A high-content screen for small-molecule regulators of epithelial cell-adhesion molecule (EpCAM) cleavage yields a robust inhibitor

Tretter, Jana Ylva; Schorpp, Kenji; Luxenburger, Elke; Trambauer, Johannes; Steiner, Harald; Hadian, Kamyar; Gires, Olivier; Niessing, Dierk

doi:10.1074/jbc.ra118.002776

Cited by 5 publications

(5 citation statements)

References 68 publications

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“…This strategy allowed reducing from around 14,000 putative hits to 3516 hits with high confidence. HTS step two, where drug potency was evaluated by dose-response curves, estimated pharmacodynamic properties of drugs that underlie the basis of most molecular interactions in biological systems 51 . Furthermore, using both human and murine cell systems in parallel safeguarded interspecies activity of molecules crucial for future preclinical and clinical trials 52 .…”

Section: Discussionmentioning

confidence: 99%

Phenotypic high-throughput screening platform identifies novel chemotypes for necroptosis inhibition

et al. 2020

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Regulated necrosis or necroptosis, mediated by receptor-interacting kinase 1 (RIPK1), RIPK3 and pseudokinase mixed lineage kinase domain-like protein (MLKL), contributes to the pathogenesis of inflammatory, infectious and degenerative diseases. Recently identified necroptosis inhibitors display moderate specificity, suboptimal pharmacokinetics, off-target effects and toxicity, preventing these molecules from reaching the clinic. Here, we developed a cell-based high-throughput screening (HTS) cascade for the identification of small-molecule inhibitors of necroptosis. From the initial library of over 250,000 compounds, the primary screening phase identified 356 compounds that strongly inhibited TNF-α-induced necroptosis, but not apoptosis, in human and murine cell systems, with EC 50 < 6.7 μM. From these, 251 compounds were tested for RIPK1 and/or RIPK3 kinase inhibitory activity; some were active and several have novel mechanisms of action. Based on specific chemical descriptors, 110 compounds proceeded into the secondary screening cascade, which then identified seven compounds with maximum ability to reduce MLKL activation, IC 50 >100 μM, EC 50 2.5-11.5 μM under long-term necroptosis execution in murine fibroblast L929 cells, and full protection from ATP depletion and membrane leakage in human and murine cells. As a proof of concept, compound SN-6109, with binding mode to RIPK1 similar to that of necrostatin-1, confirmed RIPK1 inhibitory activity and appropriate pharmacokinetic properties. SN-6109 was further tested in mice, showing efficacy against TNF-α-induced systemic inflammatory response syndrome. In conclusion, a phenotypic-driven HTS cascade promptly identified robust necroptosis inhibitors with in vivo activity, currently undergoing further medicinal chemistry optimization. Notably, the novel hits highlight the opportunity to identify new molecular mechanisms of action in necroptosis.

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Section: Discussionmentioning

confidence: 99%

Phenotypic high-throughput screening platform identifies novel chemotypes for necroptosis inhibition

et al. 2020

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“…To evaluate the diagnostic value of candidate genes, multivariate Logistic regression analysis was performed on 11 DE-mRNAs, and EPCAM and RAB30 were nally screened. EPCAM is a type I transmembrane protein that regulates cell cycle progression and differentiation [22]. The extracellular domain of EPCAM can signi cantly enhance osteogenesis of mesenchymal stem cells under differentiation conditions [23].…”

Section: Discussionmentioning

confidence: 99%

Construction of potential periodontitis-related miRNA-mRNA regulatory network

Zhang¹,

Zheng²,

Bian³

et al. 2020

Preprint

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Background MicroRNAs (miRNAs) are found to be involved in the pathogenesis of periodontitis, a major cause of tooth loss in adults. However, a comprehensive miRNA-mRNA regulatory network has still not been established. Methods One miRNA expression profile and 2 gene expression profiles were downloaded from the GEO database and analyzed using GEO2R. Candidate genes commonly appeared in differentially expressed mRNAs (DE-mRNAs) and target genes of differentially expressed miRNAs (DE-miRNAs) were selected for functional and pathway enrichment analyses using Enrichr database. Multivariate Logistic regression analysis was used to screen independent variables among candidate genes. The diagnostic values of screened genes were determined by the area under the receiver operating characteristic (ROC) curve (AUC). Results A total of 5 DE-miRNAs (4 upregulated and 1 downregulated) and 11 candidate genes (3 upregulated and 8 downregulated) were screened. After the construction of miRNA-mRNA regulatory network, 12 miRNA-mRNA pairs were identified. In the network, the upregulated genes were significantly enriched in cellular triglyceride homeostasis and positive regulation of B cell differentiation, whereas the downregulated genes were enriched in vesicle organization, negative regulation of lymphocyte and leukocyte migration. EPCAM and RAB30 were screened as risk factors of periodontitis. The combined AUC of these 2 genes was 0.896 (GSE10334) and 0.916 (GSE16134). Conclusion In this study, we established a potential periodontitis-related miRNA-mRNA regulatory network, which brings new insights into the molecular mechanisms and provides key clues in seeking novel therapeutic targets for periodontitis. In the future, more experiments need to be carried out to validate our current findings.

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“…However, relatively few pharmacological inhibitors against EpCAM have been developed. Recently, high-content screening of a small molecule compound library yielded several robust inhibitors of EpCAM signaling [118]. Indeed, several of the compounds reduced EpCAM-mediated cyclin D1 (CCND1) expression in colorectal carcinoma cell line HCT-8; however, they have yet to be tested against tumor growth [118].…”

Section: Pharmacologic Inhibitorsmentioning

confidence: 99%

“…Recently, high-content screening of a small molecule compound library yielded several robust inhibitors of EpCAM signaling [118]. Indeed, several of the compounds reduced EpCAM-mediated cyclin D1 (CCND1) expression in colorectal carcinoma cell line HCT-8; however, they have yet to be tested against tumor growth [118]. Other mechanisms of pharmacological inhibition of EpCAM have targeted EpCAM-related pathways instead.…”

Section: Pharmacologic Inhibitorsmentioning

confidence: 99%

Informed by Cancer Stem Cells of Solid Tumors: Advances in Treatments Targeting Tumor-Promoting Factors and Pathways

MacLean,

Walker,

Arun

et al. 2024

IJMS

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Cancer stem cells (CSCs) represent a subpopulation within tumors that promote cancer progression, metastasis, and recurrence due to their self-renewal capacity and resistance to conventional therapies. CSC-specific markers and signaling pathways highly active in CSCs have emerged as a promising strategy for improving patient outcomes. This review provides a comprehensive overview of the therapeutic targets associated with CSCs of solid tumors across various cancer types, including key molecular markers aldehyde dehydrogenases, CD44, epithelial cellular adhesion molecule, and CD133 and signaling pathways such as Wnt/β-catenin, Notch, and Sonic Hedgehog. We discuss a wide array of therapeutic modalities ranging from targeted antibodies, small molecule inhibitors, and near-infrared photoimmunotherapy to advanced genetic approaches like RNA interference, CRISPR/Cas9 technology, aptamers, antisense oligonucleotides, chimeric antigen receptor (CAR) T cells, CAR natural killer cells, bispecific T cell engagers, immunotoxins, drug-antibody conjugates, therapeutic peptides, and dendritic cell vaccines. This review spans developments from preclinical investigations to ongoing clinical trials, highlighting the innovative targeting strategies that have been informed by CSC-associated pathways and molecules to overcome therapeutic resistance. We aim to provide insights into the potential of these therapies to revolutionize cancer treatment, underscoring the critical need for a multi-faceted approach in the battle against cancer. This comprehensive analysis demonstrates how advances made in the CSC field have informed significant developments in novel targeted therapeutic approaches, with the ultimate goal of achieving more effective and durable responses in cancer patients.

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A high-content screen for small-molecule regulators of epithelial cell-adhesion molecule (EpCAM) cleavage yields a robust inhibitor

Cited by 5 publications

References 68 publications

Phenotypic high-throughput screening platform identifies novel chemotypes for necroptosis inhibition

Phenotypic high-throughput screening platform identifies novel chemotypes for necroptosis inhibition

Construction of potential periodontitis-related miRNA-mRNA regulatory network

Informed by Cancer Stem Cells of Solid Tumors: Advances in Treatments Targeting Tumor-Promoting Factors and Pathways

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