bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4783: Suppression of IL-6-induced metastatic potential by inhibition of mitochondrial fission in ovarian cancer cells
Authors: Lee Juwon , Han Youngjin , Song Yong Sang 
Published: September 2020

Abstract
Ovarian cancer is the fifth most common cause of cancer death among females in the United States. In tumor microenvironment, inflammation has been shown to play a key role, governing malignant properties of cancer cells such as angiogenesis, chemoresistance and metastasis. Interleukin-6 (IL-6) is a pro-inflammatory cytokine, produced in response to injury, infection, and inflammation. IL-6 can regulate growth, differentiation and gene expression of cells. In our previous study, we have shown that IL-6 was found to be present at a high level in malignant ascites when compared to those in benign ascites. Similar to many other oncogenic cytokines, IL-6 has been reported to increase malignant properties of cancer such as metastasis. Mitochondria, energy plants of cells regulating metabolic homeostasis, are structurally dynamic organelles undergoing continuous fission and fusion. In metastatic cancer cells, mitochondrial fission has been shown to be more frequent, than in the primary cancer cells. A key facilitator of mitochondrial fission is a molecule called dynamin-1-related protein 1 (Drp1) and its phosphorylation status can determine the activity of the molecule. Herein, we investigated the effect of the inflammatory cytokine IL-6 on the mitochondrial dynamics and the relationship between the changes in mitochondrial dynamics and metastatic potential of ovarian cancer cells. Treatment of IL-6 on epithelial ovarian cancer (EOC) cell lines (PA-1 and SKOV3) increased the metastatic potential, such as invasion and migration, verified by trans-well invasion and wound healing assays. IL-6 did not change the Drp1 expression at the protein level but modulated the ratio of phospho-Drp1s616 (active form) and phospho-Drp1s637 (inactive form), together with increased fission of mitochondria in the EOC cells. An increase of mitochondrial fission by IL-6 was confirmed through MitoSpy staining. Inhibition of mitochondrial fission by RNAi and a pharmacologic inhibitor attenuated metastatic potential of the EOC cells. Altogether, our study strongly suggests the role of mitochondrial fission in the metastatic potential of EOC cells induced by IL-6. The inhibition of mitochondrial fission could be a potential therapeutic approach suppressing the metastasis of EOC.
Citation Format: Juwon Lee, Youngjin Han, Yong Sang Song. Suppression of IL-6-induced metastatic potential by inhibition of mitochondrial fission in ovarian cancer cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4783.
Abstract 4790: An in situ hybridization assay reveals high mitochondrial DNA copy number in stem cell/proliferative cell compartments
Authors: CHEN JIAYU , Zheng Qizhi , Peiffer Lauren B. , Hicks Jessica L. , Haffner Michael C. , Rosenberg Avi Z. , Levi Moshe , Wang Xiaoxin , Ozbek Busra , Yegnasubramanian Srinivasan , De Marzo Angelo M. 
Published: September 2020

Abstract
Mitochondria are critical regulators of energy production, metabolism and cell death. Warburg observed that in the presence of oxygen, many cancer cells take up abundant amounts of glucose, and excrete the carbon mainly as lactate. Warburg postulated that this aerobic glycolysis was essential and universal in cancer, and was caused by mitochondrial dysfunction. Yet, more recent findings suggest that aerobic glycolysis also occurs in rapidly dividing normal cells to support anabolic reactions needed for cell growth and replication. Furthermore, a number of metabolites generated by the tricarboxylic acid cycle and the electron transport chain are also used for macromolecular synthesis in proliferating cancer and normal cells. This suggests that an increase in mitochondrial activity, especially those functions related to anabolic metabolism, may be required for cancer development and progression. One mechanism to increase mitochondrial function is through mitochondrial biogenesis, which is partly regulated by regulating mitochondrial DNA (mtDNA) copy number.
Alterations in mtDNA sequence and copy number have been implicated in various cancers. Since most prior measurements of mtDNA copy number have been performed using homogenates of complex tissues composed of multiple cell types, very little is known about cell type-specific mtDNA copy number heterogeneity. Here, we validated a quantitative in situ hybridization approach to study mtDNA levels at the single cell level in tissue sections. In hierarchically organized self-renewing tissues, higher levels of mtDNA were observed in stem/proliferative compartments than in differentiated compartments. Also, we found zonal heterogeneity of mtDNA levels in the liver that mirrors the known oxygen tension gradient. In the kidney, we observed markedly lower mtDNA levels in glomeruli and the collecting ducts compared to cortical proximal and distal tubules. This work provides visualization of mtDNA copy number in distinct cell populations in situ for the first time across normal mammalian tissues, and can serve as a foundation for quantifying cell type-specific mtDNA copy number dynamics in any relevant diseases, especially in cancers and their precursor lesions.
Citation Format: JIAYU CHEN, Qizhi Zheng, Lauren B. Peiffer, Jessica L. Hicks, Michael C. Haffner, Avi Z. Rosenberg, Moshe Levi, Xiaoxin Wang, Busra Ozbek, Srinivasan Yegnasubramanian, Angelo M. De Marzo. An in situ hybridization assay reveals high mitochondrial DNA copy number in stem cell/proliferative cell compartments [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4790.
Abstract 4802: Functional analysis of cancer associated mitochondrial genome alterations
Authors: Otto Neil 
Published: September 2020

Abstract
It is well documented that a variety of alterations can occur in the mitochondrial genome during human tumor development. These alterations often correlate with cancer phenotypes like treatment resistance and metastasis. Alterations to the mitochondrial genome may therefore be a biomarker for early detection, prognosis and/or predictor of therapeutic sensitivity. However, a method to easily and quickly create isogenic populations of human cells with mitochondrial genome specific alterations is not available. To combat this, we have developed a technology using transient expression of mitochondria targeted TAL domain nickases and TAL endonucleases (TALENs). This the first reliable method to introduce specific mutations into the mitochondrial genome of mammalian cells. We have efficiently induced mtDNA deletions in zebrafish embryos, human and mouse cell lines. In mouse NIH3T3 fibroblasts, we deleted a mitochondrial genome region corresponding to the same disease sequence seen in the human Kearns-Sayre Syndrome (KSS). The deletion resulted in a measurable phenotype including decreased mitochondria basal respiration, ATP production and maximal respiration. We are expanding this work by engineering targeted and precise mtDNA deletions in conjunction with known cancer initiating nuclear mutations in mammalian cells, as well as systemically in mice. These tools will allow us to determine the effect mitochondrial alterations have on the transformation of cancer cells harboring known cancer causing nuclear mutations. We are also developing an inducible system for expressing mitoTALENs, making it possible to create “conditional” loss of function mutations in mitochondrial genes in human cell lines, including induced pluripotent stem cells (iPSCs). This inducible model system will be highly useful in settings where the mitochondrial deletion may cause a selective disadvantage during early cellular differentiation.
Citation Format: Neil Otto. Functional analysis of cancer associated mitochondrial genome alterations [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4802.
Abstract 4815: Differential expression profiling of long noncoding RNA establishes UCA1 as a hallmark of renal medullary carcinoma
Authors: Msaouel Pavlos , Malouf Gabriel G. , Su Xiaoping , Yao Hui , Tripathi Durga N. , Gao Jianjun , Rao Priya , Bertocchio Jean-Philippe , Karam Jose , Wood Christopher G. , Walker Cheryl L. , Genovese Giannicola , Tannir Nizar M. 
Published: September 2020

Abstract
Renal medullary carcinoma (RMC) is a rare but highly lethal renal cell carcinoma (RCC) that predominantly afflicts young African Americans with sickle cell hemoglobinopathies. There are currently no approved targeted therapies for RMC and the median survival from diagnosis is 13 months despite best available treatments. New biomarkers and therapeutic targets are therefore urgently needed for this very aggressive disease. No studies to date have profiled the long noncoding RNA landscape of RMC. We performed RNA sequencing in untreated primary tumor samples obtained from 11 patients with RMC. The lncRNA expression profile of RMC was compared to that of adjacent normal kidney (n = 6 patients with RMC), upper tract urothelial carcinoma (UTUC), and closely related RCCs including collecting duct carcinoma (CDC), clear cell renal cell carcinoma (ccRCC), papillary renal cell carcinoma (pRCC), and chromophobe renal cell carcinoma (chRCC). We performed principal component analyses as well as hierarchical clustering of the lncRNA expression levels using the Pearson correlation coefficient as the distance metric and the ward's linkage rule. The read counts of each lncRNA were fitted using a negative binomial model and a Wald test was used in DESeq2 to compare expression levels. The false discovery rate (FDR) was controlled using the Benjamini and Hochberg method. We found that the lncRNA signature of RMC is clearly distinct from that of ccRCC, pRCC, and chRCC, but substantially overlaps with CDC. Urothelial cancer associated 1 (UCA1) was the highest upregulated lncRNA (305-fold increase, FDR &lt; 0.001) in RMC compared with adjacent normal kidney. This dramatic upregulation of UCA1 was similar to that seen in UTUC and not found in any of the other RCCs, including CDC. UCA1 is known to be associated with chemotherapy resistance and was previously considered to be highly specific for urothelial carcinoma. Our findings now suggest that UCA1 is also a hallmark of RMC. Future studies will focus on elucidating the functional consequences of UCA1 expression in RMC pathobiology.
Citation Format: Pavlos Msaouel, Gabriel G. Malouf, Xiaoping Su, Hui Yao, Durga N. Tripathi, Jianjun Gao, Priya Rao, Jean-Philippe Bertocchio, Jose Karam, Christopher G. Wood, Cheryl L. Walker, Giannicola Genovese, Nizar M. Tannir. Differential expression profiling of long noncoding RNA establishes UCA1 as a hallmark of renal medullary carcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4815.
Abstract 4827: N6-Methyladenosine modification of LINC00901 promotes pancreatic cancer progression
Authors: Peng Wan-Xin , Yang Liu , Mo Yin-Yuan 
Published: September 2020

Abstract
N6-methyladenosine (m6A) is one of the most prevalent modifications of mRNAs in eukaryotes. Numerous recent studies have indicated that m6A plays the critical role in multiple physiologic functions and disease processes by regulation of mRNA processing, splicing, translation, and stability. Although the majority of RNA methylation studies focus on mRNAs, little is known whether long-coding RNAs (lncRNAs) are also subject to m6A modification. Therefore, this study aims to identify m6A modified lncRNAs and their functional role in pancreatic cancer. We performed methylated RNA immunoprecipitation (MeRIP) profiling against a focused group of lncRNAs and identified five lncRNA candidates, among which the highly enriched lncRNA was LINC00901. RNA dot blot, m6A RNA methylation quantification detection and in vivo RNA pulldown combined with m6A RNA methylation Western Blot confirmed the m6A modification of LINC00901. Of note, the analysis of TCGA datasets indicated that the high level of LINC00901 is associated with the poor overall survival (OS) of pancreatic ductal adenocarcinoma (PDAC) patients. Consistent with this result, we showed that LINC00901 promotes pancreatic cancer cell growth. Importantly, we identified that YTHDF1 serves as a “reader” of the m6A modified LINC00901 and furthermore, ectopic expression of YTHDF1 causes downregulation of LINC00901, suggesting a role for m6A modification in regulation of LINC00901 expression. Experiments are underway to determine how LINC00901 promotes pancreatic cancer cell growth.
Citation Format: Wan-Xin Peng, Liu Yang, Yin-Yuan Mo. N6-Methyladenosine modification of LINC00901 promotes pancreatic cancer progression [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4827.
Abstract 4861: Loss of YAP1 activates Wnt/beta-Catenin signaling in prostate cancer
Authors: Cheng Siyuan , Yang Shu , Prieto-Dominguez Nestor , Connelly Zachary M. , Shi Yingli , Yu Xiuping 
Published: September 2020

Abstract
Prostate cancer (PCa) is the most common non-cutaneous cancer among American men. In 2019, 174,650 new patients will be diagnosed from this disease. Most patients with localized PCa could be treated successfully. However, when PCa develops into castrate-resistant prostate cancer (CRPCa), the five-year survival rate drops below 30%. Neuroendocrine prostate cancer (NEPCa) is an aggressive subtype of PCa observed in 20%-30% CRPCa patients. Understanding how NEPCa progressed from prostate adenocarcinoma could reveal novel therapeutic targets. Previous studies indicate that activation of Wnt/beta-Catenin signaling promotes the progression to CRPCa with an increased NE differentiation. Additionally, animal studies suggest that Wnt/beta-Catenin signaling is active in NEPCa. Here we explored the mechanisms that promote Wnt/beta-Catenin signaling in NEPCa. Using TRAMP mice, which develop NEPCa after castration, we assessed the expression of Wnt-related genes and found that after androgen deprivation, the expression of Wnt7A is increased in prostate derived from both wild type and TRAMP mice but the expression of DKK1, a Wnt inhibitor, is decreased in wild type prostate but not in TRAMP prostate. The elevated expression of Wnt7a in combination with the lack of induction of Dkk1 in TRAMP tumors provides a mechanism to activate Wnt/beta-Catenin signaling in these tumors. Furthermore, we found the expression of YES-associated Protein (YAP1), a key transcriptional coactivator of Hippo pathway, is lost in NEPCa, in contrast to the elevated expression of YAP1 in high-grade prostate adenocarcinoma. YAP1 knockdown activated Wnt/beta-Catenin activity in PCa PC3 and 22Rv1 cells, indicating that the loss of YAP1 expression provides another mechanism to activate Wnt/beta-Catenin signaling, promoting NEPCa phenotype in PCa cells. Funding: This study is supported by: FWCC/Foundation Legacy Funds, DOD New Investigator Award, The Office of Research SEED awards, NIH R01 AND R03.
Citation Format: Siyuan Cheng, Shu Yang, Nestor Prieto-Dominguez, Zachary M. Connelly, Yingli Shi, Xiuping Yu. Loss of YAP1 activates Wnt/beta-Catenin signaling in prostate cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4861.
Abstract 4867: Using a fragment based approach for the identification of TEAD S3 pocket binders
Authors: Delarbre Laure , Venier Olivier , Valtingojer Iris , Houtmann Jacques , Lowinski Maryse , Parent Annick , Bégassat Françoise , Lasbleiz Angelique , Seys Catherine , Bombart Philippe , Debussche Laurent , Rak Alexey 
Published: September 2020

Abstract
YAP1 and TEAD are transcriptional regulators involved in organ size control and cell proliferation. Aberrant activation of YAP1 has been reported for multiple tumor types and inhibiting the interaction of YAP1 with its partner protein TEAD is thought to be a means of targeting aberrant YAP1 activity. In order to identify small molecule inhibitors of YAP1 interaction with TEAD, we have performed a fragment-based-screening looking for ligands binding to TEAD at its interface with YAP1 (TEAD pocket S3). We have employed three techniques for fragment screening (NMR, SPR, DSF) and have cross tested positive hits identified by one technique with the respective orthogonal method. Using this strategy on Sanofi proprietary fragment libraries, three different fragment series have been identified. X-Ray co-structures with TEAD have been obtained for all three fragment series confirming binding to the S3 pocket of TEAD. Binding mode determination by X-Ray crystallography has set the stage for subsequent fragment optimization.
Citation Format: Laure Delarbre, Olivier Venier, Iris Valtingojer, Jacques Houtmann, Maryse Lowinski, Annick Parent, Françoise Bégassat, Angelique Lasbleiz, Catherine Seys, Philippe Bombart, Laurent Debussche, Alexey Rak. Using a fragment based approach for the identification of TEAD S3 pocket binders [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4867.
Abstract 4868: In vitro and in cellulo mass spectrometry study of TEAD1 palmitoylation
Authors: Buzy Armelle , Guichard Olivier , Zacchayus Jean-Luc , Venier Olivier , Courtin Olivier , Valtingojer Iris , Debussche Laurent , Mikol Vincent , Guillemot Jean-Claude 
Published: September 2020

Abstract
The HIPPO pathway plays an important role in cancer progression and one of its essential components are the transcription factor proteins from the TEAD family. The TEAD family consists of 4 different family members (TEAD1-TEAD4) who modulate gene expression through the interaction with co-activator proteins such as YAP1 and TAZ. Recent studies have shown that TEADs are partially palmitoylated. Activity and stability of TEAD proteins are regulated by this modification. Here, we attempt to better understand the palmitoylation status of TEAD proteins in cells. In a first step, we have developed LC/MS/MS-based assays for the detection of palmitoylated TEAD peptides using recombinant TEAD1 protein. Subsequently, we have set out to analyze endogenous TEAD proteins using YAP1 activated malignant mesothelioma H2052 tumor cells. H2052 tumor cells express preferentially TEAD1 protein. TEAD1 was immunoprecipitated from H2052 tumor cells and subjected to peptidic digest and LC/MS/MS analysis. Peptide mapping permitted to confirm the expected site of palmitoylation (cysteine 359 of TEAD1). In addition, novel sites of both palmitoylation and myristoylation were detected on the lysine residue 336 of TEAD1. While these hydrophobic peptides could not be recovered after in-gel digestion of immunoprecipitation enriched TEAD1 of H2052 cells, digestion performed directly on immunoprecipitation beads allowed us to recover those peptides and hence to confirm the existence of TEAD lysine-myristoylation and lysine-palmitoylation in cells. In this study, we show the development of an MS-based strategy to characterize post translational modifications directly on endogenous TEAD1 proteins present in H2052 tumor cells.
Citation Format: Armelle Buzy, Olivier Guichard, Jean-Luc Zacchayus, Olivier Venier, Olivier Courtin, Iris Valtingojer, Laurent Debussche, Vincent Mikol, Jean-Claude Guillemot. In vitro and in cellulo mass spectrometry study of TEAD1 palmitoylation [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4868.
Abstract 4871: The roles of E3/E4 ubiquitin ligase UBE4B and its targets p53 and Ku70 in neuroblastoma cell death and differentiation
Authors: Clorennec Christophe Le , Lesperance Jacqueline , Huo Yuchen , Zage Peter E. 
Published: September 2020

Abstract
Children with high-risk neuroblastoma have poor survival rates and need novel treatment strategies. UBE4B is an E3/E4 ubiquitin ligase whose gene is found in the chromosome 1p36 region commonly deleted in high-risk neuroblastoma tumors and whose expression is associated with neuroblastoma patient outcomes. However, the functional roles of UBE4B in neuroblastoma pathogenesis are not known. We hypothesized that UBE4B would interact with mediators of DNA damage and repair pathways, leading to the observed associations between UBE4B and patient outcomes. Using cell lines with stable UBE4B depletion and over-expression, we evaluated the interaction of UBE4B with the E3 ubiquitin ligase ITCH/AIP4 and the effects of UBE4B expression levels on p53 and Ku70 ubiquitination and proteasomal degradation by co-immunoprecipitation and Western blots. We also evaluated the role of UBE4B in neuroblastoma differentiation, cell death, and responses to chemotherapy and HDAC inhibition using continuous live cell imaging and Western blots. We determined that UBE4B directly interacts with ITCH/AIP4 in the last WW domain of ITCH and we also determined that Ku70 directly interacts with ITCH in the three first WW domain of ITCH. We also determined that UBE4B depletion leads to reduced polyubiquitination and degradation of both p53 and Ku70 in neuroblastoma cells. UBE4B depletion also led to reduced apoptosis induced by both HDAC inhibition with vorinostat and chemotherapy treatment with carboplatin and doxorubicin via stabilizaton of the ITCH target protein c-FLIPL leading inhibition of caspase 8 activation. Our results have identified novel interactions and novel targets for UBE4B ubiquitin ligase activity and a direct role of the UBE4B-ITCH complex in HDAC inhibitor and DNA-damage induced cell death, and suggest that the association of UBE4B expression with neuroblastoma patient outcomes may be mediated by effects on responses of patient tumors to chemotherapy.
Citation Format: Christophe Le Clorennec, Jacqueline Lesperance, Yuchen Huo, Peter E. Zage. The roles of E3/E4 ubiquitin ligase UBE4B and its targets p53 and Ku70 in neuroblastoma cell death and differentiation [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4871.