bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1963: BB102, a highly selective and potent reversible-covalent FGFR4 inhibitor effective in multiple xenograft tumor models
Authors: Wang Qi , Duan Gongping , Hua Wei , Wei Xianglong , Li Min , Wang Junheng , Zhang Xingmin 
Published: March 2024

Abstract
Introduction: Fibroblast growth factor receptor 4 (FGFR4) belongs to the FGFR family which interact with fibroblast growth factors (FGFs) to regulate cell proliferation, differentiation and apoptosis. Due to unique structural domain, FGFR4 is specifically bound by FGF19 with high affinity in the presence of co-receptor β-Klotho. Hyperactivated FGF19/FGFR4 signaling, induced by FGF19/FGFR4 overexpression and FGFR4 mutation, was reported to be associated with the growth, metastasis and invasion of various cancer. Thus, FGFR4 is considered as a novel target to treat cancer with hyperactivated FGF19/FGFR4 signaling. Here we report BB102 as a highly selective and potent reversible-covalent FGFR4 inhibitor.
Methods: BB102 was developed through structure-based drug design, and optimized by SAR analysis and medicinal chemistry iteration. Biochemical and cell assays were applied in evaluation of BB102 inhibitory activity. Preclinical studies including pharmacokinetic (PK) and toxicity were performed in rats and dogs. Xenograft tumor models in mice were conducted to demonstrate the in vivo efficacy of BB102.
Results: As a reversible-covalent FGFR4 inhibitor, about half compound of BB102 dissociated from FGFR4 kinase within 24 h, which was conducive to overcome the fast re-synthesis rate of FGFR4 (&lt; 2 h) in hepatocellular carcinoma cells. BB102 only inhibited kinase activity of FGFR4WT, FGFR4N535K and FGFR4V550L at nanomolar levels in 207 kinase panel screening. BB102 completely blocked the phosphorylation of the downstream protein ERK at 16.7 nM. BB102 exhibited strong inhibitory effects on the cell proliferation in vitro and tumor growth in mouse models inoculated with the high expression of both FGF19 and FGFR4, or FGFR4 mutated tumor cells. PK profile of BB102 showed good oral bioavailability (F&gt;70%) with a nearly linear dose-dependent exposure. The compound had no effects on the respiratory and central nervous systems in rats, cardiovascular system in Beagle dogs or hERG liability in vitro. In comprehensive safety evaluation, the Severely Toxic Dose in 10% (STD10) in rats was 300 mg/kg/day and the Highest Non-Severely Toxic Dose (HNSTD) in dogs were 100 mg/kg/day.
Conclusion: BB102 is a highly selective and potent reversible-covalent FGFR4 inhibitor with promising efficacy in high expression of both FGF19 and FGFR4, and/or FGFR4 mutated tumor models and good safety in preclinical studies. The phase I study of BB102 is in progress.
Citation Format: Qi Wang, Gongping Duan, Wei Hua, Xianglong Wei, Min Li, Junheng Wang, Xingmin Zhang. BB102, a highly selective and potent reversible-covalent FGFR4 inhibitor effective in multiple xenograft tumor models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1963.
Abstract 2001: Autocrine HGF maturation as a targetable regulatory step in cetuximab resistance in colorectal cancer
Authors: Jones Vivian Truong , Graves-Deal Ramona , Cao Zheng , Bogatcheva Galina , Ramirez Marisol A. , Harmych Sarah J. , Higginbotham James N. , Wahoski Claudia C. , Joshi Neeraj , Roland Joseph T. , Ayers Gregory D. , Liu Qi , Coffey Robert J. , Janetka James W. , Singh Bhuminder 
Published: March 2024

Abstract
Although amplifications and mutations in receptor tyrosine kinases (RTKs) act as bona fide oncogenes, in most cancers, RTKs maintain moderate expression and remain wild-type. Consequently, cognate ligands control many facets of tumorigenesis, including resistance to anti-RTK therapies. Herein, we show that the ligands for the RTKs MET and RON, HGF and HGFL, respectively, are synthesized as inactive precursors that are activated by cellular proteases. Our newly generated HGF/HGFL protease inhibitors could overcome both de novo and acquired cetuximab resistance in colorectal cancer (CRC). Conversely, HGF overexpression was necessary and sufficient to induce cetuximab resistance and loss of polarity. Moreover, HGF-induced cetuximab resistance could be overcome by the downstream MET inhibitor, crizotinib, and upstream protease inhibitors. Additionally, HAI-1, an endogenous inhibitor of HGF proteases, (i) was downregulated in CRC, (ii) exhibited increased genomic methylation that correlated with poor prognosis, (iii) HAI-1 expression correlated with cetuximab response in a panel of cancer cell lines, and (iv) exogenous addition of recombinant HAI-1 overcame cetuximab resistance in CC-HGF cells. Thus, we describe a targetable, autocrine HAI-1/Protease/HGF/MET axis in cetuximab resistance in CRC.
Citation Format: Vivian Truong Jones, Ramona Graves-Deal, Zheng Cao, Galina Bogatcheva, Marisol A. Ramirez, Sarah J. Harmych, James N. Higginbotham, Claudia C. Wahoski, Neeraj Joshi, Joseph T. Roland, Gregory D. Ayers, Qi Liu, Robert J. Coffey, James W. Janetka, Bhuminder Singh. Autocrine HGF maturation as a targetable regulatory step in cetuximab resistance in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2001.
Abstract 2003: Chromatin based targeting of a master regulatory complex in estrogen receptor epithelial ovarian cancer
Authors: Jaiswal Sushil K. , Fedkenheuer Kevin , Khamar Ronak , Tan Hua , Gotea Valer , Elkahloun Abdel , Annunziata Christina , Elnitski Laura 
Published: March 2024

Abstract
Ovarian cancer remains a significant contributor to cancer-related mortality in the United States, with low survival rates over the past decades. Hormonal therapies targeting estrogen receptors have shown limited efficacy. To explore mechanism of the inefficacy, our approach involved investigating ESR1 degradation within the higher order chromatin structure formed by Megacomplex. We identified a 1.3MDa Megacomplex containing ESR1, FOXA1 and PITX1, with colocalization at 2,440 genomic positions within the PEO4 cell line. Unlike cytoplasmic ESR1, the nuclear form was degraded minimally by ICI182780 (ICI), moreover, ESR1 incorporated into the Megacomplex macromolecule was least degraded. We hypothesized that this resistance might be due to sequestration within the Megacomplex, thereby preventing ICI from effectively targeting ESR1. Addition of a small molecule inhibitor of active chromatin, JQ1, in combination with ICI effectively enhanced the degradation of ESR1 within Megacomplex. The combination stopped nuclear occupancy of ESR1 and also degraded the cytoplasmic protein, thus breaking the ESR1 signaling circuit responsible for tumor cell proliferation. Pathway enrichment results consistently identified similar gene sets obtained independently for RNA-seq analysis after ESR1 modulation, and ChIP-seq analysis of Megacomplex bound genes. Likewise, proteins enriched within the Megacomplex macromolecule, as revealed from mass-spec analysis, returned similar pathway enrichments, directly implicating Megacomplex in ESR1 ovarian cancer gene regulation. We conclude, ESR1 in Megacomplex is resistant to ICI and sensitized by combining ICI with JQ1. This combined treatment strategy showed inhibition of cell proliferation and viability by uncovering ESR1 within the Megacomplex, thus elucidating a novel drug resistance mechanism.
Citation Format: Sushil K. Jaiswal, Kevin Fedkenheuer, Ronak Khamar, Hua Tan, Valer Gotea, Abdel Elkahloun, Christina Annunziata, Laura Elnitski. Chromatin based targeting of a master regulatory complex in estrogen receptor epithelial ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2003.
Abstract 2045: Novel TFRC humanized mouse model for drug delivery to the brain
Authors: Sonego Fabiane , Pappalardo Angela , Martin Gaëlle , Ozgür Burak , Ravn Peter , Cherifi Yacine , Sotty Florence , Isnard-Petit Patricia , Thiam Kader 
Published: March 2024

Abstract
Gliomas are the most common neoplasm in the central nervous system (CNS). Although immunotherapy has shown efficacy in several tumor types, no immunotherapies are approved by the US Food and Drug Administration for treatment of gliomas. One of the biggest challenges in using immunotherapies in the CNS is the existence of the blood-brain barrier (BBB), which limits drug delivery and the subsequent therapeutic drug efficacy. In addition to biodistribution, assessment of PK in preclinical models do not reflect the clinical findings. Conjugation of therapeutic agents with molecules binding to TRFC expressed along the BBB are being used as one of the alternative approaches to deliver drugs into the CNS. Herein, we describe a novel TFRC humanized mouse model (hTFRC), developed to enable assessment of compounds targeting human TFRC and aiming to deliver drug through the BBB. hTFRC model was developed by knock-in at the mouse endogenous locus to enable a physiological expression of TFRC. Splenocytes from hTFRC mice stimulated with labelled pHrodo Red human transferrin show internalization of human transferrin, suggesting that humanization of the transferrin receptor does not change the endocytosis process. In vivo, treatment of hTFRC mice with anti-BACE1/anti-TFRC antibodies enables the transfer of anti-BACE1 to the brain, while anti-BACE1 antibody is not. As a consequence of the drug delivery, mice treated with anti-BACE1/anti-TFRC show a marked reduction in Amyloid-β 1-40 levels in the brain. Altogether, the data suggest that hTFRC is expressed in the BBB and enables the efficient shuttling of therapeutic antibodies to the brain. While assessment of TFRC-targeting antibodies in a glioma model remains to be performed, the hTFRC model is a promising tool to assess delivery and efficacy of immunotherapies in gliomas. Moreover, the hTFRC model is also being intercrossed with the well-established hSA/FcRn humanized mouse model (Viuff et al., 2016), to enable a more translatable PK and biodistribution of compounds targeting TFRC.
Citation Format: Fabiane Sonego, Angela Pappalardo, Gaëlle Martin, Burak Ozgür, Peter Ravn, Yacine Cherifi, Florence Sotty, Patricia Isnard-Petit, Kader Thiam. Novel TFRC humanized mouse model for drug delivery to the brain [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2045.
Abstract 2060: Automation of 3D cancer spheroid assay for compound screening
Authors: Sirenko Oksana , Lim Angeline , Michlmayr Astrid , Tong Zhisong , Keidel Emilie , Spira Felix 
Published: March 2024

Abstract
Finding efficient drug combinations to treat cancer patients is critical for therapy success. Accordingly, there is a critical need to develop methods for efficient testing drug efficacy to discover new therapeutic targets. 3D cancer models are highly valuable tools for cancer research and drug development. However, the complexity of performing 3D assays remains a hurdle for adopting these methods for compound screening. To alleviate the bottlenecks that come with labor-intensive manual protocols we developed cell culture automation methods using CellXpress.ai instrument. The instrument enables automation of 2D or 3D culture for prolonged complex workflows. The CellXpress.ai provides automated plating, passaging, media exchanges, monitoring organoids, alone with compound treatment and the end point assays with high content image analysis. In the present study we describe full automation of imaging, analysis, and cell culture process which enables scaling up complex 3D cell-based assays and compound screening. As an example of 3D protocol, we automated culture and imaging of colorectal cancer 3D spheroids formed from HCT116 cell line in U-shape low attachment plates. HCT116 cells were expanded in 2D, then spheroids were formed after automated dispensing of cell suspension into U-shape 96 or 384 plates. After 48h spheroids were treated with 12 anti-cancer compounds, at multiple concentrations for 3-5 days, then stained and imaged. Cell plating, compound additions, media exchange, and staining were performed automatically by CellXpressAI instrument. During the culture spheroids were monitored using transmitted light, with immediate analysis of phenotypic changes, including inhibition of growth or spheroid disintegration. For endpoint assay spheroids were stained with combination of Hoechst nuclear stain and viability dyes Calcein AM and EtHD, then imaged, and analyzed for multiple read-outs including spheroid size, marker intensities, and live-dead cell scoring. We observed concentration-dependent inhibition of spheroid growth and cell death in response to anti-cancer compounds and evaluated effective compound concentrations for cytotoxic or cytostatic effects. Cell culture process automation for 3D assays, powered by imaging and image analysis has a potential to bring 3D biology into another level, allowing to increase throughput and reproducibility, and enabling variety of high throughput drug discovery and precision medicine applications.
Citation Format: Oksana Sirenko, Angeline Lim, Astrid Michlmayr, Zhisong Tong, Emilie Keidel, Felix Spira. Automation of 3D cancer spheroid assay for compound screening [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2060.
Abstract 2066: Measuring the activity of SMUG1 DNA glycosylase with a novel sSTRIDE-SMUG1 assay
Authors: Stepien Karolina , Sierpowski Franek , Prucsi Zsombor , Koman Szymon , Jarosz Monika , Waligorska Agnieszka , Kordon-Kiszala Magdalena , Solarczyk Kamil 
Published: March 2024

Abstract
On average, spontaneous deamination of cytosine to aberrant base uracil happens 70-200 times per human genome per day. These mutagenic U/G mispairs are, however, corrected by error-free base repair (BER) initiated by uracil-DNA glycosylases. SMUG1 (single-strand selective monofunctional uracil DNA glycosylase), as one of the major glycosylases, removes uracil and oxidized pyrimidines from DNA. In spite of the name, SMUG1 removes uracil also from double-stranded DNA. Unrepaired by SMUG1 U/G mispairs are mutagenic, and alterations in SMUG1 expression levels have been linked to cancer development. Low SMUG1 transcripts are associated with poor prognosis in breast cancer and SMUG1 loss has been shown to cause PARPi resistance in HR-deficient background. Hence, tools describing SMUG1 activities in DNA damage response may provide new approaches for cancer cell treatment. We report here the development, optimization and validation of an assay measuring the activity of SMUG1. The new assay is based on the STRIDE platform technology, which enables direct and sensitive detection of single- or double-strand DNA breaks in situ, in fixed cells. sSTRIDE-SMUG1 assay detects DNA nicks localized in close proximity to the SMUG1 protein and can thus be utilized as a direct reporter of its activity. The experiments were performed in HAP1 cells. First, in untreated cells we have shown that the sSTRIDE-SMUG1 signals constitute ca. 1-2% of total single-strand DNA breaks foci detected by the classic sSTRIDE assay variant, while the negative controls have shown that the number of false-positive foci does not exceed 15% of the total number of signals. As expected, treatment of cells with a cytotoxic nucleotide hydroxymethyl-deoxyuridine (hmdU) resulted in an increase in the number of detected sSTRIDE-SMUG1 signals. To verify the specificity of the assay, SMUG1 was knocked-down in HAP1 wild-type cells using siRNA and then the level of sSTRIDE-SMUG1 signals was measured in both untreated and hmdU-treated cells. In all tested conditions, SMUG1 expression silencing has resulted in a decrease in sSTRIDE-SMUG1 foci. We believe that the sSTRIDE-SMUG1 assay can prove to be a very useful solution to study DNA repair mechanisms involving aberrant base excision. Additionally, sSTRIDE-SMUG1 measurements may be of the utmost importance in characterization of agents inducing synthetic lethality or trapping BER intermediates.
Citation Format: Karolina Stepien, Franek Sierpowski, Zsombor Prucsi, Szymon Koman, Monika Jarosz, Agnieszka Waligorska, Magdalena Kordon-Kiszala, Kamil Solarczyk. Measuring the activity of SMUG1 DNA glycosylase with a novel sSTRIDE-SMUG1 assay [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2066.
Abstract 2119: Translational research to facilitate development of novel therapeutic combinations of letrozole for the treatment of glioblastoma
Authors: Karve Aniruddha Sunil , Gabani Bhavesh Babulal , Gadgil Sidharth Nitin , Kulkarni Shravani Parag , Gudelsky Gary A. , Desai Pankaj B. 
Published: March 2024

Abstract
Letrozole (LTZ), a third-generation aromatase inhibitor, appears to be a promising novel therapeutic for the treatment of glioblastoma (GBM). It synergistically enhances the efficacy of the DNA alkylating agent temozolomide, one of the few approved chemotherapeutics for GBM, against patient-derived GBM cells. RNA-seq analysis of tumor from recurrent GBM patients treated with LTZ prior to surgery in a “window of opportunity” phase 0/1 dose escalation clinical trial revealed that LTZ downregulates genes encoding for DNA-damage repair proteins (BRCA2) in a dose dependent manner. Our central hypothesis for this project is that combination of LTZ with targeted therapeutic agents that promote DNA damage results in synergistic effects against GBM. The objective of this study is to evaluate the effects of poly-ADP ribose polymerase (PARP) inhibitors in combination with LTZ against patient-derived GBM cells. Our secondary aim for this project is to delineate the blood-brain barrier (BBB) permeability of PARP inhibitors. Employing patient-derived G43, G75 and JHH-136 GBM lines we assessed the influence of PARP inhibitors (olaparib, pamiparib, veliparib and niraparib) in combination with LTZ on cell viability and neurosphere growth. We then utilized male CD-IGS rats (n=6; body wt., 250g) to assess plasma and brain extracellular fluid (ECF) pharmacokinetics (PK) of PARP inhibitors using cerebral microdialysis. The IC50 values for PARP inhibitors in these assays ranged from 0.06 to 11.31 µM. Treatment in combination with LTZ (0.05 µM), a non-cytotoxic concentration of LTZ in these lines, significantly decreased the IC50 values for PARP inhibitors (0.012 to 0.51 µM). Combination Index analysis indicated that this combination was strongly synergistic. Furthermore, the plasma and brain ECF PK analyses suggested that PARP inhibitors such as pamiparib and niraparib exhibit facile transport across the BBB barrier as reflected in Kp,uu,brain (ratio of AUCecf/AUCplasma, unbound) values of 0.98 and 0.45, respectively. Overall, these results provide a strong foundation for pursuing further development of combination therapy of GBM with LTZ and PARP inhibitors. A careful assessment of the clinical efficacy and safety profile of these agents will further guide shortlisting combination of LTZ for pre-clinical and clinical development.
Citation Format: Aniruddha Sunil Karve, Bhavesh Babulal Gabani, Sidharth Nitin Gadgil, Shravani Parag Kulkarni, Gary A. Gudelsky, Pankaj B. Desai. Translational research to facilitate development of novel therapeutic combinations of letrozole for the treatment of glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2119.
Abstract 212: Repression of NKX2-1 promote epithelial-mesenchymal transition and tumor neutrophil recruitment in lung cancer progression
Authors: Chiou Shih-Hwa , Tsai Ping-Hsing , Wang Mong-Lien 
Published: March 2024

Abstract
Background: Malignant transformation is associated with tumor aggressiveness and poor patient outcome. This cellular alteration is due to either activation of oncogenes or the silencing of tumor suppressor genes. NKX2-1 was identified as a lineage-specific gene downregulated in poorly differentiated lung adenocarcinoma (LUAD). However, the molecular mechanism and functional role of infiltrated immune cells in NKX2-1-negative tumors are not clearly understood.
Methods: Clinical relevance of NKX2-1 expression and disease status were evaluated by immunohistochemical analysis of LUAD tissue arrays and the overall survival analysis of TCGA databases. In vitro and in vivo experiments were applied to confirm the role and mechanisms of NKX2-1 actions in transformed cells. The secretion of chemokines was evaluated by using chemokine array. Flow cytometry and single-cell RNA sequencing analysis of mouse tumor tissue were used to demonstrate the infiltration of neutrophils in mouse models.
Results: Low expression of NKX2-1 was observed in high-grade LUAD tumors. LUAD patients who express a pattern of NKX2-1high/TGF-βlow had better survival outcomes than those with NKX2-1low/TGF-βhigh. Mechanistically, TGF-β induced the downregulation of NKX2-1 which led to enhanced epithelial-mesenchymal transition (EMT), cell motility, and CXC chemokine expression. The high release of these chemoattractants promoted the recruitment of neutrophils. Meanwhile, NKX2-1-dependent suppression of CXC chemokines gene expression may be via repressive histone methylation at the promoter region of CXC chemokines in lung cancer cells. In the in vivo syngeneic LUAD models, exacerbated neutrophil infiltration was noted. Single-cell RNA sequencing and flow cytometry both revealed that neutrophils were recruited via CXCLs/CXCR2 activation in LUAD tumors with NKX2-1 knockdown.
Conclusion: Our data showed that NKX2-1 represses tumor growth, metastasis, and neutrophil recruitment via the CXCLs/CXCR2-dependent mechanisms. Notably, targeting CXCR2 in NKX2-1-negative tumors may improve LUAD patient outcomes.
Citation Format: Shih-Hwa Chiou, Ping-Hsing Tsai, Mong-Lien Wang. Repression of NKX2-1 promote epithelial-mesenchymal transition and tumor neutrophil recruitment in lung cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 212.