bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1527: Urinary miRNAs as predictors of tumor metastasis in localized patients with clear cell renal cell carcinoma: a pilot study
Authors: Shu Xiang , Hildebrandt Michelle AT , Borges dos Reis Rodolfo , Ye Yuanqing , Gu Jian , Wood Christopher G. , Wu Xifeng 
Published: July 2016

Abstract
Introduction: 25-50% of localized renal cell carcinoma (RCC) patients will eventually develop metastasis which has a dramatic adverse effect on prognosis following treatment. Unfortunately, reliable biomarkers predicting RCC metastasis are still lacking. MicroRNAs (miRNAs) are important regulators for vital biological processes in humans. Urine miRNAs(UmiRNAs) present promising potential as biomarkers for diagnosis of urological cancers. However, no study has assessed the predictive value of UmiRNAs for RCC metastasis.
Materials and Methods: We prospectively collected pre- and post-surgery urine samples from 117 patients with localized clear cell RCC (ccRCC,T1-3 N0/x M0). UmiRNAs were extracted and purified from 250 μl urine supernatants and measured using RT-PCR. Relative quantifications (RQ) of 137 selected miRNAs were calculated by 2−ΔΔct method. The ratio of post/pre RQs were tested for the association with distant metastasis using multivariable Cox regression models.
Results: More than half of the patients were diagnosed at pT3 (53%) or grade III-IV (69%). Twenty-two patients (18.8%) had metastasis during a mean follow-up of 73 months. The levels of 34 UmiRNAs were significantly changed after nephrectomy (p&lt;0.05, 29 of 34 significantly reduced). The 50% or more reduced post-nephrectomy levels of five UmiRNAs (miR-191-5p, miR-324-3p, miR-186-5p, miR-93-5p, and miR-30b-5p) independently conferred a 2- to 4-fold increased risk of metastasis with miR-191-5p being the most significant one (hazard ratio [HR] = 4.16, 95% confidence interval [CI] = 1.38-12.58, p = 0.011). Stratified analyses by stage and Fuhrman grade revealed that the associations were most profound in pT3 patients for miR-191-5p, miR-324-3p, and miR-186-5p. Ingenuity pathway analysis showed that the predicted target genes of these five UmiRNAs were enriched in cell survival and proliferation networks.
Conclusion: Our study suggests that urinary miR-191-5p, miR-324-3p, and miR-186-5p are potential biomarkers for the prediction of metastasis in localized, particularly pT3 ccRCC patients after nephrectomy. These findings may help improve individualized patient management in the clinic.
Citation Format: Xiang Shu, Michelle AT Hildebrandt, Rodolfo Borges dos Reis, Yuanqing Ye, Jian Gu, Christopher G. Wood, Xifeng Wu. Urinary miRNAs as predictors of tumor metastasis in localized patients with clear cell renal cell carcinoma: a pilot study. [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 1527.
Abstract 1547: Circulating tumor cells from a syngeneic mouse model of hepatocellular carcinoma demonstrate epithelial-mesenchymal transition, decreased MHC I expression, and increased CCR7 expression
Authors: Huaman Jeannette A. , Muhith Ubayed , Das Dibash K. , Naidoo Michelle , Ogunwobi Olorunseun O. 
Published: July 2016

Abstract
Metastasis is the leading cause of death in cancer patients, worldwide. A better understanding of how cancer cells detach from the primary tumor, escape from immunosurveillance while in circulation, and successfully colonize distant parts of the body, is needed. Circulating tumor cells (CTCs) are excellent tools to study this process. Using a syngeneic hepatocellular carcinoma mouse model, we were able to isolate cancer cells from the primary tumor and CTCs from the blood and establish novel primary tumor-derived cell lines and novel CTC lines. Wound healing assays revealed that CTCs have greater migratory capacity than the cancer cells from the primary tumors. However, MTT assays revealed that CTCs had less proliferative capacity than the cancer cells from the primary tumors. Furthermore, molecular characterization of the cancer cells from primary tumors and CTCs using Western blotting showed a decreased expression of E-cadherin and an increased expression of fibronectin by CTCs, suggesting that epithelial-to-mesenchymal transition (EMT) takes place in CTCs in comparison to cancer cells from primary tumors. To address the question of how CTCs may evade immunosurveillance, we investigated the profile of expression of two cell surface proteins involved in immune system function: the major histocompatibility complex class I (MHCI)- required for cytotoxic T cell activation and subsequent cancer cell killing, and chemokine receptor 7 (CCR7)- exploited by cancer cells for immune evasion. Our analysis revealed that there is decreased cell surface expression of MHCI and increased expression of CCR7 in the CTCs in comparison to the cancer cells from primary tumors. These findings indicate that although CTCs are not more proliferative, they have acquired significantly increased migratory capacity than cancer cells in primary tumors. This may be explained by their demonstration of EMT. Furthermore, their loss of MHC I expression and their gain of CCR7 expression may help with evasion of killing by cytotoxic T cells and result in enhanced metastasis.
Citation Format: Jeannette A. Huaman, Ubayed Muhith, Dibash K. Das, Michelle Naidoo, Olorunseun O. Ogunwobi. Circulating tumor cells from a syngeneic mouse model of hepatocellular carcinoma demonstrate epithelial-mesenchymal transition, decreased MHC I expression, and increased CCR7 expression. [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 1547.
Abstract 155: The analysis of the genomic evolution of squamous cell carcinoma of the lung
Authors: Krause Arthur , Lorber Thomas , Perrina Valeria , Dietsche Tanja , Barrett Michael , Ruiz Christian , Bubendorf Lukas 
Published: July 2016

Abstract
Background:
Genomic tumor heterogeneity is one reason for recurrence and resistance to cancer therapy. It is still not known what leads to the relapse or the emergence of metastasis in individual patients. To contribute to answer this question, we investigated the evolutionary patterns in squamous cell carcinoma of the lung (SCC) in 10 patients with multiple matched tumors (primary vs recurrence/metastasis).
Methods:
To identify the genomic profile, frozen tissues from SCC patients were subjected to multiparameter ploidy profiling (MPP). Tumor populations were sorted based on ploidy (DNA-content using DAPI) and p40 was used to control for diploid tumor populations. Subsequent genomic profiling by array comparative genomic hybridization (aCGH) and targeted massive parallel sequencing (Ion Torrent Comprehensive Cancer Panel with an all-exon coverage of 409 cancer genes) was applied.
Results:
The usage of MPP enabled the determination of the clonal composition of the SCC and define genomic characteristics. Due to flow sorting the tumor populations were effectively isolated and existence of distinct clonal populations was indicated. Aneuploid and diploid tumors were successfully revealed using the additional marker p40. Therefore, the absence of non-malignant cells increased the precision in the genomic analyses
Conclusion:
The study uncovered the composition of human SCC in the sense of different clonal populations, which harbor private genomic aberrations and mutations confirmed by aCGH and NGS. Even if our data is still preliminary we provide evidence that applying MPP increases the precision of aCGH and sequencing analysis. Genomic analyses of sorted pure tumor population allows to reconstruct the clonal evolution and facilitates the understanding of tumor heterogeneity and their evolutionary pattern, which could have potentially an impact on therapeutic response or metastasis development.
Citation Format: Arthur Krause, Thomas Lorber, Valeria Perrina, Tanja Dietsche, Michael Barrett, Christian Ruiz, Lukas Bubendorf. The analysis of the genomic evolution of squamous cell carcinoma of the lung. [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 155.
Abstract 157: Single-cell DNA sequencing identifies a late dissemination model in metastatic colorectal cancer
Authors: Leung Marco L. , Casasent Anna , Wang Yong , Sei Emi , Navin Nicholas 
Published: July 2016

Abstract
Metastatic colorectal cancer (mCRC) is a devastating disease with only 11% 5-year survival rate for stage IV patients. The genomic basis of metastasis has been difficult to study, in part due to the extensive intratumor heterogeneity at both the primary and metastatic tumor sites. Previous studies have applied bulk next-generation sequencing (NGS) methods, which have limited ability to resolve intratumor heterogeneity. To address this problem, we developed a highly-multiplexed single cell DNA sequencing method that combines flow-sorting of single nuclei, multiple-displacement-amplification, low-input library preparation, library barcoding, targeted capture and NGS to generate high-coverage data from single tumor cells. To increase throughput, we performed targeted single-cell DNA sequencing using a panel of 201-cancer genes to analyze single cells from primary tumors and liver metastases from two mCRC patients. In each patient, we sequenced 90 single tumor cells from the primary and metastatic tumor, to delineate the clonal architecture of the tumor and reconstruct their phylogenetic lineages. In addition, we sequenced populations of tumor cells at high coverage depth (200X).
Our data identified a large number of nonsynonymous mutations that evolved in the root nodes during the earliest stages of primary tumor evolution and were maintained in all single cells during the clonal expansion of the tumor mass. We also identified a small number of mutations that were specific to the liver metastases, which are likely to play an important role in metastatic dissemination. Using the single cell data, we constructed phylogenetic trees, which revealed branched evolution at both organ sites. Collectively, our data suggest that both mCRC patients are consistent with a late-dissemination model, in which the primary tumors evolved for a long period of time prior to the dissemination of clones to distant organ sites. This model has important clinical implications, by suggesting that surgical intervention or therapeutic treatment of the primary CRC tumor can prevent metastasis, since single tumor cells do not disseminate at the earliest stages of tumor growth
Citation Format: Marco L. Leung, Anna Casasent, Yong Wang, Emi Sei, Nicholas Navin. Single-cell DNA sequencing identifies a late dissemination model in metastatic colorectal 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 157.
Abstract 1605: Cetuximab inhibits migration, invasion, metastasis and epithelial-mesenchymal transition but not proliferation via GEP100-Arf6-AMAP1 pathway in head and neck squamous cell carcinoma
Authors: Matsumoto Yoshifumi , Sakurai Hiroyuki , Kogashiwa Yasunao , Saito Koichiro 
Published: July 2016

Abstract
Background: Recently, irradiation combined with Cetuximab (Cmab) is being one of the standard therapeutic options against head and neck squamous cell carcinoma (HNSCC), especially in advanced/recurrent cases. While this concurrent chemo-radiotherapy is widely accepted and tolerated, Cmab monotherapy has been reported to be unsatisfactory to treat HNSCC, and detailed mechanism of Cmab to show such clinical efficacy is unknown.
Methods: To assess the impact of Cmab on proliferation, migration, and invasion of HNSCC, WST assay, wound healing assay, and matrigel invasion assay were incorporated in this study. Effects of Cmab on the grade of epithelial-mesenchymal transition (EMT), expression level of membrane type 1-matrix-metalloproteinase (MT1-MMP), and activity of EGFR-GEP100-Arf6-AMAP1 pathway, were evaluated using the immunocytochemistry, western blot analysis and Arf6 activation assay, respectively. Furthermore, In vivo efficacy of Cmab was evaluated in highly aggressive and metastatic HSC-3-M3 cells using orthotopic xenograft model in nude mice nu/nu.
Results: Inhibition of migration and invasion were observed in 4 HNSCC cell lines by Cmab application. However, proliferation potential of HNSCC were not affected by this chemotherapeutic agent. Increased expression of E-cadherin, with decreased expression of N-cadherin and MT1-MMP, were observed after Cmab treatment in vitro to suggest that Cmab may inhibit EMT in HNSCC. Furthermore, inhibition of Tyr 1086 phosphorylation in EGFR, as well as the inactivation of Arf6 coupled with suppression of both GEP100 and AMAP1 expressions were observed by Cmab application. These results suggested that Cmab might have the potential to suppress the activity of EGFR-GEP100-Arf6-AMAP1 pathway. Furthermore, less metastasis of the cancer was observed in the Cmab treatment group compared with control group in vivo.
Conclusions: Our study is the first to provide that Cmab could affect HNSCC through inhibition of migration and invasion of cancer cells, regardless of its anti-proliferative action. EMT inhibition after Cmab application was also clarified in this study. Furthermore, our study suggested that EGFR-GEP100-Arf6-AMAP1 pathway may have some role in the invasion and metastasis of HNSCC. Future studies to further assess the precise mechanism of Cmab against HNSCC are warranted.
Citation Format: Yoshifumi Matsumoto, Hiroyuki Sakurai, Yasunao Kogashiwa, Koichiro Saito. Cetuximab inhibits migration, invasion, metastasis and epithelial-mesenchymal transition but not proliferation via GEP100-Arf6-AMAP1 pathway in head and neck squamous cell 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 1605.
Abstract 1611: Epithelial-mesenchymal-epithelial transition induced by long term exposure to TGFB1 creates cellular heterogeneity
Authors: Oliveira Patricia , Carvalho Joana , Rocha Sara , Azevedo Mafalda , Vieira Andre F. , Ferreira Daniel , Mendes Nuno , Reis Ines , Vinagre Joao , Heravi-Moussavi Alireza , Nunes Joana B. , Lima Jorge , Máximo Valdemar , Burleigh Angela , Roskelley Calvin , Paredes Joana , Carneiro Fatima , Huntsman David , Oliveira Carla 
Published: July 2016

Abstract
Reversible and dynamic transitions between epithelial and mesenchymal cellular states (EMT/MET) contribute to cancer progression and dissemination. Whereas EMT facilitates initial steps of tumour cell detachment, MET is likely required for colonization at distant sites. Although MET is generally perceived as mirroring EMT, we hypothesize that MET entails its own set of novel and/or differentially active molecular circuitries, generating cells with features distinct from the original epithelial state.
Using an in vitro TGFβ1-induced EMT/MET model, we demonstrated that MET generates co-existing heterogeneous cell populations (Reverted-Epithelial or RE-cells) with novel phenotypic and functional properties, such as increased self-renewal, in vivo increased tumourigenicity and distinct chemoresistance properties.
Overall, our results indicate that MET is a permissive process, driving cellular plasticity towards heterogeneity and with it, creating novel biological signatures of relevance for cancer growth.
Citation Format: Patricia Oliveira, Joana Carvalho, Sara Rocha, Mafalda Azevedo, Andre F. Vieira, Daniel Ferreira, Nuno Mendes, Ines Reis, Joao Vinagre, Alireza Heravi-Moussavi, Joana B. Nunes, Jorge Lima, Valdemar Máximo, Angela Burleigh, Calvin Roskelley, Joana Paredes, Fatima Carneiro, David Huntsman, Carla Oliveira. Epithelial-mesenchymal-epithelial transition induced by long term exposure to TGFB1 creates cellular heterogeneity. [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 1611.
Abstract 1612: Overexpression or inducible expression of Snail, Zeb1 or TGFβ recapitulates epithelial-mesenchymal transition (EMT) in colorectal cancer cell lines in vitro
Authors: Bhagwat Shripad V. , Hamm Nicole E. , Hsu Yu-Chih , Qian Yuewei , Wild Robert , Lee Jonathan A. , Peng Sheng-Bin 
Published: July 2016

Abstract
Epithelial-mesenchymal transition (EMT) is one of the hallmarks of cancer and a critical process providing tumor cells with the ability to metastasize to distant sites. Several previous studies have revealed that EMT plays a crucial role in the progression and aggressiveness of colorectal cancer (CRC). Snail, Zeb1 and TGFβ are considered as master EMT genes and their overexpression has been linked to metastatic disease. Our study was conducted to establish an EMT platform in-house to determine the role of Snail, Zeb1 and TGF-β1 in mediating EMT in CRC cell lines in vitro and using them for studying drug sensitivity and target validation. Screening of six CRC cell lines for expression of epithelial (E) and mesenchymal (M) markers by western blot analysis suggested that DLD-1 and HT-29 cells are epithelial in nature and were used to generate stable overexpression and tet-inducible models. Overexpression of Snail or Zeb1 or TGF-β1 induced EMT in both DLD-1 and HT-29 colorectal cancer cells as indicated by downregulation of E-cadherin and upregulation of vimentin by western blot analysis and immunofluorescence staining. These cells which have undergone EMT also showed significant growth advantage in 3D growth on matrigel or soft agar. Stable ‘E’ and ‘M’ cells were also used to test the sensitivity of various inhibitors including erlotinib, salinomycin and chemotherapeutic agents such as paclitaxel, gemcitabine, cisplatin, oxaloplatin and 5-FU. Consistent with previous findings, erlotinib showed sensitivity only in ‘E’ cells and not ‘M’ cells. Chemotherapy agents have also shown higher sensitivity in ‘E’ cells than ‘M’ cells. In contrast, Salinomycin showed equipotent sensitivity in ‘E’ and ‘M’ cells. We have also generated tet-inducible models of Snail, Zeb1 and TGF-β1 and they behaved very similar to overexpression models. These inducible models were useful to determine whether any inhibitor treatment or knock down of specific gene (target) can block EMT. In our studies, testing of a Focal Adhesion Kinase (FAK) inhibitor or Dasatinib (SFK inhibitor) as examples showed blockade of EMT in these models. Overall, establishing several EMT models in-house enabled us to validate new targets involved in EMT, test new inhibitors whether it can block EMT or study drug sensitivity in ‘E’ vs. ‘M’ cells.
Citation Format: Shripad V. Bhagwat, Nicole E. Hamm, Yu-Chih Hsu, Yuewei Qian, Robert Wild, Jonathan A. Lee, Sheng-Bin Peng. Overexpression or inducible expression of Snail, Zeb1 or TGFβ recapitulates epithelial-mesenchymal transition (EMT) in colorectal cancer cell lines in vitro. [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 1612.
Abstract 1621: SRGAP2 expression levels may induce a biphasic metastatic phenotype in osteosarcoma cell lines
Authors: Marko Tracy , Shamsan Ghaidan , Largaespada David 
Published: July 2016

Abstract
The purpose of this study is to characterize the role of Slit-Robo GTPase-Activating Protein 2 (SRGAP2) in osteosarcoma (OS) metastasis. OS is the most common primary bone tumor and the eighth leading pediatric cancer. Despite combined therapies, treatment failure is experienced within 5 years of diagnosis by over 40% of patients, generally due to metastatic disease. Using the conditional Sleeping Beauty transposon mutagenesis system, our laboratory recently identified candidate tumor promoting genes in transgenic mice that develop OS, including the potential tumor suppressor SRGAP2 involved in regulating actin dynamics. We hypothesized that loss of SRGAP2 would increase the metastatic potential of OS cell lines in vitro and in vivo, whereas gain would decrease metastatic potential. Gene expression was amplified with a stably integrated, tetracycline-inducible over expression vector containing SRGAP2 cDNA and gene silencing was accomplished with a clustered regularly interspaced short palindromic repeat (CRISPR) targeting the gene. Studies were conducted in the well-characterized murine osteosarcoma cell lines K12 and its more aggressive derivative K7M2 and human osteosarcoma cell lines HOS and its more aggressive derivative 143B. Compared to luciferase-controls, overexpression and knockout of SRGAP2 had no effect on cell proliferation in K12, K7M2, HOS, and 143B (n = 4). A reduction of soft agar colony formation was observed in knockout cell lines but not in cell lines with SRGAP2 overexpression for K12 and 143B (n = 3); HOS cell lines did not form colonies in soft agar. Interestingly, overexpression of SRGAP2 decreased wound-healing closure in 143B and K12 lines, but increased closure time in HOS (n = 4). Tail vein injections in NRG mice are currently being investigated. Overexpression of SRGAP2 in K12 decreased development of macrometastases in lungs, whereas knockout of SRGAP2 eliminated gross detection of lung masses (n = 3). Future studies include imaging fixed cells to determine if SRGAP2 knockout and overexpression affects cell morphology and completing studies with K7M2 lines. These results suggest that SRGAP2 has a biphasic phenotype in osteosarcoma cell lines, with an optima in expression for efficient metastasis. It's dual role of deforming cell membranes independent of its ability to activate Rac GTPases and alter actin dynamics may be responsible for the observed results.
Citation Format: Tracy Marko, Ghaidan Shamsan, David Largaespada. SRGAP2 expression levels may induce a biphasic metastatic phenotype in osteosarcoma cell lines. [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 1621.