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An incomplete trafficking defect to the cell-surface leads to paradoxical thrombocytosis for human and murine MPL P106L

Fabrizia Favale, Kahia Messaoudi, Leila N. Varghese, Siham Boukour, Christian Pecquet, Vitalina Gryshkova, Jean Philippe Defour, Roxana-Irina Albu, Olivier Bluteau, Paola Ballerini, Guy Leverger, Isabelle Plo, Najet Debili, Hana Raslova, Remi Favier, Stefan N. Constantinescu and William Vainchenker

Key Points

  • MPL P106L induces thrombocytosis due to an incomplete trafficking defect that allows very low cell-surface levels.

  • The P106L mutation uncouples MPL signaling from its THPO clearance functions.

Abstract

The mechanisms behind the hereditary thrombocytosis induced by the thrombopoietin (THPO) receptor MPL P106L mutant remain unknown. A complete trafficking defect to the cell surface has been reported, suggesting either weak constitutive activity or nonconventional THPO-dependent mechanisms. Here, we report that the thrombocytosis phenotype induced by MPL P106L belongs to the paradoxical group, where low MPL levels on platelets and mature megakaryocytes (MKs) lead to high serum THPO levels, whereas weak but not absent MPL cell-surface localization in earlier MK progenitors allows response to THPO by signaling and amplification of the platelet lineage. MK progenitors from patients showed no spontaneous growth and responded to THPO, and MKs expressed MPL on their cell surface at low levels, whereas their platelets did not respond to THPO. Transduction of MPL P106L in CD34+ cells showed that this receptor was more efficiently localized at the cell surface on immature than on mature MKs, explaining a proliferative response to THPO of immature cells and a defect in THPO clearance in mature cells. In a retroviral mouse model performed in Mpl−/− mice, MPL P106L could induce a thrombocytosis phenotype with high circulating THPO levels. Furthermore, we could select THPO-dependent cell lines with more cell-surface MPL P106L localization that was detected by flow cytometry and [125I]-THPO binding. Altogether, these results demonstrate that MPL P106L is a receptor with an incomplete defect in trafficking, which induces a low but not absent localization of the receptor on cell surface and a response to THPO in immature MK cells.

  • Submitted June 17, 2016.
  • Accepted November 3, 2016.
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