bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5175: Expression profile and role of LOXL3 in astrocytomas
Authors: Laurentinho Talita de Souza , Soares Roseli da Silva , Marie Suely Kazue , Oba-Shinjo Sueli Mieko 
Published: August 2019

Abstract
Astrocytomas are the most common primary central nervous system tumors in adults. The WHO classification based on histologial characteristics according consider four malignant grades, glioblastoma (GBM) being the most malignant (grade IV). Molecular classification of GBM is based on the integration of somatic mutations, DNA methylation and specific transcript/protein expression and consider four subtypes: proneural, classic, mesenchymal and neural. Lysyl oxidase family is composed of five members (LOX, LOXL1, LOXL2, LOXL3 and LOXL4) which are enzymes responsible for catalyzing lysine-derived cross-links of collagen and elastin. The aim of the present work was to analyze the gene expression of LOXL3 in astrocytomas of different grades and different GBM molecular subtypes, the impact of LOXL3 expression level on overall survival in GBM cases of our cohort. These data were also analyzed in the TCGA database to validate our findings. LOXL3expression increased with astrocytoma malignant grades, and proved to be a prognostic factor, as GBM patients with lower LOXL3 expression presented longer survival time than those with higher expression (p&lt;0.041). Analogous data was obtained in the TCGA database, when considering temozolamide-treated GBM patients (p=0.049). Additionally, LOXL3 expression was higher in mesenchymal GBM subtype than in proneural and classic GBM subtypes in TCGA cohort (p&lt;0.001). To further understand LOXL3 functional role in gliomagenesis, LOXL3 was silenced with two distinct siRNA sequences in U87MG, a mesenchymal subtype of GBM cell line. LOXL3 down regulation was confirmed at gene and protein expression levels. Functional assays with silenced LOXL3 demonstrated a decrease in the cell proliferation (p&lt;0.05), and an increase in the apoptosis associated to temozolamide treatment (p&lt;0.05). Immunofluorescence analysis in U87MG cells showed LOXL3 colocalized with mitochondria. Interestingly, such colocalization decreased when LOXL3 was downregulated, associated to a clear morphological alteration in mitochondria shape. A rough increase in mitochondria volume was observed, suggesting fused mitochondria. In addition, such condition was associated to a 23% decrease in mitochondrial DNA copy number. Altogether, our findings suggest that LOXL3 is upregulated in GBM, with impact in prognosis and might play a role in mitochondria dynamics and/or mitophagy, modulating apoptosis. Transcriptome analysis, currently in progress, will further clarify the associated players in this signaling pathway.
Citation Format: Talita de Souza Laurentinho, Roseli da Silva Soares, Suely Kazue Marie, Sueli Mieko Oba-Shinjo. Expression profile and role of LOXL3 in astrocytomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5175.
Abstract 5184: N-CoR2 epigenetically regulates adhesion-dependent branching morphogenesis and its loss correlates with malignant progression of the mammary gland
Authors: Huang Shenq-Shyang , Weaver Valerie M. , Tsai Kelvin K.C. 
Published: August 2019

Abstract
Branching morphogenesis depends upon extracellular matrix remodeling and dynamic cell-cell and cell-extracellular matrix interactions, although how this developmental program is coordinated remains unclear. Expression profiling of mammary tissue at different stages of branching morphogenesis combined with histological and functional studies identified the histone deacetylase (HDAC) regulator, nuclear receptor co-repressor 2 (N-CoR2), as a key orchestrator of adhesion-dependent branching morphogenesis in the mammary gland. Molecular studies revealed that N-CoR2 actively represses fibronectin (FN) and thrombospondin-1 (THBS-1) promoter accessibility in an HDAC-dependent manner. Consistently, histological analysis revealed that N-CoR2 is abundantly expressed in the luminal epithelial cells localized along the outer rim of invading terminal end buds (TEB) and that its expression is lowest at sites of TEB branching bifurcation. In addition, reducing N-CoR2 expression drove the scattering of human mammary acini in vitro and promoted promiscuous branching of humanized mouse mammary tissue in vivo, suggesting that downregulation of N-CoR2 is required for branching morphogenesis in mammary gland. Furthermore, downregulation of N-CoR2 in breast cancer cells increased the expression of both FN and THBS-1 and reduced the expression of E-cadherin and enhanced the migratory capability of breast cancer cells. Intriguingly, immunohistochemical and clinical correlative analysis indicated that loss of N-CoR2 correlates with perturbed cell-cell and enhanced cell-ECM adhesion and acquisition of an invasive phenotype and local lymph node metastasis in a subset of breast carcinomas. These findings suggest N-CoR2 may epigenetically regulate adhesion-dependent branching morphogenesis in the mammary gland and that its loss could promote malignant progression of some breast cancers.
Citation Format: Shenq-Shyang Huang, Valerie M. Weaver, Kelvin K.C. Tsai. N-CoR2 epigenetically regulates adhesion-dependent branching morphogenesis and its loss correlates with malignant progression of the mammary gland [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5184.
Abstract 5193: Conditional knockout of histone chaperone FACT in mice: Consequences for normal tissues
Authors: Ito Masaki A. , Sandlesh Poorva , Goswami Imon , Prendergast Laura A. , Gurova Katerina 
Published: August 2019

Abstract
Histone chaperone FAcilitates Chromatin Transcription (FACT) is, a complex of SSRP1 and SPT16, involved in chromatin remodeling during transcription, replication, and DNA repair. FACT is selectively expressed during early stages of development and later exclusively present in the stem cell niches in adult normal tissues. It is expressed in undifferentiated aggressive tumors with poor prognosis.¹ FACT inhibition is not tolerated by most tumor, unlike normal cells. This vulnerability of tumor cells upon FACT inactivation suggests evaluating FACT inhibition as an antitumor therapeutic approach.¹ Since FACT knockout (KO) is embryonically lethal (E3.5),² study of FACT’s function in different non-tumor cells of mammalian organisms was not possible and was mostly studied in single cell organisms and in tumor cell lines. Therefore, to assess the safety of FACT inhibition as an anticancer approach, by understanding its role in normal physiology, we generated a conditional KO of Ssrp1 mouse model using CreERT2-LoxP system.³ We harvested stem cells of different lineages to better understand the role of FACT in normal mouse physiology. Using the hematopoietic stem cells (HSCs) from the bone marrow (BM), we performed BM colony formation assay. We found that FACT inactivation significantly decreased the number of colonies of HSCs. Furthermore, inhibiting FACT significantly increased caspase 3 expression in mouse adipose stem cells (ASCs), resulting in apoptosis and decreased survival. These results prompted us to comprehensively evaluate roles of FACT in stem cell development and survival. To this end, fibroblasts in a conditional knockout mouse were reprogrammed by retroviral vector encoding four pluripotency transcription factors (Oct3/4, Klf4, Sox2 and c-Myc). We have found successful generation of induced pluripotent stem (iPS) cells 14 days after reprogramming. In conclusion, FACT associates with development and/or survival of mouse HSCs and ASCs. Further evaluation using iPS cells is expected to delineate roles of FACT in various stages of stem cell development and thus normal physiology. 1. Garcia H, et al. Cell Rep, 2013 4(1) 2. Cao S, et al. Mol Cel Bio. 2003 23(15) 3. Sandlesh P et al. PLOS ONE. 2018 13(6)
Citation Format: Masaki A. Ito, Poorva Sandlesh, Imon Goswami, Laura A. Prendergast, Katerina Gurova. Conditional knockout of histone chaperone FACT in mice: Consequences for normal tissues [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5193.
Abstract 5202: Molecular mechanism of transcriptional regulation of REV7, a DNA damage response gene
Authors: Murakumo Yoshiki , Shimada Yuko , Kato Takuya , Sakurai Yasutaka 
Published: August 2019

Abstract
REV7 is involved in several biological response after DNA damage, including the double strand break repair and translesion DNA synthesis. It has bee reported that high levels of REV7 expression are observed in several malignant tumors and are associated with resistance to chemotherapeutic agents such as cisplatin. In normal tissues, REV7 expression is extremely high in germ cells compared with the other normal cells, suggesting the cell type-specific expression of REV7. However, the mechanism of regulation of REV7 expression has not been understood. To assess the mechanism of regulation of REV7 expression, we analyzed the promotor region of the human REV7 locus to identify transcriptional regulators controlling REV7 expression. The transcriptional start point of the REV7 gene was determined by 5’-RACE using human testis cDNA. About 3 kb of the upstream region of the REV7 locus was isolated by genomic PCR and the promoter activity was analyzed by the luciferase reporter assay. Using several deletion mutants, we found that there are two transcriptional activation regions in the upstream of the REV7 locus. Bioinformatic analyses revealed several motifs of transcription factor binding sites in these regions, and we focused on one molecule as a candidate for the transcription factor controlling REV7 expression. We will present and discuss our further analyses.
Note: This abstract was not presented at the meeting.
Citation Format: Yoshiki Murakumo, Yuko Shimada, Takuya Kato, Yasutaka Sakurai. Molecular mechanism of transcriptional regulation of REV7, a DNA damage response gene [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5202.
Abstract 5217: RB loss reprograms AR and E2F1 signaling in models of prostate cancer progression
Authors: Mandigo Amy C. , McNair Chris , Schiewer Matthew J. , Knudsen Karen E. 
Published: August 2019

Abstract
Prostatic adenocarcinoma (PCa) is the most frequently diagnosed non-cutaneous malignancy, and the third leading cause of cancer death in males in the United States. A crucial component to the development and progression of PCa is the activity of the androgen receptor (AR). As such, targeting the AR-signaling axis through androgen deprivation therapy (ADT) is the first line of therapy for PCa. However, cells invariably become resistant to this therapy and men relapse with the incurable form of the disease termed, castration-resistant prostate cancer (CRPC). In addition to AR, another principal component aiding in the progression of disease is the retinoblastoma tumor suppressor protein (RB). RB functions to repress tumor development by negatively regulating the activity of the E2F family of transcription factors, preventing cell cycle progression. RB is lost in roughly 30% of CRPC tumors and is sufficient to induce a CRPC phenotype in hormone-sensitive cells under ADT conditions. Analyses into the molecular significance of RB loss on disease progression identified a potential cooperation between AR and the RB-E2F1 signaling axis. Biological assessment performed in isogenic RB knockdown (i.e. hormone-sensitive and CRPC models) identified distinct functional consequences of RB loss depending on AR status and disease state. Transcriptome analysis identified divergently regulated gene signatures between disease stages in the presence of AR activation, which were not seen under ADT conditions, implicating a unique role of AR in transcriptional regulation with the loss of RB. Data to be discussed will also include further comparison of the E2F1 and AR cistromes in the absence of RB to identify the mechanism by which the AR-RB-E2F1 signaling axis function in promoting the progression of disease.
Citation Format: Amy C. Mandigo, Chris McNair, Matthew J. Schiewer, Karen E. Knudsen. RB loss reprograms AR and E2F1 signaling in models of prostate cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5217.
Abstract 5226: Non-canonical functions of TERT in glioblastoma pathobiology
Authors: Ferdosi Shayesteh R. , Bollam Saumya , Peng Sen , Taylor Brett , Gokhale Vijay , Hurley Laurence , Berens Michael , Dhruv Harshil 
Published: August 2019

Abstract
Mutations in the promotor region of TERT are the most common non-coding mutations across cancers, especially in glioblastoma (GBM), which has a TERT mutation frequency rate over 80%. In the absence of any effective molecular targeting therapy for GBM, elucidating oncogenic signaling of TERT could open new avenues in GBM treatment. Canonically, mutations of TERT, which result in TERT upregulation, maintain telomere length in the nucleus and promote indefinite proliferation of cancer cells. However, a non-canonical function of TERT in the mitochondria has recently been suggested. We screened GBM cell models against a novel small molecule inhibitor (RG1534, Reglagene Inc.) that interferes with the functionality of a mutated hTERT promoter. RG1534 selectively suppresses glioma cell viability without affecting non-transformed normal human astrocytes. More interestingly, RG1534 treatment leads to rapid apoptosis induction in glioma cell lines that does not correlate with the time course of the telomere shortening effect. We further validated this rapid apoptosis behavior in glioma cell lines using siRNA and CRISPR/Cas-9 mediated hTERT knockdown. We also measured the protein expression of TERT in subcellular fractions of glioma cell lines and demonstrated the presence of higher TERT expression in mitochondrial extract compared to the nuclear extract. Finally, using MitoSOX dye we demonstrated significant increase of ROS generation in glioma cells treated with RG1534 as compared to control. In summary, our results demonstrate that non-canonical functions of TERT may play critical role in glioma pathobiology and require more detail investigation.
Citation Format: Shayesteh R. Ferdosi, Saumya Bollam, Sen Peng, Brett Taylor, Vijay Gokhale, Laurence Hurley, Michael Berens, Harshil Dhruv. Non-canonical functions of TERT in glioblastoma pathobiology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5226.
Abstract 5234: Epigenetic control of heparanase expression through CRISPR/dCas9
Authors: Zeng Guihua , Liang Fu-Sen , Cui Lina 
Published: August 2019

Abstract
Heparanase is the only known enzyme that cleaves heparan sulfate (HS) and hence participates in the degradation and remodeling of the extracellular matrix (ECM). Pathological expression of heparanase at abnormal levels can contribute to the development of various diseases. In our research, directed by clustered regularly interspaced short palindromic repeats (CRISPR) system, the promoter region of heparanase was successfully and specifically targeted with screened and combined single guide RNAs (sgRNAs). Linking dCas9 with different histone modification enzymes (eg. P300, hHDAC3, LSD1), the expression of heparanase was successfully up- or down-regulated through acetylation, deacetylation and demethylation. This approach allows us to tune heparanase expression in various cell lines on demand, enabling the study of heparanase activity in real time.
Citation Format: Guihua Zeng, Fu-Sen Liang, Lina Cui. Epigenetic control of heparanase expression through CRISPR/dCas9 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5234.
Abstract 5241: Whole transcriptome TempO-Seq profiling of focal areas of H&E stained FFPE: Differentiation of normal colon and cancer phenotypes between donors
Authors: Trejo Christy , Imler Elliot , Babic Milos , Shepard Peter , Nagle Raymond , Yeakley Joanne , Seligmann Bruce 
Published: August 2019

Abstract
Profiling gene expression from small amounts of FFPE using methods that require extraction of RNA from the tissue, such as RNA-seq, have been problematic. We report use of a modification of the whole transcriptome TempO-Seq® assay (Yeakley et al, PLOSone, 2017; DOI:10.1371/journal.pone.0178302) to profile 2 mm2 areas of 5 μm thick FFPE sections by scraping the tissue and assaying lysates directly without extracting the RNA. Duration of fixation and the amount of sample down to a 2 mm2 area do not impact the performance of the assay. Good results can be achieved using FFPE blocks that are up to 30 years old. As a consequence of the ability to assay areas as small as 2 mm2 from H&E stained tissue, we were able to scrape and profile areas of colorectal cancer vs adjacent normal and obtain replicates within donors, and then compare between donors. The assay shows excellent reproducibility, with R2 &gt; 0.95 between biological replicates (separate samples taken from the same patient and lysed/processed separately). While the normal tissues from the patients exhibited a range of expression variability, they all clustered very tightly when analyzed via principal component analysis. The cancer tissue profile from each patient was not only substantially different from the normal tissue profile of that patient, but substantially different from all profiles within the normal tissue cluster of all patients. Furthermore, the cancer tissue profiles were separated into distinct subgroups. Changes in regulatory pathways leading to oncogenesis could be directly distinguished between patients. These results demonstrate the ability to use whole transcriptome TempO-Seq profiling to investigate molecular subtypes of colon cancer using archival FFPE samples. We are expanding the cohort of patients being profiled to investigate these signatures further, and to draw out signatures which allow direct subtype classification purely from expression data.
Citation Format: Christy Trejo, Elliot Imler, Milos Babic, Peter Shepard, Raymond Nagle, Joanne Yeakley, Bruce Seligmann. Whole transcriptome TempO-Seq profiling of focal areas of H&E stained FFPE: Differentiation of normal colon and cancer phenotypes between donors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5241.
Abstract 5245: Integrative analysis of KIF4A, 9, 18A, and 23 and their clinical significance in low-grade glioma and glioblastoma multiforme
Authors: Cho SangYeon , Kim DongWoon 
Published: August 2019

Abstract
To determine the prognostic significance of kinesin superfamily gene (KIF) expression in patients with brain cancer, including low-grade glioma (LGG) and glioblastoma multiforme (GBM), we comprehensively analyzed KIFs in 515 LGG and 595 GBM patients. Among KIFs, KIF4A, 9, 18A, and 23 showed significant clinical implications in both LGG and GBM. The mRNA and protein expression levels of KIF4A, 9, 18A, and 23 were significantly increased in LGG and GBM compared with those in the normal control groups. The mRNA expression levels of KIF4A, 9, 18A, and 23 in LGG were significantly increased in the high-histologic-grade group compared with those with a low histologic grade. Genomic analysis showed that the percent of mRNA upregulation of KIF4, 9, 18A, and 23 was higher than that of other gene alterations, including gene amplification, deep deletion, and missense mutation. In addition, LGG patients with KIF4A, 18A, and 23 gene alterations were significantly associated with a poor prognosis. In survival analysis, the group with high expression of KIF4A, 9, 18A, and 23 mRNA was significantly associated with a poor prognosis in both LGG and GBM patients. Gene Set Enrichment Analysis (GSEA) revealed that high mRNA expression of KIF4A, 18A, and 23 in LGG and GBM patients showed significant positive correlations with the cell cycle, E2F targets, G2M checkpoint, Myc target, and mitotic spindle. By contrast, high mRNA expression of KIF9 in both LGG and GBM patients was significantly negatively correlated with the cell cycle, G2M checkpoint, and mitotic spindle pathway. However, it was significantly positively correlated with EMT and angiogenesis. This study has extended our knowledge of KIF4A, 9, 18A, and 23 in LGG and GBM and shed light on their clinical relevance, which should help to improve the treatment and prognosis of LGG and GBM.
Citation Format: SangYeon Cho, DongWoon Kim. Integrative analysis of KIF4A, 9, 18A, and 23 and their clinical significance in low-grade glioma and glioblastoma multiforme [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5245.