Our long-term aim is to elucidate the cellular and molecular events underlying the early ontogeny of the hematopoietic system in the mammalian embryo. While definitive hematopoiesis in the fetus and adult is known to be multilineage, primitive hematopoiesis in the yolk sac is centered on the synthesis of erythroid cells that are critical for survival of the embryo. We recently made the seminal observation that primitive erythroblasts in the mouse embryo ultimately enucleate. However, the process and mechanisms regulating enucleation of primitive erythroid cells remain unknown and are the subject of this renewal. In the first aim of this proposal, we will test the hypothesis that primitive erythroblasts respond to chemotactic stimuli, including SDF-1, that facilitate the migration of hematopoietic stem and progenitor cells to the fetal liver. In the second aim, we will test the hypothesis that primitive erythroid cells can and do interact with fetal liver macrophage cells, using erythroblast island reconstitution and enucleation assays established in the laboratory. The adhesion molecules emp and ICAM4 each play important roles in definitive erythroid-macrophage interactions. We have obtained preliminary data that primitive erythroblasts express both emp and ICAM4 and in the third aim we will test the hypothesis that these molecules are functionally important for terminal maturation of primitive erythroblasts. Finally, we have recently discovered that primitive erythroid cells in mice lacking c-myb fail to enucleate and in the fourth aim we will test the hypothesis that this enucleation failure is erythroid cell non-autonomous. These studies of terminal erythroid maturation are facilitated by the synchronous differentiation and enucleation of primary primitive erythroid cells. Furthermore, these studies will allow us to compare and contrast the terminal maturation of primitive erythroid cells with definitive erythroid cells that mature extravascularly in erythroblast islands. A better understanding of the mechanisms responsible for terminal erythroid maturation will ultimately lead to the efficient and complete ex vivo synthesis of erythrocytes to serve an ever-expanding clinical need for red blood cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
3R01DK079361-12S2
Application #
8009927
Study Section
Erythrocyte and Leukocyte Biology Study Section (ELB)
Program Officer
Bishop, Terry Rogers
Project Start
2010-01-18
Project End
2010-12-31
Budget Start
2010-01-18
Budget End
2010-12-31
Support Year
12
Fiscal Year
2010
Total Cost
$99,875
Indirect Cost
Name
University of Rochester
Department
Pediatrics
Type
Schools of Dentistry
DUNS #
041294109
City
Rochester
State
NY
Country
United States
Zip Code
14627
Zhou, Sitong; Huang, Yu-Shan; Kingsley, Paul D et al. (2017) Microfluidic assay of the deformability of primitive erythroblasts. Biomicrofluidics 11:054112
Huang, Yu-Shan; Delgadillo, Luis F; Cyr, Kathryn H et al. (2017) Circulating primitive erythroblasts establish a functional, protein 4.1R-dependent cytoskeletal network prior to enucleating. Sci Rep 7:5164
Frame, Jenna M; Fegan, Katherine H; Conway, Simon J et al. (2016) Definitive Hematopoiesis in the Yolk Sac Emerges from Wnt-Responsive Hemogenic Endothelium Independently of Circulation and Arterial Identity. Stem Cells 34:431-44
McGrath, Kathleen E; Frame, Jenna M; Palis, James (2015) Early hematopoiesis and macrophage development. Semin Immunol 27:379-87
McGrath, Kathleen E; Frame, Jenna M; Fegan, Katherine H et al. (2015) Distinct Sources of Hematopoietic Progenitors Emerge before HSCs and Provide Functional Blood Cells in the Mammalian Embryo. Cell Rep 11:1892-904
Bulger, Michael; Palis, James (2015) Environmentally-defined enhancer populations regulate diversity of tissue-resident macrophages. Trends Immunol 36:61-2
Frame, Jenna M; McGrath, Kathleen E; Palis, James (2013) Erythro-myeloid progenitors: ""definitive"" hematopoiesis in the conceptus prior to the emergence of hematopoietic stem cells. Blood Cells Mol Dis 51:220-5
Waugh, Richard E; Huang, Yu-Shan; Arif, Binish J et al. (2013) Development of membrane mechanical function during terminal stages of primitive erythropoiesis in mice. Exp Hematol 41:398-408.e2