Transdifferentiation of erythroblasts to megakaryocytes using FLI1 and ERG transcription factors

Abstract

Platelet transfusion has been widely used to prevent and treat life-threatening thrombocytopenia; however, preparation of a unit of concentrated platelet for transfusion requires at least 4-6 units of whole blood. At present, a platelet unit from a single donor can be prepared using apheresis, but lack of donors is still a major problem. Several approaches to produce platelets from other sources, such as haematopoietic stem cells and pluripotent stem cells, have been attempted but the system is extremely complicated, time-consuming and expensive. We now report a novel and simpler technology to obtain platelets using transdifferentiation of human bone marrow erythroblasts to megakaryocytes with overexpression of the FLI1 and ERG genes. The obtained transdifferentiated erythroblasts (both from CD71+ and GPA+ erythroblast subpopulations) exhibit typical features of megakaryocytes including morphology, expression of specific genes (cMPL and TUBB1) and a marker protein (CD41). They also have the ability to generate megakaryocytic CFU in culture and produce functional platelets, which aggregate with normal human platelets to form a normal-looking clot. Overexpression of FLI1 and ERG genes is sufficient to transdifferentiate erythroblasts to megakaryocytes that can produce functional platelets.

A screen for Fli-1 transcriptional modulators identifies PKC agonists that induce erythroid to megakaryocytic differentiation and suppress leukemogenesis

“…In contrast, no significant change in the percentage of cells expressing the erythroid marker CD71 was observed (Figure 11E). In this case, leukemic cells that underwent EMC may also express CD71 [45]. As the majority of splenic cells represent erythroleukemia in these mice, the results suggest that over-activation of Fli-1 or activation of the PKC-MAPK-Fli-1 axis by A75 suppress leukemogenesis at least in part by inducing EMD.…”

Single-cell analyses reveal aberrant pathways for megakaryocyte-biased hematopoiesis in myelofibrosis and identify mutant clone-specific targets

“…Expected patterns of expression of transcription factors known to be involved in megakaryocyte and erythroid differentiation were observed (e.g. progressive increase in GATA1, GATA2), as well as antagonistic expression of two key regulators of megakaryocyte-erythroid cell fate decision FLI1 and KLF1 (Bouilloux et al, 2008;Dore and Crispino, 2011;Frontelo et al, 2007;Palii et al, 2019;Siripin et al, 2015) (Fig. 4B, 4C).…”

Towards the Manufacture of Megakaryocytes and Platelets for Clinical Application

“…The use of ESCs poses significant ethical concerns which disable them as a potential cell source. Differentiation of adipose tissue-derived stem cells (ASCs) [43][44][45] or transdifferentiation from erythrocytes [46] and fibroblasts [47,48] have been demonstrated; however, further investigations will be required prior to establishment of those cells as robust sources for PLT production. Currently, native or genetically modified iPSCs are accepted as the most promising cell source for in vitro PLT pharming due to their availability, upscalability, and low ethical concerns.…”

“…Sirpin et al [46] successfully generated PLTs from MKs that have been transdifferentiated from human bone marrow erythroblasts by the overexpression of FLI1 and ERG genes. Within 7-11 days after transduction, CD41+ MKs appeared in culture.…”