microenvironment correlated with reactive oxygen stress and regulate prostate cancer migration and metastasis. In conclusion, our studies showed that sADAM9 release by cancer-associated stromal cells could be a potential biomarker for prostate cancer progression.

Citation Format: Chia-Ling Hsieh, Shian-Ying Sung. Novel ADAM9 variant induces prostate cancer metastasis [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 2130.
Abstract 2134: IL6 and CXCL8 mediate redundant, targetable tumor-host interactions that drive osteosarcoma lung metastasis
Authors: Roberts Ryan D. , Cam Hakan , Brandolini Laura , Hinckley John M. , Gross Amy C.
Published: August 2018

Osteosarcoma (OS) kills patients through aggressive metastatic spread, primarily to lung tissues. Our lab has sought to understand the tumor-host interactions that facilitate growth of OS cells specifically within the lung. We previously identified two intercellular signaling molecules, IL6 and CXCL8, which exhibit enhanced expression when primary tumors become lung metastases. Here, we show that epigenetic changes drive aberrant expression of ΔNp63, endowing OS cells with the capacity to colonize lung tissue. IL6 and CXCL8 serve as downstream mediators induced directly by ΔNp63 and necessary for survival and proliferation within the lung. Inhibition of IL6 and CXCL8 signaling has no direct effect on the proliferation of OS cells, but IL6 inhibition has a profound effect on their ability to form colonies in soft agar. In preclinical mouse models of OS, inhibitors of IL6 and CXCL8 signaling profoundly impair the ability of OS cells to colonize lung tissue, completely preventing metastasis in some models. Co-culture studies suggest that strong paracrine interactions with bronchial epithelial and smooth muscle cells, but not macrophages or lung fibroblasts, drive increased expression of IL6 and CXCL8. These data suggest a mechanism where aberrant ΔNp63 expression within developing OS tumors generates subsets of tumor cells endowed with increased IL6 and CXCL8 expression. This triggers feed-forward paracrine loops with bronchial epithelial and smooth muscle cells that facilitate growth and survival within the lung, leading to metastasis.

Citation Format: Ryan D. Roberts, Hakan Cam, Laura Brandolini, John M. Hinckley, Amy C. Gross. IL6 and CXCL8 mediate redundant, targetable tumor-host interactions that drive osteosarcoma lung metastasis [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 2134.
Abstract 214: Genetic and transcriptional instability alters cell line drug response
Authors: Ben-David Uri , Siranosian Ben , Ha Gavin , Tang Helen , Lyons Nicholas J. , Burns Robert , Nag Anwesha , Cimini Beth , Tsvetkov Peter , Tubelli Andrew A. , Wong Bang , Thorner Aaron R. , Bittker Joshua , Meyerson Matthew , Beroukhim Rameen , Golub Todd R.
Published: August 2018

Inconsistencies in cell line-based studies jeopardize reproducibility of cancer research. Natural evolution leading to genetic and transcriptional heterogeneity within cancer cell lines may contribute to such inconsistencies. To systematically test this hypothesis, we performed comprehensive genetic and transcriptomic characterization of 27 strains of the common breast cancer cell line MCF7, and then assessed the response of those strains to 321 anti-cancer compounds. We found extensive variation in genomic profiles and responses to small molecules across strains. Genetic variation occurred at all levels – point mutations, rearrangements, and copy number changes – and affected multiple oncogenes and tumor suppressor genes. Similar findings were obtained across 23 strains of the lung cancer cell line A549, indicating that such variation is a general property of cancer cell lines. These changes resulted in substantial differences in gene expression programs, cell morphology and proliferation, and strikingly high variability in drug response. Over 75% of the compounds that exhibited strong activity in some of the strains, were completely inactive in other strains. These results indicate that genomic variation within commonly used cancer cell lines is extensive, resulting in disparate drug responses. Such variation may contribute to difficulties in conducting reproducible cell line-based studies.

Citation Format: Uri Ben-David, Ben Siranosian, Gavin Ha, Helen Tang, Nicholas J. Lyons, Robert Burns, Anwesha Nag, Beth Cimini, Peter Tsvetkov, Andrew A. Tubelli, Bang Wong, Aaron R. Thorner, Joshua Bittker, Matthew Meyerson, Rameen Beroukhim, Todd R. Golub. Genetic and transcriptional instability alters cell line drug response [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 214.
Abstract 2143: IL-6 mediated signaling regulates cytotoxic effect of Burkitt lymphoma cells by combined treatment of Danusertib and BKM120
Authors: Liu Jun , Ahn Kwang-Sung , Hong Jun-Shik , Yoon Sung-Soo
Published: August 2018

Even though Burkitt Lymphoma is relative has a good response to combined treatment, some part of outcome of chemotherapy is very poor. One of the obstacles is the chemoresistance due to several cytokines which are released from stromal cells. The effects of either Danusertib or BKM120 on Namalwa and BJAB, respectively, were different. On Namalwa the IC 50 of Danusertib was 24.64 µM (72 h) and BKM120 was 12.44 µM (72h). On BJAB the IC 50 of Danusertib was 6.63 µM (72h) and BKM120 was 1.42 µM (72h). But in the combined treatment, we found that BJAB showed less sensitive than Namalwa, the IC 50 on Namalwa was Danusertib 8.33 µM+ BKM120 2.08 µM (72h) and on BJAB was Danusertib 20.36 µM+ BKM120 5.09 µM (72h). To figure out the resistant pattern of these two cell lines we performed 3 steps treatment with 16 µM Danusertib and 4 µM BKM120, every step we culture 1.5×105/mL cells in 24hours first, and then treated with combined inhibitors 48 hours. We found that although Namalwa was sensitive to combined treatment at the first step, it was more easily to become resistant to combined treatment. To confirm the resistant pattern, co-culture system was used to identify what kinds of cytokines are involving in the chemoresistance of Burkitt lymphoma. We found that IL-6 was induced with stromal cell including inflammatory cytokines when cells were co-cultured with stromal cells. Also, IL-6 mediated signaling pathway such as JAK/STATIL-6 and AKT pathway, was activated by IL-6 and sIL-6. We suggest that IL-6 mediated signaling could mediate signaling regulates cytotoxic effect of Burkitt lymphoma cells by combined treatment of Danusertib and BKM120 as well as inflammatory effects may regulate the response of chemotherapeutic treatment.

Citation Format: Jun Liu, Kwang-Sung Ahn, Jun-Shik Hong, Sung-Soo Yoon. IL-6 mediated signaling regulates cytotoxic effect of Burkitt lymphoma cells by combined treatment of Danusertib and BKM120 [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 2143.
Abstract 2155: Modeling therapeutic response in BRAF V600-mutant melanoma using patient-derived xenografts
Authors: Feng Tianshu , Golji Javad , Li Ailing , Zhang Xiamei , Ruddy David , Stuart Darrin , Meyer Matthew
Published: August 2018

Approximately 50% of metastatic melanoma harbor oncogenic mutations of BRAF, resulting in constitutive activation of the MAPK signaling pathway. Treatment using BRAF and MEK inhibitor combination significantly improved survival in patients with BRAF V600-mutant melanoma. However, the duration of response remains limited as most patients develop resistance. Previously, our group established a collection of patient derived xenografts (PDXs), and conducted a PDX clinical trial using a 1x1x1 experimental design. The PDX clinical trial showed that BRAF V600-mutant melanoma models varied in their response to BRAF and MEK inhibitors, similar to observations in the clinical setting. Therefore, we sought to elucidate the mechanisms governing differential responses to MAPK inhibition in vivo. The PDX models were classified into complete and partial responders based on their sensitivity to BRAF and MEK inhibitors. Using this classification, we characterized the downstream signaling associated with therapeutic response. Whereas complete responders showed almost full suppression of MAPK signaling, partial responders showed heterogeneous levels of MAPK signaling upon BRAF and MEK inhibition. Ongoing work seeks to identify determinants of therapeutic response using biased and unbiased approaches, in an effort to inform combination strategies that improve survival in patients with BRAF V600-mutant melanoma.

Citation Format: Tianshu Feng, Javad Golji, Ailing Li, Xiamei Zhang, David Ruddy, Darrin Stuart, Matthew Meyer. Modeling therapeutic response in BRAF V600-mutant melanoma using patient-derived xenografts [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 2155.