bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2934: The role of Globo H in cancer cell survival
Authors: Kuo Tzer-Min , Tsai Yi-Chien , Lee Chin-Chan , Lai Jiann-Shiun 
Published: September 2020

Abstract
BACKGROUND: Globo H (GH), a glycosphingolipid (GSL) carbohydrate antigen, is found on a variety of epithelial tumors and is a promising target for cancer immunotherapy given the marked difference in expression of GH between cancer cells and normal tissues. Globo series antigens (Globo H, SSEA-4) have been implicated in playing important roles in protecting tumor cells from apoptosis, suppression of immune cell activity in the tumor microenvironment, and promoting endothelial cell angiogenesis through shedding the glycosphingolipids to immune cells and endothelial cells, respectively. Recently, apoptosis regulation by protein interaction with Globo-series GSLs in breast cancer cells was reported. This study aims to investigate the GH and anti-GH antibody effects on cancer cell survival.
METHODS: In vivo: After allografting murine B16F10 melanoma subcutaneously, C57BL/6 mice, n=5, were treated with Globo-H ceramide (GHCer) by peritumoral injection and monitored for 26 days. In vitro: GH expression on the surface of tumor cells and colony formative activity in GHCer-treated human cells were measured. Cytokine antibody array and Microwestern protein array analyses were performed in the GHCer-treated A549 cells. Cell viability in high GH–expressing lung cancer cells treated with erlotinib was assessed. Erlotinib resistance and cytokine levels in high GH–expressing tumor cells treated with anti-GH antibody were measured.
RESULTS: In vivo: GHCer-treated mice showed accelerated B16F10-tumor growth rates (3.36-fold; P&lt;0.01, n=5) with less tumor necrosis and higher proliferating cell nuclear antigen expression in the internal tumor tissues. In vitro: In GHCer-treated human lung cancer A549 and breast cancer MDA-MB-231 cells, higher colony formative activity and increased exogenous GH expression on the surface of tumor cells compared to the control group were observed, while cell proliferation was not enhanced. Up-regulated erlotinib-resistant cytokine IL-8 and FGF2 and K-RAS-related signaling were observed in A549 cells with exogenous high GH expression. Exogenously GHCer-added lung cancer cells displayed higher cell viability (37.8% increase) during erlotinib challenge, and an anti-GH antibody treatment decreased the GH effects of erlotinib resistance.
CONCLUSION: GH promotes cancer cell viability in vitro and in vivo, indicating an important role in cancer cell survival during tumor progression. Anti-GH antibody treatment was shown to be able to abolish the GH effects on erlotinib resistance of lung cancer cells in vitro.
Citation Format: Tzer-Min Kuo, Yi-Chien Tsai, Chin-Chan Lee, Jiann-Shiun Lai. The role of Globo H in cancer cell survival [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 2934.
Abstract 2958: ONC206, an imipridone derivative, induces cell death through activation of the integrated stress response in serous endometrial cancer in vitro
Authors: Zhang Yingao , Huang Yu , Yin Yajie , Fan Yali , Tucker Katherine , Staley Allison , Prabhu Varun , Allen Joshua , Zhou Chunxiao , Bae-Jump Victoria 
Published: September 2020

Abstract
Objectives: ONC206 (Oncoceutics) is a selective dopamine receptor D2 (DRD2) antagonist and an analogue of ONC201, an imipiridone currently being investigated in phase II clinical trials for serous endometrial cancer (EC). ONC206 has been shown to inhibit cell proliferation through activation of the integrated stress response (ISR) and apoptosis in brain, breast, and colorectal cancer cell lines. Thus, we investigated the efficacy of ONC206 for serous EC as well as its underlying mechanism of action.
Methods: The serous EC cell lines, ARK1 and SPEC-2, were used. Cellular proliferation was measured using MTT assay. Cleaved caspase-3 and 9, markers of apoptosis, were assessed by ELISA assays. Adhesion and invasion were assessed by laminin and wound healing assays, respectively. Relative In vitro ISR was estimated by ROS and JC-1 assays. Western immunoblotting evaluated effects of ONC206 on proteins central to cellular stress and invasion. All statistical analyses were performed using the software Prism 8 (GraphPad).
Results: ONC206 inhibited cellular proliferation in a dose-dependent manner and was more potent than ONC201 in ARK1 (IC50=0.33uM vs IC50=1.59uM) and SPEC-2 (IC50=0.24uM vs IC50=0.81uM) cell lines. Treatment with ONC206 decreased JC-1 fluorescence (p=0.001-0.01), and increased ROS production (p&lt;0.001), cleaved caspase-3 (p=0.008-0.02), and caspase-9 (p=0.02-0.04) activity in both cell lines. ONC206 increased expression of the stress proteins ATF4, PERK, and BiP. Adhesion (p=0.01-0.02) and wound healing (p&lt;0.001) were significantly impaired after treatment with ONC206 and was accompanied by decreases in proteins Slug, Snail, Vimentin, and VEGF-C. Pretreatment with stress inhibitor n-acetylcysteine (NAC) significantly attenuated the efficacy of ONC206 on ROS production (p=0.001-0.01), wound healing (p&lt;0.001), and proliferation (p=0.001-0.002) in both cell lines.
Conclusions: Our results suggest that ONC206 demonstrates nanomolar potency for the inhibition of proliferation in serous EC cell lines. Specifically, ONC206 utilizes ISR activation as a significant pathway in the propagation of its anti-proliferative and anti-metastatic effects. Thus, ONC206 may be a promising agent in serous EC which has limited treatment options.
Citation Format: Yingao Zhang, Yu Huang, Yajie Yin, Yali Fan, Katherine Tucker, Allison Staley, Varun Prabhu, Joshua Allen, Chunxiao Zhou, Victoria Bae-Jump. ONC206, an imipridone derivative, induces cell death through activation of the integrated stress response in serous endometrial cancer in vitro [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 2958.
Abstract 2979: The effect of interferon gamma on melanoma progression under MAPK inhibitor therapy
Authors: Chen Kevin , McMillan Rachel , Chatrizeh Mona , Hugo Willy 
Published: September 2020

Abstract
About half of metastatic cutaneous melanoma harbor a mutation in the BRAF protein, causing constitutive activation of the MAPK pathway. Currently, patients with such mutation are treated with MAPK pathway inhibitors (MAPKi) targeting the mutant BRAF and its downstream MEK proteins. However, within a span of 6-12 months, these patients will relapse, and their tumors will acquire resistance to this treatment. In this project, we aim to understand the role of IFN gamma (IFNg) to help us determine if the resistance phenotype differs between cells exposed with MAPKi alone versus cells treated with MAPKi and IFNg. Previous literature has reported on the development of a dedifferentiated resistance phenotype which was characterized by a downregulation of MITF and an upregulation of AXL proteins. However, the patient-derived transcriptomic data comparing paired tumors pre- and post-MAPKi therapy did not show such changes in MITF or AXL levels. We posit that the discrepancy may be because most of the earlier studies on MAPKi resistance were done in vitro or immune-suppressed in vivo models. Melanoma biopsies from patients responding to MAPKi treatment showed a substantial increase in the tumor-infiltrating T cell and other immune cells. One of the most dominant immune factors produced by the T cells is the interferon gamma (IFNg). Intriguingly, our data show that IFNg, in the presence of MAPKi, can suppress the dedifferentiation switch that is induced by MAPKi-alone. Specifically, through serial western blots, we discovered that MAPKi and IFNg reduced the previously reported AXL upregulation and MITF downregulation in four different mutant BRAF melanoma lines. RNA sequencing from the same time points revealed that melanoma treated with MAPKi and IFNg upregulate IFN-stimulated genes that are typically expressed by immune cells; this leads us to believe that some novel transcriptomic programs have been activated by the combination of MAPKi and IFNg. By solely examining one immune factor, IFNg, we discovered one potential explanation for the discrepancy in the prevalence of MAPKi-induced dedifferentiation phenotype in the patient-derived melanoma samples. We are in the midst of performing drug screening experiments on MAPKi and IFNg-resistant melanoma lines in order to discover novel therapeutic vulnerabilities that were not observed with MAPKi-alone. Finally, by taking into account the secreted factors in the tumor microenvironment, our study can provide a more accurate depiction of MAPKi resistance evolution in mutant BRAF melanoma patients.
Citation Format: Kevin Chen, Rachel McMillan, Mona Chatrizeh, Willy Hugo. The effect of interferon gamma on melanoma progression under MAPK inhibitor therapy [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 2979.
Abstract 2986: NR2F1 underlies persistence of residual disease in melanoma
Authors: Tiago Manoela , Teh Jessica L. , Purwin Timothy J. , Cai Weijia , Hollingworth Connor , Lopez-Anton Melisa , Aguirre-Ghiso Julio A. , Aplin Andrew E. 
Published: September 2020

Abstract
Despite the clinical success of targeted therapy and checkpoint inhibitors in melanoma, therapeutic responses are transient, followed by relapse that may be driven by a small subpopulation of residual or drug-tolerant cells. Understanding residual disease and metastasis mechanisms can provide clues both for developing improved versions of a drug and for guiding the selection of appropriate drug combinations for melanoma therapy. Here, we found that the well-known marker of tumor dormancy, Nuclear Receptor Subfamily 2 Group F Member 1 (NR2F1), was overexpressed in minimal disease residual cells following CDK4/6 and MEK inhibitors (CDK4/6i+MEKi) treatment in vivo. Furthermore, melanoma cells overexpressing NR2F1 were less sensitive to (CDK4/6i+MEKi) or BRAF and MEK inhibitors (BRAFi+MEKi) treatment in vitro and in vivo models, inhibiting apoptosis. Surprisingly, we did not find any evidence of decreased cell growth in our model. Using a three-dimensional tumor spheroid assay in vitro, we found the NR2F1 expression enhanced melanoma invasion following CDK4/6i+MEKi or BRAFi+MEKi treatments. The use of published RNA Seq data sets that were gathered from the GEO database and Single Cell Seq data sets from PDX melanoma samples showed that high expression of NR2F1 is enriched in the undifferentiated cell state and invasive cells, respectively. Furthermore, BRAF mutant patient sample with an acquired mutation in NRAS Q61R following BRAFi+MEKi+CDKi presented a high expression of NR2F1. Altogether, these findings suggest that NR2F1 may play a role in residual disease persistence besides known features of tumor dormancy, especially important in determining responses to dramatic changes in the environment, such as changes induced by anti-cancer therapy.
Citation Format: Manoela Tiago, Jessica L. Teh, Timothy J. Purwin, Weijia Cai, Connor Hollingworth, Melisa Lopez-Anton, Julio A. Aguirre-Ghiso, Andrew E. Aplin. NR2F1 underlies persistence of residual disease 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 2986.
Abstract 3003: MET mediates entrectinib resistance in ROS1 gene fusion positiveNSCLC
Authors: Tyler Logan , Le Anh , Nijmeh Hala , Doebele Robert 
Published: September 2020

Abstract
ROS1 gene fusions are predominantly found in non-small cell lung cancer (NSCLC), and ROS1 gene fusion-positive (ROS1+) tumors comprise 1-2% of all diagnosed lung adenocarcinomas. Currently, there are two ROS1 tyrosine kinase inhibitors (TKIs) which are US FDA approved, crizotinib and entrectinib, both of which generate tumor response rates in excess of 70% and demonstrate prolonged disease control. However, acquired resistance to TKIs is inevitable, and has been reported in ROS1+ NSCLC tumors or cancer cell models by our group and others and include ROS1 kinase domain mutations (e.g., L1951R, S1986Y/F, F2004V, L2026M, and G2032R), activation of bypass signaling pathways (e.g., EGFR, RAS, or KIT), or phenotypic transformation (such as epithelial to mesenchymal transition). In order to better understand mechanisms of acquired TKI resistance in ROS1 NSCLC, our lab utilized primary, patient-derived cancer cell models harboring these rare oncogenic kinase rearrangements. Herein, using a patient-derived NSCLC cancer cell line (CUTO28) harboring a TPM3-ROS1 fusion, we derived an entrectinib-resistant cell line (CUTO28-ER) through in vitro culture under drug selection. In these CUTO28-ER cells, we identified: (1) MET-mediated bypass signaling as an acquired resistance mechanism to entrectinib, (2) upregulation of MET signaling is accomplished via MET gene amplification, and (3) resistance could be overcome by the dual ROS1/MET inhibitor crizotinib. To our knowledge, this is the first reported case of MET-mediated acquired resistance in a ROS1+ cancer. This cell line model finding was supported by another primary, patient-derived cell line (CUTO38) harboring a CD74-ROS1 fusion; this cell line was derived from a NSCLC patient following disease progression on entrectenib in the clinic. CUTO38 is resistant to entrectinib in vitro and displays elevated MET expression, MET activation with increased phosphorylation, and dependence on MET for cell proliferation and survival. Notably, MET activation in CUTO38 was not generated by gene amplification, MET exon 14 splicing, MET gene fusion, or autocrine HGF expression, suggesting alternate means of MET activation. These findings suggest that interrogation of MET gene amplification or other MET biomarkers should be performed in ROS1+ NSCLC patients who experience disease progression on entrectinib or other ROS1 TKIs that do not inhibit MET, since there are clinically available MET inhibitors, some of which are dual ROS1/MET inhibitors, that may overcome drug resistance in these patients.
Citation Format: Logan Tyler, Anh Le, Hala Nijmeh, Robert Doebele. MET mediates entrectinib resistance in ROS1 gene fusion positiveNSCLC [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 3003.
Abstract 3016: Tucatinib inhibits CYP3A, CYP2C8 and P-gp-mediated elimination and is impacted by CYP2C8 inhibition in healthy volunteers
Authors: Topletz-Erickson Ariel R. , Lee Anthony , Sun Hao , Mayor JoAl , Walker Luke , Endres Christopher J. 
Published: September 2020

Abstract
Background: Tucatinib is a potent, highly selective HER2 tyrosine kinase inhibitor in development for the treatment of patients with HER2+ metastatic breast cancer. In vitro metabolism studies suggest that drug metabolizing enzymes CYP2C8 and CYP3A play a role in tucatinib metabolism. Tucatinib exhibits competitive inhibition of CYP2C8, CYP2C9, CYP3A, and P-gp, and metabolism-dependent inactivation of CYP3A in vitro. ONT-380-012 was a clinical drug interaction study conducted to evaluate the magnitude of potential enzyme and transporter interactions for tucatinib (both as a victim and perpetrator), and the safety of healthy subjects when administered tucatinib doses at therapeutic levels (300 mg BID).
Methods: Healthy volunteers (n=116) at multiple centers were enrolled in the study. Parts A-C evaluated the effects of a strong CYP2C8 inhibitor (gemfibrozil), a strong CYP3A inhibitor (itraconazole), and a CYP3A/CYP2C8 inducer (rifampin) on single-dose tucatinib (300 mg) PK. Parts D and E assessed the effects of steady-state tucatinib (300 mg BID) on single-dose PK of substrate probes for CYP2C8 (repaglinide), CYP2C9 (tolbutamide), CYP3A (midazolam), and P-gp (digoxin). Plasma samples were collected for PK analysis and drug concentrations were measured using validated LC-MS/MS methods.
Results: A strong CYP3A inhibitor (itraconazole) increased tucatinib AUCinf and Cmax 1.3-fold. A CYP3A/CYP2C8 inducer (rifampin) decreased tucatinib AUCinf and Cmax 48% and 37%, respectively. A strong CYP2C8 inhibitor (gemfibrozil) increased tucatinib AUCinf and Cmax 3.1- and 1.6-fold, respectively.
Tucatinib increased the AUCinf and Cmax of the CYP3A substrate (midazolam) 5.7- and 3.0-fold, respectively, the AUCinf and Cmax of the CYP2C8 substrate (repaglinide) 1.7-fold, and the AUCinf of the P-gp substrate (digoxin) 1.5-fold. Tucatinib had no impact on the PK of the CYP2C9 substrate (tolbutamide). Overall, tucatinib was well tolerated in healthy volunteers when administered 300 mg BID.
Conclusions: Together, these data indicate tucatinib is metabolized primarily by CYP2C8 and to a lesser extent via CYP3A. Tucatinib was found to be a strong inhibitor of CYP3A, a weak inhibitor of CYP2C8 and P-gp, and had no impact on CYP2C9-mediated metabolism in vivo.
Citation Format: Ariel R. Topletz-Erickson, Anthony Lee, Hao Sun, JoAl Mayor, Luke Walker, Christopher J. Endres. Tucatinib inhibits CYP3A, CYP2C8 and P-gp-mediated elimination and is impacted by CYP2C8 inhibition in healthy volunteers [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 3016.
Abstract 3024: Relationship between venetoclax concentrations and undetectable MRD in patients with chronic lymphocytic leukemia
Authors: Gopalakrishnan Sathej , Menon Rajeev , Chyla Brenda , Samineni Divya , Jacobson Amanda , Kim Su Young , Mensing Sven , Salem Ahmed Hamed 
Published: September 2020

Abstract
Minimal residual disease (MRD) is an important clinical endpoint in chronic lymphocytic leukemia (CLL) because it correlates with progression free survival and overall survival. The objective of this work is to investigate the impact of venetoclax (VEN) exposures on undetectable MRD rates in blood and bone marrow (BM) in relapsed/refractory (R/R) CLL patients receiving VEN monotherapy or combination therapy with rituximab (RTX).
Data from a total of 708 patients from four VEN monotherapy studies and one combination Phase 1b study (VEN + 6-9 cycles of RTX) in R/R CLL patients were utilized to perform this analysis. The median number of prior therapies in these studies was 3, 42% of the patients had 17p chromosomal deletion and 28% had prior B-Cell Receptor inhibitor (BCRi) therapy. Blood and BM MRD assessments were available for 70% and 38%, respectively, of patients included in the analysis. Absence of an MRD measurement was assumed as detectable MRD for the purposes of this analysis.
The average concentration (Cavg) of VEN was computed for each patient by fitting a population pharmacokinetic model to the observed VEN concentration data. The model was used to calculate area under the curve and compute Cavg accounting for VEN dose ramp-ups, reductions and interruptions. The relationships between venetoclax Cavg and undetectable MRD rates were assessed by using quartile plots. Logistic regression analyses were also conducted to quantify the exposure-response relationship.
With VEN monotherapy, the undetectable MRD rates in blood and BM increased with the increase in VEN exposures. With VEN + RTX combination treatment, the undetectable MRD rates in blood and BM showed an increase with increasing VEN average concentration, but the benefit saturated with limited increase at exposures beyond 400mg.
Logistic regression analyses indicated statistically significant (p&lt;0.01) relationships between VEN average concentrations and undetectable MRD rates in both blood and BM.
These results demonstrate that higher venetoclax concentrations are associated with higher undetectable MRD rates in both blood and BM. When VEN is combined with RTX, an increase in VEN dose beyond 400mg is unlikely to result in higher undetectable MRD rates.
Citation Format: Sathej Gopalakrishnan, Rajeev Menon, Brenda Chyla, Divya Samineni, Amanda Jacobson, Su Young Kim, Sven Mensing, Ahmed Hamed Salem. Relationship between venetoclax concentrations and undetectable MRD in patients with chronic lymphocytic 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 3024.
Abstract 3039: Essential role of IRAK4/TPL2 signaling axis in MAPK activation by oncogenic RAS and genotoxic stress
Authors: Dodhiawala Paarth B. , Khurana Namrata , Zhang Daoxiang , Cheng Yi , Li Lin , Seehra Kuljeet , Jiang Hongmei , Grierson Patrick M. , Wang-Gillam Andrea , Ruzinova Marianna B. , Lim Kian-Huat 
Published: September 2020

Abstract
Pancreatic ductal adenocarcinoma (PDAC) has a 5-year survival of 9% and effective treatment options remain elusive. Oncogenic mutations of KRAS occur in &gt;95% of PDACs and are well-established as the bona fide driver event. However, inhibition of KRAS oncoprotein or its downstream signaling cascades remains unsuccessful in PDAC patients. Furthermore, parallel survival pathways including constitutive activation of the NF-κB pathway poses an additional therapeutic barrier. Previous work from our lab showed that Interleukin-1 Receptor associated kinase 4 (IRAK4) is a major driver of NF-kB cascade in PDAC. Here, through an unbiased reverse phase protein array screen and RNA sequencing, we discovered IRAK4 controls MAPK activity downstream of KRAS. Ablation of IRAK4 completely abolishes RAS-induced transformation in human and murine cell lines. Mechanistically, we implicate a KRAS-driven IL-1β signaling loop that activates IRAK4 and uncover MAP3K8 (or TPL2/COT) as the kinase through which IRAK4 activates MEK and ERK. Suppression of TPL2 abrogates KRAS-driven MEK-ERK activity and transformed growth of PDAC cell lines. In addition, TPL2 inhibition suppresses p105/p50 NF-kB activation, a valuable phenomenon that distinguishes TPL2 inhibition from MEK inhibition. We find TPL2 inhibition synergizes with chemotherapy to suppress growth of PDAC cell lines in vitro and patient-derived xenograft tumor model in vivo. Analyses of PDAC tissue microarray showed TPL2 expression to be marginally associated with poor prognosis. Additionally, we are the first to characterize gain-of-function point mutations in TPL2 which hyperactivate MAPK and NF-kB, in part by preventing TPL2 protein degradation. Together, our study broadens the understanding of the oncogenic RAS signaling network and reveals IRAK4 and TPL2 as novel practical therapeutic targets in RAS-driven cancers.
Citation Format: Paarth B. Dodhiawala, Namrata Khurana, Daoxiang Zhang, Yi Cheng, Lin Li, Kuljeet Seehra, Hongmei Jiang, Patrick M. Grierson, Andrea Wang-Gillam, Marianna B. Ruzinova, Kian-Huat Lim. Essential role of IRAK4/TPL2 signaling axis in MAPK activation by oncogenic RAS and genotoxic stress [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 3039.