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. 2025 Jul 29:S0092-8674(25)00798-6.
doi: 10.1016/j.cell.2025.07.009. Online ahead of print.

Design of soluble Notch agonists that drive T cell development and boost immunity

Affiliations

Design of soluble Notch agonists that drive T cell development and boost immunity

Rubul Mout et al. Cell. .

Abstract

The rational design of receptor agonists to control cell signaling is an emerging strategy for developing disease therapeutics. Creating a soluble cytokine-like agonist for the Notch receptor, which regulates cell fate in embryonic and adult development, is challenging, as receptor activation requires a mechanical force that is usually mediated by cell-associated transmembrane ligands. Here, we exploit computationally designed protein complexes with precise valencies and geometries to generate soluble cytokine-like Notch agonists. These molecules promote cell-cell bridging, cluster Notch receptors at cell synapses, and activate receptor signaling. We show that these agonists drive T cell differentiation from cord blood progenitors and human induced pluripotent stem cells (iPSCs) and in bioreactor production of T cells in liquid suspension. When delivered intravenously in mice, they stimulate cytokine production, expansion of antigen-specific CD4+ T cells, and antibody class switching. These de-novo-designed ligands can be broadly applied to optimize in vitro cell differentiation and advance immunotherapy development.

Keywords: Notch signaling; T cell development; T cell immunity; computational protein design; iPSC-derived T cells; immunotherapy; protein agonist; soluble Notch activation; vaccine.

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Conflict of interest statement

Declaration of interests R.M., R.J., E.D.E., D.B., S.C.B., and G.Q.D. filed patents related to C3-DLL4 soluble Notch agonist. R.M. and G.Q.D. filed patents related to C5(15H)-DLL4 soluble Notch agonist. G.Q.D. is an advisory board member at Cell.

References

    1. Siebel C, and Lendahl U (2017). Notch Signaling in Development, Tissue Homeostasis, and Disease. Physiol Rev 97, 1235–1294. 10.1152/physrev.00005.2017. - DOI - PubMed
    1. Sprinzak D, and Blacklow SC (2021). Biophysics of Notch Signaling. Annu Rev Biophys 50, 157–189. 10.1146/annurev-biophys-101920-082204. - DOI - PMC - PubMed
    1. Luca VC, Kim BC, Ge C, Kakuda S, Wu D, Roein-Peikar M, Haltiwanger RS, Zhu C, Ha T, and Garcia KC (2017). Notch-Jagged complex structure implicates a catch bond in tuning ligand sensitivity. Science 355, 1320–1324. 10.1126/science.aaf9739. - DOI - PMC - PubMed
    1. Biktasova AK, Dudimah DF, Uzhachenko RV, Park K, Akhter A, Arasada RR, Evans JV, Novitskiy SV, Tchekneva EE, Carbone DP, et al. (2015). Multivalent forms of the Notch ligand DLL-1 enhance antitumor T cell immunity in lung cancer and improve efficacy of EGFR targeted therapy. Cancer Res 75, 4728–4741. 10.1158/0008-5472.CAN-14-1154. - DOI - PMC - PubMed
    1. Smyrlaki I, Fördős F, Rocamonde-Lago I, Wang Y, Shen B, Lentini A, Luca VC, Reinius B, Teixeira AI, and Högberg B (2024). Soluble and multivalent Jag1 DNA origami nanopatterns activate Notch without pulling force. Nat Commun 15, 465. 10.1038/s41467-023-44059-4. - DOI - PMC - PubMed

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