TY - JOUR
T1 - Tuning MPL signaling to influence hematopoietic stem cell differentiation and inhibit essential thrombocythemia progenitors
AU - Cui, Lu
AU - Moraga, Ignacio
AU - Lerbs, Tristan
AU - Van Neste, Camille
AU - Wilmes, Stephan
AU - Tsutsumi, Naotaka
AU - Trotman-Grant, Aaron Claudius
AU - Gakovic, Milica
AU - Andrews, Sarah
AU - Gotlib, Jason
AU - Darmanis, Spyros
AU - Enge, Martin
AU - Quake, Stephen
AU - Hitchcock, Ian S.
AU - Piehler, Jacob
AU - Garcia, K. Christopher
AU - Wernig, Gerlinde
N1 - Funding Information:
K.C.G. acknowledges NIH-RO1-AI51321, the Ludwig Institute, the Mathers Foundation, and HHMI for funding; G.W. acknowledges the National Heart, Lung, and Blood Institute, the Ludwig Institute, Scleroderma Research Foundation (SRF), and the Emmerson Collective. Deutsche Forschungsgesellschaft (DFG) provided salary support to T.L. and I.S.H. was supported by Cancer Research UK grant A24593 and by the University of York Technology Facility.
Publisher Copyright:
© This open access article is distributed under Creative Commons Attribution-NonCommercialNoDerivatives License 4.0 (CC BY-NC-ND).
PY - 2021/1/12
Y1 - 2021/1/12
N2 - Thrombopoietin (TPO) and the TPO-receptor (TPO-R, or c-MPL) are essential for hematopoietic stem cell (HSC) maintenance and megakaryocyte differentiation. Agents that can modulate TPO-R signaling are highly desirable for both basic research and clinical utility. We developed a series of surrogate protein ligands for TPO-R, in the form of diabodies (DBs), that homodimerize TPO-R on the cell surface in geometries that are dictated by the DB receptor binding epitope, in effect "tuning" downstream signaling responses. These surrogate ligands exhibit diverse pharmacological properties, inducing graded signaling outputs, from full to partial TPO agonism, thus decoupling the dual functions of TPO/TPO-R. Using single-cell RNA sequencing and HSC self-renewal assays we find that partial agonistic diabodies preserved the stem-like properties of cultured HSCs, but also blocked oncogenic colony formation in essential thrombocythemia (ET) through inverse agonism. Our data suggest that dampening downstream TPO signaling is a powerful approach not only for HSC preservation in culture, but also for inhibiting oncogenic signaling through the TPO-R.
AB - Thrombopoietin (TPO) and the TPO-receptor (TPO-R, or c-MPL) are essential for hematopoietic stem cell (HSC) maintenance and megakaryocyte differentiation. Agents that can modulate TPO-R signaling are highly desirable for both basic research and clinical utility. We developed a series of surrogate protein ligands for TPO-R, in the form of diabodies (DBs), that homodimerize TPO-R on the cell surface in geometries that are dictated by the DB receptor binding epitope, in effect "tuning" downstream signaling responses. These surrogate ligands exhibit diverse pharmacological properties, inducing graded signaling outputs, from full to partial TPO agonism, thus decoupling the dual functions of TPO/TPO-R. Using single-cell RNA sequencing and HSC self-renewal assays we find that partial agonistic diabodies preserved the stem-like properties of cultured HSCs, but also blocked oncogenic colony formation in essential thrombocythemia (ET) through inverse agonism. Our data suggest that dampening downstream TPO signaling is a powerful approach not only for HSC preservation in culture, but also for inhibiting oncogenic signaling through the TPO-R.
KW - Hematopoietic stem cells
KW - Megakaryopoiesis
KW - Myeloproliferative neoplasm
KW - Thrombopoietin signaling
UR - https://www.biorxiv.org/content/10.1101/2020.09.23.290593v1
UR - http://www.scopus.com/inward/record.url?scp=85098984754&partnerID=8YFLogxK
U2 - 10.1073/pnas.2017849118
DO - 10.1073/pnas.2017849118
M3 - Article
C2 - 33384332
SN - 0027-8424
VL - 118
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 2
M1 - e2017849118
ER -