bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5042: A CRISPR/Cas9 knockout screen identify novel drug resistance genes in AML
Authors: Huang Jian 
Published: July 2016

Abstract
Acute myeloid leukemia (AML) is a malignant hematopoietic disease and the most common type of acute leukemia in adults. Despite remarkable progress made in the therapy of AML, the mainstays of treatment have not significantly changed over the last 20 years. Technological advances have greatly accelerated genomic discovery and precision medicine holds promise to better tailor medications to AML with specific mutations. Importantly, one major obstacle to greater success with target therapy of leukemia is drug resistance. But the mechanisms underlying drug resistance in AML are poorly understood.
The activating mutations of FLT3 are now recognized as the most common molecular abnormality in AML and FLT3ITD mutations are found in nearly 30% of AML patients. The poor prognosis of patients harboring FLT3 mutations renders FLT3 an obvious target of therapy. Quizartinib (AC220) is a potent and selective second-generation inhibitor of FLT3. It is in clinical trials for the treatment of relapsed or refractory FLT3ITD positive and negative AML patients and as a maintenance therapy. So far, those clinical trials have showed very promising result. However, drug resistance to AC220 has also been reported through the early clinical studies. To understand the underlying mechanisms of drug resistance to AC220, we undertook an unbiased approach with a novel CRISPR pooled library to screen new genes whose loss of function confer resistance to AC220. In our screen, we identified SPRY3 and GSK3 as positive hits and the related Wnt and FGF-Ras-ERK signaling activation as the major mechanisms to cause resistance to the FLT3 inhibitor AC220. In the next step, we will explore the detailed underlying mechanisms by which SPRY3 and GSK3 mutations lead to drug resistance to AC220. We will also confirm our findings in primary AML patient samples. Taken together, our study will provide new insight into signaling pathways that contribute to the acquired resistance in AML. The knowledge learned from this study may lead to the development of more efficient therapeutic avenues for AML.
Citation Format: Jian Huang. A CRISPR/Cas9 knockout screen identify novel drug resistance genes in AML. [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 5042.
Abstract 5047: The MUC1 membrane-bound mucin increases tumor cell properties and chemoresistance in renal clear cell carcinoma
Authors: Gaudelot Kelly , Gnemmi Viviane , Bouillez Audrey , Gibier Jean-Baptiste , Fanchon Mélanie , Woszczyk Justine , Hémon Brigitte , Van Seuningen Isabelle , Aubert Sébastien , Perrais Michael 
Published: July 2016

Abstract
Introduction: MUC1, an O-glycoprotein membrane-bound mucin, is overexpressed in renal clear-cell carcinomas (CRCC) with correlation to two major prognostic factors, Tumor-Node-Metastasis stage and nuclear Fürhman grade. Previously, we have shown that (i) MUC1 was significantly overexpressed in metastatic CRCC vs non-metastatic CRCC and (ii) MUC1 is a target gene of HIF-1 transcription factor which is a part of the hypoxia pathway, the main renal carcinogenetic pathway. Furthermore, CRCC is highly resistant to common systemic chemotherapies.
Material and method: To better understand the roles of MUC1 in CRCC, we used two renal cell lines expressing MUC1 (786-O cells) or not (ACHN cells). 786-O cells were stably transfected with shRNA targeting MUC1 while ACHN cells with full-length MUC1. Proliferation, drug resistance, migration and invasion properties were studied in vitro in the different cellular clones using MTS cell proliferation assay, wound healing assay and Boyden chambers coated with Matrigel, respectively. Signaling pathways were screened by proteome profiler and western blot.
Results and discussion: We showed that MUC1 expression was associated with increased invasion and migration properties of renal carcinomatous cells and a decrease of cell-cell interactions. MUC1 overexpressing cells (i) expressed higher levels of anti-apoptotic factors and MDR genes involved in chemoresistance processes and (ii) were more resistant to chemotherapeutic drugs.
Conclusion: Our results show that MUC1 plays a role in biological properties of renal cancer cells suggesting important function for this mucin in tumour progression and chemo-resistance. Our data confirm its potential as a therapeutic target in this type of cancer.
Citation Format: Kelly Gaudelot, Viviane Gnemmi, Audrey Bouillez, Jean-Baptiste Gibier, Mélanie Fanchon, Justine Woszczyk, Brigitte Hémon, Isabelle Van Seuningen, Sébastien Aubert, Michael Perrais. The MUC1 membrane-bound mucin increases tumor cell properties and chemoresistance in renal clear 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 5047.
Abstract 5062: Hypoxia-induced amoeboid cancer cell migration
Authors: te Boekhorst Veronika , Jiang Liying , Lehmann Steffi , Zuidema Alba , Friedl Peter 
Published: July 2016

Abstract
Tumor hypoxia, by elevating hypoxia-inducible factors (HIFs), is an established inducer of epithelial-to-mesenchymal transition (EMT) and subsequent cancer cell invasion and metastasis. However, how hypoxia impacts on tumor cell invasion programs and translates into modes of dissemination of moving tumor cells remains unknown. Using breast cancer (BC) and head and neck cancer (HNC) cells which efficiently invade from multicellular spheroids into 3D fibrillar collagen, we addressed which signaling programs that control cell-cell adhesions, cell-matrix interactions and cytoskeletal dynamics and which modes of tumor cell migration are induced by hypoxia.
While both cancer models primarily invaded collectively under normoxic conditions, severe hypoxia (0.2% oxygen) or pharmacological stabilization of HIF-1 using the prolyl hydroxylase inhibitor dimethyloxalylglycine (DMOG) induced EMT-like detachment and migration of single cells into collagen, exhibiting a 2- and 4-fold increase of single BC and HNC cells, respectively. Besides infrequent spindle-shaped movement, hypoxia or DMOG predominantly induced the transition from collective to amoeboid single-cell migration, with blebby amoeboid migration as the predominant migration mode (4- and 3-fold increase for BC and HNC, respectively) next to filopodial amoeboid migration.
Hypoxia-induced amoeboid migration is characterized by variably fast and persistent migration, reaching up to several 100 μm/day, mediated by actin-rich filopodial or blebby protrusions at the leading edge. Criteria for amoeboid movement included (i) actin-containing blebs that interacted with collagen structures in a polarized fashion, (ii) weak dependence on matrix-metalloproteinase-mediated collagen remodeling, (iii) low dependence on focalized integrin-mediated matrix adhesions, and (iv) high dependence on RhoA-mediated actomyosin contraction for efficient bleb formation and migration speed. Interference with ROCK signaling using Y-27632 and myosin using blebbistatin significantly inhibited hypoxia-induced amoeboid dissemination and bleb formation. Thus, hypoxia induces plasticity of cancer invasion modes, with efficient blebby amoeboid dissemination as primary outcome. While blebby amoeboid motion is known as a rudimentary migration mode in embryonic stem cells, here we observe blebby amoeboid migration as a simple (low MMP- and integrin dependent) but efficient salvage migration mode of cancer cells under severe hypoxic stress.
Citation Format: Veronika te Boekhorst, Liying Jiang, Steffi Lehmann, Alba Zuidema, Peter Friedl. Hypoxia-induced amoeboid cancer cell migration. [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 5062.
Abstract 507: Prediction of colorectal cancer recurrence with plasma miRNAs in the ColoCare Study
Authors: Dickinson Brandon , Adams Scott V. , Ulrich Cornelia , Vickers Kathy , Wurscher Michelle , Grady William M. , Makar Karen W. 
Published: July 2016

Abstract
Background
Colorectal cancer is the third most common cancer and third most common cause of cancer-related death in the US. Recurrence of primary tumors occurs in 10-40% of patients depending on tumor stage. Early detection of recurrence can increase the treatment options for those eligible for curative therapy, which may lead to better survival. We determined the levels of multiple different plasma miRNAs and assessed their association with clinical outcomes to identify specific miRNAs that may serve as predictors of CRC recurrence.
Methods
Colorectal cancer patients (age 18-80, newly diagnosed, stage I-IV) were recruited as part of the ColoCare Study in Seattle, Washington, between 2007 and 2011. Patients who had a blood sample taken &gt;30 and &lt;365 days after surgery and prior to any recurrence were included in the study (n = 83).
Candidate miRNAs (miRNA 31, 141, 200b, 203, 16, 17-3p, 29a, 92a, 125b) were quantified using pre-designed TaqMan microRNA assays in triplicate assays. Plasma miRNA expression was normalized and expressed as copy number. Cox proportional hazards regression with robust standard errors was used to estimate hazard ratios of recurrence associated with level of each miRNA, adjusted for age, sex and tumor site and stratified by stage. No recurrences were observed in stage I patients. Statistical analyses were conducted separately for stages II-III and II-IV colorectal cancer.
Results
During a median of 1,067 days (33-2,254 days) follow-up after surgery, 20 recurrences were observed. In stage II-III patients, the presence of detectable circulating levels of miR-141 (HR = 4.30 [1.10-16.84]) or miR-203 (HR = 8.04 [1.39-46.4]) were associated with risk of recurrence. In stage II-IV patients, detectable miR-203 (HR = 3.52 [1.13-10.96]) was associated with recurrence, and elevated miR-29a levels were associated with greater risk of recurrence (HR: 1.80 per natural log unit, 95% CI: 1.06-3.04).
Conclusion
The present study identified multiple plasma miRNAs that may help to predict those patients at greatest risk of colorectal cancer recurrence. Our results require validation in order to determine the role for such plasma markers in clinical practice.
Citation Format: Brandon Dickinson, Scott V. Adams, Cornelia Ulrich, Kathy Vickers, Michelle Wurscher, William M. Grady, Karen W. Makar. Prediction of colorectal cancer recurrence with plasma miRNAs in the ColoCare 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 507.
Abstract 5076: Ras association (RalGDS/AF-6) domain family member integrates with notch signaling to regulate tumor cell migration
Authors: Perumalsamy Lakshmi R. , S. Mahalingam 
Published: July 2016

Abstract
Tumor metastasis, the spread of tumor cells to distant tissues from its site of initiation is a multi-step and multi-factorial process. Hyperactivation of RAS pathway is reported in &gt;50% of different cancers and is associated with metastasis and poor prognosis. Since Ras signaling pathway is essential for normal development, its been difficult to develop drugs against it in tumors. This led to the identification of several Ras effectors. Ras association (RalGDS/AF-6) domain family (RASSF) proteins are non-enzymatic effectors of Ras. RASSF7 is a member of the less characterized N-terminal RASSFs. Similar to other RASSFs, RASSF7 also associates with Ras. Several mutations within RASSF7 open reading frame as well as differential expression pattern were observed in various tumors. The role of RASSF7 and its downstream signaling is not yet clearly understood.
Microarray analysis for RASSF7 identified several differentially regulated genes with enrichment for pathways including cell adhesion, inflammation, actin cytoskeleton and cell-cell communication suggesting that RASSF7 might regulate cell migration. Ectopic expression of RASSF7 inhibited cell migration of multiple cancer cell lines in vitro. RT-qPCR and western blotting analysis suggest that RASSF7 can transcriptionally upregulate Notch1 expression. Knockdown of Notch1 abrogated RASSF7 mediated inhibition of cell migration. Our experiments suggest that RASSF7 alters the localization of Notch1. Cross-talk Notch signaling is a key signaling pathway regulating tumor progression across several different cancers. Collectively, our data suggests that RASSF7 impinges on Notch1 to regulate cell migration.
Citation Format: Lakshmi R. Perumalsamy, Mahalingam S. Ras association (RalGDS/AF-6) domain family member integrates with notch signaling to regulate tumor cell migration. [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 5076.
Abstract 5080: Amylase overexpression promotes ovarian cancer invasion
Authors: Mohamed Mai , Kruk Patricia 
Published: July 2016

Abstract
Introduction: Hyperamylasemia is an overexpression of amylase. Hyperamylasemia is associated with poor clinical outcome in ovarian cancer and reportedly serves as a prognostic biomarker for ovarian cancer. However, the function of hyperamylasemia in ovarian cancer is thus far unknown. Since amylase cleaves α1, 4-glycosidic bonds in carbohydrates, hyperamylasemia may play an active role in the etiology of ovarian cancer.
Purpose: The purpose of this study was to characterize amylase expression in ovarian cancer cell lines and establish an active role for hyperamylasemia in ovarian cancer.
Hypothesis: We hypothesized that secreted amylase enzymes cleave α1, 4-glycosidic bonds found in the carbohydrate shell which may result in a modified glycocalyx structure facilitating ovarian cancer cell invasion. Consequently, inhibiting amylase expression could abrogate ovarian cancer cell invasion/metastasis.
Methods: To characterize amylase isozyme expression, we assayed amylase expression in normal and cancerous ovarian epithelial cell lines by western immunoblot, qPCR, amylase assays and computational analyses. We measured the ability of amylase to promote cellular proliferation by MTS assays. We assayed invasive capacity of cultured cells using collagen coated Boyden chambers and abrogated amylase expression with either amylase siRNA or treatment with the blue-green algae, Spriulina.
Results: It was found that the amylase genes have identical or similar biochemical and bioinformatics properties: similar molecular weights, theoretical isoelectric points, disorder and hydropathy profiles, secondary structure and posttranslational modifications. In contrast to normal cell lines, ovarian cancer cell lines overexpress amylase at the messenger and protein levels and secrete high levels of metabolically active amylase. Amylase isozymes 1 and 2B were preferentially overexpressed in ovarian cancer cell lines. Inhibition of amylase abrogated ovarian cancer cell migration/invasion through collagen coated Boyden chambers which was associated with amylase-dependent alteration of extracellular carbohydrates. Further, the blue-green algae, Spirulina, was found to be a novel inhibitor of amylase.
Conclusion: Herein, we describe for the first time an active function of amylase to promote ovarian cancer cell invasion potentially mediated by amylase-dependent alteration of extracellular carbohydrates. Further, transcriptional inhibition of amylase by Spirulina demonstrates its clinical potential as an adjuvant nutraceutical therapy.
Citation Format: Mai Mohamed, Patricia Kruk. Amylase overexpression promotes ovarian cancer invasion. [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 5080.
Abstract 5086: Legumain-contained exosomes mediate remodeling of tumor extracellular matrix and protumor effects of TAMs
Authors: Shen Long , Shi Yuzhi , Xun Jing , Chen Chuanai , Du Lingfang , Tan Xiaoyue , Xiang Rong 
Published: July 2016

Abstract
It has been recognized that tumor associated macrophages (TAMs) in tumor microenvironment play a variety of pro-tumor effects during the tumor progression, while the role of extracellular matrix in mediating these effects is still unclear.
In this study, we focus on elucidating the way as well as the underlying mechanism about the involvement of an asparaginyl endopeptidase, legumain in the crosstalk between TAMs and ECM. Our data showed that co-culture of breast cancer cells 4T1 with macrophages Raw 264.7 promoted asparaginyl endopeptidase, legumain, expression with the introduction of markers of M2 - polarized macrophage. Legumain was detected in the condition medium (CM) collected from RAW 264.7 co-cultured with 4T1 cells by western blotting, with the existence of asparagine endopeptide enzyme activity assessed by enzyme activity assay. Result of western blotting showed that adding the CM of RAW 264.7 co-cultured with 4T1 cells depleted extracellular matrix (ECM) contents, fibronectin and collagen I. Transwell assay showed increased infiltration of 4T1 cells through the ECM contents after treatment with the CM of RAW 264.7 co-cultured with 4T1 cells. Furthermore, treatment with CM of RAW 264.7 co-cultured with 4T1 suppressed the adhesion of endothelial cells with ECM, promoting tube formation of endothelial cells compared with control. All the above results could be abolished by synthesized legumain inhibitor, RR-11a. Interestingly, legumain was also detected in the exosomes isolated from CM of RAW 264.7 co-cultured with 4T1. Applying legumain contained exosomes could replicate the effect of CM of RAW 264.7 co-cultured with 4T1 on the depletion of ECM contents, promotion of 4T1 infiltration and suppression of endothelial cell adhesion with ECM.
Altogether, our results suggest that legumain is overexpressed and secreted out by TAMs; TAMs exosomes contain legumain and help delivering legumain to the ECM thus exert its effect on the depletion of ECM, pro-infiltration and angiogenesis.
Citation Format: Long Shen, Yuzhi Shi, Jing Xun, Chuanai Chen, Lingfang Du, Xiaoyue Tan, Rong Xiang. Legumain-contained exosomes mediate remodeling of tumor extracellular matrix and protumor effects of TAMs. [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 5086.
Abstract 5095: Hic-5 modulation of the stromal matrix is required for mammary tumor progression
Authors: Goreczny Gregory J. , Ouderkirk Jessica , Olson Eric , Krendel Mira , Turner Christopher 
Published: July 2016

Abstract
Primary tumors grow and invade into surrounding tissues/stroma, frequently resulting in metastasis to distant organs. Tumor cell-stroma crosstalk plays a crucial, but incompletely understood role during tumor progression and metastasis. For example, increased stromal matrix density and/or rigidity correlates with poor prognosis in human breast cancer patients. Previous studies have implicated Hic-5, a focal adhesion scaffold protein, in tumor cell invasion, proliferation and metastasis. To investigate the role of Hic-5 in mammary tumor progression, Hic-5 -/- mice were generated and crossed with mice expressing Polyoma Middle T Antigen (PyMT) driven by the MMTV promoter. The PyMT mouse is a well-established model for human breast cancer progression. Tumors from the Hic-5 -/- PyMT mice demonstrated an increased latency and had reduced growth as compared to Hic-5 +/- PyMT controls. The absence of Hic-5 did not affect the progression of the tumor to a carcinoma stage, but there was a decrease in metastasis to the lungs. Immunohistochemical analysis showed that Hic-5 is primarily expressed in the cancer associated fibroblasts (CAFs) and the Hic-5 -/- PyMT tumor stroma exhibits reduced extracellular matrix (ECM) deposition. Furthermore, 3D cell-derived matrices (CDM) generated in vitro using isolated CAFs, confirmed that Hic-5 -/- PyMT CAFs do not efficiently deposit and organize ECM. The organization of the stromal matrix has been shown previously to increase the rigidity of the tumor, which in turn promotes FAK Y397 phosphorylation in the tumor cells to promote cell invasion and proliferation. Accordingly, the Hic-5 -/- PyMT tumors exhibited a reduction in FAK phospho-Y397 staining and attenuation of downstream ERK activation, suggesting that their stromal matrix is not optimally organized for tumor cell signaling. Taken together, these data demonstrate that Hic-5 expression in CAFs is important for stromal matrix organization/remodeling during tumor progression and that the loss of the organization of the ECM in the Hic-5 -/- tumor stroma may contribute to the reduced tumor growth and metastasis observed in these animals.
Supported by NIH CA163296 to CET, NS066071 to EO and Carol M. Baldwin Breast Cancer Fund to CET.
Citation Format: Gregory J. Goreczny, Jessica Ouderkirk, Eric Olson, Mira Krendel, Christopher Turner. Hic-5 modulation of the stromal matrix is required for mammary tumor progression. [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 5095.
Abstract 5096: Development of a decellularized tumor model for the evaluation of breast carcinomas
Authors: Martin Elizabeth , Pashos Nicholas , Gimble Jeffrey , Bunnell Bruce , Burow Matthew , Collins-Burow Bridgette 
Published: July 2016

Abstract
Breast cancer recurrence is a clinical manifestation of tumor progression in patients which has been shown to significantly increase mortality rates. While up to one-fifth of breast cancer patients will experience recurrence, the mechanisms underlying this process remain poorly understood. Recently, gene expression profiling of human tumor metastasis and recurrence demonstrated that the extracellular matrix (ECM)-receptor interactions pathway was enhanced. In breast carcinomas, the ECM is known to vary with cancer progression. The ECM acts as a chemical reservoir, structural component, and mediator of tumorigenesis to tumors. To date, the ECM of breast carcinomas has been evaluated within the context of a seeded cellular population, i.e. intact tumor. Here we demonstrate for the first time a mechanism to evaluate the ECM devoid of breast cancer cells. We have employed a model of tissue decellularization to breast cancer cell line derived tumors and demonstrate that tumor ECM architecture remains intact when devoid of cells. Validation of loss of cell content is evident through DNA quantification, nuclear DAPI stain, and H&E stain. Furthermore we demonstrate that structural proteins, fibronectin, collagen, and elastin are still retained within the decellularized scaffold. Applications of the tumor decellularization model will aid in the evaluation of ECM structural and molecular characteristics independent of cancer cell populations. The data presented here will enhance methods used to evaluate the cancer microenvironment and further our understanding of breast cancer recurrence, altering existing treatment strategies to extend survival.
Citation Format: Elizabeth Martin, Nicholas Pashos, Jeffrey Gimble, Bruce Bunnell, Matthew Burow, Bridgette Collins-Burow. Development of a decellularized tumor model for the evaluation of breast carcinomas. [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 5096.