Tuning of granulopoietic signaling by de novo designed agonists

Ullrich, T.; Pollmann, C.; Ritter, M.; Haaf, J.; Aghaallaei, N.; Tesakov, I.; El-Riz, M.; Maksymenko, K.; Hatskovska, V.; Kandabarau, S.; Klimiankou, M.; Lengerke, C.; Welte, K.; Hernandez Alvarez, B.; Müller, P.; Lupas, A. N.; Piehler, J.; Skokowa, J.; ElGamacy, M.

doi:10.1101/2023.11.25.568662

bioRxiv · 10.1101/2023.11.25.568662

Tuning of granulopoietic signaling by de novo designed agonists

Ullrich, T.·Pollmann, C.·Ritter, M.·Haaf, J.·Aghaallaei, N.·Tesakov, I.·El-Riz, M.·Maksymenko, K.·Hatskovska, V.·Kandabarau, S.·Klimiankou, M.·Lengerke, C.·Welte, K.·Hernandez Alvarez, B.·Müller, P.·Lupas, A. N.·Piehler, J.·Skokowa, J.·ElGamacy, M.

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Abstract

Enhancing cytokine-based therapies by systematically tuning how an agonist associates its receptor is emerging as a powerful new concept in drug discovery. Here, we report the design and characterization of agonists that tune the granulocyte-colony stimulating factor receptor (G-CSFR) activity, which is central for the proliferation and granulocytic differentiation of hematopoietic stem cells. Using design agonists, we study the impact of varying the receptor-binding affinity and dimerization geometry on receptor association, downstream signaling, and cellular response. Hence, we achieved agonists with altered signaling specificities that are hyper-thermostable, can outcompete the native ligand (G-CSF), and bias granulopoietic differentiation over triggering proliferation. Furthermore, the design agonists differentially modulate the kinetics and amplitudes of signal transduction pathways, and gene expression patterns. Unlike G-CSF, they achieve selective activation of gene sets with hematopoietic functions with minimal unwanted effects on immunomodulatory signaling. These findings demonstrate the potential of dissecting the complex G-CSFR signaling, and open up ways for new therapeutic applications for designed cytokines. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=116 SRC="FIGDIR/small/568662v3_ufig1.gif" ALT="Figure 1"> View larger version (30K): org.highwire.dtl.DTLVardef@af56e3org.highwire.dtl.DTLVardef@171920forg.highwire.dtl.DTLVardef@12c2c0aorg.highwire.dtl.DTLVardef@ff7556_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Ullrich, T., Pollmann, C., Ritter, M., Haaf, J., Aghaallaei, N., Tesakov, I., El-Riz, M., Maksymenko, K., Hatskovska, V., Kandabarau, S., Klimiankou, M., Lengerke, C., Welte, K., Hernandez Alvarez, B., Müller, P., Lupas, A. N., Piehler, J., Skokowa, J., ElGamacy, M.. 2023-11-25. Tuning of granulopoietic signaling by de novo designed agonists. https://doi.org/10.1101/2023.11.25.568662

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