Pilot Screening of TREM1 Inhibitors in Cell-Based Reporter Assays Reflecting TREM1/DAP12 Receptor Assembly and Functionality.

in ACS chemical neuroscience by Natalia Filippova, Roman Hromov, James Shi, Peter H King, Louis B Nabors

TLDR

  • A high-throughput split luciferase assay was developed to screen for TREM1 inhibitors and identify pharmacological modulators.
  • The study identified several compound scaffolds disrupting TREM1/DAP12 interaction, providing the first generation of TREM1 receptor modulators.
  • The findings have implications for understanding and treating TREM1-mediated diseases, including cancer, Parkinson's, Alzheimer's, stroke, and sepsis.

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, ICvalues, 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.

Overview

  • The study aimed to develop pharmacological modulators targeting proinflammatory TREM1 receptors on myeloid-derived cells, a new oncogenic target in cancer and other diseases.
  • The researchers established a high-throughput split luciferase assay to screen for inhibitory compounds and evaluate their dose-responses, IC values, and specificity.
  • The study identified compound scaffolds disrupting TREM1/DAP12 interaction and revealed potential mechanisms of action and binding sites using CB-Dock2 docking software.

Comparative Analysis & Findings

  • The assay was validated using ligand-dependent and -independent TREM1 inhibitory peptides, showing robust luminescence signals in the presence of luciferin.
  • Several compound scaffolds were identified that disrupted DAP12 dimerization, TREM1 dimerization, and the TREM1/DAP12 interaction.
  • The study provided the first generation of pharmacological modulators for TREM1 receptors, a significant breakthrough in understanding TREM1-mediated diseases.

Implications and Future Directions

  • The findings have implications for the development of new treatments for cancer, Parkinson's, Alzheimer's, stroke, and sepsis, as well as for understanding the role of TREM1 in these diseases.
  • Future studies can focus on optimizing the identified compounds and exploring their therapeutic potential in vivo.
  • The development of pharmacological modulators for TREM1 receptors may provide a new approach for treating TREM1-mediated diseases, offering a potential breakthrough in therapeutic strategies.
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