bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3053: Development of novel cell-based bioassays for the development of biologics targeting LAG-3 and PD-1xLAG-3
Authors: Grailer Jamison , Garvin Denise , Hartnett Jim , Fan Frank , Cong Mei , Cheng Zhi-jie Jey 
Published: August 2019

Abstract
Programmed cell death protein 1 (PD-1) and Lymphocyte-activation gene 3 (LAG-3) are immune checkpoint receptors that bind to PD-L1/PD-L2 or MHCII, respectively, on antigen presenting cells or tumor cells. PD-1 or LAG-3 activation results in the inhibition of T cell receptor (TCR) signaling and reduced T cell activation. However, co-blockade of PD-1 and LAG-3 leads to a synergistic enhancement in T cell responses, suggesting functionally divergent inhibitory signaling mechanisms between the two receptors. Preclinical studies suggest that blocking LAG-3 re-activates the immune system to kill cancer cells, and co-blockade of both PD-1 and LAG-3 lead to a synergistic increase in tumor control and survival in preclinical models. Current methods for measuring the potency of LAG-3 and PD-1xLAG-3 blocking antibodies measure cytokine production from human PBMCs upon Staphylococcal Enterotoxin B (SEB) superantigen stimulation. These protocols are tedious, highly variable, and require the use of highly toxic superantigen. Herein, we describe the development of reporter-based bioassays for the development of LAG-3 and PD-1xLAG-3 combination-targeted biologics. Engineered T effector cell lines expressing LAG-3 alone or in combination with PD-1 are activated in an antigen-dependent manner by APC cell lines. The assays are specific for LAG-3 or PD-1xLAG-3-targeted biologics and are robust as demonstrated by Design of Experiments analyses. In addition, the assays can be used to measure the relative potencies of test samples compared with reference sample in a detection range of 50% to 200% and show appropriate precision, accuracy and linearity. The assays can detect potency changes for heat-stressed samples, and are tolerant to human serum, demonstrating the potential to be further developed into neutralizing antibody (NAb) assays. Therefore, these bioassays can serve as valuable tools for the development of biologics targeting LAG-3 and PD-1xLAG-3.
Citation Format: Jamison Grailer, Denise Garvin, Jim Hartnett, Frank Fan, Mei Cong, Zhi-jie Jey Cheng. Development of novel cell-based bioassays for the development of biologics targeting LAG-3 and PD-1xLAG-3 [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 3053.
Abstract 3055: Genome-wide CRISPR/Cas9 screen for the identification of novel YAP1/TAZ modulators
Authors: Naujoks Jan , Potze Lisette , Anurin Anna , Kuehnlenz Julia , Lesche Ralf , Kamburov Atanas , Nevedomskaya Ekaterina , Steffen Andreas , Lange Martin , Nicke Barbara 
Published: August 2019

Abstract
Aberrant activation of the Hippo pathway effectors YAP1/TAZ promotes cell proliferation and tumorigenesis. To identify novel regulators of YAP1/TAZ as a possible means to treat cancer, we established a novel, FACS-based screening system monitoring YAP1/TAZ activity in MDA-MB-231 breast cancer cells. Using these cells, we performed a pooled genome-wide CRISPR/Cas9 knockout screen. We identified approximately 50 genes potentially activating YAP1/TAZ with functions in the Actin Cytoskeleton signaling, p53 signaling, cell polarity or ER stress, amongst others. Moreover, we identified about 30 potential targets which when knocked out induce activity of YAP1/TAZ. The list of hits included genes known to affect the YAP1/TAZ activity such as AJUBA, LATS2 and TEAD, demonstrating the validity of the screen. Functional validation of the novel potential YAP1/TAZ modulators will aid to the further understanding of YAP1/TAZ biology and may open the door to new therapeutic avenues for targeting YAP1/TAZ in cancer.
Citation Format: Jan Naujoks, Lisette Potze, Anna Anurin, Julia Kuehnlenz, Ralf Lesche, Atanas Kamburov, Ekaterina Nevedomskaya, Andreas Steffen, Martin Lange, Barbara Nicke. Genome-wide CRISPR/Cas9 screen for the identification of novel YAP1/TAZ modulators [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 3055.
Abstract 3059: Identification and validation of an Aspergillus nidulanssecondary metabolite derivative as an inhibitor of the Musashi1-RNA interaction
Authors: Lan Lan , Liu Jiajun , Amith Amber , Appelman Carl , Wu Xiaoqing , Li Ke , Roy Anuradha , Gowthaman Ragul , Karanicolas John , Somoza Amber , Wang Clay , Oakley Berl , Guzman Roberto De , Neufeld Kristi , Xu Liang 
Published: August 2019

Abstract
RNA-binding protein Musashi-1 (MSI1) is a key regulator of several stem cell populations. It is involved in tumor proliferation and maintenance, and it regulates target mRNAs at the translational level. The known mRNA targets of MSI1 include Numb, APC and P21WAF-1, key regulators of Notch, Wnt signaling and cell cycle progression, respectively. In this study, we aim to identify small molecule inhibitors of MSI1-mRNA interaction thus blocking the growth of cancer cells with high levels of MSI1. Using a fluorescence polarization (FP) assay, we screened small molecules from several chemical libraries for those that disrupt the binding of MSI1 to its consensus RNA binding site. Two clusters of hit compounds have been identified, one of which is composed of derivatives of secondary metabolites from Aspergillus nidulans. The top hit, Aza-9, from this cluster was further validated by surface plasmon resonance and nuclear magnetic resonance which revealed that Aza-9 binds to MSI1 directly, in the RNA binding pocket. Next, we tested whether Aza-9 has anti-cancer potential in cells and whether such activities work through MSI1. Due to poor cellular uptake, we used PEGylated liposomes to facilitate Aza-9 entry into the cells. Aza-9-lipo inhibits colon cancer cell proliferation, induces apoptosis and autophagy, and down-regulates Notch/Wnt signaling in colon cancer cell lines. In conclusion, we identified a series of potential lead compounds for inhibiting MSI1 function while establishing a framework for identifying small molecule inhibitors of RNA binding proteins using FP-based screening methodology.
Citation Format: Lan Lan, Jiajun Liu, Amber Amith, Carl Appelman, Xiaoqing Wu, Ke Li, Anuradha Roy, Ragul Gowthaman, John Karanicolas, Amber Somoza, Clay Wang, Berl Oakley, Roberto De Guzman, Kristi Neufeld, Liang Xu. Identification and validation of an Aspergillus nidulanssecondary metabolite derivative as an inhibitor of the Musashi1-RNA interaction [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 3059.
Abstract 3066: The expression of carcinoembryonic antigen-related cell adhesion molecule 6 and 8 in breast cancer
Authors: Iwabuchi Erina , Miki Yasuhiro , Takagi Kiyoshi , Onodera Yoshiaki , Shibahara Yukiko , Ishida Takanori , Suzuki Takashi , Sasano Hironobu 
Published: August 2019

Abstract
Carcinoembryonic antigen-related cell adhesion molecule (CEACAM) 6 is a protein reported to be involved in tumor cell proliferation in breast cancer. The presence of intratumoral CEACAM8 positive neutrophils has been reported as an independent prognostic factor in both renal cell and hepatocellular cancer. In addition, CEACAM6 and 8 form a heterodimer; however, the possible interactions of these two proteins remain unclear in breast cancer. Therefore, in this study we first immunolocalized CEACAM6 and 8 in surgical pathology specimens of breast cancer. We examined 109 breast cancer and 20 metastatic breast cancer patients in this study. We then established MCF-7 cells (CEACAM6 positive) stably transfected with CEACAM8 and analyzed the interaction between CEACAM6 and CEACAM8 using proximity ligation assay (PLA) and further evaluated cell proliferation, invasion, and adhesion. Results of immunohistochemical analysis revealed that the number of CEACAM6 positive breast cancer cells was significantly higher in patients with low histological grade and stage also in CEACAM8 positive cells. The number of CEACAM6 and 8 double positive breast cancer cells was significantly higher in patients with low stage. In addition, CEACAM6 and 8 interaction was detected using the PLA through high expression of both in MCF-7 cells. Furthermore, their co-expression promoted cell adhesion of MCF-7 cells to the monolayer of endothelial cells but suppressed both cell invasion and proliferation. In metastatic breast cancer, CEACAM6 status was significantly higher in bone metastases than in lung metastases. To the best of our knowledge, this is the first study demonstrating the expressions of CEACAM6 and 8 in breast cancer. Results of our present study also indicated that both CEACAM6 and CEACAM8 could be involved in less aggressive subtypes of breast cancer possibly trough inhibition of vascular invasion and proliferation. In addition, results indicated that CEACAM6 was involved in breast cancer metastases to the bone but further studies are required to clarify the clinical significance of their interactions in breast cancer patients.
Citation Format: Erina Iwabuchi, Yasuhiro Miki, Kiyoshi Takagi, Yoshiaki Onodera, Yukiko Shibahara, Takanori Ishida, Takashi Suzuki, Hironobu Sasano. The expression of carcinoembryonic antigen-related cell adhesion molecule 6 and 8 in breast cancer [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 3066.
Abstract 3072: GFH018, a novel TGF-βRI inhibitor, for the treatment of advanced solid tumors
Authors: Hu Gang , Zhao Rong , Zhou Fusheng , Lan Jiong , Zheng Biao , LU Qiang , Lu Jianyu , Wu Lifang , Sun Fei , Hu Lihong , Han Shuhua , Tong Haijun , Xia Yuanfeng , Li Wei , Ma Liping , Chen Ying , Ding Charles , Chan Chichung , Chen Shuhui 
Published: August 2019

Abstract
Background: The transforming growth factor-β (TGF-β) signaling pathway has been shown to play an important role in various cellular processes including growth inhibition, cell migration, tumor invasion, epithelial-mesenchymal transition, and immune-suppression. TGF-βs are often overexpressed in various types of cancers, and the anti-TGF-β therapy has been pursued intensively for cancer treatment.
Results: GFH018 is a potent, selective inhibitor of TGF-βRI with a kinase inhibition activity IC50 of 70.5 nM. Our studies demonstrated that GFH018 could inhibit TGF-β-induced SMAD phosphorylation and migration of tumor cells. GFH018 is a potent immune regulator in that it could inhibit in vitro Treg cell induction and reverse M2 phenotype to M1, leading to increased pro-inflammatory cytokine production. In addition, GFH018 exhibited a significant inhibitory effect on in vitro angiogenesis formation assay. Importantly, GFH018 has shown promising anti-tumor efficacy in multiple in vivo mouse syngeneic models as a monotherapy as well as in combination with an anti-PD-(L)1 antibody. Following oral administration, GFH018 demonstrated fast absorption, and high oral bioavailability in preclinical animal species including mouse, rat, and dog. Results from the toxicology studies in rat and dog have also supported further development of GFH018.
Conclusion: GFH018 has the potential to be developed as a monotherapy or combinational therapy with checkpoint inhibitors for advanced solid tumor patients. The first-in-human clinical trial is scheduled in 2019.
Citation Format: Gang Hu, Rong Zhao, Fusheng Zhou, Jiong Lan, Biao Zheng, Qiang LU, Jianyu Lu, Lifang Wu, Fei Sun, Lihong Hu, Shuhua Han, Haijun Tong, Yuanfeng Xia, Wei Li, Liping Ma, Ying Chen, Charles Ding, Chichung Chan, Shuhui Chen. GFH018, a novel TGF-βRI inhibitor, for the treatment of advanced solid tumors [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 3072.
Abstract 3081: DZD9008, an oral, wild type selective EGFR inhibitor for the treatment of non-small-cell lung cancer with Exon20 insertion and other activating mutations
Authors: Xu Yan , Zhang Lin , Zhu Lifang , Wang Yingchun , Wang Mei , Yang Zhenfan 
Published: August 2019

Abstract
Background: Several EGFR TKIs have been approved for the treatment of non-small cell lung cancer (NSCLC) with L858R mutation, Exon 19 deletion, and T790M mutations. However, for patients with uncommon mutations, such as EGFR or HER2 Exon20ins, there lacks safe and effective therapy. All Exon20ins inhibitors under clinical development have a negative selectivity profile, inhibiting wild type EGFR more potently than mutant ones. Consequently, severe wild type EGFR-driven toxicities have been observed in patients. DZD9008 is an oral, potent, irreversible, wild type-selective EGFR TKI against EGFR or HER2 Exon20ins and other mutations. The present study shows anti-tumor activity of DZD9008 in tumor cell lines and xenograft models.
Materials and Methods: The enzyme assay was performed by incubating DZD9008 with recombinant enzymes at 2 mM ATP concentrations. The cellular activity of DZD9008, including phosphorylation of EGFR or HER2 and cell proliferation, was evaluated in a panel of cell lines expressing wild type EGFR, mutant EGFR or HER2, using MSD assay and CellTiter-Glo assays. DZD9008 in vivo anti-tumor activity was evaluated in both cell line-derived (CDX) and patient-derived xenograft (PDX) models, carrying EGFR single or double mutations, Exon20ins mutations, and wild type EGFR.
Results: The enzymatic IC50 of DZD9008 in mutant EGFR ranges from 0.4 to 2.1 nM. DZD9008 downregulated pEGFR with IC50 ranging from 1 to 22 nM in a panel of tumor cell lines expressing EGFR L858R, Exon19del, L858R/T790M, various Exon20ins or uncommon mutations. Similar activity against pHER2 was observed in tumor cells with HER2 Exon20ins mutation, with IC50 at 7 nM. In contrast, DZD9008 was less potent in modulating pEGFR in tumor cells expressing wild type EGFR, with IC50 greater than 80 nM. In cell proliferation assays, DZD9008 suppressed cell proliferation with GI50 of 1 to 60 nM in tumor cells carrying EGFR L858R, Exon19del, L858R/T790M, various Exon20ins or uncommon mutations. In CDX and PDX models carrying EGFR Exon19del single mutation, L858R/T790M double mutations, and PDX models harboring G719S/L861Q or Exon20ins, DZD9008 induced dose dependent tumor growth inhibition and regression. Good PK/PD relationship was established across these tumor models.
Conclusion: DZD9008 is a potential EGFR TKI for NSCLC patients with EGFR or HER2 Exon20ins and other activating mutations.
Citation Format: Yan Xu, Lin Zhang, Lifang Zhu, Yingchun Wang, Mei Wang, Zhenfan Yang. DZD9008, an oral, wild type selective EGFR inhibitor for the treatment of non-small-cell lung cancer with Exon20 insertion and other activating mutations [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 3081.
Abstract 3083: A novel hedgehog signaling inhibitor for targeting pancreatic ductal adenocarcinoma
Authors: Khire Atul , Zhong Yi , Rajanala Kalpana , Iacobuzio-Donahue Christine , Resh Marilyn D. 
Published: August 2019

Abstract
Sonic Hedgehog (Shh) signaling is deregulated in pancreatic cancer and can drive tumor onset and progression. Previous attempts to block Shh signaling used Smoothened (Smo) inhibitors, which target canonical Shh signaling. These approaches met with limited success, due to development of resistance, and contributions from non-canonical, Smo-independent signaling pathways that operate within tumor epithelial cells. Here, we show that Hedgehog acyltransferase (Hhat), the enzyme responsible for palmitoylation of Shh, is a novel target for inhibition of Shh signaling in human and mouse pancreatic cancer cells and tumors. TDI-003410, a small molecule inhibitor of Hhat recently developed by our group, reduced proliferation of AsPC1 cells, a human pancreatic cancer cell line, in vitroand in subcutaneous xenografts in mice in vivo. The mechanism is via induction of a G2/M cell cycle arrest leading to apoptosis. We next used cell lines isolated from tumors formed in KPC (K-RasG12D/p53R172H) mice, a genetically engineered mouse model of pancreatic cancer.TDI-003410 inhibited the growth of KPC cell linesand also reduced Gli1 levelsin vitro. Neither LDE225, a Smo inhibitor, nor SAG, a Smo activator, had any effect on KPC cell proliferation or Gli1 levels in vitro. In orthotopic xenografts using KPC cells implanted into the pancreas, TDI-3410 treatment significantly reducedin vivotumor burden. No significant changes in CD31 or Trichrome (collagen) stainingwith TDI-3410 were detected in KPC orthotopic tumor tissue. However, Gli-1 levels in tumor tissue from TDI-3410 treated mice were significantly reduced compared to the vehicle controls. These findings provide proof of concept for the potential therapeutic efficacy of Hhat inhibition in pancreatic cancer. The concept of using an Hhat inhibitor is unique: Hhat is a novel target that hits upstream, directly at hedgehog ligands. This approach can inhibit both canonical and non-canonical (Smo-independent) as well as autocrine and paracrine Shh signaling.
Citation Format: Atul Khire, Yi Zhong, Kalpana Rajanala, Christine Iacobuzio-Donahue, Marilyn D. Resh. A novel hedgehog signaling inhibitor for targeting pancreatic ductal adenocarcinoma [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 3083.
Abstract 3088: Farnesyl/geranylgeranyl transferase dual inhibitor thwarts mutant KRas-driven patient-derived pancreatic tumors
Authors: Kazi Aslamuzzaman , Xiang Shengyan , Yang Hua , Chen Liwei , Kennedy Perry , Ayaz Muhammad , Fletcher Steven , Cummings Christopher , Lawrence Harshani , Beato Francisca , Yang Ya'an , Kim Michael P. , Delitto Andrea , Underwood Patrick , Fleming Jason B. , Trevino Jose , Hamilton Andrew D. , Sebti Said M. 
Published: August 2019

Abstract
Although mutant KRas is a significant driver of pancreatic oncogenesis and resistance to therapy, there are no KRas inhibitors available for these patients. Farnesyltransferase inhibitors (FTIs) were developed as potential anticancer drugs because KRas requires farnesylation for its membrane localization and cancer-causing activity. However, KRas becomes geranylgeranylated and active when cancer cells are treated with FTIs. In this study, we designed a Ras C-terminal mimetic dual Farnesyl/geranylgeranyltransferase-1 (GGT-1) inhibitor, FGTI-2734, to overcome the geranylgeranylation-dependent resistance to FTIs. Immunofluorescence, cellular fractionation, and gel shift assays showed that FGTI-2734, but not the selective FTI-2148 and GGTI-2418, inhibited membrane localization of KRas in mt KRas pancreatic, lung, and colon human cancer cells. FGTI-2734 inhibited the growth in mice of mt KRas-dependent but not -independent human tumors, indicating its selectivity for mt KRas-driven cancers. Importantly, FGTI-2734 inhibited the in vivo growth of xenografts derived from four pancreatic cancer patients with mt KRas (two G12D, and two G12V) tumors. In addition, FGTI-2734 was highly effective at inhibiting, in three-dimensional co-cultures with chemotherapy resistance-promoting pancreatic stellate cells, the viability of primary and metastatic mutant KRas (G12D, G13D, and G12V) tumor cells derived from 8 pancreatic cancer patients. Finally, FGTI-2734 suppressed oncogenic pathways mediated by Akt, mTOR, and cMyc while upregulating p53 and inducing apoptosis in patient-derived xenografts in vivo. Thus, the development of this novel dual FT and GGT-1 inhibitor overcomes a major hurdle in KRas resistance, thwarting the growth of patient-derived mutant KRas-driven xenografts from pancreatic cancer patients, and as such it warrants further advanced preclinical and clinical studies.
Citation Format: Aslamuzzaman Kazi, Shengyan Xiang, Hua Yang, Liwei Chen, Perry Kennedy, Muhammad Ayaz, Steven Fletcher, Christopher Cummings, Harshani Lawrence, Francisca Beato, Ya'an Yang, Michael P. Kim, Andrea Delitto, Patrick Underwood, Jason B. Fleming, Jose Trevino, Andrew D. Hamilton, Said M. Sebti. Farnesyl/geranylgeranyl transferase dual inhibitor thwarts mutant KRas-driven patient-derived pancreatic tumors [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 3088.
Abstract 3093: Prognostic value of novel immune basal subtypes in muscle invasive bladder cancer
Authors: Choi Woonyoung , Sundi Debasish , Lee I-ling , Siefker-Radtke Arlene , Dinney Colin , Czerniak Bogdan , Kim Seungchan , McConkey David 
Published: August 2019

Abstract
Using gene expression profiling we previously reported the existence of 3 molecular subtypes (basal, p53-like and luminal) characterized by distinct gene expression patterns and clinical outcomes in muscle-invasive bladder cancer (MIBC). Among the subtypes, basal tumors were associated with advanced stage at presentation and shorter survival in the absence of neoadjuvant chemotherapy (NAC). Our recent analysis identified two novel immune subtypes within the basal tumors (basal immune signature enriched - “BIE” and basal immune signature suppressed - “BIS”) using immune infiltration markers that are known to have prognostic value in multiple solid tumors. The analysis revealed that BIE had significantly improved survival outcomes while BIS was associated with the worst survival outcomes among all subtypes in TCGA MIBC data (n=408). Survival outcomes in BIE tumors were validated in an independent cohort (GSE48075, p &lt;0.05). The characterization of the immune landscape in our novel subtypes using TCGA’s recent pan-cancer immunogenomics data showed that the BIE subtype was significantly enriched with ‘INF -γ dominant’ immune subtype out of the 6 TCGA immune subtypes (wound healing, IFN-γ dominant, inflammatory, lymphocyte depleted, immunologically quiet and TGF-β dominant, p &lt; 0.05). Since immune activation has been associated with response to chemotherapy in other tumor types, we explored the potential significance of the novel immune subtypes using public data consisting of patients who received NAC followed by radical cystectomy (GSE52219 and GSE69795). In these cohorts, chemotherapy was active as measured by pathological downstaging rates (≤ pT1 at cystectomy). Based on pathological downstaging, BIE was the most responsive to chemotherapy while BIS and p53-like tumors were resistant to chemotherapy (p &lt; 0.05). We also used the recently developed CIBERSORT tool to estimate stromal cell infiltration in the NAC cohort. The results predicted that the BIE subtype was highly enriched with CD8+ T cells and NK cells (p &lt;0.001). Taken together, our results reveal the existence of novel immune subtypes within basal tumors with distinct molecular features and survival outcomes in MIBC. The clinical implications of these subtypes will be further evaluated.
Citation Format: Woonyoung Choi, Debasish Sundi, I-ling Lee, Arlene Siefker-Radtke, Colin Dinney, Bogdan Czerniak, Seungchan Kim, David McConkey. Prognostic value of novel immune basal subtypes in muscle invasive bladder cancer [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 3093.