bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2964: Saffron has enhanced anti-proliferative effect against colorectal cancer cells with deficient DNA mismatch repair
Authors: Amin Amr , Soleimani Akbar , Murali Chandraprabha , Beresova Lucie , Shoraka Hamid , Brim Hassan , Ashktorab Hassan 
Published: September 2020

Abstract
Background: Colorectal cancer (CRC) is one of the most common cancers worldwide. A subtype of CRC is associated with defective mismatch repair (dMMR) genes. Saffron has many potentially protective roles against colon malignancy, however, these roles in the context of of dMMR have not been explored. In this study, we aimed to study the effects of saffron in CRC cell lines with dMMR.
Methods: Saffron crude extracts and specific compounds (crocin, crocetin and safranal) were used in colorectal cancer cell lines HCT116, HCT116+3 (inserted MLH1), HCT116+5 (inserted MH3) and HCT116+3+5 (Inserted MLH1 and MSH3). CDC25b, p-H.H2AX, TPDP1, and GAPDH were analyzed by Western blot. Cell cycle was analyzed by FACS. Proliferation and cytotoxicity were analyzed by MTT. Wound and migration assays were also performed.
Results: Saffron crudes restricted the proliferation in colon cells with deficient MMR (HCT116) compared to proficient MMR. Wound healing assay indicates that deficient MMR cells are doing better than proficient MMR cells. The expression of CDC25b and TDP1 were upregulated in proficient MMR cell compared to deficient MMR cell, while H2AX was significantly upregulated in both cells type particularly at &gt;10 mg/ml saffron in a concentration-dependent manner (after normalization with GADPH as control).The reduction in cellular proliferation and motility might be due to cell cycle arrest at G2-phase rather than to apoptosis. The major active saffron compounds, safranal and crocin reproduced total saffron crudes effects but with lesser outcome.
Conclusion: Saffron's anti-proliferative and healing effects is significant in cells with deficient MMR MLH1 and MSH3 genes. This effect may have a therapeutic implications and benefits for the CRC MSI tumors that are generally not recommended for 5FU-based treatment.
Citation Format: Amr Amin, Akbar Soleimani, Chandraprabha Murali, Lucie Beresova, Hamid Shoraka, Hassan Brim, Hassan Ashktorab. Saffron has enhanced anti-proliferative effect against colorectal cancer cells with deficient DNA mismatch repair [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 2964.
Abstract 2995: Dissecting renal cell carcinoma vulnerabilities: Using CRISPR/Cas9 to study resistance to glutaminase inhibition
Authors: Raczka Aleksandra M. , Reynolds Paul A. 
Published: September 2020

Abstract
The aim of this study was to identify genes, which when knock out confer resistance to a novel glutaminase inhibitor in Renal Cell Carcinoma (RCC). RCC represents around 3% of all malignancies worldwide, with its most common type, clear cell RCC (ccRCC), making up around 70-80% of all RCC. The most frequent genomic alteration in ccRCC involves loss of VHL gene in &gt;90% of ccRCC cases. Large-scale cancer genomics sequencing studies have identified several driver genes beyond VHL, particularly PBRM1 (40%), SETD2 (15%), and BAP1 (10%). RCC is characterised by significant tumour heterogeneity and inherent (in 25-30% cases) or acquired resistance to available chemotherapeutics. Glutamine addiction is a potential new therapeutic target for RCC as it has been recently shown not only to rely on glutamine for energy generation and maintenance of redox homeostasis, but also de novo pyrimidine synthesis. CB-839 is a small, orally administered reversible inhibitor of human kidney-type glutaminase (GLS). CB-839 is currently in early phase clinical trials and shows promising results. Given the significant incidence of resistance to previously approved therapies, we have applied a genome-scale CRISPR/Cas9 approach in a cell culture model of ccRCC (786-0 cell line) to identify candidate genes, which when knocked down confer resistance to CB-839. Next generation sequencing data analysis of drug-selected sgRNA library representation from two timepoints was performed using the MAGeCKFlute bioinformatics workflow. We conducted the screen in two biological replicates and verified successful genome-scale sgRNA library coverage at baseline in both cases. As a result, we identified candidate genes and pathways that are implicated in tumour metabolism, autophagy and metastasis. Additionally, this analysis identified KD genes, which in combination with CB-839 are synthetic lethal. We have prioritized the top 12 candidate genes, which we are validating in four cell lines (786-0, A498, A704 and Caki-1). After establishing CRISPR/Cas9 single gene knockdown (KD) cell lines, KD is confirmed using RT-qPCR and Western Blot, followed by functional analysis. KD cells were treated with increasing concentrations of the drug in parallel with a cell line containing a scrambled sgRNA to detect growth changes between the two. After in vitro validation, we performed ex vivo validation of the candidate hits. Our data show that the genome-scale CRISPR/Cas9 approach is effective in identifying candidates modulating drug resistance to CB-839.
Citation Format: Aleksandra M. Raczka, Paul A. Reynolds. Dissecting renal cell carcinoma vulnerabilities: Using CRISPR/Cas9 to study resistance to glutaminase inhibition [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 2995.
Abstract 3102: Analysis of plasma and serum as a source of circulating tumor DNA for hotspot mutations detection in liquid biopsy
Authors: Pittella-Silva Fabio , Chin Yoon Ming , Chan Hui Ting , Nagayama Satoshi , Miyauchi Eisaku , Low Siew-Kee , Nakamura Yusuke 
Published: September 2020

Abstract
Blood-based analysis of ctDNA is a promising tool for cancer screening, drug selection, detection of minimum residual disease, monitoring release/recurrence and response to treatment. Although plasma is widely used to obtain ctDNA, biorepositories worldwide possess highly-valuable serum samples collection and comparative studies on the use of serum vs plasma as ctDNA source are still essential. Here we analyzed cell free DNA (cfDNA) from matched plasma and serum samples from healthy donors, colorectal and lung cancer patients to evaluate the differences of serum and plasma on mutation detection efficacy and reproducibility by targeted NGS. We initially addressed the difference of both analytes on the quality of cfDNA recovery and on sequencing performance. We then compared mutation detection using matched samples from healthy individuals spiked-in with standard oligonucleotides including common mutations. The amount of cfDNA was more abundant in serum than plasma and the presence of longer DNA fragments was significantly higher in serum samples. Spiked-in mutated nucleotides were detected in both plasma and serum samples, but allele frequencies (AF) were approximately half in serum samples compared with plasma samples, suggesting ctDNA from serum is more diluted in DNAs of noncancerous origins. We further confirmed this difference in detection efficiency using matched plasma and serum samples from cancer patients and found that mutations with low AF are more frequently missed in serum samples. In addition, concordance with tissue-biopsy mutations found at diagnosis was better in plasma samples. Finally, we examined the use of fragment size selection as an alternative tool to improve mutation detection in serum samples. Our findings clearly demonstrated that the use of serum in retrospective studies should consider the limitations for detecting low AF mutations and indicate the advantages of plasma samples for prospective clinical applications of liquid biopsy.
Citation Format: Fabio Pittella-Silva, Yoon Ming Chin, Hui Ting Chan, Satoshi Nagayama, Eisaku Miyauchi, Siew-Kee Low, Yusuke Nakamura. Analysis of plasma and serum as a source of circulating tumor DNA for hotspot mutations detection in liquid biopsy [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 3102.
Abstract 3270: Dual blockade of PD-L1 and LAG-3 using a bispecific antibody improves anti-tumor immunity
Authors: Ni Haiqing , Qiu Yajing , Jing Hua , Zhang Pan , Chen Bingliang , Wu Zhihai , Zhou Shuaixiang , Yu Michael , Liu Junjian 
Published: September 2020

Abstract
Background: Anti-PD-1/PD-L1 treatments are effective in a fraction of patients with advanced malignancies. However, the majority of patients still do not respond. Resistance to cancer immunotherapy can be mediated by additional immune checkpoints. Lymphocyte-activation gene 3 (LAG-3) is a member of the Ig superfamily and participates in immune suppression. Combination of anti-PD-1 and anti-LAG-3 showed synergistic anti-tumor effect in preclinical mouse models and early clinical trials. Here, we develop IBI323, an anti-PD-L1/LAG-3 bispecific antibody, which preserves the dual pathway blockade function while processes new biological functions.
Material & Methods: The binding affinity of IBI323 to antigens were measured by Fortebio and confirmed at cellular level using PD-L1 overexpressing cells and primary T cell. IBI323 was also assessed by ELISA for PD-1 and LAG-3 pathway blocking function. IBI323 mediated PD-L1+ and LAG-3+ cells bridging was evaluated by flow cytometry. Anti-tumor activity of IBI323 was evaluated in human PD-L1 knock-in mice bearing PD-L1 overexpressing MC38 tumors. Besides tumor growth, the frequencies and quality of the tumor-specific T cells were assessed by flow cytometry. In addition, florescence labeled antibodies was used for detecting the biodistribution of IBI323 in vivo.
Results: IBI323 bound to PD-L1 and LAG-3 with high affinity (KD &lt;1 nm). At cellular level, IBI323 bound to PD-L1 overexpressing cells and LAG-3+ T cells in a similar manner to parental antibodies. IBI323 was able to block PD-1/PD-L1 and LAG3/MHCII interactions simultaneously. Moreover, bridging between PD-L1+ cells and LAG-3+ cells was observed in the presence of IBI323 in vitro. Correspondingly, IBI323 was mainly distributed at the location of PD-L1+ tumor in vivo. In mice harboring established PD-L1+ MC38 tumor, IBI323 treatment was superior to combination of individual anti-PD-L1 and LAG-3 antibodies. The better activity of IBI323 treatment was reflected by an enhanced number of tumor-specific CD8+ cytotoxic T cells.
Conclusions: We developed a novel bispecific antibody against PD-L1 and LAG-3, which resulted in dramatic tumor growth inhibition. The anti-tumor efficacy of IBI323 was better than either monotherapy or anti-PD-L1 and LAG-3 combination treatment. Based on the encouraging preclinical results, IBI323 could soon be evaluated in clinical trials.
Citation Format: Haiqing Ni, Yajing Qiu, Hua Jing, Pan Zhang, Bingliang Chen, Zhihai Wu, Shuaixiang Zhou, Michael Yu, Junjian Liu. Dual blockade of PD-L1 and LAG-3 using a bispecific antibody improves anti-tumor immunity [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 3270.
Abstract 3361: Discovery and preclinical characterizations of a humanized anti-claudin 18.2 antibody SPX-101
Authors: Zhu Guidong , Ma Jichun , Ye Jingdong , Qin Jingdong , Cai Yi 
Published: September 2020

Abstract
Claudin 18.2 (CLDN 18.2) is a major component of the tight junctions and is highly expressed in several cancers including gastric (80%), esophageal (80%), pancreatic (60%) and lung cancers, but only expressed at very low levels in healthy tissues. The chimeric IgG1 CLDN 18.2 antibody zolbetuximab significantly improved PFS and OS in gastric cancer patients in a CLDN 18.2-dependent manner on the background of EOX. This research aimed to produce humanized anti-claudin18.2 antibodies with enhanced activities and improved pharmaceutical characteristics compared to known CLDN 18.2 antibodies. We used several immunization methods to produce mouse anti-CLDN18.2 antibodies with high affinity and excellent selectivity against CLDN 18.1. Humanization of a selected clone led to a humanized anti-CLDN18.2 antibody (h533o) with significantly improved binding affinity and cellular activity in ADCC and CDC assays compared to zolbetuximab. The binding affinity of h533o was relatively insensitive to elevated temperatures or acidity, suggesting the robustness of target engaging ability in tumor microenvironment. Maturation and selection by the off-rate approach, using phage libraries that systematically modified the CDR regions, identified multiple antibodies with comparable affinity and selectivity. Mutational scan of the Fc region in h533o identified SPX-101 with reduced ADA effect in mice while retaining other favorable properties. SPX-101 suppressed tumor growth in several xenograft mouse models including NUGC4 and HEK293 cells that stably expressed CLDN18.2. In summary, SPX-101 represents a humanized anti-CLDN18.2 antibody with high affinity and specificity, high effector cell-mediated cytotoxicity, low immunogenicity, and in vivo tumor control efficacy in xenograft rodent models.
Citation Format: Guidong Zhu, Jichun Ma, Jingdong Ye, Jingdong Qin, Yi Cai. Discovery and preclinical characterizations of a humanized anti-claudin 18.2 antibody SPX-101 [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 3361.
Abstract 3366: In vivo efficacy of a PD-L1 targeted, antigen seeding engineered toxin body
Authors: Ramos Hilario J. , Brieschke Brigitte , LeMar Sara , Dekker Joseph D. , Iberg Aimee , Robinson Garrett L. , Sarkar Asis , Anand Banmeet , Singh Melissa M. , Zhao Jay , Higgins Jack P. , Willert Erin K. 
Published: September 2020

Abstract
Engineered toxin bodies (ETBs) comprised of a proprietarily engineered Shiga-like Toxin A subunit (SLTA) genetically fused to antibody-like binding domains work through novel mechanisms of action and can force internalization, self-route through intracellular compartments to the cytosol, and induce potent cell-kill via the enzymatic and permanent inactivation of ribosomes. Our PD-L1 targeted ETB, MT-6402, includes antigen seeding technology (AST), and additionally delivers a viral antigen for presentation in complex with MHC-I to resident viral-specific cytotoxic T lymphocytes (CTLs). The fusion protein provides a powerful, dual mechanism of action to specifically target and destroy PD-L1 positive tumor and inhibitory immune cells. In vitro testing of human tumor cell lines has demonstrated that cell surface PD-L1 expression is required for activity of the PD-L1 targeted ETBs. MT-6402 demonstrated effective depletion of PD-L1 expressing cells across a panel of immortalized cancer cell lines from multiple origins, including lung, skin, breast, and ovary. In a co-culture assay of PD-L1 target cells and antigen specific T-cells, PD-L1 targeted MT-6402 delivers a viral antigen to the target cells and can activate cytokine secretion and T-cell mediated killing. Additionally, MT-6402 treatment of human peripheral blood mononuclear cells (PBMCs) leads to selective depletion of PD-L1 positive cells (IFN-γ stimulated monocytes) in the absence of depletion of PD-L1 low/negative lymphocytes. Murine models, including patient derived xenografts, have demonstrated that the PD-L1 targeted ETBs are efficacious in vivo. Additional in vivo tumor models to further describe the effect of MT-6402 are being explored. Exploratory studies in a primate model have shown that MT-6402 can be tolerated at levels shown to induce T cell activation (type 1 cytokines). MT-6402 delivers a powerful and unique dual mechanism of action. The combination of a PD-L1 specific direct cell kill and redirection of a robust effector T cell response to the tumor has potential benefit in solid tumor indications, including in the relapsed setting, when disease has progressed after checkpoint and/or other therapies.
Citation Format: Hilario J. Ramos, Brigitte Brieschke, Sara LeMar, Joseph D. Dekker, Aimee Iberg, Garrett L. Robinson, Asis Sarkar, Banmeet Anand, Melissa M. Singh, Jay Zhao, Jack P. Higgins, Erin K. Willert. In vivo efficacy of a PD-L1 targeted, antigen seeding engineered toxin body [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 3366.
Abstract 3451: Non-genetic RTK-mediated cetuximab resistance in colorectal cancer offers multiple targets for therapeutic intervention
Authors: Graves-Deal Ramona , Bogatcheva Galina , Damalanka Vishnu , Klampfer Lidija , Janetka James W. , Coffey Robert J. , Singh Bhuminder 
Published: September 2020

Abstract
Using 3D type I collagen cultures of human colorectal cancer (CRC) cell line HCA-7 derivatives (CC, SC, CC-CR), we previously identified that activation of receptor tyrosine kinases (RTKs) MET and RON contributed to resistance to anti-EGFR monoclonal antibody, cetuximab. CC cells are sensitive to cetuximab, while SC and CC-CR cells are resistant in 3D. Both de novo and acquired modes of cetuximab resistance in SC and CC-CR, respectively, could be overcome by crizotinib, a multi-RTK inhibitor that also targets MET and RON. In fact, we now show that the cetuximab/crizotinib combination leads to synergistic growth inhibition in both SC and CC-CR. In addition to steady-state upregulation of MET and RON phosphorylation in SC cells, we also observe upregulation of ERBB3 phosphorylation after cetuximab addition, indicating a need for monitoring RTK activity during treatment. We have also identified a set of KRAS wild-type human CRC patient-derived tumor xenografts (PDXs) that show increased MET/RON phosphorylation. As noted previously, addition of the MET ligand (HGF) and ERBB3 ligand (NRG1) induced cetuximab resistance in CC cells and HGF-induced cetuximab resistance was overcome by crizotinib addition. We now show that MET/RON-dependent cetuximab resistance is due to autocrine production of their cognate ligands HGF/HGFL, respectively. For these studies, we employed selective inhibitors of the proteases (HGFA, Matriptase, and Hepsin) that mediate processing and maturation of HGF/HGFL; these inhibitors (ZFH7116 and VD2173) were able to overcome cetuximab resistance in SC and CC-CR cells. Thus, RTK ligand processing is an additional therapeutic target to counter cetuximab resistance in CRC.
Citation Format: Ramona Graves-Deal, Galina Bogatcheva, Vishnu Damalanka, Lidija Klampfer, James W. Janetka, Robert J. Coffey, Bhuminder Singh. Non-genetic RTK-mediated cetuximab resistance in colorectal cancer offers multiple targets for therapeutic intervention [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 3451.
Abstract 3513: Genome-wide scan for novel prostate cancer susceptibility loci with clinical consequence utilizing increased genetic drift of the Finnish population
Authors: Sipeky Csilla , Tammela Teuvo L. , Auvinen Anssi , Schleutker Johanna 
Published: September 2020

Abstract
Background: Prostate cancer (PC) is the second most common cancer in men worldwide with a rising prevalence. However, only limited number of clinically actionable variants are known. The objective of this study is to identify novel PC susceptibility loci with potential clinical relevance utilizing the increased genetic drift and reduced selective pressure of the recently bottlenecked powerful resource of the Finnish population.
Methods: We meta-analysed &gt;185.000 germline single nucleotide polymorphisms (SNPs) in 2738 PC cases and 2401 controls of Finnish origin in the Collaborative Oncological Gene-Environment Study (COGS) using Illumina iSelect custom SNP genotyping platform.
Results: Altogether 160 variants remain significant on GWAS level (p&lt;5 × 10−8) in 18 different gene regions. Out of the 160 identified PC susceptibility hits 78 (49%) are novel variants not reported earlier in association with PC. PC associated signals are in close proximity to one another forming genetic hot-spots on chromosomes 8, 7, 11, 17 and 19. The strength of association varies for 101 PC risk variants (OR 1.24-1.86) and 59 protective SNPs (OR 0.59-0.80). The strongest risk gene is CASC8 (OR=1.86; p=1.214 × 10−11). The most significant signals were found at the 8p21.2 chromosomal region (p&lt;7.6 × 10−16). Functional annotation identified a missense coding SNP at KLK3 gene to be a candidate causal variant. We showed evidence for association both with early-onset PC (OR 1.73; P=0.02), high serum PSA level at diagnosis (OR 1.36; P=0.024) and with aggressive PC (Gleason score ≥8; OR 1.44; P=0.02) at 17q21.
Conclusion: The results facilitate risk stratification for screening, clinical studies, treatment choices and functional research of PC
Citation Format: Csilla Sipeky, Teuvo L. Tammela, Anssi Auvinen, Johanna Schleutker. Genome-wide scan for novel prostate cancer susceptibility loci with clinical consequence utilizing increased genetic drift of the Finnish population [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 3513.