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Pilot Screening of TREM1 Inhibitors in Cell-Based Reporter Assays Reflecting TREM1/DAP12 Receptor Assembly and
Natalia Filippova1, Roman Hromov2,3, James Shi4
1Department of Neurology, University of Alabama at Birmingham, Birmingham, Alabama 35233, United States.
Abstract:
Proinflammatory TREM1 receptors expressed on myeloid-derived cells have recently been recognized as a new oncogenic target in cancer, including gliomas. They are established chemotherapeutic targets in neurodegenerative Parkinson's and Alzheimer's diseases, and they also contribute to stroke and sepsis severities. TREM1 activation requires the TREM1/DAP12 interaction for receptor clustering and signal transduction coordinated by TREM1 ligands. Here, we established the quantitative cell-based high-throughput split luciferase assays of DAP12 dimerization, TREM1 dimerization, and TREM1/DAP12 interaction that allow screening of the inhibitory compounds with quantitative dose-responses, IC50 values, and specificity evaluation. The assays are based on the reconstitution of firefly luciferase activity during DAP12 dimerization, TREM1 dimerization, and TREM1/DAP12 interaction, leading to robust luminescence signals in the presence of luciferin. The ligand-dependent and -independent SCHOOL TREM1 inhibitory peptides were utilized for assay validation. Our pilot screen identified several compound scaffolds disrupting DAP12 dimerization, TREM1 dimerization, and the TREM1/DAP12 interaction. The compound potential mechanisms of action and binding sites in the TREM1 and DAP12 complexes were revealed using CB-Dock2 docking software. To our knowledge, this is the first report providing the first generation of pharmacological modulators for TREM1 receptors.
Insights
Researchers developed new assays to screen for compounds that inhibit TREM1 receptors, which are involved in cancer and neurodegenerative diseases. This work identifies the first generation of potential TREM1-targeting drugs.
Area of Science:
- Immunology
- Pharmacology
- Oncology
Background:
- Proinflammatory TREM1 receptors on myeloid cells are implicated in cancer (e.g., gliomas), neurodegenerative diseases (Parkinson's, Alzheimer's), stroke, and sepsis.
- TREM1 receptor activation depends on TREM1/DAP12 interaction for signal transduction, making this complex a potential therapeutic target.
Purpose of the Study:
- To establish quantitative, high-throughput screening assays for identifying inhibitors of TREM1 receptor function.
- To develop the first generation of pharmacological modulators targeting the TREM1 receptor pathway.
Main Methods:
- Development of cell-based, high-throughput split luciferase assays to measure DAP12 dimerization, TREM1 dimerization, and TREM1/DAP12 interaction.
- Assay validation using known TREM1 inhibitory peptides.
- Pilot screening of compound libraries and computational docking (CB-Dock2) to predict mechanisms of action and binding sites.
Main Results:
- Successful establishment of robust, quantitative assays for TREM1 pathway components.
- Identification of several compound scaffolds that disrupt DAP12 dimerization, TREM1 dimerization, and the TREM1/DAP12 interaction.
- Initial characterization of potential drug candidates and their binding interactions within the TREM1/DAP12 complex.
Conclusions:
- The developed assays provide a powerful tool for screening TREM1 inhibitors.
- This study reports the first generation of pharmacological modulators for TREM1 receptors, offering new therapeutic possibilities for associated diseases.
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