bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5266: Continuous trifluridine treatment in colorectal cancer cell lines- investigating its potential
Authors: Tsunekuni Kenta , Matsuoka Kazuaki , Kobunai Takashi , Takechi Teiji , Eguchi Hidetoshi , Mori Masaki , Doki Yuichiro , Ishii Hideshi
Published: September 2020

Background: Trifluridine (FTD) is a key component of the novel, orally-administered, antitumor drug trifluridine/tipiracil, and is approved for treatment of patients with metastatic colorectal and gastric/GEJ cancers refractory to standard chemotherapy. FTD, an antineoplastic thymidine analogue, is incorporated into the genomic DNA of tumor cells, thereby triggering cell death. An international, double-blind, placebo-controlled Phase III study confirmed the efficacy of trifluridine/tipiracil in patients previously treated with standard chemotherapy. However, the clinical benefit of trifluridine/tipiracil in the first-line metastatic setting still remains unknown. Therefore, we analyzed the sensitivity for FTD, 5-FU, L-OHP, and SN-38 in parental and three FTD long-term treated cell lines.

Methods: Three human colorectal cancer cell lines (DLD-1, HCT-116, and RKO) and their respective FTD long-term treated cell lines (treated by step-wise increase in concentrations of FTD starting at 1 µM and ending at 400 µM over a 5 month period) were used. Sensitivity assays using crystal violet staining for FTD, 5-FU, L-OHP, and SN-38 and wound healing assay to assess the migration capacity were performed in these cell lines. Further, mRNA was purified and the resulting microarray was utilized to analyze expression profiles.

Results: Continuous FTD long-term treated cell lines were resistant to FTD. In the sensitivity assays, the IC50 values in all three continuously FTD treated cell lines increased by >22-fold for FTD, by <2-fold for 5-FU and L-OHP, and by <9-fold for SN-38 (2 to 8.2-fold in three cell lines) compared to those in parental cell lines. We found that FTD long-treated cells had weaker migration ability than parental cells in all three cell lines. Additionally, HIF1A related glycolytic mRNA was downregulated in FTD long-treated cell lines compared to that in parental cell lines.

Conclusions: Although long-term treatment with FTD induced resistance in colorectal cancer cell lines, no cross-resistance to other drugs (5-FU, L-OHP) and partial resistance to SN-38 that are used in standard chemotherapy was observed. Therefore, FTD is expected to be effective even after treatment with standard chemotherapy. Although FTD/TPI is currently used in the later line, it is suggested from this basic study that FTD/TPI may be applicable to take to the front line therapy.

Citation Format: Kenta Tsunekuni, Kazuaki Matsuoka, Takashi Kobunai, Teiji Takechi, Hidetoshi Eguchi, Masaki Mori, Yuichiro Doki, Hideshi Ishii. Continuous trifluridine treatment in colorectal cancer cell lines- investigating its potential [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 5266.
Abstract 5313: TT-00420, a novel multiple kinase inhibitor to treat TNBC
Authors: Peng Peng , Li Guoyu , Qiang Xiaoyan , Tian Yuwei , Yu Qi , Marangoni Elisabetta , Wu Frank
Published: September 2020

TT-00420, a clinical stage spectrum-selective multiple kinase inhibitor, has been shown to be very efficacious against TNBC cell lines and in vivo models in our previous study. TT-00420 simultaneously inhibits Aurora A/B, RTKs involved in angiogenesis, JAK kinase family as well as CSF1R in kinase profiling assay. In breast cancer cell panel screening, TT-00420 specifically inhibits the proliferation across different subtypes of TNBC, while leaving ER/PR+ and Her2+ cells intact. Incubation of TNBC cell line HCC1806 with TT-00420 results in dose-dependent down-regulation of EGFR, TWIST1, SNAI2, FOSL1, and MYC, a cluster of genes collectively known as “Achilles cluster” essential for TNBC cell growth and proliferation. TT-00420 exhibits great in vivo efficacy in a panel of TNBC PDX models. In the mechanism study, we reveal that the inhibition of Aurora A or B lays the cornerstone of pan-TNBC inhibition capability of TT-00420, and the inhibition of angiogenesis pathway or JAK family or EGFR contributes to potency and activity of TT-00420 against TNBC at various degrees. Treatment of a mouse syngeneic TNBC model with TT-00420 normalizes the vessel conditions in tumors, which is consistent with TT-00420's capability to enhance the anti-tumor immune response both alone or combined with PD-1 blockade. Taken together, TT-00420 represents a novel synergistic inhibitory mechanism to treat TNBC.

Citation Format: Peng Peng, Guoyu Li, Xiaoyan Qiang, Yuwei Tian, Qi Yu, Elisabetta Marangoni, Frank Wu. TT-00420, a novel multiple kinase inhibitor to treat TNBC [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 5313.
Abstract 5315: BPI-28592, a next-generation TRK kinase inhibitor that potently inhibits primary and resistant NTRK mutations
Authors: Fu Bang , Yan Dan , Liu Lingling , Guo Jing , Huang Jisheng , Ma Tianyi , Li Yinlong , Ren Wei , Chen Jie , Liu Xiangyong , Lan Hong , Ding Lieming , Wang Jiabing
Published: September 2020

Oncogenic NTRK gene fusions occur at very low frequency in common cancers but high frequency in rare tumors including childhood malignances. The 1st-generation TRK inhibitors, e.g., larotectinib and entrectinib, have been used successfully to treat cancers carrying the NTRK fusions with excellent response rate and long duration. Nonetheless, duration of response may eventually be limited by acquired resistance mutations, such as TRKA-G595R or TRKC-G623R. We aimed at identifying a 2nd-generation inhibitor which can cover the primary TRK fusions and overcome the common resistant mutants induced by the 1st -generation TRK inhibitors. Using structural modeling to guide rational drug design, we discovered BPI-28592 as a 2nd-generation TRK inhibitor with desired potency and good drug-like properties. BPI-28592 is of a novel structure and possessed sub-nanomolar potency against a variety of primary TRK fusions. Importantly, the molecule exhibits low nanomolar inhibitory activity against acquired mutants commonly identified in patients who are refractory to 1st-generation TRK inhibitors, including TRKA-G595R TRKA-F589L, TRKA-G667C, and TRKC-G623R, etc. BPI-28592 showed a highly selective profile in a Kinome scan of 468 individual kinases. In cell-based assays, BPI-28592 dose-dependently inhibited proliferation of primary as well as resistant TRK mutant cell lines, blocked TRK protein phosphorylation and downstream signaling. Furthermore, BPI-29592 demonstrated in vivo efficacy in xenograft models derived from primary and resistant TRK mutant cells. BPI-28592 exhibited a profile of high oral exposure across multiple pre-clinical species, favorable ADME properties, and wide therapeutic window in pre-clinical toxicology studies. In conclusion, we discovered BPI-28592 as a novel, potent and selective next-generation TRK kinase inhibitor. The molecule has completed IND-enabling studies and is planned to enter Phase 1 clinical development in early 2020.

Citation Format: Bang Fu, Dan Yan, Lingling Liu, Jing Guo, Jisheng Huang, Tianyi Ma, Yinlong Li, Wei Ren, Jie Chen, Xiangyong Liu, Hong Lan, Lieming Ding, Jiabing Wang. BPI-28592, a next-generation TRK kinase inhibitor that potently inhibits primary and resistant NTRK mutations [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 5315.
Abstract 5327: KPG-121, a novel CRBN modulator, potently inhibits growth of metastatic castration resistant prostate cancer as a single agent or in combination with androgen receptor signaling inhibitors both in vitro and in vivo
Authors: Ge Chuansheng , Zhang Lei , Liao Baisong
Published: September 2020

Prostate cancer is the common cancer in men and the second leading cause of cancer-related deaths in men. This disease depends on dihydrotestosterone (DHT) which activates the androgen receptor (AR) signal. Endocrine therapy (including castration surgery, anti-androgen deprivation therapy or combined androgen blockade therapy) is a preferred treatment for prostate cancer. The treatment is very effective for the majority of patients at the initial stage, but after a median period of treatment for 14 to 30 months, most patients will gradually develop metastatic castration resistant prostate cancer (mCRPC) with a median survival of less than 20 months, and urgently need a further treatment. Androgen receptor antagonists in combination with anti-androgen drugs are currently the most effective drug treatment to manage mCRPC where castration has failed. Enzalutamide, Abiraterone Acetate, Apalutamide and Darolutamide belong to this class of androgen receptor or androgen synthesis inhibitors. Here we show that KPG-121, a small molecule immunomodulator, significantly enhances the therapeutic effects of Enzalutamide, Abiraterone Acetate, Apalutamide and Darolutamide in pre-clinical mCRPC tumor models. The phase 1 trials were initiated to evaluate the safety, PK profile and preliminary efficacy of the combinatorial therapy in the patients with metastatic castration resistant prostate cancer in the US (IND 134979). KPG-121 is a novel generation of Lenalidomide analogue. KPG-121 is structurally similar to lenalidomide but with stronger binding with CRBN target, enhanced immunomodulatory activity, better anti-proliferative and anti-angiogenic properties compared to lenalidomide. Results from in vivo metastatic castration resistant prostate cancer xenograft models demonstrated that KPG-121 at tested daily dose levels ranging from 1.0 mg/kg to 10 mg/kg in combination with Enzalutamide, Abiraterone Acetate, Apalutamide or Darolutamide significantly improved anti-tumor efficacy compared to the AR signaling inhibitors alone. In addition, KPG-121 showed favorable DMPK profiles and potentially manageable toxicity in pre-clinical IND enabling study.

Citation Format: Chuansheng Ge, Lei Zhang, Baisong Liao. KPG-121, a novel CRBN modulator, potently inhibits growth of metastatic castration resistant prostate cancer as a single agent or in combination with androgen receptor signaling inhibitors both in vitro and in vivo [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 5327.
Abstract 5345: Heterogeneous tumor PSMA expression represents a resistance mechanism to PSMA-targeted radioligand therapy
Authors: Lueckerath Katharina , Current Kyle , Meyer Catherine , Magyar Clara , Mona Christine E. , Almajano Joel , Slavik Roger , Stuparu Andreea D. , Cheng Chloe , Dawson David , Radu Caius , Czernin Johannes
Published: September 2020

Radioligand therapy (RLT) with prostate-specific membrane antigen (PSMA)-targeting ligands is effective in ~50% of patients with advanced prostate cancer (PC). Causes for RLT failure are not well understood. The prevalent PSMA heterogeneity in PC might contribute to RLT failure. Here we investigate the relationship between RLT efficacy and PSMA levels per cell, and PSMA heterogeneity. PC cells expressing different levels of PSMA (RM1-PSMA low, medium, high, or RM1-YFP that do not express PSMA), or a mix of PSMA− and PSMA+ cells (RM1-YFP/ RM1-PSMA-high; PC3/ PC3-PIP) at various ratios were subcutaneously injected into NSG mice. Mice were treated with 177Lu- or 225Ac-PSMA617 (RLT) and tumor growth was monitored. In a subset of mice, radioligand uptake (γ-counting), DNA damage and PSMA expression (anti-53BP1 and -PSMA immunohistochemistry, respectively) were quantified in tumors resected 2 days after RLT. Increasing PSMA levels and fractions of PSMA+ cells improved RLT efficacy in both, the RM1 and PC3 model. PSMA expression correlated with radioligand uptake into the tumor and the degree of DNA damage and. Treatment with 225Ac-PSMA617 (vs. 177Lu-PSMA617) improved RLT outcomes and tended to enhance the differences in therapeutic efficacy between experimental groups. Taken together, we demonstrate in mouse models of PC that optimal anti-tumor efficacy of RLT hinges on homogenously high target expression. Although PSMA-RLT is effective even in tumors with low PSMA levels or with a small number of PSMA+ cells, low or heterogeneous PSMA expression might result in undertreatment and selection of treatment resistant clones. Systematic assessment of intra- and inter-lesion PSMA heterogeneity is currently not feasible clinically; however, this issue might be addressed by individual patient dosimetry to optimize safely delivered maximal tumor doses. Clinical studies designed to determine intra- and inter-lesion PSMA heterogeneity and to optimize PSMA-RLT for each patient are highly warranted.

Citation Format: Katharina Lueckerath, Kyle Current, Catherine Meyer, Clara Magyar, Christine E. Mona, Joel Almajano, Roger Slavik, Andreea D. Stuparu, Chloe Cheng, David Dawson, Caius Radu, Johannes Czernin. Heterogeneous tumor PSMA expression represents a resistance mechanism to PSMA-targeted radioligand therapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5345.
Abstract 5367: Identification and clinical value of the circulating tumor cells (CTCs) in the colorectal cancer
Authors: Hamid Faysal-Bin , Islam Farhad , Lu Cu-Tai , Matos Marco , Cheng Tracie , Gopalan Vinod , Lam Alfred King-yin
Published: September 2020

The number of circulating tumor cells (CTCs) hase been found as a promising predictive and diagnostic biomarker of colorectal cancer (CRC). The purpose of the study was to enumerate and characterize CTCs from the CRC patients and evaluate the correlation with the clinical stages.Peripheral blood from 57 CRC patients and 6 healthy donors were subjected to isolate CTC by a negative selection (EasySepTM) method. The CTCs were detected by the immunofluorescence staining with the tumor cell markers such as epithelial cell adhesion molecule (EPCAM) and cytokeratin (CK) antibodies and counted them. The extent of positive expressions of EPCAM and cytokeratin (CK) proteins were used to characterize the CTCs and the subpopulations were correlated with the tumor stages of the CRC patients. P<0.05 was considered as statistically significant.CTCs were detected in 72% (41/57) of the CRC patients. Comparing to the healthy donors (HD), the CTC count was higher (p=0.0062) in the CRC patients. In addition, CTC clusters were found in 32% (18/57) of patients with CRC while total CTCs were significantly higher than the cluster count (p=0.001). Among the CRC patients, CTCs were more abundant in the advanced stages compared to the early stages (p=0.0019). Moreover, we found the length of the individual CTCs were significantly larger than the CTCs in the cluster (p<0.0001) and significantly correlated (p=0.036) with the tumor stages. However, we identified the EPCAMPosCkPos subclones in 38 (93%) out of 41 CTC positive cases and a median of 8.0 (range: 0-60) CTCs were counted. In contrast, EPCAMPosCkNeg subpopulations were detected in 27 (66%) patients and the median CTC count was 2.0 (range: 0-18). Interestingly, EPCAMPosCkPos CTC subclones were significantly higher (p=0.011) than EPCAMPosCkNeg subclones, and the number of EPCAMPosCkPos CTCs was merely associated (p=0.0068) with the tumor stages. We conclude that the morphological and phenotypic features of the CTCs are closely relevant to the clinical stages, which could provide vital information to improve the patient stratification and treatment selection.

Citation Format: Faysal-Bin Hamid, Farhad Islam, Cu-Tai Lu, Marco Matos, Tracie Cheng, Vinod Gopalan, Alfred King-yin Lam. Identification and clinical value of the circulating tumor cells (CTCs) in the colorectal 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 5367.
Abstract 5403: Targeting death-receptors in therapy resistant progressive neuroblastoma: A preliminary insight
Authors: Somasundaram Dinesh Babu , Maher Andrew , Aravindan Sheeja , Herman Terence S. , Aravindan Natarajan
Published: September 2020

Neuroblastoma is the most common cancer in infants and accounts for 9.1% of childhood cancer-related deaths. Despite the current standard of care (SOC), drug-resistant neuroblastoma significantly compromises patient survival. Hence it is critical to identify new and less toxic therapeutic approaches for better patient outcomes. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), which mediates its effect through the activation of the death receptor (DR) 4 and 5 are found on many cell types and are the main pathway for apoptosis. These can death receptors can kill most tumor cell types expressing TRAIL-receptor 1. Mining RNA-Seq data across multiple cohorts of neuroblastoma patients indicated that high levels of DR4 and DR5 significantly associated with poor clinical outcomes (OS, EFS, RFS). Further, examining the surface expression of DR4 and DR5 by immunohistochemistry using anti-human rabbit monoclonal antibodies on custom-archived TMAs for 100 NB patients revealed profound correlation with poor OS and RFS. Immunoblot analysis for DR4 and DR5 level in a panel of stage 4 patient-derived cell-lines during diagnosis and from progressive disease after therapy displayed high-levels of death receptor expression in progressive disease. Preliminary co-culture investigations with genetically engineered mesenchymal stem cells (MSCs) expressing tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) mimicking targeted delivery showed heightened differentiation and death (limited dilution tumorosphere assay) of therapy resistant cells. Further these studies exhibited inhibition of tumor cell migration and invasion. The results demonstrate the benefit of targeted TRAIL delivery in in the treatment of therapy resistant neuroblastoma and further imply that the genetically engineered MSCs could serve as a clinically valuable tool for improved therapeutic strategy in this setting.

Citation Format: Dinesh Babu Somasundaram, Andrew Maher, Sheeja Aravindan, Terence S. Herman, Natarajan Aravindan. Targeting death-receptors in therapy resistant progressive neuroblastoma: A preliminary insight [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 5403.
Abstract 5408: Chloroquine inhibits PARP expression in hepatoblastoma
Authors: Pihlajoki Marjut , Eloranta Katja , Kyrönlahti Antti , Heikinheimo Markku
Published: September 2020

Hepatoblastoma is the most common pediatric liver malignancy accounting 1 % of all pediatric malignancies. Despite advances in medical treatment and surgical techniques, the prognosis for advanced metastatic hepatoblastoma remains poor, underscoring the need for new therapeutic modalities. Chloroquine (CQ), a drug used to treat malaria and rheumatologic diseases, has been shown to inhibit the growth and survival of various cancer types. In this study we examined the antineoplastic activity of CQ on HB cells. Established human HB cell line (HUH6) cells were cultured in presence of clinically acceptable concentrations of CQ. Morphology, viability, and apoptosis were assessed after 48 h and 96 h of CQ treatment showing decreased cell survival and increased caspase 3/7 activity representing induction of apoptosis. Metabolomic profiling of CQ treated HUH6 cells demonstrated significant decrease in NAD+ and aspartate concentration. Furthermore, RT-qPCR based Death Pathway Finder array was conducted to elucidate the molecular mechanisms underlying the CQ effect showing downregulated expression of PARP1 and PARP2. Quantitative western blotting was performed to validate differentially expressed genes at the protein level. Taken together, CQ treatment inhibits cell survival in HB cells in vitro, presumably by disturbing NAD+/NADH balance, impairing aspartate biosynthesis, and reducing PARP expression. CQ thus holds potential as a new treatment modality in HB management.

Citation Format: Marjut Pihlajoki, Katja Eloranta, Antti Kyrönlahti, Markku Heikinheimo. Chloroquine inhibits PARP expression in hepatoblastoma [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 5408.
Abstract 5469: iCVA- A knowledge-based cancer variation annotation application
Authors: Kumar Gaurav , Kelly Melanie , Fortina Paolo , Ertel Adam
Published: September 2020

A typical genomic sequencing experiment results in a long list of statistically significant variant candidates among protein-coding genes without any unifying biological theme. This leads to a daunting task of identifying the causal variants and genes to accurately diagnose the disease for clinical or research utility. While a number of downstream tools exist to aid in cancer genome annotation and interpretation, including Clinical Interpretations of Variants in Cancer (CIViC), ClinGen, Database of Curated Mutations (DoCM), Oncotator, and Variant Interpretation for Cancer (VIC), these only provide selective information (somatic/germline/gene-variant specific) or are time/resource consuming. To address this challenge, we developed a software called iCVA using the following methodology: 1) we obtained a priori defined gene sets for different cancer-inducing mechanisms such as pro-oncogenesis, tumor-suppression, DNA repair, angiogenesis, inflammation, metabolism, hypoxia, cell cycle and immune system. 2) genomic variation data were integrated from 1000 Genome, Exome Sequencing Project, Exome Aggregation Consortium (ExAC), ClinVar, dbSNP, GNOMAD, The Cancer Genome Atlas (TCGA), the Catalogue of Somatic Mutations in Cancer (COSMIC) and the International Cancer Gene Census (ICGC), and re-classified into a consensus classification according to ACMG guidelines. 3) Finally, we devised a reporting system to process mutation data from a sequencing experiment to utilize the harmonized pathway and mutational information integrated in a local database to identify and classify cancer-specific gene mutations. The iCVA reporting system classifies mutations as germline or somatic and then sub-classifies these into different cancer-specific mechanisms, including those known to be targetable through existing therapies. To our knowledge, iCVA is the first tool to provide a comprehensive report on cancer variants in a simplified and faster manner, to accelerate the genomic characterization of cancer samples analyzed by high-throughput DNA sequencing.

Citation Format: Gaurav Kumar, Melanie Kelly, Paolo Fortina, Adam Ertel. iCVA- A knowledge-based cancer variation annotation application [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 5469.
Abstract 5518: Tumor infiltrating B cells and tertiary lymphoid structures are associated with tumor response to PD-L1 blockade in NSCLC
Authors: Patil Namrata S.
Published: September 2020

Inhibitors of the programmed cell death-1/programmed cell death ligand1 (PD-1/PD-L1) signaling axis have demonstrated sustained efficacy in NSCLC, however there are gaps in our understanding of the mechanisms of how PD-1/ PD-L1 blockade improve patient outcomes, especially as they relate to long term survival. Differential gene expression analysis in tumors from NSCLC patients who derived >12 months OS benefit on atezolizumab monotherapy showed high expression of B cell transcripts in the randomized phase II POPLAR and phase III OAK studies (> median, HR: 0.61, 95% CI 0.47-0.78, and HR=0.54, CI 0.42-0.7, p<0.001 respectively). Immunofluorescence imaging confirmed the presence of CD79+ B cells in these tumors. High expression of CD79A was associated with the presence of tertiary lymphoid structures (TLS) that in turn were associated with atezolizumab survival benefit. B cell receptor sequencing of patient biopsies pre and post atezolizumab treatment suggest clonal expansion of tumor infiltrating plasma cells in patients responsive to atezolizumab. Collectively, these results suggest that NSCLC tumor patients enriched for CD79A+ B cells and IgG+ plasma cells derive greater overall survival benefit from atezolizumab and emphasize the potential of these genes as actionable biomarkers in NSCLC.

Citation Format: Namrata S. Patil. Tumor infiltrating B cells and tertiary lymphoid structures are associated with tumor response to PD-L1 blockade in NSCLC [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 5518.
Abstract 5594: Inmune gene expression by nCounter in mucinous adenocarcinoma lung cancer
Authors: Lladó Ruth Román , Esteban Cristina Aguado , Giménez-Capitán Ana , Daisuke Ueda , Ito Masaoki , Tsutani Yasuhiro , Miyata Yoshihiro , Teixidó Cristina , Reguart Noemí , Okada Morihito , Rodríguez Sonia , Balada Ariadna , Aldeguer Erika , Ramirez Santiago Viteri , Cao Maria Gonzalez , Aguilar Andrés , Rosell Rafael , Vila Miguel Angel Molina
Published: September 2020

Introduction: Although immune checkpoint inhibitors have become a standard of care for multiple cancer types; currently used biomarkers, such as PD-L1 expression, MSI or TMB, fail to identify all responders. In addition, little is known about markers of response to immunotherapy in some infrequent histological subtypes, such as mucinous adenocarcinoma of the lung. Gene expression signatures incorporating not only PD-L1 but also other immune modulators can have a better predictive power, particularly in these histological subtypes.

Methods: A cohort of 165 baseline samples from with stage I-IV non-small cell lung cancer (NSCLC) patients was retrospectively analyzed, 55 stage I-IIIA mucinous adenocarcinomas (MA), 31 stage I-IIIA nonmucinous adenocarcinomas (NMA) and 79 stage IIIB-IV NMAs. RNA was extracted from FFPE tumor samples and the expression levels of a 7-gene immune signature (CD4, CD8A, FOXP3, GZMM, INFG, CD274 and PDCD1) were determined using a nCounter panel. Krustal-Wallis test was used for statistical comparisons.

Results: No significant differences were observed in the expression levels of FOXP3 and PDCD1 (codifying forkhead box P3 and PD-1) between the three groups analyzed. In contrast, GZMM, IFNG and CD4 expression (codifying granzyme M, interferon gamma and CD4) was significantly higher in early stage MA compared to stage I-IIIA or stage IIIB-IV NMA. CD8A expression was significantly higher in early stage tumors, independently of the presence of mucinous component. In addition, the levels of CD274 mRNA (codifying PD-L1) were significantly higher in early stage MA compared to advanced stage NMA. Finally, among the stage I-IIIA MA, 27/55 (49%) were KRAS-mutant, but they did not show significant differences with KRAS-wt MAs in the expression of any of the genes tested.

Conclusions: Mucinous adenocarcinoma of the lung shows a different pattern of expression of immune-related genes, which may influence response to immunotherapy.

Citation Format: Ruth Román Lladó, Cristina Aguado Esteban, Ana Giménez-Capitán, Ueda Daisuke, Masaoki Ito, Yasuhiro Tsutani, Yoshihiro Miyata, Cristina Teixidó, Noemí Reguart, Morihito Okada, Sonia Rodríguez, Ariadna Balada, Erika Aldeguer, Santiago Viteri Ramirez, Maria Gonzalez Cao, Andrés Aguilar, Rafael Rosell, Miguel Angel Molina Vila. Inmune gene expression by nCounter in mucinous adenocarcinoma 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 5594.
Abstract 564: Futibatinib (TAS-120) plus chemotherapy demonstrates a synergistic effect across various FGFR-deregulated cancer cell lines and xenograft models
Authors: Sootome Hiroshi , Miura Akihiro , Komori Toshiharu , Hirai Hiroshi
Published: September 2020

Introduction: Futibatinib, a selective, irreversible fibroblast growth factor receptor (FGFR) 1-4 inhibitor, has antitumor activity in FGFR-deregulated cancer models. The potential synergistic antitumor effects of targeted therapy plus chemotherapy (chemo) was evaluated by combining futibatinib with cytotoxic agents in in vitro and in vivo models.

Methods: Gastric, breast, lung, endometrial, and bladder cancer cell lines with various FGFR aberrations were treated with futibatinib and cytotoxic agents (5-FU, paclitaxel, cisplatin, or gemcitabine) individually or in combination. In vitro cytotoxicity of futibatinib was assessed by PARP cleavage activity. Any combination effect on cell growth was determined by the Bliss independence method, combination index (CI; CalcuSyn software), or isobologram analysis. The antitumor effects of futibatinib plus S-1 or paclitaxel and futibatinib plus gemcitabine or cisplatin were evaluated in gastric FGFR2-amplified SNU-16 and endometrial FGFR2-N549K-mutated AN3CA mouse xenograft models, respectively, by measuring relative tumor volume (RTV).

Results: A synergistic effect on cell growth inhibition was observed with futibatinib plus chemo in SNU-16 cells (CIs at ED90 were 0.50 [5-FU], 0.71 [paclitaxel], 0.76 [cisplatin], and 0.29 [gemcitabine]). Increased PARP cleavage and decreased cell growth were observed in SNU-16 cells treated with futibatinib plus 5-FU vs monotherapy. Synergized cell growth inhibition occurred in various FGFR-deregulated cancer cell lines. In xenograft models, vs monotherapy, a significant reduction (P<0.01) in RTV occurred with futibatinib plus S-1, paclitaxel (Table), or cisplatin and a numerically greater reduction with futibatinib plus gemcitabine.

Table.Relative tumor volume in SNU-16 mouse xenografts treated with futibatinib or paclitaxel or combination therapyTreatment (mg/kg/d)Relative tumor volumeDay 0Day 4Day 8Day 11Day 15Control11.562.373.344.58Futibatinib (5)a11.321.952.773.83Futibatinib (15)a11.211.822.453.16Paclitaxel (60)b10.480.410.410.59Futibatinib + paclitaxel (5 + 60)c10.470.270.170.14Futibatinib + paclitaxel (15 + 60)c10.440.220.120.07aDays 1-14 (oral).bDay 1 (intravenous).cDays 1-14 + Day 1.

Conclusions: Futibatinib showed a synergistic antitumor effect in vitro and in vivo when combined with various cytotoxic agents. These data are promising and warrant further investigation.

Citation Format: Hiroshi Sootome, Akihiro Miura, Toshiharu Komori, Hiroshi Hirai. Futibatinib (TAS-120) plus chemotherapy demonstrates a synergistic effect across various FGFR-deregulated cancer cell lines and xenograft models [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 564.
Abstract 5644: Claudin 18.2 X 4-1BB bispecific antibody induced potent tumor inhibition through tumor-specific 4-1BB activation
Authors: Jiang Wenqing , Fang Lei , Wang Zhengyi , Guo Taylor B. , Park Eunyoung , Sung Eunsil , Jung Jaeho
Published: September 2020

Background

Claudin 18.2 (CLDN18.2) is a gastric-specific membrane protein. In the healthy tissue, CLDN18.2 expression is restricted to the short-lived differentiated cells of gastric mucosa as a component of tight junction with limited accessibility of antibody treatment. However, it was ectopically expressed at significant levels in a variety of primary lesion and metastases of epithelial tumor entities, including gastric, pancreatic, esophageal, and lung adenocarcinoma cells. Taken together, CLDN18.2 was believed to be an ideal tumor antigen for immunotherapy. Here, we generated and described a bispecific antibody TJ-CD4B which could redirect and activate T cells to CLDN18.2 positive tumor cells by engaging 4-1BB antigen, thus positioned as a potential novel therapy for gastric cancer and other CLDN18.2 positive tumors.

Methods

TJ-CD4B was evaluated for its antigen binding through protein-based binding and cell based binding assay. The cell-based function of TJ-CD4B was further evaluated in a 4-1BB signaling reporter assay and primary human T cell assay. The in vivo efficacy was validated in 4-1BB humanized C57BL/6 mice transferred with colon cancer cell line MC38 overexpressed with human CLDN18.2. Percentage of tumor infiltrating and peripheral CD8+ T cells was evaluated by FACS analysis.

Results

TJ-CD4B showed stronger binding capability to CLDN18.2 when compared with benchmark monoclonal antibody against CLDN18.2 (IMAB362). Functional evaluation of TJ-CD4B indicated the activation of 4-1BB signaling was solely dependent on CLDN18.2 expression on the cell. Furthermore, TJ-CD4B showed superior 4-1BB activity than benchmark 4-1BB monoclonal antibody (Urelumab) in the presence of cells expressing a wide range of CLDN18.2 level including CLDN18.2-low cells. Using a humanized 4-1BB mouse model, TJ-CD4B showed strong tumor growth inhibition (TGI) of CLDN18.2 expressing tumor cells. Meanwhile, we also observed TJ-CD4B increased tumor infiltrating lymphocytes while it had no impact on peripheral lymphocytes. Drug developability assessment indicated that TJ-CD4B was a reasonably developable molecule to be taken forward.

Conclusions

Our data showed that CLDN18.2-targeted 4-1BB bispecific antibody, TJ-CD4B, displayed potent anti-tumor activity only in tumor microenvironment, possibly minimizing the risk of peripheral toxicity. These compelling data supported the further development of TJ-CD4B, which aims to enter clinical phase in early 2021.

Citation Format: Wenqing Jiang, Lei Fang, Zhengyi Wang, Taylor B. Guo, Eunyoung Park, Eunsil Sung, Jaeho Jung. Claudin 18.2 X 4-1BB bispecific antibody induced potent tumor inhibition through tumor-specific 4-1BB activation [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 5644.
Abstract 5666: A noninvasive approach for early prediction of therapeutic benefit from immune checkpoint inhibition for lung cancer
Authors: Nabet Barzin Y. , Esfahani Mohammad S. , Hamilton Emily G. , Chabon Jacob J. , Moding Everett J. , Rizvi Hira , Steen Chloe B. , Chaudhuri Aadel A. , Liu Chih Long , Hui Angela B. , Stehr Henning , Goljenola Linda , Jin Michael C. , Jeon Young-Jun , Tseng Diane , Merghoub Taha , Neal Joel W. , Wakelee Heather A. , Padda Sukhmani K. , Ramchandran Kavitha J. , Das Millie , Bonilla Rene F. , Yoo Christopher , Chen Emily L. , Ko Ryan B. , Newman Aaron M. , Hellmann Matthew D. , Alizadeh Ash A. , Diehn Maximilian
Published: September 2020

Although treatment of non-small cell lung cancer (NSCLC) with immune checkpoint inhibitors (ICI) can produce remarkably durable responses, most patients develop early disease progression. Furthermore, initial response assessment by conventional imaging is often unable to identify which patients will achieve durable clinical benefit (DCB). Here, we analyze 211 samples from 99 patients and demonstrate that pre-treatment circulating tumor DNA (ctDNA) and circulating immune profiles are independently associated with DCB. We further show that ctDNA dynamics after a single ICI infusion can identify the majority of patients who will achieve DCB. Integrating these determinants, we describe an entirely noninvasive multi-analyte assay (DIREct-On, Durable Immunotherapy Response Estimation by immune profiling and ctDNA- On-treatment) that robustly predicted DCB, and that was validated in two independent cohorts (AUC = 0.89-0.93, PPV = 92-100%, HR = 0.04-0.11). Taken together, these results demonstrate that integrated ctDNA and circulating immune cell profiling can provide accurate, noninvasive, and early forecasting of ultimate outcomes for NSCLC patients receiving ICI.

Citation Format: Barzin Y. Nabet, Mohammad S. Esfahani, Emily G. Hamilton, Jacob J. Chabon, Everett J. Moding, Hira Rizvi, Chloe B. Steen, Aadel A. Chaudhuri, Chih Long Liu, Angela B. Hui, Henning Stehr, Linda Goljenola, Michael C. Jin, Young-Jun Jeon, Diane Tseng, Taha Merghoub, Joel W. Neal, Heather A. Wakelee, Sukhmani K. Padda, Kavitha J. Ramchandran, Millie Das, Rene F. Bonilla, Christopher Yoo, Emily L. Chen, Ryan B. Ko, Aaron M. Newman, Matthew D. Hellmann, Ash A. Alizadeh, Maximilian Diehn. A noninvasive approach for early prediction of therapeutic benefit from immune checkpoint inhibition for 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 5666.
Abstract 5678: Novel glucocorticoid receptor degrading bifunctional molecules as therapeutics in castration-resistant prostate cancer
Authors: Link James T. , Blazanin Nicholas , Deribe Yonathan Lissanu
Published: September 2020

While the majority of men with prostate cancer will experience modest morbidity, advanced castration-resistant prostate cancer (CRPC) remains a devastating disease with significant mortality and limited treatment options. Recently approved anti-androgen therapies such as enzalutamide have shown promising clinical responses. Unfortunately, patients frequently develop resistance to these therapies. Signaling through the glucocorticoid receptor (GR) has emerged as a primary mechanism of resistance to enzalutamide in CRPC suggesting that GR is an attractive therapeutic target. However, existing small molecule antagonists of GR suffer from partial agonist activity leading to activation of a subset of GR target genes which limits their anti-tumor efficacy. To overcome this problem, we have developed bifunctional small molecules or proteolysis targeting chimeras (PROTACs) that induce potent and specific degradation of GR. Using iterative structure-activity relationship (SAR) studies, we have developed a novel chemical linker that imparted superior properties to PROTACs. These novel GR PROTACs induce profound degradation of GR. Using proteomic analysis and quantification of more than 8000 proteins, we show that our GR PROTAC is highly selective for GR. GR PROTACs inhibited proliferation of enzalutamide-resistant prostate cancer cells, have favorable pharmacokinetic properties and are active in vivo. Importantly, GR PROTACs work synergistically with enzalutamide to inhibit growth of prostate cancer xenografts in a castration-resistant mouse model of prostate cancer. Mechanistically, we identified improved suppression of AR and GR target genes in the combination of GR PROTAC and enzalutamide. More importantly, the combination was able to suppress crucial developmental pathways implicated in prostate cancer resistance to therapy and tumor dormancy. Our novel GR PROTACs highlight the power of targeted protein degradation to deliver excellent chemical probes and potentially impactful cancer therapeutics.

Citation Format: James T. Link, Nicholas Blazanin, Yonathan Lissanu Deribe. Novel glucocorticoid receptor degrading bifunctional molecules as therapeutics in castration-resistant prostate 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 5678.
Abstract 5698: Novel conditionally active bispecific T cell engagers targeting solid tumors
Authors: Cugnetti Ana Paula G. , Liu Haizhen , Wang Jing , Xing Charles , Wheeler Christina , Lucas Matthew , Chang Cathy , Frey Gerhard , Boyle William J. , Short Jay M.
Published: September 2020

The use of CD3/target bispecific molecules has great therapeutic potential in oncology. This potential, however, is diminished due to serious on-target drug-related toxicities, restricting the development of bispecific T cell engagers to targets that have limited expression in normal tissues. Conditionally Active Biologic (CAB) technology is a proprietary platform that generates antibodies which have no or very little binding to the target antigen in healthy tissue (normal physiological conditions), but have strong binding in the context of diseased tissues (tumor microenvironment) based on the glycolytic tumor metabolism (including Warburg effect). Using our CAB technology, we have developed CD3/target bispecific antibodies that bind to recombinant CD3 and to CD3 expressing cells under in vitro tumor microenvironment conditions, but not in physiologic conditions (CAB CD3). CAB CD3 bispecific antibodies were generated against various well-established tumor associated antigens (TAA), including EpCAM, B7-H3, HER-2 and EGFR. All of these molecules play important roles in cancer biology and are attractive targets for the development of therapeutic bispecific antibodies. In addition, we have also developed CAB TAA / CAB CD3 bispecific antibodies. In vitro and in vivo efficacy data as well as toxicology studies in non-human primates for CAB-T cell engagers will be presented. Our data demonstrates that CAB bispecific antibodies are efficacious in vivo and have lower toxicity comparing to non-CAB bispecific antibodies. The CAB technology will allow the generation of a new class of T-cell engagers with increased safety margin and therapeutic index in the clinic.

Citation Format: Ana Paula G. Cugnetti, Haizhen Liu, Jing Wang, Charles Xing, Christina Wheeler, Matthew Lucas, Cathy Chang, Gerhard Frey, William J. Boyle, Jay M. Short. Novel conditionally active bispecific T cell engagers targeting solid tumors [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 5698.
Abstract 5708: Deciphering Warburg effect: hypoxia inhibits tumor cell differentiation through reducing acetyl-CoA generation and chromatin accessibility
Authors: Ye Jiangbin , Li Yang , Gruber Joshua J. , Litzenburger Ulrike M. , Zhou Yiren , Miao Yu R. , LaGory Edward L. , Li Albert M. , Hu Zhen , Hart Lori S. , Maris John M. , Chang Howard Y. , Giaccia Amato J.
Published: September 2020

The Warburg effect is a metabolic hallmark of all cancer cells, characterized by increased glucose uptake and glycolysis for lactate generation. The generation and excretion of lactate would appear be a waste of carbon backbone and energy that is needed for proliferation. It was proposed by Warburg that the cause and consequence of the Warburg effect were the injury of respiration and cell dedifferentiation, respectively. One common factor that damages mitochondrial respiration is hypoxia, which is a metabolic stress that blocks cell differentiation and promotes cancer progression. The underlying mechanism by which this occurs is poorly understood, and no effective therapeutic strategy has been developed to overcome this resistance to differentiation. Using a neuroblastoma (NB) differentiation model, we have discovered that hypoxia represses the differentiation induced by retinoic acid (RA) as demonstrated by loss of neuron differentiation markers and changes in cell morphology, associated with reduction of global histone acetylation, that are caused by the induction of pyruvate dehydrogenase kinases (PDKs). PDKs phosphorylate pyruvate dehydrogenase (PDH), thereby blocking pyruvate entry into the TCA cycle, reducing acetyl-CoA generation, and promoting the Warburg effect. Genetic and pharmaceutical inhibition of PDK restores histone acetylation and NB cell differentiation morphology. Acetate supplementation restores histone acetylation, along with differentiation markers expression and neuron differentiation. In addition, ATAC-Seq analysis demonstrated that hypoxia treatment significantly reduces chromatin accessibility at RAR/RXR binding sites, which can be restored by acetate supplementation. These findings suggest that (1) combining RA and acetate supplementation represents a potentially effective therapeutic strategy for neuroblastoma treatment; (2) diverting pyruvate away from acetyl-CoA generation is a key mechanism by which the Warburg effect blocks cell differentiation.

Citation Format: Jiangbin Ye, Yang Li, Joshua J. Gruber, Ulrike M. Litzenburger, Yiren Zhou, Yu R. Miao, Edward L. LaGory, Albert M. Li, Zhen Hu, Lori S. Hart, John M. Maris, Howard Y. Chang, Amato J. Giaccia. Deciphering Warburg effect: hypoxia inhibits tumor cell differentiation through reducing acetyl-CoA generation and chromatin accessibility [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 5708.
Abstract 5712: Nutrient scarcity confers breast cancer brain metastasis sensitivity to serine synthesis pathway inhibition
Authors: Ngo Bryan , Kim Eugenie , Doll Sophia , Bustraan Sophia , Luengo Alba , Davidson Shawn M. , Ali Ahmed , Ferraro Gino , Kang Diane , Ni Jing , Liang Roger , Plasger Ariana , Kastenhuber Edward R. , Eskandari Roozbeh , Bacha Sarah , Sriram Roshan , Stein Benjamin D. , Bakhoum Samuel F. , Mullarky Edouard , Snuderl Matija , Mainolfi Nello , Suri Vipin , Friedman Adam , Manfredi Mark , Sabatini David M. , Jones Drew , Yu Min , Zhao Jean J. , Jain Rakesh K. , Heiden Matthew G. Vander , Mann Matthias , Cantley Lewis C. , Pacold Michael E.
Published: September 2020

The metabolic milieu of the brain is severely deprived of nutrients, including the amino acids serine and its catabolite glycine. The metabolic rewiring required for tumor cells to survive in the nutrient-limited environment of the brain and the metabolic vulnerabilities this confers are poorly understood.

Here we demonstrate that cell-intrinsic de novo serine synthesis is a major determinant of triple-negative breast cancer (TNBC) brain metastasis. Whole proteome comparison of TNBC cells that differ in their capacity to colonize the brain reveals that 3-phosphoglycerate dehydrogenase (PHGDH), which catalyzes the rate-limiting step of glucose-derived serine synthesis, is the most significantly upregulated protein in cells that efficiently metastasize to the brain. Expression of catalytically active PHGDH in a non-brain trophic cell line promoted brain metastasis. Furthermore, genetic silencing or pharmacological inhibition of PHGDH attenuated brain metastasis burden in mice.

These findings indicate that nutrient availability determines serine synthesis pathway dependence in brain metastasis, and suggest that PHGDH inhibitors may be useful in the treatment of patients with cancers that have spread to the brain.

Citation Format: Bryan Ngo, Eugenie Kim, Sophia Doll, Sophia Bustraan, Alba Luengo, Shawn M. Davidson, Ahmed Ali, Gino Ferraro, Diane Kang, Jing Ni, Roger Liang, Ariana Plasger, Edward R. Kastenhuber, Roozbeh Eskandari, Sarah Bacha, Roshan Sriram, Benjamin D. Stein, Samuel F. Bakhoum, Edouard Mullarky, Matija Snuderl, Nello Mainolfi, Vipin Suri, Adam Friedman, Mark Manfredi, David M. Sabatini, Drew Jones, Min Yu, Jean J. Zhao, Rakesh K. Jain, Matthew G. Vander Heiden, Matthias Mann, Lewis C. Cantley, Michael E. Pacold. Nutrient scarcity confers breast cancer brain metastasis sensitivity to serine synthesis pathway 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 5712.