A strategy to design protein-based antagonists against type I cytokine receptors

PLoS Biol. 2024 Nov 26;22(11):e3002883. doi: 10.1371/journal.pbio.3002883. eCollection 2024 Nov.

Abstract

Excessive cytokine signaling resulting from dysregulation of a cytokine or its receptor can be a main driver of cancer, autoimmune, or hematopoietic disorders. Here, we leverage protein design to create tailored cytokine receptor blockers with idealized properties. Specifically, we aimed to tackle the granulocyte-colony stimulating factor receptor (G-CSFR), a mediator of different types of leukemia and autoinflammatory diseases. By modifying designed G-CSFR binders, we engineered hyper-stable proteins that function as nanomolar signaling antagonists. X-ray crystallography showed atomic-level agreement with the experimental structure of an exemplary design. Furthermore, the most potent design blocks G-CSFR in acute myeloid leukemia cells and primary human hematopoietic stem cells. Thus, the resulting designs can be used for inhibiting or homing to G-CSFR-expressing cells. Our results also demonstrate that similarly designed cytokine mimics can be used to derive antagonists to tackle other type I cytokine receptors.

MeSH terms

  • Crystallography, X-Ray / methods
  • Cytokines / metabolism
  • Drug Design
  • HEK293 Cells
  • Hematopoietic Stem Cells / drug effects
  • Hematopoietic Stem Cells / metabolism
  • Humans
  • Leukemia, Myeloid, Acute / drug therapy
  • Leukemia, Myeloid, Acute / metabolism
  • Protein Binding
  • Protein Engineering / methods
  • Receptors, Granulocyte Colony-Stimulating Factor* / metabolism
  • Signal Transduction / drug effects

Substances

  • Receptors, Granulocyte Colony-Stimulating Factor
  • Cytokines

Grants and funding

This project has received funding from the M. Schickedanz Kinderkrebsstiftung (ME, JS), Deutsche Forschungsgemeinschaft (DFG: No 500215849 to ME, JS; and PI 405/15 to JP), José Carreras Leukämie-Stiftung (DJCLS 10 R/2024 to ME, JS), and internal funds from the Max Planck Society (TU, MDH). The funders did not play any role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.