bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 6059: Targeting the anthracycline metabolizing enzyme AKR1C3 in adipocytes to improve cytotoxicity
Authors: Paharkova Vladislava , Orgel Etan , Neely Michael , Louie Stan. , Mittelman Steven 
Published: September 2020

Abstract
Introduction: We have reported that mouse and human adipocytes take up and metabolize the anthracycline, daunorubicin (DNR), reducing its concentration in the adipocyte microenvironments. This may contribute to anthracycline resistance for cancers, which reside in adipocyte rich environments such as omentum and bone marrow. Adipocytes express several carbonyl- and aldo-keto reductases (CBRs and AKRs) which metabolize and inactivate anthracyclines, and it is unclear which of these might be important targets to improve treatment outcome.
Experimental Procedures: We knocked out AKR1C3 in the human preadipocyte cell line Chub S7 using CRISPR/Cas9 gene editing technology. We chose AKR1C3 first as it is one of the overexpressed enzymes in adipocytes with the highest anthracycline-metabolizing activity. We delivered ribonucleoprotein complexes of CRISPR-Cas9 enzyme plus guide RNAs by nucleofection. Then we established single-cell derived clones and tested for successful KO by Western blot. Finally, we quantified adipocyte lysate AKR activity using a colorimetric assay based on NADPH-dependent reduction of phenanthrenequinone.
Data Summary: We chose three Chub S7 preadipocyte clones that demonstrated successful AKR1C3 knockout based on almost undetectable protein expression by western blot. AKR activity was significantly reduced in all three clones; control preadipocytes had 44±4.7, while clones had activity of 28±7.3, 33±3.4, and 35±5.7 pmol/min/µg (p&lt;0.05 for all comparisons, n=3 for each assay).
Conclusions: These findings demonstrate that AKR1C3 knockout can be successfully done in human preadipocytes using a CRISPR-Cas9 system. Knockout of AKR1C3 significantly reduces overall aldoketoreductase activity in preadipocyte cells. This implies that a substantial portion of preadipocyte AKR activity is dependent on the isoenzyme, AKR1C3. Future testing is needed to determine whether AKR1C3 KO will reduce the clearance of anthracyclines from the cancer microenvironment, and may represent a treatment target to enhance anthracycline cytotoxicity.
Citation Format: Vladislava Paharkova, Etan Orgel, Michael Neely, Stan. Louie, Steven Mittelman. Targeting the anthracycline metabolizing enzyme AKR1C3 in adipocytes to improve cytotoxicity [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 6059.
Abstract 6123: An indole compound MA-35 attenuates tumorigenesis in an inflammation-induced colon cancer model
Authors: Kanehara Keigo , Ohnuma Shinobu , Naitoh Takeshi , Uuno Michiaki , Abe Takaaki 
Published: September 2020

Abstract
Backgrounds: In inflammatory bowel disease (IBD), chronic inflammation results in the development of colon cancer known as colitis-associated cancer (CAC), that is associated with tumor necrosis factor-α (TNF-α) signaling. In addition, intestinal fibrosis in IBD patients is a common clinical complication, which is promoted by transforming growth factor β1 (TGF-β1). In our previous study, mitochonic acid (MA-35), derivative of indole-3-acetic acid known as a plant hormone auxin, showed anti-TNF-α effect by inhibiting IκB kinase (IKK) phosphorylation and anti-TGF-β1 effect by inhibiting Smad3 phosphorylation in a renal fibrosis mouse model.
Aim: To identify the possible mechanism of anti-tumor effects and anti-fibrotic effects of MA-35 using azoxymethane (AOM)/ dextran sulfate sodium (DSS)-induced CAC mouse model.
Methods and Results: MA-35 was orally administered every day for 70 days in an AOM/DSS model with a feeding tube. There was no difference in weight loss between the AOM/DSS group and the AOM/DSS+MA-35 group, however the disease activity index score and the survival rate were improved by MA-35. MA-35 also blocked progression of the anemia and shortening of the colon induced by AOM/DSS. MA-35 surprisingly reduced the formation of tumors in the colon. In the microscopic evaluation, MA-35 reduced inflammation and fibrosis in area with dysplasia. Furthermore, the TNF-α mRNA level in the colon tended to be reduced and the interleukin 6, TGF-β1 and fibronectin 1 mRNA levels in the colon were significant reduced by MA-35. In western blotting, MA-35 inhibited IKK phosphorylation, and both NF-κB p65 phosphorylation and TNF-α as the downstream signals for p-IKK in human colorectal cancer cell line HT-29. Similarly, MA-35 inhibited Smad2/3 phosphorylation, and both fibronectin 1 and TGF-β1 as the downstream signals for p-Smad2/3 in HT-29.
Conclusions: MA-35 inhibited the development of CAC by reducing inflammation and fibrosis in an AOM/DSS mouse model by inhibiting both TNF-α signaling and TGF-β1 signaling. MA-35 may be a potent therapeutic agent for inhibiting the development of CAC in IBD patients.
Citation Format: Keigo Kanehara, Shinobu Ohnuma, Takeshi Naitoh, Michiaki Uuno, Takaaki Abe. An indole compound MA-35 attenuates tumorigenesis in an inflammation-induced colon cancer model [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 6123.
Abstract 6154: H3K27M gliomas are characterized by a stall in the epithelial-mesenchymal transition
Authors: CHENEY ALLISON R. , Sanders Lauren M. , Seninge Lucas , Beale Holly C. , Kephart Ellen Towle , Pfeil Jacob , Learned Katrina , Lyle A. Geoffrey , Bjork Isabel , Haussler David , Salama Sofie R. , Vaske Olena M. 
Published: September 2020

Abstract
Pediatric diffuse midline gliomas are lethal cancers, the majority of which harbor the H3 p.K27M mutations. Although it has potential implications on the treatment of diffuse midline glioma as a disease driver; the timing, cell type of origin, and effect of the H3 p.K27M mutation on early embryonic brain development is poorly understood. The purpose of our study is to elucidate the molecular mechanisms by which the histone H3 p.K27M mutation drives tumorigenesis of pediatric diffuse midline gliomas using the analysis of genomic datasets. Here, we performed differential RNA sequencing gene expression analysis of a cohort of H3K27M and H3 wild type (WT) pediatric diffuse midline gliomas, revealing that genes in the epithelial-mesenchymal transition (EMT) pathway were significantly differentially expressed between the mutant and WT tumors. Several EMTs are required for normal brain development. Overall, pre-EMT genes, including the master regulator of EMT SNAI1, were overexpressed in H3K27M tumors compared to the WT tumors, while post-EMT genes were underexpressed. We hypothesized that the H3 p.K27M mutation may lead to gliomagenesis by inducing a stall in the EMT in early brain development. To test this hypothesis, we examined published single-cell RNA sequencing data from pediatric diffuse midline gliomas alongside similar data from organoid models of neural development, collected from multiple developmental timepoints. This analysis revealed transcriptional similarities between H3K27M and pre-EMT neural stem cells. Currently, we are investigating the expression of EMT markers in H3K27M and WT pediatric glioma primary cell lines, using Western blotting, RT-PCR, and CRISPRi screening. In conclusion, we observed aberrant expression of genes involved in EMT in H3K27M pediatric gliomas. Our observations are consistent with a model in which the p.H3K27M mutation is associated with a pre-EMT cell phenotype, potentially due to an arrest in the EMT pathway or de-differentiation of mature astrocytes.
Citation Format: ALLISON R. CHENEY, Lauren M. Sanders, Lucas Seninge, Holly C. Beale, Ellen Towle Kephart, Jacob Pfeil, Katrina Learned, A. Geoffrey Lyle, Isabel Bjork, David Haussler, Sofie R. Salama, Olena M. Vaske. H3K27M gliomas are characterized by a stall in the epithelial-mesenchymal transition [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 6154.
Abstract 6157: Hypoxia increases motility of neuroblastoma cells induced by the neuropeptide Y/Y5 receptor pathway and exacerbates their metastatic potential
Authors: Abualsaud Nouran , Caprio Lindsay , Hong Sung Hyeok , Galli Susana , Kitlinska Joanna 
Published: September 2020

Abstract
Neuroblastoma (NB) is a pediatric malignancy that originates from precursors of sympathetic neurons and expresses neuronal proteins, such as neuropeptide Y (NPY) and its receptors. NB cells constitutively express NPY Y2 receptor (Y2R), which maintains their proliferation and tumor vascularization. However, pro-apoptotic conditions additionally induce expression of Y5 receptor (Y5R), which acts as a survival factor for tumor cells. High Y5R expression was also observed in angioinvasive NB cells surrounding blood vessels, suggesting a role for this receptor in NB dissemination. Indeed, in NB patients, high serum levels of NPY associated with the presence of metastases. Hence, the goal of this study was to identify mechanisms that may underlie potential pro-metastatic effects of NPY. We have found that NPY, acting via Y5R, stimulates NB cell motility and acts as a chemotactic factor for these cells in a concentration-dependent manner. Hypoxia, a known condition promoting metastasis, increased expression of Y5R in SK-N-BE(2) and SK-N-AS NB cell lines. Consequently, hypoxic NB cells had increased motility and a chemotactic response to NPY. This increase in NPY-driven migration was fully blocked by Y5R antagonist. Pre-incubation of NB cells in hypoxia before orthotopic injections into adrenal glands of SCID/bg mice exacerbated metastasis in xenografts from both NB cell lines. In mice bearing non-MYCN amplified SK-N-AS tumors, the hypoxia-driven metastases were located in soft tissues, while MYCN-amplified SK-N-BE(2) xenografts derived from hypoxic cells exhibited enhanced dissemination to lungs. The latter pattern of metastases is also observed in NB patients with MYCN amplification. Further studies are required to determine if the hypoxia-driven increase in NB metastasis is Y5R-dependent. Altogether, our data support the role of tumor hypoxia and NPY/Y5R axis in NB dissemination. Since this pathway is also activated during chemotherapy and promotes NB cells resistance to treatment, NPY/Y5R may also be involved in the secondary dissemination of the recurrent tumors. If the role of Y5R in NB metastasis is validated in preclinical models, this NPY receptor may become a potential therapeutic target preventing the disease dissemination.
Citation Format: Nouran Abualsaud, Lindsay Caprio, Sung Hyeok Hong, Susana Galli, Joanna Kitlinska. Hypoxia increases motility of neuroblastoma cells induced by the neuropeptide Y/Y5 receptor pathway and exacerbates their metastatic potential [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 6157.
Abstract 6229: MDM2 inhibitors sensitize testicular cancer models to mTORC1/2 inhibitors and cisplatin
Authors: de Jong Steven , de Vries Gerda , Meersma Gert Jan , Suurmeijer Albert J.H. , van Vugt Marcel A.T.M. , Gietema Jourik A 
Published: September 2020

Abstract
Testicular cancer (TC) is one of the most common solid tumors in young men that can effectively be treated with cisplatin-based chemotherapy. However, 10% of those wild type p53 expressing TC patients will not survive their disease. The aim of this study was to identify effective combinations with MDM2 inhibitors in TC models. A panel of cisplatin sensitive and -resistant human TC cell lines and TC patient-derived xenograft (PDX) models have been treated with MDM2 inhibitors (Nutlin-3a, RG7388), cisplatin and AKT/mTORC1/2 inhibitors (MK2206, AZD8055 and INK128). Survival, apoptosis, and clonogenic assays were performed. Transcriptional activity of p53 was determined using chromatin immunoprecipitation (CHIP) followed by quantitative PCR of target promoters. Mechanisms of apoptosis induction were studied by measuring mitochondrial priming, siRNA approaches and BH3 mimetics. We found that MDM2 inhibitors sensitized TC cell lines to cisplatin and mTORC1/2 inhibition. Nutlin-3a in combination with cisplatin or AZD8055 treatment induced caspase 3 and PARP cleavage and reduced clonogenic outgrowth. Enriched p53 binding to the promoter regions of known activators of extrinsic and intrinsic apoptosis, FAS, PUMA and NOXA, was observed in response to Nutlin-3a or cisplatin. BH3 profiling of mitochondria and siRNA mediated silencing identified PUMA and NOXA as key pro-apoptotic proteins in TC cell lines. Interestingly, addition of the BH3 mimetics ABT-737 and A-1210477, mimicking PUMA and NOXA respectively, to combination treatments shifted the balance towards apoptosis as well. In vivo experiments showed reduced tumor growth especially in the wild type p53 PDX model TC1 treated with RG7388/cisplatin, which was reflected in p53 activation and increased caspase-3 activity. No potentiating effect of this combination was observed in the p53 wild type PDX model TC4. In conclusion, we have demonstrated that targeting of MDM2 in combination with chemotherapy can be very effective in TC models. Our results warrant further investigation of this drug combination in the treatment of TC. Supported by a grant from the Dutch Cancer Society (RUG 2014-6691)
Citation Format: Steven de Jong, Gerda de Vries, Gert Jan Meersma, Albert J.H. Suurmeijer, Marcel A.T.M. van Vugt, Jourik A Gietema. MDM2 inhibitors sensitize testicular cancer models to mTORC1/2 inhibitors and cisplatin [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 6229.
Abstract 6232: Preclinical efficacy of CIGB-300, an anti-CK2 peptide, on breast cancer metastasic colonization
Authors: Gottardo Maria F. , Sidabra Johanna E. , Capobianco Carla S. , Garona Juan , Rodriguez Silvio Perea , Aonso Daniel F. , Farina Hernan G. 
Published: September 2020

Abstract
CK2 is a serine/threonine kinase that is overexpressed in breast cancer and its inhibition is associated to reduced tumor growth and disease progression. With more than three hundred substrates, this kinase is involved in a large number of cellular functions, such as replication, cell proliferation, differentiation, regulation of apoptosis and angiogenesis. CIGB-300 is an antitumor peptide with a novel mechanism of action, since it binds to the phospho-acceptor domain of CK2 substrates impairing the correct phosphorylation by the enzyme. Our aim was to evaluate the potential therapeutic benefits of CIGB-300 on breast cancer disease using experimental models with translational relevance. We demonstrated that CIGB-300 reduces breast cancer cell growth in MDA-MB-231, MCF-7 and F3II cells, exerting a pro-apoptotic action. We found that CIGB-300 increased Bax and decreased Bcl-2 expression, so the intrinsic pathway was involved in the proapoptotic effect of CIGB-300 in breast cancer cells. Moreover, CIGB-300 inhibits the CK2-mediated phosphorylation of B23, such phosphorylation impairment was correlated with the ability of CIGB-300 to induce nucleolar disassembly. Also, we investigate whether CIGB-300 affects the sensitivity of tumor cells to chemotherapeutic agents. We evaluated the effects of the combination of CIGB-300 and cisplatin on cell viability and apoptosis. We demonstrated that CIGB-300 sensitize breast cancer cells to cisplatin, reducing cell viability and increasing apoptosis. In vivo, the effect on experimental breast cancer lung metastasis was evaluated after surgical removal of primary F3II tumors or after tail vein injection of tumor cells. Systemic CIGB-300 treatment inhibited breast cancer colonization of the lung, reducing the size and number of metastatic lesions. The present preclinical study establishes for the first time the efficacy of CIGB-300 on breast cancer. These encouraging results suggest that CIGB-300 could be used for the management of breast cancer as an adjuvant therapy after surgery, limiting tumor metastatic spread and thus protecting the patient from distant recurrence.
Citation Format: Maria F. Gottardo, Johanna E. Sidabra, Carla S. Capobianco, Juan Garona, Silvio Perea Rodriguez, Daniel F. Aonso, Hernan G. Farina. Preclinical efficacy of CIGB-300, an anti-CK2 peptide, on breast cancer metastasic colonization [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 6232.
Abstract 6275: Targeting RRM2 by triapine sensitizes glioblastoma to radiotherapy
Authors: Corrales-Guerrero Sergio , Bell Erica H. , Chakravarti Arnab , Shen Changxian , Williams Terence M. 
Published: September 2020

Abstract
Glioblastoma multiforme (GBM) remains characterized by poor clinical outcomes and limited treatment options. Currently, the standard of care for patients with this disease is surgery with radiation therapy and concurrent and adjuvant temozolomide. However, most patients succumb to the disease within the first 15 months after diagnosis. Therefore, effective treatment regimens are desperately needed. Exacerbated cell proliferation as well as repair of DNA damage induced by chemoradiation requires a constant supply of dNTPs. Ribonucleotide reductase (RNR) is the key enzyme for the synthesis of dNTPs and is a complex formed by two subunits: RRM1 and RRM2
3-aminopyridine-2-carboxaldehyde thiosemicarbazone (3AP - commercially Triapine) is a molecule that potently inhibits RRM2 by affecting its active site, and has been shown to cross the brain-blood barrier (Jiang et al. 2006. PMID: 16834759). We herein report that triapine induces DNA replication stress and sensitizes GBM cells to radiation. Analyses of available datasets including TCGA show that RRM2 is overexpressed in GBM and lower grade glioma (LGG), and a higher expression of RRM2 associates with worse overall survival and disease-free survival of GBM and LGG patients. Nevertheless, RRM2 levels do not change between initial and recurrent tumors (Wang et al. 2017. PMID: 28697342). Using human GBM cell lines LN229, LN18 and U251, we have performed cytotoxicity assays and clonogenic radiation assays. The IC50 of triapine in these cell lines are between 1-2 μM. We find that triapine significantly radiosensitizes LN229, LN18 and U251 cells. Consistent with the role of RRM2 in dNTPs biosynthesis, triapine treatment of GBM cells induces robust phosphorylation of CHK1 at S345, which is accompanied by elevated gamma H2AX. Importantly, we reveal that triapine promotes ionizing radiation induced phosphorylation of CHK2 at T68. Acknowledging the increasing concerns about using established cell lines for GBM studies, we are currently moving to validate our results in primary GBM cell lines. Our results demonstrate that RRM2 is overexpressed in glioblastoma and targeting RRM2 with triapine enhances radiation-induced DNA damage leading to radiosensitization of glioblastoma cells. Our findings suggest that triapine is a promising agent for the sensitization of glioblastoma to radiotherapy.
Citation Format: Sergio Corrales-Guerrero, Erica H. Bell, Arnab Chakravarti, Changxian Shen, Terence M. Williams. Targeting RRM2 by triapine sensitizes glioblastoma to radiotherapy [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 6275.
Abstract 6288: A high-throughput in vitro characterization method of CAR T cells using image cytometry
Authors: Chan Leo Li-Ying , Maldini Colby , Love Andrea , Tosh Kevin , Gayout Kevin , Smith Tim , Riley James 
Published: September 2020

Abstract
Since the FDA approval of two CAR T cell therapies, Kymriah (Tisagenlecleucel, Novartis) and Yescarta (Axicabtagene ciloleucel, Kite) for CD19+ malignancies, there has been a significant interest for developing new CARs targeting other disease indications. As such, the ability to simultaneously monitor manufacturing criteria and functional characteristics of multiple CAR T cell products would likely be critical to accelerate candidate therapies into clinical trials. Here, we demonstrate that image-based cytometry using the Celigo Image Cytometer (Nexcelom Bioscience LLC) yields high-throughput measurements of CAR T cell proliferation and size throughout the cell-manufacturing process, and captures the kinetics of antigen-specific CAR T cell-mediated killing in vitro. The data acquired and analyzed by the Celigo Image Cytometer were congruent with results from conventional instruments when tested contemporaneously, such as the Multisizer 3 (Beckman Coulter) which monitored the properties of CAR T cell expansion, and a luciferase-based assay which measured the cytotoxic potential of CAR T cells. More importantly, the use of bright field and fluorescent imaging by the Celigo Image Cytometer provided kinetic measurements and rapid data acquisition and analysis which were direct advantages over the conventional technologies described herein. Together, the Celigo Image Cytometer enabled fast, reproducible, and high-throughput assessment of CAR T cell manufacturing criteria and effector function, which could greatly facilitate the identification and evaluation of novel CARs with therapeutic potential.
Citation Format: Leo Li-Ying Chan, Colby Maldini, Andrea Love, Kevin Tosh, Kevin Gayout, Tim Smith, James Riley. A high-throughput in vitro characterization method of CAR T cells using image cytometry [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 6288.