The continuous life-long production of all mature blood cells in the circulation and hematopoietic tissues is contingent on hematopoietic stem cells (HSC). These rare cells reside in the bone marrow of the adult and are the clinically important cells in transplantation therapies for blood-related diseases and leukemias. The manipulation of limited numbers of adult HSCs has been difficult, since ex vivo expansion of these cells has not yet been achieved. Despite the identification and use of many hematopoietic growth factors, HSCs can at best be maintained, not expanded. We hypothesized that at some early stage in ontogeny HSCs are generated and expanded in a unique embryonic microenvironment. An understanding of the developmental processes leading to HSC emergence in the embryo should provide novel insights and improved methods for the ex vivo expansion of HSCs for clinical use. To this end, our long-term research objective has been to determine the cellular and molecular mechanisms by which HSCs are generated/expanded within the mammalian embryo. We have shown that the mouse aorta-gonad-mesonephros (AGM) region autonomously generates the first fully potent HSCs in close association with the aortic endothelium. We have demonstrated that Runxl and GATA-2 transcription factors are required for the generation and expansion of these HSCs. However, more factors have yet to be identified. In various preliminary studies of the AGM microenvironment, IL-3, BMP-4 and hedgehog factors have been implicated as effectors of AGM HSC emergence, but these factors must be further characterized. Thus, we will examine each factor for its role in the induction and expansion of HSCs in the midgestation mouse aorta. Using classical embryologic methods, we will deliver factors to whole cultured mouse embryos by bead/cell pellet implantation and transgenesis, to determine their effect on AGM HSCs. In similar factor manipulation experiments, we aim to establish the lineage relationships of precursors to AGM HSCs by dye marking of aortic endothelium. We will also investigate the long-term fate of the HSCs generated in the mouse AGM by Cre-lox recombination technology. If indeed embryonic aorta-derived HSCs migrate and colonize the adult bone marrow, our insights into their generation and expansion should lead to novel methods for clinical HSC expansion and thus, improve transplantation therapies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37DK054077-13
Application #
7895619
Study Section
Hematopoiesis Study Section (HP)
Program Officer
Bishop, Terry Rogers
Project Start
1998-06-01
Project End
2011-03-31
Budget Start
2010-04-01
Budget End
2011-03-31
Support Year
13
Fiscal Year
2010
Total Cost
$245,662
Indirect Cost
Name
Erasmus Medical Center
Department
Type
DUNS #
411761687
City
Rotterdam
State
Country
Netherlands
Zip Code
3015 -GE
Kauts, Mari-Liis; Rodriguez-Seoane, Carmen; Kaimakis, Polynikis et al. (2018) In Vitro Differentiation of Gata2 and Ly6a Reporter Embryonic Stem Cells Corresponds to In Vivo Waves of Hematopoietic Cell Generation. Stem Cell Reports 10:151-165
Eich, Christina; Arlt, Jochen; Vink, Chris S et al. (2018) In vivo single cell analysis reveals Gata2 dynamics in cells transitioning to hematopoietic fate. J Exp Med 215:233-248
Kaimakis, Polynikis; de Pater, Emma; Eich, Christina et al. (2016) Functional and molecular characterization of mouse Gata2-independent hematopoietic progenitors. Blood 127:1426-37
Crisan, Mihaela; Solaimani Kartalaei, Parham; Neagu, Alex et al. (2016) BMP and Hedgehog Regulate Distinct AGM Hematopoietic Stem Cells Ex Vivo. Stem Cell Reports 6:383-95
Li, Zhuan; Vink, Chris S; Mariani, Samanta A et al. (2016) Subregional localization and characterization of Ly6aGFP-expressing hematopoietic cells in the mouse embryonic head. Dev Biol 416:34-41
Crisan, Mihaela; Dzierzak, Elaine (2016) The many faces of hematopoietic stem cell heterogeneity. Development 143:4571-4581
Kauts, Mari-Liis; Vink, Chris S; Dzierzak, Elaine (2016) Hematopoietic (stem) cell development - how divergent are the roads taken? FEBS Lett 590:3975-3986
Crisan, Mihaela; Kartalaei, Parham Solaimani; Vink, Chris S et al. (2015) BMP signalling differentially regulates distinct haematopoietic stem cell types. Nat Commun 6:8040
Marks-Bluth, Jonathon; Khanna, Anchit; Chandrakanthan, Vashe et al. (2015) SMAD1 and SMAD5 Expression Is Coordinately Regulated by FLI1 and GATA2 during Endothelial Development. Mol Cell Biol 35:2165-72
Solaimani Kartalaei, Parham; Yamada-Inagawa, Tomoko; Vink, Chris S et al. (2015) Whole-transcriptome analysis of endothelial to hematopoietic stem cell transition reveals a requirement for Gpr56 in HSC generation. J Exp Med 212:93-106

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