bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1788: A high-throughput chemical genetic screen reveals SALL4-induced metabolic vulnerabilities in cancer
Authors: Tan Justin L. , Li Feng , Yeo Joanna Z. , Yong Kol Jia , Bassal Mahmoud A. , Ng Guo Hao , Lee May Yin , Leong Chung Yan , Tan Hong Kee , Wu Chan-Shuo , Liu Bee Hui , Chan Hon Man , Tan Zi Hui , Chan Yun Shen , Wang Siyu , Lim Zhi Han , Toh Tan Boon , Hooi Lissa , Low Kia Ngee , Ma Siming , Kong Nikki R. , Stein Alicia J. , Wu Yue , Thangavelu Matan T. , Suzuki Atsushi , Periyasamy Giridharan , Asara John M. , Dan Yock Young , Bonney Glenn K. , Chow Edward K. , Lu Guo-Dong , Ng Huck Hui , Kanagasundaram Yoganathan , Ng Siew Bee , Tam Wai Leong , Chai Li , Tenen Daniel G. 
Published: September 2020

Abstract
Transcription factors are important drivers of cancer but the development of therapeutics against these factors has had limited success. We developed a stringent high-throughput chemical genetic screening platform to identify compounds that target oncogenic transcription factor SALL4 dependency in liver cancer. The platform comprises SALL4 low- and high-expressing endogenous and engineered isogenic liver cancer cell lines. Unexpectedly, from screening 21,575 natural product extracts, the top hits were four oxidative phosphorylation inhibitors that selectively reduced SALL4-dependent cell viability. The ATP synthase inhibitor oligomycin suppressed SALL4-expressing cancer in lung and liver cancer cell culture models, and in patient-derived xenograft models of liver cancer. Oligomycin also synergized with sorafenib, the standard-of-care targeted therapy in liver cancer, to effectively suppress SALL4-driven tumorigenesis in vivo. When aberrantly expressed in cancer, SALL4 binds ~50% of mitochondrial genes, including many oxidative phosphorylation genes, to predominantly upregulate their expression. SALL4 upregulation also alters the levels of oxidative phosphorylation-related metabolites and functionally increases oxidative phosphorylation activity. Application of our endogenous/isogenic transcription factor-screening platform revealed a therapeutically actionable oxidative phosphorylation vulnerability in SALL4-expressing cancers.
Citation Format: Justin L. Tan, Feng Li, Joanna Z. Yeo, Kol Jia Yong, Mahmoud A. Bassal, Guo Hao Ng, May Yin Lee, Chung Yan Leong, Hong Kee Tan, Chan-Shuo Wu, Bee Hui Liu, Hon Man Chan, Zi Hui Tan, Yun Shen Chan, Siyu Wang, Zhi Han Lim, Tan Boon Toh, Lissa Hooi, Kia Ngee Low, Siming Ma, Nikki R. Kong, Alicia J. Stein, Yue Wu, Matan T. Thangavelu, Atsushi Suzuki, Giridharan Periyasamy, John M. Asara, Yock Young Dan, Glenn K. Bonney, Edward K. Chow, Guo-Dong Lu, Huck Hui Ng, Yoganathan Kanagasundaram, Siew Bee Ng, Wai Leong Tam, Li Chai, Daniel G. Tenen. A high-throughput chemical genetic screen reveals SALL4-induced metabolic vulnerabilities in 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 1788.
Abstract 1796: Requirement for LIM kinases in acute myeloid leukemia
Authors: Jensen Patrizia , Carlet Michela , Kress Jana , Weber Andrea , Brunner Ines , Slabicki Mikolaj , Grill Gregor , Weisemann Simon , Cheng Ya-Yun , Jeremias Irmela , Scholl Claudia , Fröhling Stefan 
Published: September 2020

Abstract
Acute myeloid leukemia (AML) is a highly aggressive disease which, in spite of extensive research in the last decades, is still treated with conventional chemotherapeutic regimens in most cases. Recent approvals of FLT3 and IDH1/2 inhibitors have highlighted the promise of targeted approaches for personalized AML therapy. However, the majority of AML driver mutations currently remain “undruggable”, making the search for additional molecular targets imperative. LIM kinases 1 and 2 (LIMK1/LIMK2), which are primarily known for regulating actin dynamics via phosphorylation of the actin-depolymerizing factor cofilin, have previously been implicated in progression and metastasis formation of several epithelial tumors. However, their function in leukemia, and AML in particular, has not been addressed in detail. Using high-throughput RNA interference (RNAi) screens in AML cell lines, we identified LIMK1 as a potential novel molecular target for AML treatment. High LIMK1 expression was significantly correlated with shorter survival of AML patients, and was associated with NPM1 and DNMT3A mutations, MLL rearrangements, and elevated HOX gene expression. RNAi- and CRISPR/Cas9-mediated suppression, as well as pharmacologic inhibition of LIMK1 and its close homolog LIMK2, reduced colony formation and decreased proliferation due to slowed cell cycle progression of MLL-rearranged AML cell lines and patient-derived xenograft samples. This was accompanied by morphologic changes indicative of myeloid differentiation. Transcriptome analysis showed upregulation of several tumor suppressors as well as downregulation of HOXA9 targets and mitosis-associated genes in response to LIMK1 suppression, providing a potential mechanistic explanation for the anti-leukemic phenotype. Finally, we observed a mutual regulation between LIMK1 and CDK6, a kinase known to be involved in the differentiation block of MLL-rearranged AML, and addition of the CDK6 inhibitor palbociclib further enhanced the anti-proliferative effect of LIMK1 depletion. Our data therefore suggest that LIM kinases are promising targets for AML therapy.
Citation Format: Patrizia Jensen, Michela Carlet, Jana Kress, Andrea Weber, Ines Brunner, Mikolaj Slabicki, Gregor Grill, Simon Weisemann, Ya-Yun Cheng, Irmela Jeremias, Claudia Scholl, Stefan Fröhling. Requirement for LIM kinases in acute myeloid leukemia [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 1796.
Abstract 18: The CXCR4/CXCL12 axis is a target of a polyphenol extract from olive oil processing with potential cancer prevention and interception activities
Authors: Gallazzi Matteo , Festa Marco , Sansone Clementina , Corradino Paola , Noonan Douglas , Albini Adriana 
Published: September 2020

Abstract
The olive mill wastewater polyphenol extracts exhibit chemopreventive properties in lung cancer cell lines, acting on the CXCR4/CXCL12 axis.
Lung cancer still is a leading cause of cancer death worldwide. Although avoidance of tobacco use and smoking cessation will have the greatest impact on lung cancer development, (chemo)prevention could prove to be very effective, particularly in high risk individuals. Diverse diet-derived compounds have been reported to exert antioxidant, anti-proliferative and anti-angiogenic effects. Olive oil production involves the generation of an aqueous phase, defined as olive mill wastewater (OMWW), a polluting effluent rich in soluble polyphenols. We established a polyphenol rich, purified extract, from OMWW, that we termed A009. The effect of A009 on cell proliferation was determined by the crystal violet assay, while induction of apoptosis was measured by flow cytometry. The ability of the A009 to regulate the cell migration/invasion axis CXCR4/CXL12 and the production of pro angiogenic factors (VEGF, CXCL8, CCL2), was investigated by flow cytometry. The sprouting assay on Matrigel was used to functional verify the ability of A009 to limit invasive properties. Hydroxytyrosol, the most abundant polyphenols present in our A009, was used as reference molecules in all the assays performed.
We found that the A009 limit A549 and H1560 cell proliferation, in a dose and time dependent manner. These effects were associated with induction of apoptosis at 24 and 48 hours of treatment, that was higher in H1650, as compared to A549 cells. The A009 extracts were effective in inhibiting the production of CXCR4 and CXCL12 in both the A549 and H1650 cells, accompanied with reduction in H1650 cell capability to generate invasive sprouts. in a Matrigel layer. Finally, A009 interfere with the production of VEGF, CXCL8 and CCL2 in both the A549 and H1650 cells.
Our preliminary results suggest that A009 exhibit promising preventing and interceptive properties on lung cancer cell lines in vitro.
Citation Format: Matteo Gallazzi, Marco Festa, Clementina Sansone, Paola Corradino, Douglas Noonan, Adriana Albini. The CXCR4/CXCL12 axis is a target of a polyphenol extract from olive oil processing with potential cancer prevention and interception activities [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 18.
Abstract 1805: The therapeutic potential of 5-Azacitidine plus various BH3 mimetics in melanoma
Authors: Dart Chiara , Mukherjee Nabanita , Amato Carol , Van Gulick Robert J. , MacBeth Morgan , Couts Kasey , Turner Jacqueline , Bagby Stacey , Fujita Mayumi , Robinson William , Shellman Yiqun 
Published: September 2020

Abstract
Treatment options for melanoma have progressed in recent years, with the rise of targeted therapy and immunotherapy. Options are still limited for patients that are BRAF wildtype, treatment refractory, and patients with non-sun exposed melanoma subtypes that typically do not respond to standard therapies. Therefore, it is necessary to explore other biological processes that may provide new therapeutic approaches. One of most appealing is the BCL-2 family of apoptotic/anti-apoptotic system that is effective in leukemias, with BH3 mimetics venetoclax alone or in combination with the hypomethylating agent 5-azacitidine. We examined similar combinations on the group of eleven patient-derived and commercially available melanoma cell lines. The BH3 mimetics we tested include MCL1 inhibitors (S63845, AMG 176, AZD5991), an inhibitor of BCL2, BCLXL, and BCLW (navitoclax/ABT-263), a BCL2 inhibitor (venotoclax/ABT-199), and a BCLXL inhibitor (A1331852). Our data showed that treatments with MCL1 inhibitors plus 5-azacitidine are effective in reducing melanoma cell viability. S63845 was the most potent compound when combined with 5-azacitidine, with nine out of eleven (~80%) melanoma cell lines achieving less than 50% viability at the highest dose. There was an overall weaker response to treatment with 5-azacitidine in combination with ABT-263 or A1331852, with only 54% and 45% of cell lines responding at the highest dose, respectively. Response to ABT-199 with 5-Azacitidine was highly limited, 18% of cell lines responded at the highest concentration. Our data demonstrate that treatment with S63845 in combination with 5-azacitidine is highly effective in cell lines from a variety of melanoma subtypes, including the rare uveal and mucosal subtypes. In vivo and mechanistic experiments are planned. The novel combination of the MCL1 inhibitor S63845 and 5-azacitidine may be a promising alternative therapeutic target in melanoma.
Citation Format: Chiara Dart, Nabanita Mukherjee, Carol Amato, Robert J. Van Gulick, Morgan MacBeth, Kasey Couts, Jacqueline Turner, Stacey Bagby, Mayumi Fujita, William Robinson, Yiqun Shellman. The therapeutic potential of 5-Azacitidine plus various BH3 mimetics in melanoma [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 1805.
Abstract 1832: A real-time extracellular ATP assay to identify modulators of the innate immune response
Authors: Niles Andrew L. , Kupcho Kevin R. , Lazar Dan F. , Cali James J. 
Published: September 2020

Abstract
Damage-associated molecular patterns (DAMPs) liberated or exposed by dying cells after chemotherapeutic treatment augment innate immune responses and improve clinical outcomes as a result of the immunogenic cell death (ICD) phenotype. Extracellular ATP (eATP) is a potent DAMP that acts as a chemotactic agent for dendritic cells and as an immunomodulator for fine-tuning the inflammatory response. Current in vitro methods for identifying eATP inducers are procedurally difficult and laborious and are hampered by the instability of eATP in cell culture environments. To overcome these challenges, we have developed an easy-to-use, homogeneous, bioluminescent chemistry that can be introduced with test articles to measure dose-dependent ATP release kinetics and magnitudes of response in real time. We tested the assay using U20S and U937 cells dosed with serial dilutions of doxorubicin, idarubicin, mitoxantrone, and romidepsin. Luminescent data were continuously collected throughout the 24h exposure in 5- or 10- minute increments using a conventional multimode plate reader with CO2-independent medium, or with readers equipped with atmospheric control. When coupled with parallel cell health assays designed to measure either viability or loss of membrane integrity, the data can reveal agents that potentiate ATP release during early or late apoptosis. Our real-time data suggest the potency of eATP release is dependent upon cell model and agent and can be reproducibly and robustly measured in 96 and 384 well environments with kinetic resolution not possible by other methods. This new eATP measurement workflow may help define the capacity of new chemical entities to induce apoptosis and immunogenic cell death.
Citation Format: Andrew L. Niles, Kevin R. Kupcho, Dan F. Lazar, James J. Cali. A real-time extracellular ATP assay to identify modulators of the innate immune response [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 1832.
Abstract 1840: NUC-1031 causes release of DAMPs and upregulates PD-L1 expression in lung cancer cells
Authors: Bré Jennifer , Read Oliver J. , Harrison David J. 
Published: September 2020

Abstract
Background: The nucleoside analog gemcitabine is used for the treatment of patients with lung cancer. Its cytotoxic effect is largely attributed to generation of the active metabolite dFdCTP, which causes DNA damage. NUC-1031, a ProTide transformation of gemcitabine, is specifically designed to overcome the key cancer resistance mechanisms associated with poor survival following gemcitabine treatment. NUC-1031 has shown high response rates in clinical studies, and efficacy in patients who were refractory to or had relapsed on gemcitabine. Certain cytotoxic agents are known to promote the release of damage-associated molecular patterns (DAMPs) from cancer cells, including cell surface exposure of calreticulin (CRT) and re-localization of nuclear high mobility group box protein 1 (HMGB1). DAMPs enhance cancer cell recognition by the immune system, promoting immunogenic cell death (ICD). Conversely, studies have shown that cancer cells can upregulate programmed death-ligand 1 (PD-L1) as an adaptive, protective response to cell stress resulting from drug exposure, potentially leading to evasion from immune cell-mediated cytotoxicity. This study investigates the effect of NUC-1031 on the release of DAMPs and modulation of PD-L1 in lung cancer cells.
Methods: Non-small cell lung cancer (NSCLC) cell lines were treated with NUC-1031 for 24h and cytotoxicity was assessed at 96h post-treatment to determine half-maximal inhibitory concentrations (IC50). Cells were treated with increasing concentrations of NUC-1031 and cell surface expression of CRT and PD-L1 were analyzed by flow cytometry and complemented with RT-qPCR. IFN-γ was used as a positive control for PD-L1 induction. Localization of HMGB1 was assessed by immunofluorescence microscopy.
Results: NUC-1031 induced the release of HMGB1 from the nucleus and increased cell surface expression of CRT in a dose-dependent manner. We confirmed that IFN-γ strongly and uniformly induced cell surface PD-L1 (270% increase). By comparison, NUC-1031-induced cell surface expression of PD-L1 was modest (67% increase). This effect was dose-dependent, with no further increase in PD-L1 expression above 1 μM.
Conclusions: NUC-1031 is a potent cytotoxic agent that directly causes DNA damage through generation of dFdCTP. In addition, these data demonstrate the NUC-1031 promotes adaptive changes that alter the recognition of cancer cells by the immune system. The release of DAMPs by NUC-1031 and increase in PD-L1 is predicted to engender an immune response, which might be further potentiated by immune checkpoint inhibition.
Citation Format: Jennifer Bré, Oliver J. Read, David J. Harrison. NUC-1031 causes release of DAMPs and upregulates PD-L1 expression in lung 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 1840.
Abstract 1848: NUC-3373 induces ER stress and the release of damage associated molecular patterns in colorectal cancer cells
Authors: McKissock Fiona G. , Read Oliver J. , Harrison David J. 
Published: September 2020

Abstract
Background: NUC-3373 is a ProTide transformation of fluorodeoxyuridine monophosphate (FUDR-MP), the active anti-cancer metabolite of 5-fluorouracil (5-FU). NUC-3373 can overcome key cancer resistance mechanisms and the rapid degradation known to impact 5-FU's clinical utility. Clinical studies are currently investigating NUC-3373 as monotherapy (solid tumors) and in combination with agents commonly used to treat patients with colorectal cancer (CRC). Previous studies revealed ultrastructural adaptations in CRC cell lines exposed to NUC-3373 that were indicative of endoplasmic reticulum (ER) stress. NUC-3373 was also shown to induce the formation of long-lasting ternary thymidylate synthase (TS) complexes. The induction of ER stress may promote the release of damage-associated molecular patterns (DAMPs) which have the potential to evoke immunogenic cell death (ICD). This study examines the ability of NUC-3373 to induce both ER stress and the release of DAMPs in CRC.
Methods: Immunoglobulin-binding protein (BiP) was used as a marker of ER stress induction. BiP and TS (free and ternary complex) expression were measured by Western blot and quantified using Licor Odyssey. TS was knocked down using TYMS-targeting siRNA and lipofectamine-based transfection. DAMPs, active release of nuclear high mobility group box protein 1 (HMGB1) and cell surface exposure of calreticulin (CRT), were assessed by fluorescence microscopy and flow cytometry respectively.
Results: NUC-3373 resulted in the rapid formation of TS ternary complexes, which was associated with ER stress as assessed by BiP protein expression. siRNA knockdown studies confirmed that TS ternary complex formation is necessary for the induction of ER stress. A strong correlation between BiP and TS ternary complex formation (R2=0.97) was observed. Furthermore, NUC-3373 induced the release of DAMPs, with increased expression of CRT at the cell surface and concomitant loss of nuclear HMGB1.
Conclusions: NUC-3373, a potent inhibitor of TS activity, also induces ER stress, upregulates cell surface CRT, and decreases nuclear HMGB1 in CRC cells. The aforementioned DAMPs have been shown to cause ICD. In addition to being an effective cytotoxic agent, these findings suggest that NUC-3373 has the potential to evoke a host immune response and enhance the clinical utility of immunotherapy.
Citation Format: Fiona G. McKissock, Oliver J. Read, David J. Harrison. NUC-3373 induces ER stress and the release of damage associated molecular patterns in colorectal 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 1848.
Abstract 185: Longitudinal aqueous humor sampling reflects treatment response in retinoblastoma patients
Authors: Polski Ashley , Xu Liya , Prabakar Rishvanth K. , Kim Jonathan W. , Kuhn Peter , Cobrinik David , Hicks James , Berry Jesse L. 
Published: September 2020

Abstract
Purpose: The aqueous humor (AH) liquid biopsy for retinoblastoma (RB) facilitates in vivo sampling of tumor-derived cell-free DNA (cfDNA). We previously demonstrated that tumor-derived somatic copy number alterations (SCNAs) in AH predict poor ocular salvage. However, whether AH sampling can be used to monitor clinical response throughout therapy remains unknown. In this retrospective study, we hypothesize that SCNA amplitude (amp) and tumor fraction (TFx) of RB cfDNA in the AH correspond to treatment response over time.
Methods: Eyes that required intravitreal chemotherapy (IVM) for tumor seeding and had ≥3 sequential AH samples extracted during active IVM therapy and/or at secondary enucleation between 2015-2019 were included. Shallow whole-genome sequencing of AH cfDNA was performed to assess RB SCNA amp, and ichorCNA software estimated cfDNA TFx based on SCNAs present. Eyes without SCNAs in any sample were removed from final analysis. For each eye, clinical treatment response (regression vs progression) at each sample timepoint relative to the initial sample timepoint was determined from clinical records. Mann-Whitney U test was used for statistical analyses.
Results: Twenty eyes of 20 patients had ≥3 AH extractions during the study period; 6 eyes had no SCNAs and were excluded from final analysis. Disease progression at any timepoint was associated with significantly higher SCNA amp (1.65±0.34) and TFx (0.57±0.36) than disease regression (SCNA amp 1.22±0.34, P=0.0002; TFx 0.29±0.34, P=0.01). Relative increases in SCNA amp (Δamp 1.19±0.35) and TFx (ΔTFx 3.68±5.27) from start to end of longitudinal AH sampling were associated with overall disease progression, whereas relative decreases in SCNA amp (Δamp 0.78±0.08; P=0.005) and TFx (ΔTFx 0.55±0.52; P=0.006) were associated with overall disease regression.
Conclusions: We demonstrate that increases and decreases in AH-derived SCNA amp and TFx during therapy tend to correspond to clinical progression and regression of disease, respectively. Our findings suggest that longitudinal AH evaluation throughout active RB therapy may provide a novel quantitative method of monitoring RB treatment response.
Citation Format: Ashley Polski, Liya Xu, Rishvanth K. Prabakar, Jonathan W. Kim, Peter Kuhn, David Cobrinik, James Hicks, Jesse L. Berry. Longitudinal aqueous humor sampling reflects treatment response in retinoblastoma patients [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 185.