SynNotch Receptors for Visualizing Immunoreceptor Force Transmission and Downstream Signaling In Vivo
Year: 2026
Type: article
Abstract: Immunoreceptors experience forces that modulate their activities; however, demonstrating this in vivo has been limited by technical challenges. As a first step toward meeting this challenge, we adapted a synthetic Notch (SynNotch) receptor system to report force transmission through immunoreceptors in vivo by replacing the native ligand-binding domain with a receptor-specific antibody and rewiring Notch signaling to drive EGFP or luciferase expression. We expressed SynNotch on Jurkat cells targeting CD40 or T cell receptor (TCR) and characterized their activation in coculture with B or T cells, defining the requirements, optimal conditions, and kinetics of activation. Using complementary mechanobiology approaches, we quantified the exogenous force required for reporter activation and verified that activation depends on forces generated by receptor-expressing sender cells rather than SynNotch-expressing receiver cells. By implanting sensors and targets into immunocompromised mice, we visualized mechanically activated reporter expression on CD40 and TCR-targeting SynNotch cells in vivo. Furthermore, CD40 and TCR signaling was amplified when the receptor bore force against mechanical support from immobilized ligand, indicating that force functions as biologically relevant co-stimulus. Together, our results establish mechanically activated SynNotch reporters as a useful strategy for detecting receptor-associated mechanical signaling across 2D coculture, 3D organoid, and in vivo systems.
Source: Advanced Science
DOI: 10.1002/advs.77318
Language: English
Subfield: Cell Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $6,730
Funders: National Science Foundation, Emory University, National Research Foundation, National Institutes of Health, National Cancer Institute, National Institute of General Medical Sciences, National Institute of Biomedical Imaging and Bioengineering, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Common Fund
Awards: G157881262, Exploiting the Mechanobiology of PD-1 for Cancer Immunotherapy, G1868178788, Hydrogel-Based Aged Immune Organoids to Study Epigenetics and Trajectory of B Cells, G2520018028, Hydrogel-based lymphoid tissues for generation of activated human B cells and delivery in vivo, G2876031932, G2969719073, Lymphoid Experimental Therapeutics Platform to Study Cooperative Signaling inHuman Lymphomas, G3908227681, Noninvasive and predictive biomarkers of organ transplant rejection, G5421118785, G5957808327, G6983735952, G7727228102, G7940966131, G8568536931, Dysregulated mechanoimmunology of epigenetics-driven lymphomas, Center on Probes for Molecular Mechanotechnology, T32 Research Training Program in ImmunoEngineering, Finding Sleeping Beauty: T Cell Biosensors for Dormant Cancer Detection, Hydrogel-based Organoids of African-American Lymphomas to Study B Cell Receptor Pathway Inhibitors
OpenAlex ID: W7204204886