TREM2 hit discovery using temperature-related intensity change (TRIC) technology: A proof-of-concept high-throughput
Natalie Fuchs1, Katarzyna Kuncewicz2, Farida El Gaamouch1
1Molecular Imaging Innovations Institute (MI3), Department of Radiology, Weill Cornell Medicine, 1300 York Avenue, New York, NY, 10065, United States.
Abstract:
Triggering receptor expressed on myeloid cells 2 (TREM2) is an immunomodulatory receptor implicated in both neurodegenerative diseases and cancer. Depending on the context, TREM2 agonists or inhibitors hold therapeutic potential. To date, the majority of TREM2-targeted strategies have centered on monoclonal antibodies (mAbs), which face limitations such as poor tissue penetration and potential immunogenic side effects. To overcome these challenges and expand the chemical space for TREM2-targeting agents, we developed a high-throughput screening (HTS) platform to identify novel small molecule TREM2 binders. Using temperature-related intensity change (TRIC) technology in a 384-well plate format (NanoTemper Dianthus), we screened two focused compound libraries comprising over 1,200 molecules. From this screen, 18 preliminary hits (1.44 % hit rate) were identified and subsequently validated by dose-response binding studies using microscale thermophoresis (MST), yielding four validated hits (0.32 % hit rate) with binding affinities in the high to medium micromolar range (e.g., T2337, KD = 22.4 µM). The binding of the top hit, T2337, was further validated using surface plasmon resonance (SPR). Additionally, we assessed the functional activity of all four validated hits in a cellular assay measuring TREM2-mediated Syk phosphorylation in HEK293 cells co-expressing human TREM2 and its adaptor protein DAP12. These findings establish a robust and scalable platform for the discovery of small molecule TREM2 modulators and serve as a proof-of-concept for broader HTS campaigns targeting TREM2.
Insights
Researchers developed a high-throughput screening platform to discover small molecule binders for Triggering Receptor Expressed on Myeloid Cells 2 (TREM2), overcoming limitations of antibody therapies for neurodegenerative diseases and cancer.
Area of Science:
- Biochemistry
- Immunology
- Pharmacology
Background:
- Triggering Receptor Expressed on Myeloid Cells 2 (TREM2) is a key immunomodulatory receptor in neurodegenerative diseases and cancer.
- Current TREM2-targeting therapies, primarily monoclonal antibodies, have limitations including poor tissue penetration and immunogenicity.
- Novel small molecule modulators are needed to expand therapeutic strategies for TREM2-related conditions.
Purpose of the Study:
- To establish a high-throughput screening (HTS) platform for identifying novel small molecule TREM2 binders.
- To overcome the limitations associated with existing antibody-based TREM2-targeting agents.
- To validate the functional activity of identified small molecule TREM2 binders.
Main Methods:
- Utilized temperature-related intensity change (TRIC) technology on a NanoTemper Dianthus platform for HTS.
- Screened over 1,200 compounds from focused libraries.
- Validated hits using microscale thermophoresis (MST) and surface plasmon resonance (SPR), and assessed functional activity via a cellular Syk phosphorylation assay.
Main Results:
- Identified 18 preliminary TREM2 binder hits from over 1,200 screened compounds.
- Validated four small molecule hits with high to medium micromolar binding affinities.
- Confirmed functional activity of validated hits in a cell-based assay measuring TREM2-mediated Syk phosphorylation.
Conclusions:
- Developed and validated a robust, scalable HTS platform for small molecule TREM2 modulator discovery.
- Demonstrated the feasibility of identifying functional small molecule TREM2 binders.
- Provided a proof-of-concept for future HTS campaigns targeting TREM2 for therapeutic development.
