Tuning cytokine receptor signaling by re-orienting dimer geometry with surrogate ligands.
Most cell-surface receptors for cytokines and growth factors signal as dimers, but it is unclear whether remodeling receptor dimer topology is a viable strategy to "tune" signaling output. We utilized diabodies (DA) as surrogate ligands in a prototypical dimeric receptor-ligand system, the cytokine Erythropoietin (EPO) and its receptor (EpoR), to dimerize EpoR ectodomains in non-native architectures. Diabody-induced signaling amplitudes varied from full to minimal agonism, and structures of these DA/EpoR complexes differed in EpoR dimer orientation and proximity. Diabodies also elicited biased or differential activation of signaling pathways and gene expression profiles compared to EPO. Non-signaling diabodies inhibited proliferation of erythroid precursors from patients with a myeloproliferative neoplasm due to a constitutively active JAK2V617F mutation. Thus, intracellular oncogenic mutations causing ligand-independent receptor activation can be counteracted by extracellular ligands that re-orient receptors into inactive dimer topologies. This approach has broad applications for tuning signaling output for many dimeric receptor systems.
Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, CA 94305-5345, USA; Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305-5345, USA.
Ignacio
I
Moraga
Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305-5345, USA; Department of Pathology, Division of Hematopathology, Stanford University School of Medicine, Stanford, CA 94305-5345, USA.
Gerlinde
G
Wernig
Division of Biophysics, Department of Biology, University of Osnabrück, 49076 Osnabrück, Germany.
Stephan
S
Wilmes
Ludwig Institute for Cancer Research and de Duve Institute, Université catholique de Louvain, 1200 Brussels, Belgium.
Vitalina
V
Gryshkova
Division of Biophysics, Department of Biology, University of Osnabrück, 49076 Osnabrück, Germany.
Christian P
CP
Richter
Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305-5345, USA; Department of Internal Medicine, Division of Hematology, Stanford University School of Medicine, Stanford, CA 94305-5345, USA.
Wan-Jen
WJ
Hong
Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305-5345, USA.
Rahul
R
Sinha
Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, CA 94305-5345, USA; Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305-5345, USA.
Feng
F
Guo
DiscoveRx, 42501 Albrae Street, Fremont, CA 94538, USA.
Hyna
H
Fabionar
Primity Bio, 3350 Scott Boulevard, Suite 6101, Santa Clara, CA 95054, USA.
Tom S
TS
Wehrman
Primity Bio, 3350 Scott Boulevard, Suite 6101, Santa Clara, CA 95054, USA.
Peter
P
Krutzik
Department of Computational Biology, Department of Computer Science, University of Oxford, Oxford OX1 3QD, UK.
Samuel
S
Demharter
Institut Gustave Roussy, INSERM U1009, 94805 Villejuif, France.
Isabelle
I
Plo
Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305-5345, USA.
Irving L
IL
Weissman
Department of Computational Biology, Department of Computer Science, University of Oxford, Oxford OX1 3QD, UK.
Peter
P
Minary
Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305-5345, USA; Department of Internal Medicine, Division of Hematology, Stanford University School of Medicine, Stanford, CA 94305-5345, USA.
Ravindra
R
Majeti
Ludwig Institute for Cancer Research and de Duve Institute, Université catholique de Louvain, 1200 Brussels, Belgium.
Stefan N
SN
Constantinescu
Division of Biophysics, Department of Biology, University of Osnabrück, 49076 Osnabrück, Germany.
Jacob
J
Piehler
Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, CA 94305-5345, USA; Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305-5345, USA. Electronic address: kcgarcia@stanford.edu.
K Christopher
KC
Garcia
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D018360:Crystallography, X-Ray; D019281:Dimerization; D004921:Erythropoietin / Q000378:metabolism; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D017354:Point Mutation; D015202:Protein Engineering; D017467:Receptors, Erythropoietin / Q000819:agonists / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D016415:Sequence Alignment; D015398:Signal Transduction
D000911:Antibodies, Monoclonal; D017467:Receptors, Erythropoietin; D004921:Erythropoietin; C507924:JAK2 protein, human; D053614:Janus Kinase 2