FLI1 level during megakaryopoiesis affects thrombopoiesis and platelet biology

• Published in: Blood 2017-06, Vol.129 (26), p.3486-3494
• Main Authors: Vo, Karen K., Jarocha, Danuta J., Lyde, Randolph B., Hayes, Vincent, Thom, Christopher S., Sullivan, Spencer K., French, Deborah L., Poncz, Mortimer
• Format: Article
• Language: English
• Subjects: Animals; Blood Platelets - metabolism; Cell Differentiation; Cell Line; Humans; Induced Pluripotent Stem Cells; Jacobsen Distal 11q Deletion Syndrome - pathology; Megakaryocytes - cytology; Mice; Mice, SCID; Platelets and Thrombopoiesis; Proto-Oncogene Protein c-fli-1 - blood; Thrombopoiesis
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood-2017-02-770958

Friend leukemia virus integration 1 (FLI1), a critical transcription factor (TF) during megakaryocyte differentiation, is among genes hemizygously deleted in Jacobsen syndrome, resulting in a macrothrombocytopenia termed Paris-Trousseau syndrome (PTSx). Recently, heterozygote human FLI1 mutations have been ascribed to cause thrombocytopenia. We studied induced-pluripotent stem cell (iPSC)–derived megakaryocytes (iMegs) to better understand these clinical disorders, beginning with iPSCs generated from a patient with PTSx and iPSCs from a control line with a targeted heterozygous FLI1 knockout (FLI1+/−). PTSx and FLI1+/− iMegs replicate many of the described megakaryocyte/platelet features, including a decrease in iMeg yield and fewer platelets released per iMeg. Platelets released in vivo from infusion of these iMegs had poor half-lives and functionality. We noted that the closely linked E26 transformation-specific proto-oncogene 1 (ETS1) is overexpressed in these FLI1-deficient iMegs, suggesting FLI1 negatively regulates ETS1 in megakaryopoiesis. Finally, we examined whether FLI1 overexpression would affect megakaryopoiesis and thrombopoiesis. We found increased yield of noninjured, in vitro iMeg yield and increased in vivo yield, half-life, and functionality of released platelets. These studies confirm FLI1 heterozygosity results in pleiotropic defects similar to those noted with other critical megakaryocyte-specific TFs; however, unlike those TFs, FLI1 overexpression improved yield and functionality. •Paris-Trousseau syndrome is solely a result of FLI1 hemizygous deletion, with ETS1 levels being normal.•Elevated FLI1 levels in megakaryocytes do not interfere with and may enhance megakaryopoiesis.