bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3600: MET-mediated STING activation regulates EMT and NF-ΚB pathway in non-small cell lung cancer
Authors: Mun Seungchan , Han Bogyeong , Ahn HyunKyung , Koh Jaemoon , Kim Sehui , Chung DooHyun , Jeon YoonKyung 
Published: April 2023

Abstract
The cyclic GAMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway is known to have anti-tumor activity by activating the innate immune response against cytosolic double-stranded DNA (dsDNA). The regulating mechanism of STING remains unclear, and a pro-tumorigenic effect of STING pathway was recently reported. In this study, we investigated the expression of STING by MET and a novel role of the STING pathway in non-small cell lung cancer (NSCLC). The Cancer Genome Atlas (TCGA) and Cancer Cell Line Encyclopedia (CCLE) analysis showed that cGAS/STING pathway is enriched and STING expression is higher in MET-overexpressing tumors. Western blot for NSCLC cells and immunohistochemistry for NSCLC tumor tissues showed positive correlation between MET and STING expression in NSCLC cells. MET-overexpression and a MET agonist (HGF) treatment increased STING expression and CXCL10 and IFNβ expression and secretion, whereas MET-knockdown and a MET inhibitor (capmatinib) treatment decreased STING expression in NSCLC cells. In MET-positive and STING-positive cells, a STING agonist (poly-dAdT and diABZI) treatment led to increased activation of STING signaling pathway. In EBC-1 cells, MET knockdown decreased STING expression and STING-agonist-induced STING-pathway activation and cytokine production. To further investigate the role STING in NSCLC, EBC-1 cells were transfected with STING siRNA and submitted to RNS-seq. Gene set enrichment analysis revealed that several pathways were affected by STING expression. Epithelial-mesenchymal transition (EMT) and TNFα-signaling via NF-κB signaling were significantly downregulated in STING-knockdowned cells. In vitro experiments using EBC-1 cells, STING overexpression increased SLUG and TWIST1 expression and migration of cells, whereas STING knockdown decreased SLUG and TWIST1 expression and migration of cells. In summary, this study demonstrated that MET and STING expression were positively correlated with each other in NSCLC and MET overexpression increased STING expression and subsequent activation upon STING agonist stimulation. In addition, STING contributed to EMT in NSCLC.
Citation Format: Seungchan Mun, Bogyeong Han, HyunKyung Ahn, Jaemoon Koh, Sehui Kim, DooHyun Chung, YoonKyung Jeon. MET-mediated STING activation regulates EMT and NF-ΚB pathway in non-small cell lung cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3600.
Abstract 3629: Calcium signalling and the breast cancer microenvironment
Authors: Bong Alice H. , Gieniec Krystyna A. , Sadras Francisco , Robitaille Mélanie , Roberts-Thomson Sarah J. , Davis Felicity M. , Monteith Gregory R. 
Published: April 2023

Abstract
Calcium is a key regulator of a variety of pathways important in breast cancer progression, including those involved in cell proliferation and invasiveness. Calcium signaling is implicated in processes relevant to the breast cancer tumor microenvironment and altered calcium influx is a feature of cancer associated fibroblasts. In these studies, simultaneous use of genetically encoded calcium indicators with distinct fluorescence spectral properties (GCaMP6m or JRCaMP1b), were used to explore the cross-talk between breast cancer cells and fibroblasts via calcium signaling and changes in different cellular compartments. HMF3S human fibroblast cells co-cultured with breast cancer cells expressing GCaMP6m or JRCaMP1b were assessed in 2D and 3D co-culture using automated epi-fluorescence or confocal microscopy. Mitochondrial calcium levels were assessed using targeted GCaMP6m (2mtGCaMP6m) in HMF3S cells expressing JRCaMP1b. Results showed distinct spatial and temporal changes in free calcium levels between both fibroblasts and breast cancer cells and the mitochondria and cytosol during activation. These methodological advances provide an opportunity to better understand calcium signaling in cancer associated fibroblasts. These techniques will also help identify the calcium channels and pumps that could be targeted to manipulate the breast cancer tumor microenvironment to inhibit pathways important in breast cancer progression.
Citation Format: Alice H. Bong, Krystyna A. Gieniec, Francisco Sadras, Mélanie Robitaille, Sarah J. Roberts-Thomson, Felicity M. Davis, Gregory R. Monteith. Calcium signalling and the breast cancer microenvironment. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3629.
Abstract 3672: RNA epigenetic regulation of cholesterol metabolism in neuroblastoma
Authors: Bansal Mohit , Gupta Anamika , Ding Han-Fei 
Published: April 2023

Abstract
Pseudouridylation (conversion of uridine to pseudouridine) is a common epigenetic modification of RNA which increases its structural stability. RNA pseudouridylation is catalyzed by the family of pseudouridine synthases (PUS). The functional significance and biological relevance of mRNA pseudouridylation are poorly understood. Here we report that the oncogene MYCN, which drives the development of high-risk neuroblastoma, transcriptionally upregulates the PUS1 gene. Further, examination of gene expression profiling data from neuroblastoma patients (the SEQC dataset) reveals that higher PUS1 expression is associated with poor prognosis. The transcriptional upregulation of PUS1 expression in MYCN amplified cell lines is of functional significance, as our studies reveals that knockdown of PUS1 expression in neuroblastoma cell lines suppresses cell survival and proliferation. Our RNA-seq analysis reveals that a key downstream target of PUS1 is cholesterol metabolism. PUS1 knockdown reduces the expression of key enzymes in the mevalonate pathway for cholesterol synthesis. Mechanistically, PUS1 may targets SREBP2, a master regulator of cholesterol metabolism responsible for transcriptional upregulation of enzymes for cholesterol biosynthesis. We showed that PUS1 expression is essential for maintaining SREBP2 expression in MYCN-amplified neuroblastoma cells and for statin-induced feedback activation of the mevalonate pathway, a commonly used system for assessing the transcriptional activity of SREBP2. These findings suggest that a function of PUS1 in sustaining the proliferation of neuroblastoma cells is to promote cholesterol metabolism, which could be exploited as a therapeutic strategy against high-risk neuroblastoma.
Citation Format: Mohit Bansal, Anamika Gupta, Han-Fei Ding. RNA epigenetic regulation of cholesterol metabolism in neuroblastoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3672.
Abstract 3695: Mitochondrial uncoupling reverses reductive carboxylation in cancer
Authors: Jiang Haowen , He Clifford Jiajun , Li Albert M. , He Bo , Li Yang , Ye Jiangbin 
Published: April 2023

Abstract
When electron transport chain (ETC) activity is inhibited upon hypoxia or when cultured in suspension, reductive carboxylation plays an essential role in supporting tumor growth through converting α-ketoglutarate (α-KG) to citrate, which is converted to acetyl-coenzyme A (acetyl-CoA) for lipogenesis. However, there is no effective way to target reductive carboxylation for tumor therapy. In this study, we use the mitochondrial uncoupler niclosamide ethanolamine (NEN) and BAM15 to reverse reductive carboxylation in cancer cells. Uncoupler treatment activates ETC and increases the cellular NAD+/NADH ratio. Using U-13C-glutamine and 1-13C-glutamine tracers, we discovered that uncoupler treatment accelerates the oxidative TCA cycle and blocks reductive carboxylation, particularly under hypoxia and spheroid culture condition. Uncoupler treatment significantly reduces tumor cell proliferation, survival and sphere formation. In addition, the intracellular acetyl-CoA levels are significantly decreased by NEN treatment. Furthermore, lipidomics analysis showed that NEN treatment reduced fatty acid abundance and altered the lipid composition under both normoxia and hypoxia. Together, these data demonstrate mitochondrial uncoupling redirects the α-KG flux from reductive carboxylation back to the oxidative TCA cycle, highlighting that the ETC activity and NAD+/NADH ratio is the key switch that determines the metabolic fate of α-KG. Reversing reductive carboxylation could be one key mechanism that mitochondrial uncoupler inhibits tumor growth.
Citation Format: Haowen Jiang, Clifford Jiajun He, Albert M. Li, Bo He, Yang Li, Jiangbin Ye. Mitochondrial uncoupling reverses reductive carboxylation in cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3695.
Abstract 3697: Modulation of Nicotinamide N-Methyl Transferase (NNMT) expression in TNBC is associated with altered cell membrane cholesterol levels
Authors: Chee Woei-Yaw , Dolcen Deniz Nesli , Bennett Lynda , Conzen Suzanne D. 
Published: April 2023

Abstract
Introduction: Cancer cells often develop a program of metabolic adaptability and epigenetic remodelling to promote gene expression changes causing cancerous cell proliferation and invasion. The metabolic-epigenetic axis as a an underlying mechanism of carcinogenesis could lead to new approaches to cancer treatment. Nicotinamide N-Methyl Transferase (NNMT) is a metabolic enzyme that catalyzes the transfer of a methyl group from S-Adenosyl methionine (SAM) to nicotinamide, thereby decreasing the overall methyl pools for RNA, histone and DNA methylation. High NNMT expression has been previously associated with a poor outcome in TNBC.
Hypothesis: We hypothesized that high versus low NNMT activity and resulting oncogenic gene expression in TNBC may alter histone methylation is association with altered metabolic pathway activation.
Methods: We used two TNBC cell lines models, MDA-MB 231 and SUM159, to study the association between high versus low NNMT activity and metabolic pathway activation. Surprisingly, we identified gene expression pathways suggestive of increased free cholesterol biosynthesis gene expression with low NNMT, and then used biochemical assays and filipin to measure free cholesterol in both total high and low NNMT TNBC cell lysates and relative cholesterol expression in the High vs Low NNMT cell membranes.
Results: In vitro, NNMT knockdown resulted in increased cholesterol synthetic pathway enzyme expression in both TNBC cell lines. In vivo studies also showed that protein expression of cholesterol synthesis enzymes was also increased in NNMT-depleted MDA-MB 231 xenografted tumors. Interestingly, the increased cholesterol level upon NNMT depletion was mainly localized at the plasma membrane via Filipin III staining, suggesting a possible association with less cellular membrane fluidity and a decreased oncogenic phenotype. In ongoing experiments, we are evaluating whether changes in histone methylation in high vs low NNMT cells are associated with differential expression of cholesterol biosynthetic genes, and free cholesterol production. We hypothesize that NNMT might epigenetically decrease the methylation of histones regulating cholesterol production in TNBC.
Citation Format: Woei-Yaw Chee, Deniz Nesli Dolcen, Lynda Bennett, Suzanne D. Conzen. Modulation of Nicotinamide N-Methyl Transferase (NNMT) expression in TNBC is associated with altered cell membrane cholesterol levels. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3697.
Abstract 3702: Serine promotes aggressive phenotype in renal cancer
Authors: Karki Suman , Kundu Anirban , Brinkley Garret , Nam Hyeyoung , Goliwas Kayla , Deshane Jessy , Sudarshan Sunil 
Published: April 2023

Abstract
Tumor cells are known to have increased demand for nutrients to support proliferation. Among the most highly utilized nutrients by tumor cells are nonessential amino acids (NEAAs). While the role of NEAAs in supporting tumor growth is well-established, their role in supporting aggressive phenotypes such as migration and invasion is poorly understood. Yet, the major cause of morbidity from malignancies such as renal cell carcinoma (RCC) is metastasis. Through an unbiased approach, we assayed the role of NEAAs in promoting the migration of RCC cells. We demonstrate that exogenous serine and glutamine are essential and sufficient to support aggressive phenotypes in RCC. Moreover, the availability of both NEAAs is critical for the expression of the proinvasive transcription factor SNAIL. Our data converge on the role of these two NEAAs in supporting the translation of this factor. Perturbations that limit the availability and/or synthesis of either result in translational reprogramming through the integrated stress response (ISR) that leads to loss of SNAIL1 expression and attenuates aggressive phenotypes. Our findings associate nutrient status to the ISR including ER stress. Collectively, our study highlights new insights for NEAA metabolism in supporting aggressive tumor phenotypes and could reveal novel approaches to mitigate metastasis by targeting NEAA availability and/or utilization.
Citation Format: Suman Karki, Anirban Kundu, Garret Brinkley, Hyeyoung Nam, Kayla Goliwas, Jessy Deshane, Sunil Sudarshan. Serine promotes aggressive phenotype in renal cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3702.
Abstract 371: Integrin-mediated lipolysis by pancreatic cancer-derived extracellular vesicles in cancer-associated cachexia
Authors: Shibata Chikako , Otsuka Motoyuki , Fujishiro Mitsuhiro 
Published: April 2023

Abstract
Introduction: Cancer-associated cachexia (CAC) is a life-threating condition frequently observed in cancer patients, especially those with pancreatic cancer. CAC is a multifactorial disease, with the notable characteristic of systemic lipolysis. Although humoral factors have been implicated in its pathogenesis, critical factors remain to be determined. Since even localized pancreatic cancer frequently induce lipolysis systemically, we examined the molecular mechanisms in vitro and in vivo based on the hypothesis that extracellular vesicles (EVs) systemically released from pancreatic cancer cells induce lipolysis.
Procedures: EVs derived from the supernatants of cultured pancreatic cancer cell lines and from the sera of pancreatic cancer patients were characterized. The effects of EVs on adipocytes were determined using mature human adipocytes induced from human adipose-derived mesenchymal stem cells. To confirm the in vitro findings, lipolysis induction was examined in mice by intravenous injection of EVs. EV characteristics inducing lipolysis were determined. Those characteristics were confirmed by knockout studies and examining specifically isolated pancreatic cancer-derived EVs from heterogenous EV population in patients’ sera.
Results: Lipolysis was triggered by specific EVs from pancreatic cancer cell line via the cAMP inside and uptake of more EVs into adipocytes was crucial for higher levels of lipolysis. In vivo studies, database search, and knockout studies revealed that integrin β1 and integrin α6 on EVs were the determinants for the attachment and uptake of the EVs to adipocytes, resulting in inducing more lipolysis. There was no difference in the percentage of these integrins positive EVs between heterogenous EV population in cancer patients’ sera and EVs in cancer-free controls’ sera. However, pancreatic cancer cell-derived EVs specifically isolated by targeting CA19-9, an aberrant glycosylation, from heterogenous EV population in patients’ sera, confirmed that pancreatic cancer-derived EVs have more such integrins.
Conclusions: Cancer cell-derived EVs in the sera of pancreatic cancer patients have more ITGB1 and ITGA6 levels, which may explain the systemic induction of lipolysis in cancer cachexia from the early stages and could be novel therapeutic targets against CAC. In addition, specific isolation of cancer-derived EVs in sera from heterogenous EV population is crucial for elucidating cancer-related pathogenesis.
Citation Format: Chikako Shibata, Motoyuki Otsuka, Mitsuhiro Fujishiro. Integrin-mediated lipolysis by pancreatic cancer-derived extracellular vesicles in cancer-associated cachexia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 371.
Abstract 3726: Histone acetylation axis in the control of RNA Pol2 clusters
Authors: Gryder Berkley E. , Chin Diana , Kim Hyunmin , Osman Issra 
Published: April 2023

Abstract
Core regulatory transcription factors (CR TFs) orchestrate the placement of super enhancers (SEs) to activate transcription of cell-identity specifying gene networks and are critical in promoting cancer. We defined the core regulatory circuitry of fusion positive rhabdomyosarcoma (FP-RMS, a cancer of childhood) in primary tumors and cell lines, which includes PAX3-FOXO1 (P3F), MYOD1, SOX8, MYCN and others. To find chemical probes able to selectively inhibit CR TF transcription, we screened the Structural Genomics Consortium epigenetic probe set by RNA-seq. We found that chemical probes along the acetylation-axis, and not the methylation-axis, are able to cause selective disruption of CR TF transcription. Inhibitors of HDACs (acetylation erasers), BRD4 (acetylation readers) and CBP/p300 (acetylation writers) were all able to selectively halt CR TF transcription. For HDACs, this raised a conundrum: why would too much histone acetylation, an active chromatin mark, stop transcription at CR TFs? ChIP-seq showed that CR TFs build SEs that have the largest quantities of histone acetylation and the enzymes that write acetylation (i.e., p300), yet paradoxically also harbor the highest amounts of the opposing histone deacetylases (HDACs). To investigate the architectural effects of disabling HDACs and causing hyper acetylation, we developed Absolute Quantification of Architecture (AQuA) HiChIP, revealing erosion of native SE contacts at CR TFs, and extensive aberrant contacts. This did not cause an elongation defect, but rather removed RNA Pol2 from core regulatory genetic elements and eliminated RNA-Pol2 phase condensates in 20 minutes. We further dissected the contribution of HDAC isoforms using a set of HDAC selective inhibitors, finding HDAC1/2/3 are co-essential to CR transcription. Using HAT inhibitors/degraders, we discovered a profound dependence on CBP/p300 for clustering of Pol2 loops that connect P3F to its target genes. In the absence of CBP/p300, Pol2 long range enhancer loops collapse, Pol2 accumulates in CpG islands and fails to exit the gene body. These results reveal a potential novel axis for therapeutic interference with P3F in FP-RMS and clarify the molecular relationship of P3F and CBP/p300 in sustaining active Pol2 clusters essential for oncogenic transcription. Overall, our data reveals a SE-specific need for balancing histone acetylation states to maintain SE architecture, Pol2 clustering in 3D, and CR TF transcription.
Citation Format: Berkley E. Gryder, Diana Chin, Hyunmin Kim, Issra Osman. Histone acetylation axis in the control of RNA Pol2 clusters. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3726.
Abstract 3777: MicroRNA-inducible CRISPR/Cas9 for cell type-specific genome regulation in cancer
Authors: Oh Seung Ja , Shin Cheol-Hee , Lee Ji Min , Lee Juyong , Park Su Chan 
Published: April 2023

Abstract
MicroRNA-dependent mRNA decay plays an important role in gene silencing by facilitating posttranscriptional and translational repression. Inspired by this intrinsic nature of microRNA-mediated mRNA cleavage, here, we describe a microRNA-targeting mRNA as a switch platform called mRNA bridge mimetics to regulate the translocation of proteins. We applied the mRNA bridge mimetics platform to Cas9 protein to confer it the ability to translocate into the nucleus via cleavage of the nuclear export signal called CRISPR Self Check-In. This system performed programmed gene editing in vitro and in vivo. Combinatorial treatment with cisplatin and miR-21-EZH2 axis-targeting CRISPR Self Check-In improved sensitivity to chemotherapeutic drugs in vivo. Using the endogenous microRNA mediated mRNA decay mechanism, our platform is able to remodel a cell’s natural biology to allow the entry of precise drugs into the nucleus, devoid of non-specific translocation. The mRNA bridge mimetics strategy is promising for applications in which the reaction must be controlled via intracellular stimuli and modulates Cas9 proteins to ensure safe genome modification in diseased conditions.
Citation Format: Seung Ja Oh, Cheol-Hee Shin, Ji Min Lee, Juyong Lee, Su Chan Park. MicroRNA-inducible CRISPR/Cas9 for cell type-specific genome regulation in cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3777.