bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2164: In-licensing and knockout validation of monoclonal hybridomas to improve antibody reproducibility
Authors: Heaton Sam , Koch Sofia , Ge Jiangyan , Rizzo Joshua , Prater Michael , Dreja Hanna , Hamilton Bruce , Solache Alejandra 
Published: August 2019

Abstract
Antibodies are the most common tools in basic science and clinical research. Over the past several years there has been an ongoing discussion around antibody validation in light of poor experimental reproducibility between laboratories. This is in part due to the cross-reactivity between antibodies and off-target proteins and the variability between different antibody batches. The resulting experimental irreproducibility leads to wasted resources and compromises the advancement of science. Abcam partners with various research institutes and biotech companies to in-license hybridoma cell lines producing monoclonal antibodies. Through this in-licensing program, Abcam has taken steps to help ensure target-specificity of valuable clones. Our in-licensing program utilizes our ISO9001-accredited production method to ensure each batch performs in-line with previous batches. Where possible, all in-licensed antibodies are also tested in knockout (KO) material to confirm the specificity of the in-licensed clone and subsequent batches. Through our partnership with Horizon Discovery to test antibodies with KO cells, target genes are mutated via CRISPR-Cas9 within a haploid cell line, which results in a frameshift and a complete loss of gene expression. KO cell lines provide a true negative control for antibody validation that allows us to confirm the specificity of clones we in-license and produce. Alongside KO, we are making use of our expertise in recombinant antibodies to explore the conversion of selected in-licensed hybridomas to recombinant antibodies. A key issue with monoclonals is degradation of the clone over time, leading to inconsistent batches and irreproducible results. By sequencing and cloning the antibody, we expect to guarantee continued specificity and reproducibility; therefore, ensuring our licensors see long term impact and value generation for their innovations. Here we present KO data for several in-licensed published clones, validated in both western blot and immunocytochemistry. The antibodies have been tested in KO and wild-type cells to confirm their specificity. By providing researchers with reliable and specific antibodies that work as intended, we hope to minimize wasted resources and improve reproducibility. In addition to our KO data, we present data from the recombinant conversion of clone NAT105 to PD1 and show comparable, if not superior, performance when compared with the hybridoma.
Citation Format: Sam Heaton, Sofia Koch, Jiangyan Ge, Joshua Rizzo, Michael Prater, Hanna Dreja, Bruce Hamilton, Alejandra Solache. In-licensing and knockout validation of monoclonal hybridomas to improve antibody reproducibility [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2164.
Abstract 2167: Antibody glycoengineering for drug delivery applications
Authors: Sehgal Drishti Navin , Kalscheuer Stephen , Panyam Jayanth 
Published: August 2019

Abstract
Antibody-drug conjugates (ADC) hold considerable promise as anticancer agents. A critical determinant of the effectiveness of ADCs is the chemistry that is used to conjugate the payload. Currently used approaches include primarily conjugation to either side-chain amine or carboxylic acid groups or conjugation to thiols. Because these reactions are not site specific and not easily controlled, these chemistries can result in reduced affinity for the target antigen. Further, these conjugation reactions lack selectivity and can result in heterogeneous mixtures of products that differ in the sites and stoichiometry of modification. We investigated a glycoengineering strategy that enables the introduction of artificial azide groups in the antibody without affecting their antigen affinity. This is based on the observation that glycosyltransferases can incorporate non-natural sugars (e.g., azido mannose) at different sites on an IgG molecule. The azide groups in these artificial sugars are then available to react with alkynes through copper-catalyzed ‘click’ chemistry or with strained alkynes such as dibenzyl cyclooctyne (DBCO) allowing for biorthogonal, copper-free ‘click’ chemistry. Because the sugars are added reproducibly and at a site that does not affect antigen binding, the glycoengineering technology would overcome problems associated with traditional conjugation strategies. Using this approach, azide groups were introduced in anti-CD133 and anti-perlecan antibodies. Further, the azide groups were available to react with various DBCO conjugates including fluorophores, drug molecules and nanoparticles. Importantly, the addition of artificial sugar and subsequent azide-alkyne reaction did not affect the affinity of the antibody for the target antigen. Conjugation of a cytotoxin to the antibody resulted in enhanced cell kill in vitro and efficacy in vivo. We plan on further improving the effectiveness of this approach by investigating cleavable linker technology in combination with the glycoengineering strategy. We expect that this strategy will prove to be a unique platform technology that will have a significant impact on antibody-based therapeutics.
Citation Format: Drishti Navin Sehgal, Stephen Kalscheuer, Jayanth Panyam. Antibody glycoengineering for drug delivery applications [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2167.
Abstract 2179: Antitumor activity of JNJ-63576253 (TRC253), a small molecule antagonist of F877L mutant and wild-type androgen receptor
Authors: Bush Tammy L. , Ndikuyeze Georges Habineza , Bignan Gilles , Branch Jonathan , Ondrus Janine , Shi Yifan , Luistro Leopoldo , Hastings James , Erhardt Joseph , Hickson Ian , Patel Shefali , Connolly Peter , Zhang Zhuming , Bischoff James , Rupnow Brent , Gottardis Marco , Packman Kathryn 
Published: August 2019

Abstract
Androgen receptor (AR) antagonists have transformed prostate cancer patient care by targeting a key nodal point in tumor cell signaling. However, despite the impressive clinical activity of first- and second-generation antiandrogens, acquired resistance frequently emerges. Point mutations in the ligand-binding domain of AR, such as phenylalanine to leucine at position 877 (ARF877L), account for 10-20% of resistance. Such mutations are characterized by receptor activation, rather than inhibition, by first- and second-generation antiandrogen therapeutics.
JNJ-63576253 is a potent, high affinity competitive binder of wild type and mutant AR, including F877L. JNJ-63576253 blocks AR nuclear translocation, AR binding to DNA, and AR-dependent transcription. JNJ-63576253 inhibits the proliferation of androgen receptor driven prostate cancer cell lines, including those bearing ARF877L.
In the Hershberger assay in male Sprague Dawley rats, oral administration of JNJ-63576253 inhibited androgen sensitive organ (ASO) development in a dose-dependent manner. In male SHO mice bearing LNCaP xenografts with either wild-type or ARF877L, daily treatment with 30 mg/kg JNJ-63576253 treatment resulted in statistically significant antitumor activity, whereas second-generation antiandrogen enzalutamide had no antitumor efficacy in the LNCaP ARF877L mutant model.
Janssen and Tracon Pharma have entered a strategic licensing collaboration, whereby Tracon possesses exclusive rights for clinical development of JNJ-63576253 (now called TRC253). Tracon has entered TRC253 into Ph1/2A clinical evaluation in metastatic castration-resistant prostate cancer patients.
Citation Format: Tammy L. Bush, Georges Habineza Ndikuyeze, Gilles Bignan, Jonathan Branch, Janine Ondrus, Yifan Shi, Leopoldo Luistro, James Hastings, Joseph Erhardt, Ian Hickson, Shefali Patel, Peter Connolly, Zhuming Zhang, James Bischoff, Brent Rupnow, Marco Gottardis, Kathryn Packman. Antitumor activity of JNJ-63576253 (TRC253), a small molecule antagonist of F877L mutant and wild-type androgen receptor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2179.
Abstract 2181: A novel CXCR4 antagonist strongly mobilizes hematopoietic stem cells in vivo
Authors: Fang Xiao , Meng Qian , Fang Xiong , Mao Yujia , Xu Yan , An Jing , Huang Ziwei 
Published: August 2019

Abstract
Introduction: The transplantation of hematopoietic stem cells (HSCs) has been the standard of care for the treatment of chemotherapy sensitive relapsed acute leukemia. HSCs mainly reside in the bone marrow with only few HSCs circulating in the peripheral blood (PB), due to homing effect mediated by the interaction of chemokine receptor CXCR4 expressed on HSCs with chemotactic ligand SDF-1α rich in the bone marrow. It is clinically desirable to enhance the number of HSCs in the PB for collection during the transplantation procedure. Since SDF-1α/CXCR4 axis and interaction mediates the homing and retention of HSCs in the bone marrow, our laboratory has recently discovered and developed a novel small molecule, HFX51116, that is capable of binding CXCR4 and blocking its interaction with SDF-1α, raising the possibility that this compound can be used for releasing HSCs from the bone marrow to the PB useful for the transplantation of HSCs.
Method and Results: In vitro, there was almost no toxicity of HFX51116 at high concentrations in bone marrow stromal cells that were originally extracted from rats. In addition, HFX51116 was stable in rat serum. In vivo, we tested the short-term and long-term mobilization efficacy of HFX51116. For the short-term efficacy study, we determined the dynamic change of efficacy at different times and doses of HFX51116 in HSC mobilization assays. Furthermore, different strains of mice, such as C57/BL6, C3H/HEJ and DBA/2, were sensitive to HFX51116. HFX51116, when used in combination with G-CSF, had higher efficacy than G-CSF alone. For the long-term repopulation study, we employed the CD45.1/CD45.2 competitive assay to demonstrate that the HSCs mobilized by G-CSF and HFX51116 had the repopulation ability when compared with the bone marrow cells.
Conclusion: Our studies have demonstrated high HSC mobilization efficacy of HFX51116 in vivo and the promise of this new lead compound for further preclinical and clinical studies.
Citation Format: Xiao Fang, Qian Meng, Xiong Fang, Yujia Mao, Yan Xu, Jing An, Ziwei Huang. A novel CXCR4 antagonist strongly mobilizes hematopoietic stem cells in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2181.
Abstract 2192: Concurrent Tumor Treating Fields (TTFields) and dexamethasone decrease proliferation and clonogenicity of patient-derived glioblastoma cells in vitro
Authors: Michelhaugh Sharon K. , Kiousis Sam , Mittal Sandeep 
Published: August 2019

Abstract
Background: TTFields are FDA-approved for treatment of newly-diagnosed and recurrent glioblastoma and extend progression-free survival and overall survival in newly-diagnosed glioblastoma patients. Dexamethasone is administered perioperatively to GBM patients to reduce tumor-associated edema and increased intracranial pressure. In addition, high dose corticosteroids may be required during TTFields therapy. In this study, a patient-derived GBM cell line from a newly-diagnosed glioblastoma patient was treated in vitro with TTFields ± dexamethasone followed by proliferation and clonogenic assays.
Methods: This study was approved by the Institutional Review Board and written consent obtained from the patient. A single-cell suspension was generated from tumor tissues obtained from the newly-diagnosed patient. Cells were cultured in DMEM/F12 media with 10% fetal bovine serum and gentamicin. Prior to TTFields initiation, cells were plated on plastic coverslips (1×104cells/coverslip; n=7-8/group) and incubated overnight. Then, TTFields were applied at 200 kHz with a field intensity of ~1.6 V/cm for 14 days. Cells were treated with TTFields with or without 50 µM dexamethasone. At the end of the TTFields application, proliferation was assessed with XTT assay and cells were harvested and replated for the clonogenic assay (n=3/group). XTT data and clonogenic assay cell counts were analyzed by one-way ANOVA with Tukey’s post-hoc test.
Results: In patient-derived glioblastoma cells treated with TTFields and dexamethasone for 14 days, proliferation was significantly decreased compared to the dexamethasone-only and control groups (0.483±0.30; 1.33±0.18; 1.42±0.11, respectively, ANOVA p&lt;0.001). Similarly, cell counts in the clonogenic assay were markedly decreased in the TTFields ± dexamethasone group compared to dexamethasone-only (1184±425.9; 46464±4580; p&lt;0.05).
Conclusions: In glioblastoma cells from a newly-diagnosed patient, concurrent TTFields with dexamethasone produced a ~three-fold reduction of proliferation and reduced clonogenicity compared to control groups.
Citation Format: Sharon K. Michelhaugh, Sam Kiousis, Sandeep Mittal. Concurrent Tumor Treating Fields (TTFields) and dexamethasone decrease proliferation and clonogenicity of patient-derived glioblastoma cells in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2192.
Abstract 2193: AXL inhibitor TP-0903 attenuates AXL-TGFbeta Hippo signaling axis in lung adenocarcinoma cells
Authors: Taverna Josephine Amalia , Hung Chia-Nung , Lin Chun-Lin , Osmulski Pawel , Chen Meizhen , Wang Chiou-Miin , Lucio Nicholas L. L. , Kirma Nameer , Chou Chih-Wei , Gaczynska Maria E. , Nazarullah Alia , Wade Mark , Mouritsen Lars , Huang Tim 
Published: August 2019

Abstract
Background: Non-small cell lung cancer (NSCLC) is a molecularly heterogeneous disease with a high propensity for drug resistance and metastasis. AXL, a member of the Tyro3-AXL-Mer family of receptor tyrosine kinases, is a central regulator of epithelial-to-mesenchymal transition (EMT) and enables tumor cells to invade and acquire drug resistance. AXL is overexpressed in lung tumors, correlates positively with tumor invasion, drug resistance, and negatively predicts overall survival. We mechanistically interrogate the effects of AXL inhibitor TP-0903 on EMT in lung adenocarcinoma cells using transcriptomic and proteomic profiling.
Methods: Atomic force microscopy, Western blot analysis, RNA sequencing and mass cytometry (CyTOF) were all used to scrutinize the biomechanical properties, phenotypic, transcriptomic and proteomic profiles of A549 cells treated with 40nM TP0903 or shAXL knockdown.
Results: TP-0903 attenuates total AXL/AXL phosphorylation and blunts transcriptional responses to TGFβ-Hippo signaling by disrupting the transcriptional complexes formed by SMAD2/3, SMAD4, YAP1 and TAZ. AXL knockdown or TP-0903 reverses EMT phenotype and reduces migration potential in A549 and H2009 adenocarcinoma cell lines. CyTOF data also identified resistant clones that overexpress TGFβ receptor II, TAZ protein and display hybrid EMT phenotypes.
Conclusions: We are the first to report the interplay between AXL and TGFβ-Hippo signaling axis. TP-0903 study agent has excellent therapeutic promise in NSCLC and we speculate that TP-0903 drug can target epithelial to mesenchymal transitional states in lung cancer cells possibly through the inhibition of the AXL-TGFβ-Hippo signaling axis.
Citation Format: Josephine Amalia Taverna, Chia-Nung Hung, Chun-Lin Lin, Pawel Osmulski, Meizhen Chen, Chiou-Miin Wang, Nicholas L. L. Lucio, Nameer Kirma, Chih-Wei Chou, Maria E. Gaczynska, Alia Nazarullah, Mark Wade, Lars Mouritsen, Tim Huang. AXL inhibitor TP-0903 attenuates AXL-TGFbeta Hippo signaling axis in lung adenocarcinoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2193.
Abstract 22: Design and synthesis of substituted tetrahydroisoquinoline derivatives as anti-angiogenic and anti-breast cancer agents
Authors: Redda Kinfe Ken , Gangapuram Madhavi , Eyuinni Suresh 
Published: August 2019

Abstract
Angiogenesis is the formation of new blood vessels and plays a critical role in the development, invasion and metastasis of breast cancer pathogenesis. Vascular Endothelial Growth Factor (VEGF) is one of the important angiogenic factors secreted by tumor cells. Overexpression of VEGF significantly increases tumor growth and angiogenesis in a murine model of breast cancer. Inhibition of VGEF pathway by limiting new blood vessels formation has proven to be a valuable approach in reducing tumor growth. During the last two decades, diversified small molecules were evaluated and developed as anti-angiogenic agents. Trabectedin (ET-743; Yondelis) is a tetrahydroisoquinoline alkaloid derived from a Caribbean tunicate Ecteinascidia turbinata and has a potent in vitro and in vivo antitumor activity, including prostate and breast cancers. Trabectedin significantly inhibited the mRNA and protein expression levels of VEGF in all breast cancer cells tested. Previously, we developed a series of tetrahydroisoquinoline (THIQs) derivatives and their in-vitro anti-proliferative activities were tested against MCF-7, MDA-MB-231 human breast cancer cell lines and Ishikawa human endometrial adenocarcinoma cell lines. These THIQ derivatives have the same core structure as that of Trabectedin alkaloid and are considered potential anti-angiogenic agents. Novel substituted THIQ derivatives were designed and synthesized in our laboratory. N-amination of substituted isoquinolines by the aminating agent, O-mesytelenesulfonylhydroxylamine and treated with acyl chlorides led to the formation of ylides. The ylides were reduced using sodium borohydride to yield the desired substituted tetrahydroisoquinolines in moderate to good yields. The synthesized compounds were characterized using NMR and elemental analysis. Of the synthesized compounds, fifteen compounds were screened for anti-angiogenesis activity in the Eli Lilly’s Open Innovation Drug Discovery Program (OIDD). Five compounds exhibited greater than 50% inhibition of the tube area at 10µM in the primary single point assay. These five compounds were further screened at different concentrations to obtain dose response curves and IC50 Values. Compound 4-ethyl-N-(3,4-dihydroisoquinolin-2(1H)-yl) benzamide showed an IC50 Value of 1.72 µM.
This research was supported by the National Center for Research Resources and the National Institute of Minority Health and Health Disparities of the National Institutes of Health through Grant Number 8 G12MD007582-28.
Citation Format: Kinfe Ken Redda, Madhavi Gangapuram, Suresh Eyuinni. Design and synthesis of substituted tetrahydroisoquinoline derivatives as anti-angiogenic and anti-breast cancer agents [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 22.
Abstract 2207: The dual role of BI 2536, a small-molecule inhibitor that targets PLK1, in induction of apoptosis and attenuation of autophagy in neuroblastoma cells
Authors: Li Zhiheng 
Published: August 2019

Abstract
Neuroblastoma(NB) is the most common extra-cranial solid tumor in childhood with the overall survival less than 40%. Polo-like kinase 1 (PLK1) is a serine/threonine-protein kinase expressed during mitosis and overexpressed in multiple cancers, including neuroblastoma. We found the PLK1 expression related to the outcome of NB patients. BI2536, a small molecule inhibitor against PLK1, significantly reduced cell viability in a panal of NB cell lines, with IC50 less than 100 nM. PLK1 inhibition by BI 2536 treatment induced cell cycle arrest at G2/M phase and cell apoptosis in NB cells. Realtime PCR array revealed the PLK1-regulated genes, such as BIRC7, TNFSF10, LGALS1 and DAD1 et al. Moreover, autophagy activity was investigated in the NB cells treated with BI 2536. BI 2536 treatment in NB cells increased LC3-II puncta formation and LC3-II expression. Formation of autophagosome induced by BI 2536 was observed by TEM. However, BI2536 abrogated the autophagic flux in NB cells by reducing SQSTM1/p62 expression and AMPKαT172 phosphorylation. These results provide new clues for the molecular mechanism of cell death induced by BI 2536 and suggest that BI 2536 may act as new candidate drug for neuroblastoma.
Citation Format: Zhiheng Li. The dual role of BI 2536, a small-molecule inhibitor that targets PLK1, in induction of apoptosis and attenuation of autophagy in neuroblastoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2207.
Abstract 2214: Development and validation of PredicineATLAS, a 600-gene liquid biopsy panel for simultaneous detection of TMB and genomic alterations in Immuno-Oncology
Authors: Zhao Zhixin , Chang Amy , Cheung Tak , Xie Feng , Montesinos Carlos , Wang Amy Xiaohong , Zhou Kemin , Jia Shidong , Yu JIanjun , Du Pan 
Published: August 2019

Abstract
Introduction: Tumor mutation burden (TMB) is a measurement of the number of somatic mutations carried by tumor cells. Microsatellite instability (MSI) reflects the genetic hypermutability. Both TMB and MSI have been shown to be effective genomic biomarkers for predicting response to anti PD-1/PD-L1 immunotherapy. Whole exome sequencing (WES) is a traditional means to estimate TMB and MSI from tumor tissue, however, it is challenging to translate it into blood or urine due to its high expense and low tumor cell percentage. In the present study, we developed a 600-gene liquid biopsy TMB panel, PredicineATLAS, to measure genomic alterations and estimate TMB and MSI using human body fluids.
Methods: The PredicineATLAS panel contains 600 genes and covers 2.4Mb genomic regions (1.4 Mb of exonic coding regions). It was specifically designed for immuno-oncology, covering comprehensive immune- and cancer-related genes, biomarkers and MSI sites that are associated with anti PD-1/L1 therapy response. Reference materials with known genomic alterations and tumor cell lines were titrated with different concentrations to mimic different tumor contents for TMB and MSI measurement. Matched tissue and blood were used in validation study.
Results: PredicineATLAS panel-based TMB strongly correlated with the WES-TMB using TCGA dataset. Analytical validation showed that PredicineATLAS NGS assay is highly robust and sensitive for detection of genomic alterations. Using matched tissue and blood sample set, blood-based PredicineATLAS panel-TMB and MSI correlate with its counterpart derived from tissue WES.
Conclusions: PredicineATLAS liquid biopsy assay was developed and validated using reference materials and clinical samples. It detects genomic alterations and accurately estimate TMB and MSI, providing valuable genomic information to guide immuno-oncology therapy and clinical drug development.
Citation Format: Zhixin Zhao, Amy Chang, Tak Cheung, Feng Xie, Carlos Montesinos, Amy Xiaohong Wang, Kemin Zhou, Shidong Jia, JIanjun Yu, Pan Du. Development and validation of PredicineATLAS, a 600-gene liquid biopsy panel for simultaneous detection of TMB and genomic alterations in Immuno-Oncology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2214.