Proteasome function is required for platelet production
Dallas S. Shi, … , Dean Y. Li, Andrew S. Weyrich
Dallas S. Shi, … , Dean Y. Li, Andrew S. Weyrich
Published July 25, 2014
Citation Information: J Clin Invest. 2014;124(9):3757-3766. https://doi.org/10.1172/JCI75247.
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Research Article Hematology

Proteasome function is required for platelet production

Abstract

The proteasome inhibiter bortezomib has been successfully used to treat patients with relapsed multiple myeloma; however, many of these patients become thrombocytopenic, and it is not clear how the proteasome influences platelet production. Here we determined that pharmacologic inhibition of proteasome activity blocks proplatelet formation in human and mouse megakaryocytes. We also found that megakaryocytes isolated from mice deficient for PSMC1, an essential subunit of the 26S proteasome, fail to produce proplatelets. Consistent with decreased proplatelet formation, mice lacking PSMC1 in platelets (Psmc1fl/fl Pf4-Cre mice) exhibited severe thrombocytopenia and died shortly after birth. The failure to produce proplatelets in proteasome-inhibited megakaryocytes was due to upregulation and hyperactivation of the small GTPase, RhoA, rather than NF-κB, as has been previously suggested. Inhibition of RhoA or its downstream target, Rho-associated protein kinase (ROCK), restored megakaryocyte proplatelet formation in the setting of proteasome inhibition in vitro. Similarly, fasudil, a ROCK inhibitor used clinically to treat cerebral vasospasm, restored platelet counts in adult mice that were made thrombocytopenic by tamoxifen-induced suppression of proteasome activity in megakaryocytes and platelets (Psmc1fl/fl Pdgf-Cre-ER mice). These results indicate that proteasome function is critical for thrombopoiesis, and suggest inhibition of RhoA signaling as a potential strategy to treat thrombocytopenia in bortezomib-treated multiple myeloma patients.

Authors

Dallas S. Shi, Matthew C.P. Smith, Robert A. Campbell, Patrick W. Zimmerman, Zechariah B. Franks, Bjorn F. Kraemer, Kellie R. Machlus, Jing Ling, Patrick Kamba, Hansjörg Schwertz, Jesse W. Rowley, Rodney R. Miles, Zhi-Jian Liu, Martha Sola-Visner, Joseph E. Italiano Jr., Hilary Christensen, Walter H.A. Kahr, Dean Y. Li, Andrew S. Weyrich

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Figure 1

Pharmacologic inhibition of the proteasome induces thrombocytopenia in mice by decreasing platelet production.

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Pharmacologic inhibition of the proteasome induces thrombocytopenia in m...
(A) Mice were treated with a bolus of bortezomib (Bort) or vehicle (Veh), and platelet counts and platelet proteasome activity were measured at the indicated times. Data are mean ± SEM of 6 experiments. (B) Mouse platelets were labeled in vivo with Dylight 488, as described in Methods. In parallel, the mice were treated with a bolus of bortezomib or vehicle, and the percentage of labeled platelets was determined at 24, 48, and 96 hours after treatment. Data are mean ± SEM of 6 independent experiments. (C) Mouse platelets were depleted in the presence of bortezomib or its vehicle, as described in Methods. The percentage of platelets relative to baseline control (0 hours) is shown. Data are mean ± SEM of 5 independent experiments. Note that A–C are derived from separate experiments. *P < 0.05 vs. vehicle; #P < 0.05 vs. 0 hours.