Zinc-dependent multimerization of mutant calreticulin is required for MPL binding and MPN pathogenesis

Rivera, J.F., Baral, A.J., Nadat, F., Boyd, G., Smyth, R., Patel, H., Burman, E.L., Alameer, G., Boxall, S.A., Jackson, B.R., Baxter, E.J., Laslo, P., Green, A.R., Kent, D.G., Mullally, A. and Chen, E. 2021. Zinc-dependent multimerization of mutant calreticulin is required for MPL binding and MPN pathogenesis. Blood Advances. 5 (7), pp. 1922-1932. https://doi.org/10.1182/bloodadvances.2020002402

TitleZinc-dependent multimerization of mutant calreticulin is required for MPL binding and MPN pathogenesis
TypeJournal article
AuthorsRivera, J.F., Baral, A.J., Nadat, F., Boyd, G., Smyth, R., Patel, H., Burman, E.L., Alameer, G., Boxall, S.A., Jackson, B.R., Baxter, E.J., Laslo, P., Green, A.R., Kent, D.G., Mullally, A. and Chen, E.
Abstract

Calreticulin (CALR) is mutated in the majority of JAK2/MPL-unmutated myeloproliferative neoplasms (MPNs). Mutant CALR (CALRdel52) exerts its effect by binding to the thrombopoietin receptor MPL to cause constitutive activation of JAK-STAT signaling. In this study, we performed an extensive mutagenesis screen of the CALR globular N-domain and revealed 2 motifs critical for CALRdel52 oncogenic activity: (1) the glycan-binding lectin motif and (2) the zinc-binding domain. Further analysis demonstrated that the zinc-binding domain was essential for formation of CALRdel52 multimers, which was a co-requisite for MPL binding. CALRdel52 variants incapable of binding zinc were unable to homomultimerize, form CALRdel52-MPL heteromeric complexes, or stimulate JAK-STAT signaling. Finally, treatment with zinc chelation disrupted CALRdel52-MPL complexes in hematopoietic cells in conjunction with preferential eradication of cells expressing CALRdel52 relative to cells expressing other MPN oncogenes. In addition, zinc chelators exhibited a therapeutic effect in preferentially impairing growth of CALRdel52-mutant erythroblasts relative to unmutated erythroblasts in primary cultures of MPN patients. Together, our data implicate zinc as an essential cofactor for CALRdel52 oncogenic activity by enabling CALRdel52 multimerization and interaction with MPL, and suggests that perturbation of intracellular zinc levels may represent a new approach to abrogate the oncogenic activity of CALRdel52 in the treatment of MPNs.

JournalBlood Advances
Journal citation5 (7), pp. 1922-1932
ISSN2473-9529
2473-9537
Year2021
PublisherAmerican Society of Hematology
Publisher's version
File Access Level
Open (open metadata and files)
Digital Object Identifier (DOI)https://doi.org/10.1182/bloodadvances.2020002402
Publication dates
Published online05 Apr 2021
Published in print13 Apr 2021

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