bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract A03: Inhibition of the histone/protein acetyltransferase, Tip60, impairs Foxp3 acetylation and Treg function and thereby promotes antitumor immunity
Authors: Hancock Wayne W. , Wang Liqing , Newick Kheng , Beeler Aaron , Albelda Steven M. 
Published: May 2016

Abstract
Although the important role of Foxp3+ Tregs in suppressing immune responses to solid tumors is well accepted, to date, there has been little success in targeting this population other than by their transient depletion or “secondary” modulation by mAbs to CTLA-4. In contrast, we have undertaken a mechanism-driven approach to Treg targeting based on our ongoing research into the post-translational modifications that regulate Foxp3, such as its acetylation by one or more histone/protein acetyltransferase (HAT) enzymes. Our analysis of each of the main HAT families has shown that members of the Myst family have the greatest contribution, with Tip60 deletion leading to lethal autoimmunity within 3 weeks of birth. We have now translated these insights into the development and testing of a new pharmacologic inhibitor of Tip60 (Tip60i) that binds to the Ac-CoA site of Tip60. First, submicromolar levels of this Tip60i impaired Treg function in vitro but preserved conventional T cell proliferation. Second, Tip60 administration in vivo (10 mg/kg/d, 14 d) revoked Treg-dependent allograft tolerance induced by CD40L mAb plus donor splenocyte transfusion, and restored T cell-dependent cardiac allograft rejection. These studies illustrate the ability of the Tip60i to impair Treg functions while preserving protective effector T cell functions. Third, treatment of immunocompetent mice bearing established TC1 lung adenocarcinomas (which we have shown is a Treg-sensitive tumor) with Tip60i (1 mg/kg/d) significantly impaired tumor growth (p&lt;0.001). While further medicinal chemistry of this compound is underway, including modification with phthalimide tags, our data provide the crucial first evidence that Tip60 is a druggable and potent target for modulation of Foxp3+ Treg function in vitro and in vivo, and show that Tip60 inhibition can enhance host anti-tumor immunity.
Citation Format: Wayne W. Hancock, Liqing Wang, Kheng Newick, Aaron Beeler, Steven M. Albelda. Inhibition of the histone/protein acetyltransferase, Tip60, impairs Foxp3 acetylation and Treg function and thereby promotes antitumor immunity. [abstract]. In: Proceedings of the Fourth AACR International Conference on Frontiers in Basic Cancer Research; 2015 Oct 23-26; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2016;76(3 Suppl):Abstract nr A03.
Abstract A16: The crosstalk between Indoleamine 2,3 Dioxygenase (IDO) and cancer stem cells in murine melanoma: A potential modality of local control
Authors: Kocherla Marsha , Christianson Blake , Walker James , Qin Xu , Yu Jack C. , Baban Babak 
Published: May 2016

Abstract
Introduction: Indoleamine 2,3-dioxygenase (IDO), the rate-limiting enzyme of tryptophan catabolism, has been strongly associated with the progression of malignancy and poor survival in melanoma patients. As a result, IDO is a leading target for interventions aimed at restoring melanoma immune surveillance. IDO, which is produced by cells including dendritic cells, macrophages, stem cells, and most tumor cells, potently suppresses immune responses by depleting local tryptophan and allowing the accumulation of tryptophan metabolites. However, the actual cellular and molecular mechanisms underlying the immunoregulatory function of IDO remain largely unknown. The relatively newly described cancer stem cells (CSCs) are tumor-initiating cells (TICs) that often survive therapy and give rise to second-line tumors by self-renewal and differentiation into multiple cell types. CSCs are proposed to persist in tumors as a distinct population and to cause relapse and metastasis by giving rise to new tumors. Therefore, development of specific therapies targeted at CSCs holds hope for improvement of survival and quality of life of cancer patients, especially those with metastatic disease. Methods: C57/BL6 mice were injected with B16F10 murine melanoma cell line (1 x 106 cells/mouse, subcutaneous). Tumor growth monitored. At 3mm, tumors were injected with an IDO Inhibitor (1 MT) (Group 1) twice weekly (20 mg/mouse per injection). Controls (Group 2) received vehicle only (PBS), Group 3 weekly Cisplatin (5mg/kg), and Group 4 both Cisplatin and 1MT (IDO inhibitor). Once the first tumor reached 12mm in size, the mice were sacrificed and tumor cells subjected to flow-cytometry, and statistical analysis, and p-values &lt;0.05 considered significant. Results: Our data showed for the first time that IDO was expressed by CSCs in a murine melanoma model. These IDO-expressing CSCs may be capable of suppressing T lymphocyte infiltration into the tumor microenvironment, which was reversed by the IDO inhibitor, 1-methyl-tryptophan. Conclusion: Our findings offer an important new line of evidence that interventional targeting of IDO activity could be used to restore tumor immunity by relieving IDO-mediated immune suppression of CSCs in the tumor microenvironment as well as in tumor cells themselves.
Citation Format: Marsha Kocherla, Blake Christianson, James Walker, Xu Qin, Jack C. Yu, Babak Baban. The crosstalk between Indoleamine 2,3 Dioxygenase (IDO) and cancer stem cells in murine melanoma: A potential modality of local control. [abstract]. In: Proceedings of the Fourth AACR International Conference on Frontiers in Basic Cancer Research; 2015 Oct 23-26; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2016;76(3 Suppl):Abstract nr A16.
Abstract A17: The CEBPD transcription factor: A signaling hub for promotion of cancer cell stemness
Authors: Kuppusamy Balamurugan , Mendoza-Villaneuva Daniel , Summers Glenn , Sharan Shikha , Sterneck Esta 
Published: May 2016

Abstract
Our previous studies showed that the transcription factor CCAAT/enhancer binding protein delta (CEBPD) promotes tumor metastasis in a mouse model of mammary tumorigenesis (MMTV-Neu) (Balamurugan et al., EMBO J, 2010). Here we show that CEBPD serves as a signaling hub, which integrates and amplifies signaling pathways that promote stemness in breast cancer and glioblastoma as well as in embryonic carcinoma stem cells. This work identifies a mechanism by which hypoxia and inflammation promote cancer stem-like cells (CSCs), which are implicated in tumor progression, metastasis, and therapy resistance.
We have shown in our earlier studies that CEBPD promotes inflammatory signaling and hypoxia adaptation by inhibiting the expression of the F-box protein FBXW7, a bona fide tumor suppressor (Balamurugan et al., EMBO J 2010; Nature Communications 2013). Because FBXW7 can target the stem cell factor Notch for degradation, we hypothesized that CEBPD can promote cell stemness. We tested this hypothesis using a variety of established breast and glioblastoma tumor cell lines/xenografts, primary mouse mammary tumor cells and tissues, recent patient-derived glioblastoma cell lines, as well as embryonic stem/cancer cell lines. Deletion or depletion of CEBPD reduced the number of stem cells as determined by surface markers, sphere formation, limiting dilution xenografts, and by expression of stemness genes. Mechanistic studies in culture models show that CEBPD is expressed in CSCs and connects and amplifies both hypoxia (HIF-1) and inflammation (IL-6/STAT3) pathways to increase Notch1 activity and cell stemness. Furthermore, we found that CEBPD also directly activates the expression of stemness associated genes such as OCT4, Sox2, KLF4, Myc, Nanog, CD44 and CD133. Taken together, this study provides novel insights into the molecular mechanisms that promote cancer cell stemness, and identifies a central role for CEBPD in integrating HIF-1α,
IL-6/STAT3, and Notch1 signaling, while also activating expression of stemness factors. These results strongly suggest that pharmacological inhibition of CEBPD signaling may effectively target CSCs.
Citation Format: Balamurugan Kuppusamy, Daniel Mendoza-Villaneuva, Glenn Summers, Shikha Sharan, Esta Sterneck. The CEBPD transcription factor: A signaling hub for promotion of cancer cell stemness. [abstract]. In: Proceedings of the Fourth AACR International Conference on Frontiers in Basic Cancer Research; 2015 Oct 23-26; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2016;76(3 Suppl):Abstract nr A17.
Abstract A18: The tankyrase inhibitor G007-LK inhibits small intestine Lgr5+ stem cell proliferation without altering tissue morphology
Authors: Norum Jens Henrik , Skarpen Ellen , Brech Andreas , Kuiper Raoul , Waaler Jo , Krauss Stefan , Sørlie Therese 
Published: May 2016

Abstract
Tankyrase enzymes play crucial roles in the Wnt signaling pathway, involved in the regulation of intestinal stem cells and tissue homeostasis. Aberrant Wnt signaling is known to cause intestinal cancers and the tankyrase inhibitor G007-LK has previously been shown to inhibit tumor growth in an APC-mutant colorectal cancer xenograft model.
We have used in vivo models to address the effect of G007-LK on small intestine tissue homeostasis. The mice were treated for up to 3 weeks with the tankyrase inhibitor, without affecting the body mass or observed clinical condition of the mice. H&E staining of fixed small intestine tissue sections showed no significant differences in the general tissue morphology between control and G007-LK treated mice. In the presence of G007-LK, lineage tracing from the Lgr5+ intestinal stem cells was significantly reduced compared to control mice. When the G007-LK treatment was stopped two days prior to initiation of lineage tracing, there were no significant difference in the number of traced cells between the mice pretreated with G007-LK and untreated control group. Immunohistochemistry (IHC) staining for the proliferation marker Ki67 showed a slight reduction in the number of positive cells in the small intestine of the G007-LK treated compared to control mice. As expected, in the crypt base cells there was a reduction of nuclei staining positive for β-catenin in the G007-LK treated compared to control mice, suggesting repressed Wnt/β-catenin signaling.
Taken together, our data show that the tankyrase inhibitor G007-LK is well tolerated by the mice. The tankyrase inhibitor reduced the proliferative activity of the small intestine Lgr5+ stem cells, without altering the general morphology or function of the tissue.
Citation Format: Jens Henrik Norum, Ellen Skarpen, Andreas Brech, Raoul Kuiper, Jo Waaler, Stefan Krauss, Therese Sørlie. The tankyrase inhibitor G007-LK inhibits small intestine Lgr5+ stem cell proliferation without altering tissue morphology. [abstract]. In: Proceedings of the Fourth AACR International Conference on Frontiers in Basic Cancer Research; 2015 Oct 23-26; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2016;76(3 Suppl):Abstract nr A18.
Abstract A27: An in vivo gain-of-function screening identified the Williams-Beuren syndrome gene GTF2IRD1 as a mammary tumor promoter
Authors: Huo Yongliang , Su Yinghao , Cai Qiuyin , Macara Ian 
Published: May 2016

Abstract
Precision medicine has shown great efficacy in oncology, but its broad application requires a full understanding of the cancer genome. We have performed the first gain-of-function screening in mice to functionally identify candidates from a “druggable” gene set for their roles in breast cancer. We transduced mammary gland stem/progenitor cells isolated from MMTV-ErbB2 transgenic mice with a lentiviral cDNA library and implanted them into the cleared fat pads of syngeneic recipients. The transplanted mammary gland cells formed a normal mammary ductal tree within 6 weeks. After another 8 months latency, small lesions were detected along the mammary ducts in mice harboring cells that had been transduced with the lentiviral library. We isolated genomic DNA from these lesions to amplify any lentiviral cDNA sequences within the lesions and identified individual genes by sequencing. Gain-of-function of five genes, including RET, GTF2IRD1, ADORA1, LARS2, and DPP8, was found to promote mammary tumor growth consistently. We further studied one novel tumor promoting gene, transcription factor GTF2IRD1. GTF2IRD1 regulates the expression of multiple genes in the mammary gland system, including OPN, TβR2, and BMPR1b. In tissue culture, high levels of GTF2IRD1 inhibited cell proliferation but increased the colony formation ability of the un-transformed mammary gland cell line MCF10A. In mice, the mis-regulation of GTF2IRD1, OPN, TβR2, and BMPR1b, also individually promoted mammary cancer development. Silencing of TβR2 delayed the incidence of tumor formation by GTF2IRD1. We also found that GTF2IRD1 is highly expressed in human breast tumors, correlating with high tumor grades and poor prognosis.
In conclusion, we performed the first gain-of-function screening in vivo for breast cancer promoting genes. All candidate genes from this study are potential drug targets to treat cancer. We showed that GTF2IRD1 promotes breast cancer development in mice through regulation of its downstream targets, and correlates clinically with high tumor grades and poor prognosis. We believe our in vivo screening method is readily expandable to whole genome annotation of cancer genes.
Citation Format: Yongliang Huo, Yinghao Su, Qiuyin Cai, Ian Macara. An in vivo gain-of-function screening identified the Williams-Beuren syndrome gene GTF2IRD1 as a mammary tumor promoter. [abstract]. In: Proceedings of the Fourth AACR International Conference on Frontiers in Basic Cancer Research; 2015 Oct 23-26; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2016;76(3 Suppl):Abstract nr A27.
Abstract A46: Cellular apoptosis susceptibility (CAS) regulates integrin-beta1 and is required for tumor cell migration and invasion in hepatocellular carcinoma (HCC)
Authors: Winkler Juliane , Sticht Carsten , DiGuilio Amanda , Eiteneuer Eva Maria , Holzer Kerstin , Rössler Stephanie , Gretz Norbert , Breuhahn Kai , Schirmacher Peter , Ori Alessandro , Singer Stephan 
Published: May 2016

Abstract
Nuclear transport is an essential, highly selective process in eukaryotic cells which guarantees the bidirectional communication between the nucleus and the cytoplasm. Emerging evidence suggests that aberrant expression of nuclear transport factors may contribute to cancer formation and progression. We could recently show that the nuclear exporter Cellular apoptosis susceptibility (CAS) is overexpressed in hepatocellular carcinoma (HCC), the functional implications of which remain, however, not completely understood. Here, we integrated proteomics, transcriptomics and functional assays with patient data to further characterize the role of CAS in HCC. We found that CAS depletion in HCC cells led to deregulation of integrins, particularly integrin-beta1, and resulted in reduced cell migration and invasion. Moreover, we determined that high expression levels of CAS in HCC cases were associated with macroangioinvasion and poorer patient outcome. Our data indicate that CAS is linked to integrin signaling and correlates with an aggressive HCC phenotype.
Citation Format: Juliane Winkler, Carsten Sticht, Amanda DiGuilio, Eva Maria Eiteneuer, Kerstin Holzer, Stephanie Rössler, Norbert Gretz, Kai Breuhahn, Peter Schirmacher, Alessandro Ori, Stephan Singer. Cellular apoptosis susceptibility (CAS) regulates integrin-beta1 and is required for tumor cell migration and invasion in hepatocellular carcinoma (HCC). [abstract]. In: Proceedings of the Fourth AACR International Conference on Frontiers in Basic Cancer Research; 2015 Oct 23-26; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2016;76(3 Suppl):Abstract nr A46.
Abstract B05: Invadopodium formation is enriched in perivascular niches and leads to cancer cell intravasation
Authors: Bergman Aviv , Condeelis John , Gligorijevic Bojana 
Published: May 2016

Abstract
Invadopodia are actin-rich protrusions used for cancer cell movement across obstacles. In the past, they were mostly studied in cell cultures, where they form in contact with extracellular matrix. Our past work has suggested that elimination of key molecular components of invadopodia in cancer cells can lead to elimination of metastasis.
In the current study, we use high resolution multiphoton microscopy to understand how the tumor microenvironment controls formation of invadopodia, followed by cancer cell locomotion, intravasation and metastasis. We demonstrate that the motile cancer cells in breast carcinoma in vivo can exhibit either invadopodium-related, slow locomotion or invadopodium-independent rapid locomotion along collagen fibers. These two phenotypes occur in spatially separate tumor regions, the location of which can be predicted from measurements of microenvironmental features. For example, cancer cells form invadopodia predominantly in perivascular niches, regions surrounding major blood vessels and characterized by cross-linked collagen fibers and dense macrophages. Invadopodia in vivo extend towards blood vessels and their density decreases with blood vessel distance.
We observe that only cells with invadopodia are capable of intravasation, while fast locomotion results in cell dispersal throughout tumor. Further, either invadopodium-specific TkS5 knock-down or short-term inhibition of metalloproteases eliminate invadopodia and stop dissemination to lung. Interestingly, external modulation of collagen cross-linking also inhibits invadopodia, switching the cancer cell phenotype to fast locomotion.
Our results suggest that the perivascular niche is the site of invadopodium formation and cancer cell intravasation. Moreover, invadopodia and hence metastasis, can be inhibited by external modulation of the microenvironment. In summary, the presence of invadopodia is a great candidate for a novel early-metastasis marker, while their elimination via microenvironment modulation has therapeutic potential.
Citation Format: Aviv Bergman, John Condeelis, Bojana Gligorijevic. Invadopodium formation is enriched in perivascular niches and leads to cancer cell intravasation. [abstract]. In: Proceedings of the Fourth AACR International Conference on Frontiers in Basic Cancer Research; 2015 Oct 23-26; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2016;76(3 Suppl):Abstract nr B05.
Abstract B17: Role of Sall2 transcription factor under genotoxic stress: A transcriptomic approach
Authors: Farkas Carlos A. , Fuentes Francisco , Rebolledo Boris , Makova Kateryna , Nekrutenko Anton , Castro Ariel , Pincheira Roxana 
Published: May 2016

Abstract
The induction of genotoxic stress by chemotherapy is one of the first line treatments to eliminate neoplastic cells through apoptosis or programmed cell death. Our laboratory has characterized the Sall2 transcription factor, conserved among vertebrates, as a stress-response molecule, and demonstrated both in vitro and in vivo its requirement in the cell death-response caused by genotoxic stress. Few transcriptional targets of Sall2 are known, thus, additional targets most likely remain to be identified. We isolated mouse embryonic fibroblasts (MEFs) from a murine model deficient in Sall2 and from its normal counterpart, and use them to characterize the Sall2-dependent transcriptional response upon genotoxic stress using RNA-seq experiments. Under control conditions we identified ninety-six Sall2-dependent genes involved in immune system, inflammation, cell adhesion and the response to wounding. Under genotoxic stress one hundred twenty six genes were differentially expressed and dependent on Sall2, these genes are mainly involved in immunity, development and cell-cycle regulation. Finally, we performed validation of the analysis using qPCR. With this experimental outline, we are expecting to gain insights on how Sall2 contributes to the transcriptional program under normal and genotoxic stress-induced conditions.
Citation Format: Carlos A. Farkas, Francisco Fuentes, Boris Rebolledo, Kateryna Makova, Anton Nekrutenko, Ariel Castro, Roxana Pincheira. Role of Sall2 transcription factor under genotoxic stress: A transcriptomic approach. [abstract]. In: Proceedings of the Fourth AACR International Conference on Frontiers in Basic Cancer Research; 2015 Oct 23-26; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2016;76(3 Suppl):Abstract nr B17.
Abstract B18: A simple method to prepare oligonucleotide-conjugated antibodies for multiplex protein detection
Authors: Gong Haibiao , Holcomb Ilona , Ooi Aik , Wang Xiaohui , Majonis Daniel , Unger Marc , Ramakrishnan Ramesh 
Published: May 2016

Abstract
With rapid advances in nucleic acid detection technologies, genomics studies are being conducted in highly multiplexed formats. However, multiplex analysis is still a daunting task in the proteomics field; it would be of great interest to use genomics tools for protein detection. One way to accomplish this is to use oligonucleotide-conjugated antibodies to bind to specific protein targets, and to monitor conjugated-oligonucleotide levels rather than to measure protein levels directly. In this report we describe a simple method for preparing oligonucleotide-conjugated antibodies and discuss this method’s application in oligo extension reaction (OER) for multiplex protein detection. Conjugation is based on strain-promoted alkyne-azide cycloaddition (the Cu-free click reaction), in which the antibody is activated with a dibenzocyclooctyne (DBCO) moiety and linked covalently with an azide-modified oligonucleotide. The reaction condition and purification process were optimized to achieve maximum yield and best performance in the functional test. The OER assay employs a pair of antibody binders (two antibodies, each conjugated with its own oligonucleotide) developed for each protein target. The two oligonucleotides contain a six-base complementary region at their 3′ prime ends to allow annealing and extension by DNA synthesis enzymes to form a DNA template. The template is then detected by qPCR on the Biomark HD system. Distinct oligonucleotide sequences are assigned to different antibody binders to enable multiplex protein detection. When tested using recombinant proteins, some antibody binders, such as those specific to CSTB, MET, EpCAM and CASP3, had an estimated limit of detection (LOD) of about 0.06 pg/mL and a dynamic range of ≥5 logs. These antibody binders were used in multiplex protein detection in single cells using the C1™ system. The results in individual cells showed the expression of similar protein levels for CSTB and CASP3, but highly varied protein levels for EpCAM, CCNB1 and MUC1. This simple method for the preparation of oligonucleotide-conjugated antibodies is essential for the development of affordable multiplex protein assays, which has the potential to accelerate proteomics research.
Citation Format: Haibiao Gong, Ilona Holcomb, Aik Ooi, Xiaohui Wang, Daniel Majonis, Marc Unger, Ramesh Ramakrishnan. A simple method to prepare oligonucleotide-conjugated antibodies for multiplex protein detection. [abstract]. In: Proceedings of the Fourth AACR International Conference on Frontiers in Basic Cancer Research; 2015 Oct 23-26; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2016;76(3 Suppl):Abstract nr B18.