bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5204: Targeting OGG1 arrest cancer cell proliferationby mitochondrial dysfunction and replication stress
Authors: Benitez-Buelga Carlos , Visnes Torkild , Sanjiv Kumar , Mortusewicz Oliver , Berglund Ulrika Warpman , Helleday Thomas 
Published: September 2020

Abstract
Altered oncogene expression in cancer cells causes lost redox homeostasis resulting in oxidative DNA damage, e.g., 8-oxoguanine (8-oxoG), repaired by base excision repair (BER). PARP1 coordinates BER, and relies on the upstream 8-oxoguanine-DNA glycosylase (OGG1) to recognise and excise 8-oxoG. Here we hypothesize that OGG1 may represent an attractive target to exploit reactive oxygen species (ROS) in cancer. Although OGG1 depletion is well tolerated in non-transformed cells, we report here that OGG1 depletion obstruct A3 T-cell lymphoblastic acute leukemia growth in vitro and in vivo, validating OGG1 as a potential anti-cancer target. In line with this hypothesis, we show OGG1 inhibitors (OGG1i) target a wide range of cancer cells, with a favourable therapeutic index to non-transformed cells. Mechanistically, OGG1i and shRNA depletion cause lost mitochondrial function leading to S-phase damage, replication stress and lost proliferation or cell death, representing a novel mechanistic approach to target cancer. This study adds OGG1 to the list of BER factors, e.g., PARP1, as potential targets for treatment of cancer.
Citation Format: Carlos Benitez-Buelga, Torkild Visnes, Kumar Sanjiv, Oliver Mortusewicz, Ulrika Warpman Berglund, Thomas Helleday. Targeting OGG1 arrest cancer cell proliferationby mitochondrial dysfunction and replication 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 5204.
Abstract 5206: Quantifying DNA-protein crosslinks induced by bifunctional alkylating agents in cultured human leukemia cells
Authors: Cerrato Hailey L. , Marchant Sam W. , Naccara Allie H. , LaJoie Thomas J. , Michaels Shawnya , Brown Dennis , Rice Kevin P. 
Published: September 2020

Abstract
Bifunctional alkylating agents, such as cisplatin and several nitrosoureas, have clinical utility as antineoplastic drugs largely due to their DNA alkylating and crosslinking activity. Other drug candidates such as laromustine (VNP-40101M) and dianhydrogalacticol (VAL-083) have similar DNA-alkylating activities and have shown success in clinical trials against hard-to-treat cancers such as acute myelogenous leukemia and glioblastoma multiforme. While these agents' interstrand and instrastrand DNA crosslinking activities are widely cited as the critical cytotoxic event, their capacity to crosslink DNA to chromosomally-affiliated proteins may also contribute to their anticancer profile. Indeed, DNA-protein crosslinks (DPCs) are highly toxic phenomena that are difficult for cells to repair and severely compromise gene transcription and DNA replication. Herein, we assess concentration-dependent DPC formation by a wide variety of bifunctional alkylating agents in HL-60 human leukemia cells using two parallel approaches. In one approach, protein-bound DNA was quantified using PicoGreen after precipitating proteins from lysates of drug-treated cells and compared to total DNA concentration. In another, chromosomal DNA was extracted from lysates of drug-treated cells then treated with fluorescein isothiocyanate, which modifies primary amines on proteins. Protein concentration was measured by fluorescence and compared to that of the extracted DNA. The nitrogen mustard mechlorethamine and the chloroethyl nitrosoureas carmustine and lomustine were the most potent agents, yielding significant DPC formation at clinically relevant doses (10−5 M). Cisplatin and laromustine yielded DPCs at similar concentrations, but to a lesser extent. Dianyhydrogalacticol, epichlorohydrin, and diepoxybutane, which all rely on epoxide functional groups for attachment to cellular nucleophiles, required at least an order of magnitude higher drug concentrations for significant DPC observation. These data suggest bifunctional alkylating agents featuring halide leaving groups are most apt to crosslink DNA to nuclear proteins and may offer insights into patterns of cytotoxicity and clinical utility for this important class of chemotherapeuric agents.
Citation Format: Hailey L. Cerrato, Sam W. Marchant, Allie H. Naccara, Thomas J. LaJoie, Shawnya Michaels, Dennis Brown, Kevin P. Rice. Quantifying DNA-protein crosslinks induced by bifunctional alkylating agents in cultured human leukemia 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 5206.
Abstract 5215: Inhibition of PIM kinases targets synthetic vulnerabilities and enhances antigen presentation in B cell lymphoma
Authors: Mondello Patrizia , Tarantelli Chiara , Cascione Luciano , Arribas Alberto J. , Rinaldi Andrea , Younes Anas , Bertoni Francesco 
Published: September 2020

Abstract
The PIM kinases are highly expressed in activated B-cell (ABC) diffuse large B-cell lymphoma (DLBCL). Oncogenic cooperation between PIMs and MYC has been demonstrated. Transgenic mice co-expressing Eμ-PIM and Eμ-MYC showed accelerated lymphomagenesis. Conversely, knockdown of PIMs dramatically decreased cMYC levels and lowered tumor incidence. Based on these preclinical data, a treatment strategy aiming at disrupting the oncogenic cooperation between PIMs and MYC may improve the outcome of DLBCL. Therefore, we treated a panel of DLBCL cell lines with increasing dose of the clinically relevant pan-PIM inhibitor (PIMi) AZD1208 (from 0.1 to 10μM) for 48 hours (Hrs), which resulted in a dose-dependent growth inhibition with a stronger efficacy in ABC DLBCL cell lines. The analysis of a CRISPR loss-of-function screening in three ABC (LY3, TMD8, HBL1) and three GCB (SUDHL-4, Pfeiffer, BJAB) DLBCL cell lines (Reddy et al, 2017) showed that PIM2 silencing led to significantly decreased viability irrespective of cell-of-origin, suggesting that this oncogene is essential for cell proliferation in DLBCLs. To identify the genes through which PIMs drive the lymphoma phenotype we performed gene expression profiling using 4 ABC DLBCL cell lines (RIVA, TMD8, SUDHL-2, U2932) treated with either DMSO or AZD1208 at 1μM for 4, 8 and 12 Hrs. We observed induction of 3,439 genes whereas 2,473 genes were downregulated. Gene pathway analysis showed that AZD1208 led to downregulation of genes regulated by MYC, including its known downstream p53 and NFKB target genes. On the other hand, AZD1208 treatment broadly induced MHC class II and antigen presentation genes as well as PI3K/AKT, cell cycle and glutaminase genes. Using a high-throughput screening approach, we found that the inhibitors of cell cycle (such as the BCL2 inhibitor venetoclax/ABT199 and the PLK4 inhibitor CFI-400945) and of glutaminase (CB839) enhanced the antiproliferative effect of AZD1208, whereas combinations with the PI3K/AKT/mTOR inhibitors had negligible synergistic effect. In conclusion, our study revealed previously unknown mechanisms of action of PIM inhibitors and provides a framework for future combination strategies.
Citation Format: Patrizia Mondello, Chiara Tarantelli, Luciano Cascione, Alberto J. Arribas, Andrea Rinaldi, Anas Younes, Francesco Bertoni. Inhibition of PIM kinases targets synthetic vulnerabilities and enhances antigen presentation in B cell lymphoma [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 5215.
Abstract 522: Optimization of novel anti-4-1BB antibodies on Biocytogen's in vivo drug screening platform
Authors: Guo Chaoshe , Zhu Yu , Chen Yunyun , Yang Yi , Li Zhihong 
Published: September 2020

Abstract
4-1BB (CD137, tumor necrosis factor receptor superfamily 9) is an inducible costimulatory receptor expressed on activated T and natural killer (NK) cells. 4-1BB ligation on T cells triggers a signaling cascade that results in upregulation of anti-apoptotic molecules, cytokine secretion, and enhanced effector function. In dysfunctional T cells that have a decreased cytotoxic capacity, 4-1BB ligation demonstrates a potent ability to restore effector functions. On NK cells, 4-1BB signaling can increase antibody-dependent cell-mediated cytotoxicity. Agonistic monoclonal antibodies targeting 4-1BB have been developed to harness 4-1BB signaling for cancer immunotherapy.
We have interests in developing novel 4-1BB antibodies for boosting cancer patients' own immunity in light of other successful immune checkpoint modulators. We developed a cohort of 4-1BB specific antibodies using the classic hybridoma technology. Then, we took the advantages of Biocytogen's humanized mice and screened the purified antibodies in B-h4-1BB mice bearing MC-38 tumor cells. We found some antibodies that are as potent as Urelumab in vivo. We then humanized the antibody to obtain YH004. To determine the subtype of the antibody, we grafted the antigen-binding domain onto various human Fc backbones. Interestingly, we found that the human IgG1 isotype was more effective against tumors than the other isotypes in B-4-1BB mice with MC38 tumors and Treg depletion were noted in tumor-infiltrating lymphocyte analysis using the same murine model. In addition, we proved that the synergistic anti-tumor activity of YH004 in combination with pembrolizumab in B-hPD-1/h4-1BB mice. In brief, the 4-1BB antibody was validated in humanized mice to provide a favorable reference for future clinical evaluation.
Citation Format: Chaoshe Guo, Yu Zhu, Yunyun Chen, Yi Yang, Zhihong Li. Optimization of novel anti-4-1BB antibodies on Biocytogen's in vivo drug screening platform [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 522.
Abstract 523: Advanced engineering using CAR-T display libraries: Ultra-high throughput functional screen
Authors: Youssef Sawsan , Chou Joyce , Pineda Devin , Ives Sarah , Chiou Valerie , Buerckert Jean-Philippe , Newland Raymond , Daraeikia Shahrad , Liao Sindy , Jones Chelsea , Pettus Christina , Salas Jessica , Padilla Emelia , Smith Christopher , Day Giles , Maurer David , Waddell Ian , Glanville Jacob 
Published: September 2020

Abstract
CAR-T-cell therapies represent a new area of synthetic biology success in cancer immunotherapy. However, the complexity of their adverse events related to their activity are still a major challenge. Using our in-house fully human computationally optimized phage display library, we established a novel cell-based ultra-high throughput panning and screening CAR-T platform. Our proprietary CAR-T vector is designed as a series of cassettes incorporating our SuperHuman phage library with an optimal backbone. This allows us to design CAR-T with desired properties, including: Range of Affinities, Humanness, Epitope Diversity, Thermostability, Immunogenicity, Tonic activation, Toxicity, and Functional Activity. Our library's encoding ensures a single CAR-T construct per one reporter cell. Moreover, to confirm CAR-T-specific-activation, we incorporated three different markers that will be upregulated simultaneously. We used CD19 and BCMA to validate our platform selecting target-specific functionally active scFv candidates against both targets, which are selective to activation conditions such as tonic signaling, and tissue specific silencing. Collectively we show that our platform can be readily used to characterize CAR-T in high-speed and - throughput fashion.
Citation Format: Sawsan Youssef, Joyce Chou, Devin Pineda, Sarah Ives, Valerie Chiou, Jean-Philippe Buerckert, Raymond Newland, Shahrad Daraeikia, Sindy Liao, Chelsea Jones, Christina Pettus, Jessica Salas, Emelia Padilla, Christopher Smith, Giles Day, David Maurer, Ian Waddell, Jacob Glanville. Advanced engineering using CAR-T display libraries: Ultra-high throughput functional screen [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 523.
Abstract 524: Discovery of NKp46 antibody for NK cell engager using common light chain phage display approach
Authors: Yang Teddy , Zhu Seng , Gao Jing , Yao Jingyun , Gong Xumin 
Published: September 2020

Abstract
NK cells express activating receptors such as CD16a (FcγRIIIA), NKG2D, SLAM family members and the natural cytotoxicity receptors (NCRs). Full activation of NK cells require co-engagement of different activating receptors. NKp46, a member of NCRs, is a 46 kDa glycoprotein and belongs to immunoglobulin (Ig) superfamily, NKp46 is frequently expressed on tumor infiltrating lymphocytes. Activation of NKp46 with antibody triggers not only NK cell cytotoxicity but also cytokine release. Taken together, NKp46 is a promising target for anti-tumor therapy. Here we described the discovery of fully human NKp46 antibody by phage display approach with common light chain and naïve human B cell library. Bispecific antibody has been researched for many years and very limited number of bispecific antibodies are currently in the market. Heavy and light chain miss pairing is the bottleneck of bispecific antibody discovery and development. To overcome the hurdle, we generated a fully human common light chain phage display library with two commonly used light chain sequences combined with heavy chain fragment from human naïve scFv library. Two light chain sequences were selected based on the approved antibodies sequences and the expression levels. The phage library was then used for panning against recombinant human NKp46 extracellular domain protein and CHOK1 cells overexpressing human NKp46. After conversion to full IgG format, 4 candidates bind specifically to NKp46 and cross react with both human and cyno NKp46. Affinities of the candidates were comparable to bench mark antibodies as determined by FACS. To summarize, we have generated fully human common light chain phage display library that can be used for bispecific antibody discovery. The anti-NKp46 antibodies discovered can be further developed for NK cell engager and NK cell redirecting bispecific antibody.
Citation Format: Teddy Yang, Seng Zhu, Jing Gao, Jingyun Yao, Xumin Gong. Discovery of NKp46 antibody for NK cell engager using common light chain phage display approach [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 524.
Abstract 5240: Effective inhibition of TMPRSS2-ERG positive prostate cancer cells by a new formula of ERGi-USU-6
Authors: Eldhose Binil , Xavier Charles P. , Pandrala Mallesh , Malhotra Sanjay V. , Dobi Albert 
Published: September 2020

Abstract
Background: Prostatic adenocarcinoma is among the leading cause of cancer-related deaths among men in the United States. The Erythroblast Transformation-Specific-Related Gene, ERG is dormant in normal prostate epithelium. Due to gene fusions, the ERG oncogene is frequently activated by androgenic signals in prostate cancer. ERG disrupts the normal differentiation, promotes epithelial-mesenchymal transition, migratory and invasive properties of cancer cells. Approximately 35% of metastatic castration-resistant prostate cancers harbor ERG oncogene. Due to the failure of androgen axis directed therapies, there is an urgent need to develop inhibitors to targeting prostate cancer-causing genes, such as ERG. We have identified a potent small-molecule inhibitor ERGi-USU that is remarkably selective for inhibiting the growth of ERG positive cancer cells through direct binding to the RIOK2 atypical kinase, a putative upstream regulator of ERG. We completed a structure-activity relationship (SAR) study and compound development resulting in the potent derivative, ERGi-USU-6. Our current objective is to improve the therapeutic properties of ERGi-USU-6, by new salt formulations.
Methods: Evaluation of the five selected salt formulations of ERGi-USU-6 were performed by assessing the growth of ERG positive prostate cancer cell line (VCaP) and by monitoring ERG and RIOK2 protein levels. Selectivity was assessed by monitoring the growth, endogenous ERG and RIOK2 levels in normal ERG positive human umbilical vein derived endothelial cells (HUVEC). The IC50 values for cell growth, compared to the parental compound were calculated in a 12-step dilution range performed in triplicates and independently repeated three times. Cell growth was measured by quantitative Cell Glow assay monitoring viable cells using Perkin Elmer Envision assay instrument and protein levels were quantified by measuring bioluminescence in IBright instrument in a wide dynamic range.
Results: One new salt formula with improved activity were identified, demonstrating improved cell growth (IC50=89nM vs. parental IC50=139), ERG protein and RIOK2 protein inhibition. None of the salt formulas of ERGi-USU-6 showed any effect on the primary cultures of the ERG positive normal endothelial cells (HUVEC) in the effective concentration range.
Conclusion: The first evaluation of salt formulas of ERGi-USU-6 may open the possibilities for preclinical assessments of this remarkably cancer-selective compound.
Citation Format: Binil Eldhose, Charles P. Xavier, Mallesh Pandrala, Sanjay V. Malhotra, Albert Dobi. Effective inhibition of TMPRSS2-ERG positive prostate cancer cells by a new formula of ERGi-USU-6 [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 5240.
Abstract 5262: Identifying the role of AXL in fusion kinase inhibitor resistant non-small cell lung cancer
Authors: Peters Tara , Dimou Anastasios , Le Anh , Doebele Robert 
Published: September 2020

Abstract
Chromosomal rearrangements involving ALK, ROS1, RET and others generate novel fusion kinases which comprise approximately 10% of the oncogenic drivers in lung adenocarcinoma. Several tyrosine kinase inhibitors (TKIs) are approved or in development for each of these oncogenes and have shown marked clinical improvement for patients, but resistance inevitably occurs, a significant proportion of which is not currently druggable. AXL is a receptor tyrosine kinase that is often upregulated in resistant cancers in response to both chemotherapy and targeted therapies such as EGFR inhibitors. AXL can promote resistance to targeted therapy by signaling through the MAPK and Akt pathways or heterodimerizing with other receptors to maintain growth and survival signaling. However, the role of AXL in mediating resistance to ALK, ROS1, and RET TKIs is not well understood. In screening our unique panel of patient-derived lung adenocarcinoma cell lines harboring ALK, ROS1 or RET gene fusions, we found elevated AXL expression was correlated with resistance to cognate TKIs, as well as mesenchymal features including spindle morphology and vimentin expression. Combining the AXL inhibitor R428 with TKI treatment, such as crizotinib for ALK and ROS1 driven cell lines, was effective at inhibiting colony formation and wound healing in fusion kinase inhibitor resistant cell lines. Combination treatment downregulates Akt phosphorylation in resistant cells, and we found the resistant cell lines were also more sensitive to PI3K inhibition than cell lines that are still sensitive to their specific TKI. Interestingly, we also identified AXL cross talk with other receptor tyrosine kinases. In a RET inhibitor resistant model expression of both AXL and EGFR are elevated and the receptors interact, which can be disrupted upon EGF stimulation, to drive resistance. Collectively, we show that AXL upregulation is a common marker of TKI resistance among ALK, ROS1, and RET fusion kinase driven lung adenocarcinoma models, and provide rationale for AXL as a clinical target for combination therapy to overcome drug resistance.
Citation Format: Tara Peters, Anastasios Dimou, Anh Le, Robert Doebele. Identifying the role of AXL in fusion kinase inhibitor resistant non-small cell lung 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 5262.
Abstract 5264: Selective toxicity of MX-106-4C, a survivin inhibitor, in P-glycoprotein-mediated multidrug resistant colon cancer
Authors: Lei Zi-Ning , Wu Zhongzhi , Teng Qiu-Xu , Xiao Min , Li Wei , Wurpel John N. , Chen Zhe-Sheng 
Published: September 2020

Abstract
One of the major challenges in colon cancer chemotherapy is multidrug resistance (MDR), which is typically mediated by the overexpression of ATP-binding cassette (ABC) transporters, particularly P-glycoprotein (P-gp, ABCB1, MDR1). A number of P-gp inhibitors have been developed, however, none of these compounds have improved chemotherapeutic efficacy due to undesirable pharmacokinetic profiles or adverse effects, resulting in limited clinical success. Therefore, alternative approaches are urgently needed to circumvent MDR cancer. In previous study, a series of synthesized analogs of MX-106, as anti-cancer drugs targeting survivin, exhibited collateral sensitivity (CS) effect to P-gp overexpressing MDR colon cancer cells as well as ABCB1 gene transfected cells, reflected by more than 10-fold cytotoxic effect in P-gp positive MDR cell lines compared to drug sensitive cell lines. Among the analogs, MX-106-4C was identified as the leading compound with the most potent selective toxicity to P-gp overexpressing cells. MX-106-4C-induced CS effect was observed in both intrinsic and acquired P-gp overexpressing colon cancer cells, which was only partially reversed with the presence of a P-gp inhibitor. Nevertheless, this CS effect was abolished in ABCB1-knockout cells, indicating that the selective cytotoxicity was P-gp expression dependent, but only partially related to P-gp function. Furthermore, we found that MX-106-4C did not significantly affect P-gp ATPase activity or drug accumulation and efflux in P-gp-overexpressing cells. In P-gp overexpressing colon cancer cells, short-term (up to 72 h) incubation of MX-106-4C significantly down regulated P-gp expression at transcriptional level but not protein level, whereas long-term (14 d) incubation of MX-106-4C significantly down regulated P-gp protein expression and re-sensitized MDR colon cancer cells to doxorubicin. These findings suggested an indirect interaction and regulation between MX-106-4C and P-gp. Further study revealed that the selective cytotoxic effects of MX-106-4C were associated with cell cycle arrest at G1 phase and apoptosis through the downregulation of CDK4. Overall, this study demonstrates that MX-106-4C selectively kills P-gp positive MDR colon cancer cells and indirectly regulates P-gp, which provides a clue for CS compound design and a novel strategy to obviate P-gp-mediated colon cancer MDR by re-sensitizing heterogeneous tumors with CS agents.
Citation Format: Zi-Ning Lei, Zhongzhi Wu, Qiu-Xu Teng, Min Xiao, Wei Li, John N. Wurpel, Zhe-Sheng Chen. Selective toxicity of MX-106-4C, a survivin inhibitor, in P-glycoprotein-mediated multidrug resistant colon 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 5264.