Summary
This research project aims to investigate the role of erythroblastic island macrophages (EBI Mφ) in normal erythropoiesis and how their dysfunction, particularly due to the JAK2V/F mutation, contributes to disordered erythropoiesis in polycythemia vera (PV).
What they want
The project proposes two main aims: Aim 1 will define the mechanisms by which EBI Mφ support normal erythropoiesis, involving the generation of conditional knockout mice with selective deletion of Epor, Stat5, Igf1, Kitl, or Trf in Mφ, and examining their effects on red cell parameters and EBI formation both in vivo and in vitro. This aim also includes examining gene knockdown effects in human EPOR+ Mφ using an in vitro co-culture system. Aim 2 will define the mechanisms by which JAK2V/F in EBI Mφ contributes to PV pathogenesis, by examining the effects of JAK2 inhibitors on PV mouse models, assessing if selective deletion of Trfc, Igf1, or Kitl in Mφ or Igf1r and cKit in erythroid cells can alleviate erythrocytosis, and using chemical inhibitors to interfere with specific signaling axes. The project will also compare findings in mouse EBI Mφ to human BM EBI Mφ from PV patients.
Technical requirements
- Generation of conditional knockout mice with selective deletion of Epor, Stat5, Igf1, Kitl, or Trf in Mφ
- In vivo and in vitro examination of red cell parameters, erythroblastic island (EBI) formation, and cellular/molecular changes in EBI Mφ and erythroid cells
- In vitro co-culture system for human erythropoiesis using human EPOR+ Mφ
- Studies using JAK2 inhibitors on PV mouse models (Jak2V/F expressed in Mφ)
- Selective deletion of Trfc, Igf1, or Kitl in Mφ in mice to alleviate erythrocytosis
- Selective deletion of Igf1r and cKit in erythroid cells to alleviate erythrocytosis
- Use of chemical inhibitors to interfere with Igf1-Igf1r axis or Scf-cKit axis
- Analysis of human bone marrow EBI Mφ from PV patients
