bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4469: Disruption of ATG9A-dependent basal autophagy sensitizes cancer cells to innate inflammatory signaling
Authors: Jayatunge Dasun N. , Tsang Tsz-Ming , McEwan Colten M. , Andersen Joshua L. 
Published: April 2023

Abstract
Pathogen-derived nucleic acids are recognized by pattern recognition receptors (PRRs), including cGAS and RIG-I, which trigger the innate immune response by activating IRF3-mediated interferon gene expression. In tumors, high levels of chromosomal instability can also lead to DNA fragments leaking into the cytosol to trigger PRR-mediated interferon response, resulting in pro-inflammatory cytokine secretion from the tumor. This could potentially expose tumor cells to immune attack. However, through mechanisms that are not completely understood, tumor cells dampen the interferon response to escape immune recognition. Previous work demonstrated that loss of ATG9A, but not ATG5, increased inflammatory signaling through the STING-IRF3 cascade, suggesting that perhaps an autophagy-independent function of ATG9A regulates inflammation. Here we found that loss of ATG9A or other upstream regulators of basal autophagy (e.g., ATG101), but not core LC3 lipidation machinery (ATG5 or ATG7), sensitizes cells to dsDNA-induced IRF3 activation and interferon gene expression. We also found that loss of ATG9A or ATG101, but not ATG5 or ATG7, increases the basal activity of the ubiquitin-sensing, IRF3-targeted kinase TBK1, which increases further upon dsDNA treatment. In these ATG9A and ATG101 deficient cells, TBK1 is clustered around large p62-positive condensates that previously been shown to contain large accumulations of LC3. Our preliminary data suggest a model in which ATG9A deletion causes the accumulation of LC3-positive membrane at p62 condensates, which may act as a platform for inflammatory signaling. Our current work focuses on identifying the upstream pathways and core mechanism that regulates IRF3 activation in ATG9A-deficient cells and exploiting this mechanism to improve anti-tumor immunity.
Citation Format: Dasun N. Jayatunge, Tsz-Ming Tsang, Colten M. McEwan, Joshua L. Andersen. Disruption of ATG9A-dependent basal autophagy sensitizes cancer cells to innate inflammatory signaling. [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 4469.
Abstract 447: Genetic and pharmacological inhibition of TUT4 and TUT7 sensitizes AML cells to BCL-2 inhibition
Authors: Fyfe Corey , Fiorentino Gabriella , Steinbaugh Michael J. , Campos Joana , Lawson Hannah , Durko Jozef , O'Carroll Donal , Kranc Kamil , Harvey Darren , Ghosh Shomir , Campbell Robert M. 
Published: April 2023

Abstract
The terminal uridyltransferases TUT4/7 (ZCCHC11/6) are important mediators of RNA stability, RNA decay, histone mRNA regulation, viral RNA sensing, and miRNA biogenesis. Regulation of RNA processing pathways can be mediated by a number of RNA-binding proteins (RBPs), and dysfunctional RNA processing has implications as a therapeutic target in leukemia and lymphoma. We performed genetic and pharmacological perturbation of the activity of terminal uridyltransferases TUT4/7 (ZCCHC11/6) in MLL-rearranged acute myeloid leukemia (AML). A combination of catalytic inactivation and knockout by CRISPR/Cas9 of TUT4 and TUT7, respectively, were used to abrogate uridylation in THP-1 cells, with enzyme function measured by a cellular miRNA uridylation assay. Inhibition of uridylation in THP-1 cells reduced proliferation, induced changes in cell cycle progression, and enhanced sensitivity to the BCL-2 inhibitor, venetoclax, alone and in combination with azacitidine. BCL-2 expression was found to be reduced at the mRNA and protein levels, while increases in expression of BCL-2 regulating miRNAs miR139-5p and miR494-3p were observed. GSEA performed on uridylation deficient cells highlighted changes in cell cycle regulation, mitochondrial metabolism, and cell death pathways relative to unedited controls. These effects could be recapitulated with pharmacological inhibition using sub-inhibitory doses of a novel, selective inhibitor of TUT4/7, TS-002266, while the weakly active enantiomer TS-002455 did not show combination activity. Treatment of primary AML with combination of TS-002266 and venetoclax improved activity over venetoclax alone in a number of primary AML samples. Thus, we propose that TUT4/7 and BCL-2 inactivation may be a therapeutic strategy in leukemia.
Citation Format: Corey Fyfe, Gabriella Fiorentino, Michael J. Steinbaugh, Joana Campos, Hannah Lawson, Jozef Durko, Donal O'Carroll, Kamil Kranc, Darren Harvey, Shomir Ghosh, Robert M. Campbell. Genetic and pharmacological inhibition of TUT4 and TUT7 sensitizes AML cells to BCL-2 inhibition [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 447.
Abstract 4583: An essential role for Cmtr2 in endothelial cell function and vascular development
Authors: Yermalovich Alena , Mohsenin Zarin , Giotti Bruno , Wheeler Douglas , Xu Kelly , Tsankov Alexander , Meyerson Matthew 
Published: April 2023

Abstract
Cancer genome sequencing studies have identified multiple novel tumor suppressor genes, including the FTSJD1 or CMTR2 gene that is subject to statistically significant levels of both nonsense and frameshift mutations in lung adenocarcinoma. However, these analyses are insufficient to determine the importance and modes of action of the gene in the disease. Follow-up functional studies are essential to elucidate these gene functions. The protein product of CMTR2 has been described as an mRNA cap methyltransferase, which O-methylates the ribose of the second guanine residue of the 5’ cap, likely serving to modify efficiency of transcript processing, translation and stability, but the details of how and why specific transcripts undergo methylation by the enzyme, as well as its role in oncogenesis, remain to be elucidated. In this study we report the first comprehensive analysis of the physiologic consequences of Cmtr2 deficiency in knockout (KO) mice and primary human cells. Constitutive deletion of Cmtr2 results in embryos that die in midgestation with defects in embryo size, placental malformation and yolk sac vascularization. Endothelial-specific deletion of Cmtr2 in mice also results in vascular defects and perinatal lethality, suggesting that vascular development is a key function of Cmtr2 in animals. In summary, our study reveals that Cmtr2 plays a critical role in vascular development, maturation and remodeling. Further studies of CMTR2 function may provide insight not only into lung cancer biology but also to its broader roles in ontogeny.
Citation Format: Alena Yermalovich, Zarin Mohsenin, Bruno Giotti, Douglas Wheeler, Kelly Xu, Alexander Tsankov, Matthew Meyerson. An essential role for Cmtr2 in endothelial cell function and vascular development. [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 4583.
Abstract 4592: In vivo efficacy evaluation of anti-human VEGFR2 antibodies in humanized B-hVEGFR2 mice
Authors: Zhao Lei , Shang Chengzhang , Soto Rebecca , Liu Chonghui , Yu Zhaoxue 
Published: April 2023

Abstract
Angiogenesis, vascular development, vascular permeability, and embryonic hematopoiesis is partly regulated by VEGF binding to VEGFR2, which forms receptor dimers and activates downstream signaling pathways. In addition to developmental cues, VEGFR2 is also associated with disease progression for psoriasis, rheumatoid arthritis, diabetic retinopathy, and more significantly, tumor growth and drug resistance. Given this, inhibition of VEGFR2 signaling is actively being studied, and multiple VEGFR2 inhibitors have entered various phases of clinical studies and have shown promising results. To evaluate in vivo efficacy of anti-human VEGFR2 antibodies, we generated a human VEGFR2 knock-in (B-hVEGFR2) mouse model by replacing the murine Vegfr2 extracellular domain sequence with the corresponding human sequence. We validated human VEGFR2 gene and protein expression in B-hVEGFR2 mice by RT-PCR and flow cytometry, respectively. Furthermore, we observed an anti-tumor effect with anti-human VEGFR2 antibodies in homozygous B-hVEGFR2 mice subcutaneously implanted with MC38 colon cancer cells. Taken together, we established a novel preclinical B-hVEGFR2 mouse model for evaluation of VEGFR2-targeted immunotherapy.
Citation Format: Lei Zhao, Chengzhang Shang, Rebecca Soto, Chonghui Liu, Zhaoxue Yu. In vivo efficacy evaluation of anti-human VEGFR2 antibodies in humanized B-hVEGFR2 mice. [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 4592.
Abstract 4695: Expansion of the extracellular proximity labeling (ePL) technique to novel matrisome targets
Authors: Rich Joshua , Gurung Sadeechya , Kaur Sukhbir , Roberts David , Stetler-Stevenson William , Peeney David 
Published: April 2023

Abstract
The extracellular matrix (ECM) is a complex and dynamic meshwork of extracellular macromolecules that plays a key role in cancer progression and metastasis. The matrisome encompasses the genes encoding ECM and ECM-associated secretory proteins, and elucidating the protein-protein interactions (PPIs) of the matrisome is a promising area of study in the quest for a better understanding of cancer pathology and therapeutics. Proximity labeling, a powerful technique for investigating protein interactomes based on enzyme-fusion constructs and affinity chromatography, has considerable potential in detailing the protein-protein interactions of the matrisome. We have recently had success in developing a proximity labeling approach based on fusion constructs containing the biotinylating enzymes BioID2 and TurboID. We apply this approach to gain new insights into the interactome of the matrisome protein tissue inhibitor of metalloproteinase 2 (TIMP2), a protein found to have antitumorigenic potential. At present, we seek to expand this approach to other ECM protein targets, including tissue inhibitor of metalloproteinase 3 (TIMP3) and thrombospondin-1 (THBS-1). TIMP3 has been shown to possess anti-tumor properties, with loss of TIMP3 function associated with poor prognosis in multiple cancers. THBS-1 is a large (129 kDa) ECM protein that is relatively promiscuous with diverse functions in the tumor microenvironment. Expression of THBS-1 is frequently lost during oncogenesis, and such loss is associated with poor prognosis in some cancers. We describe how ePL can be expanded to these matrisome targets in different cellular systems (adherent vs. suspension) and show that ePL assays can be performed with stable and transient transgene expression. We show that ePL is a suitable method for studying PPIs in novel matrisome targets and expect to apply the technique to further targets in the near future.
Citation Format: Joshua Rich, Sadeechya Gurung, Sukhbir Kaur, David Roberts, William Stetler-Stevenson, David Peeney. Expansion of the extracellular proximity labeling (ePL) technique to novel matrisome targets. [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 4695.
Abstract 4701: Spatial resolution of tumor and immune cell lineages in the hypoxic microenvironment of pancreatic ductal adenocarcinoma (PDAC)
Authors: Arvidson Lisa , Prioli Reginaldo , Jensen Samuel , Smith Michael J. , White Katie O. , Heil-Chapdelaine Richard A. , Huang Chi-Chou , Phan Tuan H. , Sasaki Hideki , Angus-Hill Melinda L. 
Published: April 2023

Abstract
Understanding how the tumor microenvironment (TME) evolves during tumorigenesis and therapeutic response is crucial to developing personalized treatments with the goal of improving cancer therapy. With robust and comprehensive multiplexed imaging technologies, immune biomarker antibodies can be used to interrogate immune cell lineages and structures. When combined with specific oncology biomarker antibodies, this approach can capture the immune response within the TME in a variety of neoplasms. The Cell DIVETM Multiplex Imaging Solution allows probing and imaging of dozens of biomarkers on a whole single tissue section using an iterative staining and dye inactivation workflow. The broad portfolio of robust IHC-validated antibodies from Cell Signaling Technology (CST) enables the detection of key proteins in the TME, allowing immune cell detection and phenotyping in tissue. Here, we demonstrate multiplexed Cell DIVE imaging using a novel CST panel to probe pancreatic ductal adenocarcinoma (PDAC). These biomarker antibodies define the immune cell landscape in the hypoxic tumor. Development of the antibody panel required minimal optimization, enabled the identification of complex cell types and revealed their cell-to-cell interactions within the tumor microenvironment. The availability of cell type specific biomarkers, combined with the ability to interrogate using multiplexed tissue imaging, provides unprecedented and novel insights and spatial resolution of immune cell populations with many cell types in the TME.
Citation Format: Lisa Arvidson, Reginaldo Prioli, Samuel Jensen, Michael J. Smith, Katie O. White, Richard A. Heil-Chapdelaine, Chi-Chou Huang, Tuan H. Phan, Hideki Sasaki, Melinda L. Angus-Hill. Spatial resolution of tumor and immune cell lineages in the hypoxic microenvironment of pancreatic ductal adenocarcinoma (PDAC). [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 4701.
Abstract 4776: NRF2 is overexpressed in esophageal adenocarcinoma and its targeting sensitizes tumor cells to cisplatin through induction of ferroptosis and apoptosis
Authors: Ballout Farah , Lu Heng , Chen Zheng , Hu Tianling , Chen Lei , El-Rifai Wael , Peng Dunfa 
Published: April 2023

Abstract
Background: Esophageal adenocarcinoma (EAC) is the predominant type of esophageal cancer in the United States, with the 5-year survival rate is below 20%. EAC develops through Barrett’s esophagus (BE)-dysplasia-carcinoma cascade. Gastroesophageal reflux disease (GERD), where acidic bile salts refluxate into the esophagus, is the main risk factor for the development of BE and its progression to EAC. The NFE2-related factor 2 (NRF2) is the master cellular antioxidant regulator, involving in many cancer hallmarks.
Methods and Results: Using western blotting and immunohistochemistry technologies, we detected high NRF2 protein levels in EAC cell lines and primary EAC tissues, as compared with normal esophagus and non-neoplastic Barrett’s esophagus samples. The knockdown of NRF2 using NRF2 specific siRNAs significantly increased oxidative stress in response to cellular stress stimuli by bile salts or cisplatin. This was associated with an increase in DNA damage and inhibition of EAC cell growth. Brusatol, a NRF2 inhibitor, significantly inhibited NRF2 transcriptional activity and downregulated NRF2 target genes. We discovered that in addition to inducing apoptosis, Brusatol alone or in combination with CDDP induced significant lipid peroxidation and ferroptosis as evidenced by reduced xCT and GPX4 expression, two known ferroptosis markers. Moreover, the combination of Brusatol and CDDP significantly inhibited EAC tumor xenografts growth in vivo. We confirmed the in vitro data showing ferroptosis as an important mechanism in the xenografted tumors treated with Brusatol or Brusatol and CDDP combination using IHC staining.
Conclusion: Our data support the role of NRF2 in protecting against stress-induced apoptosis and ferroptosis in EACs. Targeting NRF2 in combination with platinum therapy can be an effective strategy for eliminating cancer cells in EAC.
Citation Format: Farah Ballout, Heng Lu, Zheng Chen, Tianling Hu, Lei Chen, Wael El-Rifai, Dunfa Peng. NRF2 is overexpressed in esophageal adenocarcinoma and its targeting sensitizes tumor cells to cisplatin through induction of ferroptosis and apoptosis. [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 4776.
Abstract 478: A novel dual-acting RAS/β-catenin inhibitor activates antitumor immunity to potentiate cancer immunotherapy
Authors: Gazi Md Yeashin , Okoko Ogacheko , Wang Xin , Brandle Caitlin , Ding Zhi-chun , Mitra Amit , Ramirez-Alcantara Veronica , Chen Xi , Keeton Adam , Maxuitenko Yulia , Piazza Gary , Zhou Gang 
Published: April 2023

Abstract
Previous research suggests that the antineoplastic activity of sulindac is mediated by a cyclooxygenase (COX) independent mechanism involving dual inhibition of RAS and β-catenin signaling. ADT030 is a novel non-COX inhibitory sulindac-derivative designed to target RAS and phosphodiesterase 10 (PDE10) to block MAPK/AKT signaling and activate cGMP/PKG signaling to suppress oncogenic β-catenin transcription, respectively. ADT-030 potently and selectively inhibits growth and induces apoptosis of a variety of cancer cell lines and has robust antitumor activity in multiple mouse tumor models by oral administration. However, the impact of ADT-030 on the tumor microenvironment (TME) has not been examined. Here we report the antitumor activity and immunomodulatory effect of ADT-030 in preclinical models. We show that oral administration of ADT030 significantly inhibited the growth of multiple types of murine tumors in immunocompetent mice. In a metastatic breast tumor model (4T1), ADT-030 not only delayed the growth of the primary tumor but also reduced tumor metastasis, leading to improved mouse survival. Immunological analysis revealed marked reduction of MDSCs in mice, and increased T cell tumor infiltration after ADT-030 treatment. The antitumor activity of ADT030 was diminished in immunodeficient mice or immunocompetent mice depleted of CD8+ T cells, suggesting the dependence of ADT-030 on host immunity. Moreover, the combination of ADT-030 and anti-PD-1 antibody therapy led to further improved tumor growth control and mouse survival. Bulk and single-cell RNAseq is being conducted to determine the impact of ADT-030 on the cell compositions and transcriptomic changes in the TME. Altogether, our study provides insight into the mechanism of action of ADT-030 and underscores the potential of ADT-030 as a monotherapy or in combination with immunotherapies for RAS and Wnt/β-catenin driven cancers.
Citation Format: Md Yeashin Gazi, Ogacheko Okoko, Xin Wang, Caitlin Brandle, Zhi-chun Ding, Amit Mitra, Veronica Ramirez-Alcantara, Xi Chen, Adam Keeton, Yulia Maxuitenko, Gary Piazza, Gang Zhou. A novel dual-acting RAS/β-catenin inhibitor activates antitumor immunity to potentiate cancer immunotherapy [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 478.
Abstract 4812: Inhibition of Smurf2 E3 ubiquitin ligase by heclin and its analogues enhances HIF-1α expression and transcriptional activity in normoxia or hypoxia
Authors: Zhao Shuai , El-Deiry Wafik S. 
Published: April 2023

Abstract
Hypoxia-inducible factors (HIFs) act as transcription factors and play an essential role in cellular and systemic responses to low oxygen environments. HIF-1α expression is induced in acute hypoxia typically through failure of its ubiquitination and degradation mediated by Von Hippel-Lindau (VHL). Previously we discovered a non-canonical mechanism where the SMAD specific E3 ubiquitin protein ligase 2 (Smurf2) promotes the ubiquitination of HIF-1α and reduces HIF-1α level in HCT116 colorectal cancer cells. Smurf2 is a HECT-type ubiquitin ligase known to interact with Smad proteins, leading to their ubiquitination and proteasomal degradation. Overexpression of Smurf2 decreased HIF-1α expression in HCT116 and SW480 colorectal cancer cells under hypoxia as well as in RCC4 VHL-deficient kidney renal clear cell carcinoma cells under normoxia. Treatment with MG132 at least partially rescued the expression of HIF-1α following Smurf2 overexpression, indicating involvement of proteasome-dependent degradation in Smurf2-mediated HIF-1α destabilization. Knockdown of SMURF2 increased HIF-1α expression under normoxia in HCT116 and RCC4 cells. To investigate the effect of Smurf2 inhibition, we tested a selective reversible inhibitor of HECT E3 ubiquitin ligases, heclin, along with its analogues, PYR-41, C646 and 4E1RCat. Treatment with heclin increased HIF-1α expression in HCT116 under hypoxia as well as in SW480 and RCC4 cells under normoxia in a dose-dependent manner. PYR-41 elevated the level of HIF-1α level in HCT116 cells under hypoxia, in RCC4 cells under normoxia and in SW480 cells under both normoxia and hypoxia. To examine the effect on HIF transcriptional activity, we performed luciferase reporter assay using plasmids containing the hypoxia response element (HRE) from the PGK1 promoter and the VEGF promoter, respectively. PYR-41 induced transcriptional activity on PGK1-HRE in both HCT116 and SW480 cells. 4E1RCat robustly activated transcription on both VEGF-HRE and PGK1-HRE in HCT116 and SW480 cells. In summary, Smurf2 targets HIF-1α for ubiquitination and degradation independently of oxygen concentration and inhibition of Smurf2 to stimulate HIF activity under normoxia or hypoxia may be beneficial in pathological circumstances featuring anemia and hypoxemia.
Citation Format: Shuai Zhao, Wafik S. El-Deiry. Inhibition of Smurf2 E3 ubiquitin ligase by heclin and its analogues enhances HIF-1α expression and transcriptional activity in normoxia or hypoxia. [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 4812.