bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract LB-282: Two different strategies of delivery CRISPR/Cas9 system to gene edit rs4644 SNP in LGALS3 gene
Authors: Corrado Alda , Lepori Irene , Miglietta Simona , Batoni Simone , Vitiello Marianna , Evangelista Monica , Chisci Elisa , Giovannoni Roberto , Poliseno Laura , Gemignani Federica , Landi Stefano 
Published: July 2017

Abstract
Galectin-3 is a glycoprotein of 31KDa with a chimeric structure. It is encoded by a single gene, LGALS3, located on chromosome 14. Previous studies showed a relation between the single nucleotide polymorphism (SNP) (rs4644, c.191C&gt;A, p.Pro64His) and the risk of cancer. In literature, some data show different results indicating that SNP could have different roles in base on the tissues. The aim of our study is to create isogenic cell lines that differ only for SNP permitting further studies about the function of SNP in different cancers. The “generation” of the cell lines is performed with a Clustered Regularly Interspaced Short Palindromic Repeats-associated Endonuclease 9 (CRISPR/Cas9) system. We have used two different strategies to deliver the system. In the first strategy, our aim was the reduction of the problem of efficiency of transfection that could depend on different cell lines. In fact, we have used the lentivirus method to deliver all components of the CRISPR/Cas9 on Nthy-Ori (normal thyroid tissue) and HCT 116 +/+ (colorectal cancer tissue) cells. Two different vectors (pCW-Cas9 and pLXgRNA) are used to build up the inducible CRISPR/Cas9 system and HR410PA-1 vector (transfected by Lipofectamine 3000) is used to knock-in. HR410PA-1 is a particular vector that facilities the homologous recombination (HR) to repair the double strand break. Moreover, to improve HR, we use also Scr7, which is an inhibitor of DNA IV ligase. In the second approach, we have used the double nickase system using two modified Cas9 vectors (SpCas9D10a) that produce only a single strand break. In this way, the problem of off-targets is notably reduced. In this approach, the knock-in is due to or single-stranded oligodeoxynucleotides (ssODNs) or HR410PA-1 vector. The first results show that some of the transfected cells are edited in a correct way. In the further steps, we will isolate the “positive clones” using the selection cassette in HR410PA-1 vector (GFP protein and/or puromycin) or the single cell dilution assay (1 cell/well) for the transfected cells with ssODNs.
Citation Format: Alda Corrado, Irene Lepori, Simona Miglietta, Simone Batoni, Marianna Vitiello, Monica Evangelista, Elisa Chisci, Roberto Giovannoni, Laura Poliseno, Federica Gemignani, Stefano Landi. Two different strategies of delivery CRISPR/Cas9 system to gene edit rs4644 SNP in LGALS3 gene [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-282. doi:10.1158/1538-7445.AM2017-LB-282
Abstract LB-286: ARID1A loss impairs enhancer-mediated gene regulation and drives colon cancer in mice
Authors: Mathur Radhika , Alver Burak H. , Roman Adrianna K. San , Wilson Boris G. , Wang Xiaofeng , Agoston Agoston T. , Park Peter , Shivdasani Ramesh , Roberts Charles W. 
Published: July 2017

Abstract
Genes encoding subunits of SWI/SNF (BAF) chromatin remodeling complexes are collectively mutated in ~20% of all human cancers, but the mechanism by which these mutations promote tumorigenesis is unclear. In this study, we investigate the tumor suppressor role of ARID1A, the subunit of SWI/SNF complexes that is the most frequent target of mutations. We develop an inducible Arid1a-knockout mouse model and find that mice develop invasive ARID1A-deficient colon adenocarcinoma. These cancers show prominent mucinous differentiation and tumor-infiltrating lymphocytes, features associated particularly with microsatellite-instable (MSI) human colorectal cancer - the subtype with the highest frequency of inactivating ARID1A mutations (37-39%). Although deregulation of Wnt signaling components (APC/β-catenin) is commonly utilized to model intestinal tumorigenesis, Arid1a-knockout mice reflect human colorectal cancer with greater accuracy. These mice do not show deregulation of Wnt signaling components, thus demonstrating a novel pathway to colon tumorigenesis that is independent of established models. Chromatin immunoprecipitation sequencing reveals that SWI/SNF complexes are targeted primarily to enhancers, where they function in coordination with transcription factors to activate gene expression. ARID1A loss impairs the targeting of SWI/SNF complexes to thousands of enhancers, which subsequently lose activity - showing reduced levels of H3K27ac and expression of nearest genes. Residual SWI/SNF complexes in ARID1A-deficient cells bind enhancers that remain active; these complexes contain ARID1B, the subunit that is mutually exclusive with ARID1A and has been identified as a therapeutic target in ARID1A-deficient cancers. Enhancers associated with developmental gene expression programs are most affected by ARID1A loss, both in the mouse colonic epithelium and in human colon cancer cells. Collectively, these findings represent an advance in colon cancer modeling and implicate enhancer-mediated gene regulation as the principal tumor suppressor function of ARID1A.
Citation Format: Radhika Mathur, Burak H. Alver, Adrianna K. San Roman, Boris G. Wilson, Xiaofeng Wang, Agoston T. Agoston, Peter Park, Ramesh Shivdasani, Charles W. Roberts. ARID1A loss impairs enhancer-mediated gene regulation and drives colon cancer in mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-286. doi:10.1158/1538-7445.AM2017-LB-286
Abstract LB-317: Identification of a novel preclinical candidate for CDK7 inhibition
Authors: Satyam Leena K. , Poddutoori Ramulu , Mukherjee Subhendu , Marappan Sivapriya , Gopinath Sreevalsam , Basavaraju Aravind , Kaza Lakshmi Narayana , Pothuganti Manoj Kumar , Nayak Shilpa , C Nandish , A Amith , MV Ravindra , Babu Dabbeeru Madhu , A Nagaraju , Tgore Suraj , Antony Thomas , Pandit Chetan , Ramachandra Murali , Chelur Shekar , Daginakatte Girish , Samajdar Susanta 
Published: July 2017

Abstract
Cyclin-dependent kinase 7 (CDK7) is an important constituent of the cellular transcriptional machinery, where it phosphorylates the C-terminal domain (CTD) of RNAP polymerase II (RNAPII). Because many tumor types are critically dependent on transcription for maintenance of their oncogenic state, pharmacological modulation of CDK7 kinase activity is considered as an approach to treat cancer. Multiple series of covalent CDK7 inhibitors were identified by iterative medicinal chemistry efforts and SAR based approach. These compounds were optimized towards attaining good physicochemical properties, high potency, good selectivity and desirable pharmacokinetic profile to achieve anti-tumor activity. We have now identified a pre-clinical candidate AU-BGB-002 which is highly potent in inhibiting CDK7 in biochemical as well as cellular assays while fully efficiently engaging the target. In a panel of kinases, AU-BGB-002 shows selectivity for CDK7. A panel of cell lines derived from a diverse set of indications are sensitive to AU-BGB-002. AU-BGB-002 exhibits excellent drug-like characteristics including solubility, permeability, metabolic stability and good oral bioavailability. When tested in a xenograft model, AU-BGB-002 treatment resulted in dose dependent tumor growth inhibition in AML xenograft model with tumor stasis at a dose of 10 mg/kg. Potent inhibiton of tumor growth was accompanied by complete target engagement and suppression of pS5RNAPII RNAPolII Ser5 phosphorylation in a parallel PK-PD study. Efficacy studies in additional xenograft models, advanced DMPK and toxicity studies are ongoing for this compound. In summary, we have identified a novel and selective CDK7 covalent inhibitor candidate with desirable drug-like properties that shows excellent efficacy in an AML xenograft model. Findings presented here support further development of AU-BGB-002 for the treatment of cancer.
Citation Format: Leena K. Satyam, Ramulu Poddutoori, Subhendu Mukherjee, Sivapriya Marappan, Sreevalsam Gopinath, Aravind Basavaraju, Lakshmi Narayana Kaza, Manoj Kumar Pothuganti, Shilpa Nayak, Nandish C, Amith A, Ravindra MV, Dabbeeru Madhu Babu, Nagaraju A, Suraj Tgore, Thomas Antony, Chetan Pandit, Murali Ramachandra, Shekar Chelur, Girish Daginakatte, Susanta Samajdar. Identification of a novel preclinical candidate for CDK7 inhibition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-317. doi:10.1158/1538-7445.AM2017-LB-317
Abstract LB-321: Cytoplasmic DEDD promotes G1- to S-phase cell cycle transition
Authors: Schmidt Keon R. , Ni Yingjia , Zhang Siyuan 
Published: July 2017

Abstract
Death Effector Domain-Containing DNA-binding protein (DEDD), a highly conserved and ubiquitous protein, has been studied primarily for its role as a scaffold protein in cell death pathways. DEDD resides predominantly in the cytoplasm, but translocates to the nucleus to propagate the terminal apoptotic signal. Overexpression of nuclear DEDD, not cytoplasmic DEDD, has been shown to be sufficient to induce cytotoxic effects. This suggests that DEDD may have functional importance beyond directing apoptotic signals. Further, the cytoplasmic function of DEDD has yet to be characterized. Other DED-containing proteins, such as FADD, have been shown to interact with cyclin/cyclin-dependent kinase cell cycle regulatory molecules. Here, we show that DEDD is implicated in cell cycle regulation and can assume a pro- proliferative function when overexpressed in the cytoplasm. To glean phenotypic changes in cell cycle transitions, we utilized a two-dimensional approach with an EdU proliferation label and a propidium iodine DNA content label differentiated via flow cytometry analysis. Based on these labels, it can be determined whether a cell is in the G1-, S-, or G2-phase of the cell cycle. We have identified that overexpression of DEDD leads to an increase in S-phase cells when compared to control suggesting an easier transition through the G1/S checkpoint. Conversely, a knockdown of DEDD leads to a decrease in S-phase cells and an increase in G1-phase cells when compared to control indicating G1 arrest. Gain-of-function flow cytometry analysis was repeated by overexpressing a DEDD construct with disrupted nuclear localization sequences. The resulting phenotype mirrored that of the overexpressed wild-type DEDD construct. Thus, these results suggest that DEDD may be interacting with regulatory molecules regulating the G1/S checkpoint of the cell cycle and that the change in G1- to S-phase transition dynamics in response to manipulated DEDD expression is not attributable to the nuclear function of DEDD. Further studies will focus on elucidating the specific molecular interactions through which DEDD can carry out this novel cell cycle regulatory function.
Citation Format: Keon R. Schmidt, Yingjia Ni, Siyuan Zhang. Cytoplasmic DEDD promotes G1- to S-phase cell cycle transition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-321. doi:10.1158/1538-7445.AM2017-LB-321
Abstract LB-322: MicroRNA-130a suppresses breast cancer cell migration and invasion by targeting FOSL1 and upregulating ZO-1
Authors: Chen Xiaowei , Zhao Min , Li Yuhuang , Dai Mushui 
Published: July 2017

Abstract
FOSL1 is frequently overexpressed in multiple types of human cancers including invasive breast cancers and implicated in cancer invasion and metastasis. However, how FOSL1 is overexpressed in cancers remains to be elucidated. Several microRNAs (miRNAs) have been shown to target FOSL1 and are downregulated in human cancers. Here, we report that miR-130a is a novel FOSL1 targeting miRNA. Using gene expression microarray analysis, we found that FOSL1 is among the most up-regulated genes in cells transfected with miR-130a inhibitors. Transient transfection-immunoblot, RNA-immunoprecipitation (IP) and luciferase reporter assays revealed that miR-130a directly targets FOSL1 mRNA at its 3’-UTR. Overexpression of miR-130a significantly reduced the levels of FOSL1 in invasive breast cancer MDA-MB-231 and Hs578T cell lines and suppresses their migration and invasion. This inhibition can be rescued by ectopic expression of miR-130a-resistant FOSL1. Interestingly, we show that overexpression of miR-130a increased the levels of tight-junction protein ZO-1 while inhibition of miR-130a reduced the levels of ZO-1, via regulating the expression of FOSL1. We further show that miR-130a expression is significantly reduced in cancer tissues from triple-negative breast cancer (TNBC) patients, correlating significantly with the upregulation of FOSL1 expression, compared to non-TNBC tissues. Together, our results reveal that miR-130a directly targets FOSL1 and suppresses FOSL1-mediated inhibition of ZO-1, thus inhibiting cancer cell migration and invasion, in TNBCs.
Note: This abstract was not presented at the meeting.
Citation Format: Xiaowei Chen, Min Zhao, Yuhuang Li, Mushui Dai. MicroRNA-130a suppresses breast cancer cell migration and invasion by targeting FOSL1 and upregulating ZO-1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-322. doi:10.1158/1538-7445.AM2017-LB-322
Abstract LB-325: Tumor suppressor effects of miR-17-92a cluster miRNAs on prostate cancer
Authors: Chakrabarti Ratna , Ottman Richard , Hasan Md Faqrul 
Published: July 2017

Abstract
MicroRNAs in miR-17-92a cluster are transcribed from a polycistronic transcription unit C13orf25 that generates six mature miRNAs, miR-17, miR-18a, miR-19a, miR- 19b, miR-20a. These miRNAs are reported to be overexpressed in colon and lung cancers, however, we noted a loss of expression of all members of this miR cluster in prostate cancer tissues compared to uninvolved tissues and in aggressive prostate cancer cells. In this study, we show that replenishment of miR-17-92a cluster as a whole showed antiproliferative effects, reduced activation of AKT and MAP kinases, delayed tumorigenicity and reduced tumor growth. Analysis of the expression of mRNA targets showed reduced expression of cell cycle regulatory proteins cyclin D1 and SSH1; and LIMK1 and FGD4 of RhoGTPase signaling pathway through direct targeting. miR-17-92 cluster expression also inhibited EMT via reduced cell migration and expression of mesenchymal markers while elevating expression and surface localization of the epithelial marker E-Cadherin. miR-17-92a miRNAs improved sensitivity of androgen dependent LNCaP 104-S prostate cancer cells to anti-androgen drug Casodex, AKT inhibitor MK-2206 2HCl, and docetaxel. The androgen refractory PC-3 cells also showed increased sensitivity to docetaxel, MK-2206 2HCl and Aurora kinase inhibitor VX680 upon ectopic expression of miR-17-92a cluster miRNAs. RNA seq analysis of prostate cancer cells expressing miR-17-92a cluster revealed altered expression of a number of oncogenic and tumor suppressor mRNAs, and noncoding RNAs, which may have implications in the altered phenotype of these cells. Our data demonstrate a tumor suppressor effect of miR-17-92a cluster miRNAs in prostate cancer cells and restoration of expression of these miRNAs has a therapeutic benefit for both androgen-dependent and -independent prostate cancer cells.
Note: This abstract was not presented at the meeting.
Citation Format: Ratna Chakrabarti, Richard Ottman, Md Faqrul Hasan. Tumor suppressor effects of miR-17-92a cluster miRNAs on prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-325. doi:10.1158/1538-7445.AM2017-LB-325
Abstract LB-334: xCT promotes malignant phenotypes in EGFR-expressing glioma
Authors: Tsuchihashi Kenji , Okazaki Shogo , Yoshikawa Momoko , Seishima Ryo , Sampetrean Oltea , Onishi Nobuyuki , Wakimoto Hiroaki , Furnari Frank , Baba Eishi , Akashi Koichi , Saya Hideyuki , Nagano Osamu 
Published: July 2017

Abstract
Malignant glioma such as glioblastoma is a cancer still difficult to treat. System xc(-) is composed of xCT and CD98hc subunits and functions as a plasma membrane antiporter for the uptake of extracellular cystine in exchange for intracellular glutamate, which has been reported to be involved in malignant phenotypes of glioma cells. However, the underlying mechanism of xCT regulation in glioma is not elucidated. Here we show that the epidermal growth factor receptor (EGFR) interacts with xCT and thereby promotes its cell surface expression and function in human glioma cells. EGFR-expressing glioma cells manifested both enhanced antioxidant capacity as a result of increased cystine uptake as well as increased extracellular glutamate which promotes glioma matrix invasion. Imaging mass spectrometry also revealed that brain tumors formed in mice by human glioma cells stably overexpressing EGFR contained higher levels of reduced glutathione compared with those formed by parental cells. Targeted inhibition of xCT suppressed the EGFR-dependent enhancement of antioxidant capacity in glioma cells as well as tumor growth and invasiveness. Our findings propose that xCT is a promising therapeutic target in EGFR-overexpressing malignant glioma.
Citation Format: Kenji Tsuchihashi, Shogo Okazaki, Momoko Yoshikawa, Ryo Seishima, Oltea Sampetrean, Nobuyuki Onishi, Hiroaki Wakimoto, Frank Furnari, Eishi Baba, Koichi Akashi, Hideyuki Saya, Osamu Nagano. xCT promotes malignant phenotypes in EGFR-expressing glioma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-334. doi:10.1158/1538-7445.AM2017-LB-334
Abstract PL01-03: Earlier detection as a key to lower cancer death rates
Authors: Vogelstein Bert 
Published: July 2017

Abstract
Two disciplines within cancer research—secondary prevention and cancer therapeutics—have historically been viewed as distinct. I will argue that these two disciplines are intertwined and that their joint pursuit is the optimal way to reduce cancer deaths. At present, the preferred way to treat cancers, in fact the only way that generally cures patients, is surgery. It is therefore widely recognized that the detection of cancers prior to the onset of metastasis is vital to reducing cancer deaths. What is less widely recognized is that the detection of metastatic cancers can also be life-saving—as long as this detection occur priors to the formation of bulky metastatic disease. Therapies that only modestly extend life when administered to patients with measurable metastases can be curative when administered to patients with metastatic disease that is present but undetectable by imaging. Such therapies include conventional cytotoxic drugs as well as genome-targeted agents and immunotherapeutics. A new paradigm for designing clinical trials to test such agents in adjuvant settings will be described. Genome-based diagnostic approaches for detecting cancers earlier will be discussed, as these are integral to the design of the next generation of adjuvant trials. In aggregate, these issues support the importance of modulating the focus of cancer research and treatment from patients with advanced disease towards patients with less-advanced disease. They also highlight the importance of developing novel diagnostic approaches that will allow the detection of smaller tumors.
Citation Format: Bert Vogelstein. Earlier detection as a key to lower cancer death rates [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr PL01-03. doi:10.1158/1538-7445.AM2017-PL01-03
Abstract PL04-01: G-quadruplexes and DNA dynamics
Authors: Balasubramanian Shankar 
Published: July 2017

Abstract
DNA is structurally dynamic in ways that have a consequence for biologic processes. A class of four-stranded DNA structure, called the G-quadruplexes, can be formed from G-rich single-stranded DNA. There is now a good level of understanding of the structural biophysics of G-quadruplexes along with a large body of work on the design and synthesis of small organic molecules that can target such structures. Some such molecules have been found to have antiproliferative effects on human cancer cells along with specific effects on the activity of certain genes, raising intriguing hypotheses about their existence and function in cells. Recent results from my laboratory provide new insights into when and where such non-Watson-Crick structures may exist in cellular DNA, together with a perspective on whether this structural motif in DNA presents new opportunities to think about intervention and future therapeutics.
Citation Format: Shankar Balasubramanian. G-quadruplexes and DNA dynamics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr PL04-01. doi:10.1158/1538-7445.AM2017-PL04-01
Abstract SY06-02: Macrophage-targeted cancer immunotherapy
Authors: Ries Carola 
Published: July 2017

Abstract
Combining immunotherapies that harness cytotoxic T cells to fight cancer has become a major effort in treating metastatic cancer patients. We present data on an example of cancer immunotherapy combination targeting innate immune cells, in particular tumor-associated macrophages (TAMs), that is currently being tested in the clinic. Combination of a TAM-depleting anti-CSF-1R antibody with an activating anti-CD40 agonist demonstrated synergistic antitumoral efficacy resulting in complete tumor remission in preclinical mouse cancer models. Ex vivo RNA sequencing of sorted TAM revealed that only the combination treatment induced a transient, profound repolarization of TAMs towards an immunostimulatory M1 phenotype. However, these TAM populations are ultimately eliminated. The central role of macrophages was further substantiated by complete loss of therapeutic benefit when TAMs were depleted prior to combinational treatment. Furthermore, rechallenge and depletion experiments demonstrated that tumor rejection is finally mediated by an antitumoral T-cell response. In colorectal cancer patients coexpression of human CSF1R and CD40 on TAMs was detected, providing the basis to explore this combination in the clinic.
Citation Format: Carola Ries. Macrophage-targeted cancer immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr SY06-02. doi:10.1158/1538-7445.AM2017-SY06-02