It is thought that within the marrow there are at least two anatomically and physiologically distinct hematopoietic niches;the osteoblastic and the vascular. Within these niches, numerous molecules and secreted factors have been implicated in the strictly regulated process of hematopoiesis. Nevertheless, a thorough understanding of the subpopulations of HSC that interact with each niche and the physiological role of each niche type in the regulation of stem cell hierarchy and function is still missing. Here, we will identify the cellular interactions that occur within each of the niches at the phenotypic, functional, and transcriptional levels, using as a distinct paradigm the developmental process whereby the marrow acquires the ability to support hematopoiesis. Experiments to be performed will employ a physiologically relevant large animal model, high resolution imaging/capture technology, sensitive transcriptome sequencing, and functional studies, which combined provide us with the unique ability to study and manipulate the interactions of human HSC and specific niche cells in vivo, thereby unraveling the role played by each discrete microenvironment in the establishment and maintenance of human hematopoiesis. The overall hypothesis of these studies is that the developmental process whereby the fetal marrow acquires the ability to support hematopoiesis can be used as a model for understanding both the interactions that occur between HSC and the cells comprising the marrow niches, and the role of these niche elements in the initiation/maintenance of hematopoiesis. We further hypothesize that this knowledge will provide the necessary tools for manipulating these niches to facilitate the engraftment and accelerate hematopoietic recover of donor HSC after HSCTx. We propose the following Specific Aims:1)Define the steps and key cellular players in the emergence of the fetal BM hematopoietic niche and in the onset of marrow hematopoiesis, as a tool to understand the cellular interactions that occur both within and amongst the vascular and the osteoblastic niches as they arise, and characterize the nature of the hematopoietic cells each supports, thus delineating the role each plays in the maintenance and expansion of the stem cell pool to drive lifelong hematopoiesis;2) Determine whether the vascular/perivascular niches present within the nascent marrow not only serve as a supportive site for lodging of HSC arriving from the fetal liver, but also harbor cells with the potential to contribute to the establishment of hematopoiesis;and 3) Investigate whether the composition of the bone and vascular niches can be manipulated by specific populations of donor-derived cells to increase the levels of engraftment and/or accelerate reconstitution of donor derived HSC following allogeneic transplantation. Success of the proposed studies would have significant implications in the isolation and expansion of primitive HSC and the understanding of the origin of hematologic malignancies, and could significantly improve the safety and success of clinical HSC transplantation and potentially allow its application to a wider range of diseases affecting human patients.

Public Health Relevance

Using fetal development as a model to gain a thorough understanding of the interactions that occur between hematopoietic stem cells and the cells that comprise the bone marrow microenvironmental niches will provide the necessary tools for manipulating these niches to facilitate the engraftment of donor HSC and accelerate hematopoietic recover following HSC transplantation. This would significantly improve the safety and success of clinical HSC transplantation and potentially allow its application to a wider range of diseases affecting human patients.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL097623-04
Application #
8309395
Study Section
Special Emphasis Panel (ZHL1-CSR-W (M1))
Program Officer
Thomas, John
Project Start
2009-08-28
Project End
2013-07-31
Budget Start
2012-08-01
Budget End
2013-07-31
Support Year
4
Fiscal Year
2012
Total Cost
$366,300
Indirect Cost
$118,800
Name
Wake Forest University Health Sciences
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
937727907
City
Winston-Salem
State
NC
Country
United States
Zip Code
27157
Jeanblanc, Christine; Goodrich, Angelina Daisy; Colletti, Evan et al. (2014) Temporal definition of haematopoietic stem cell niches in a large animal model of in utero stem cell transplantation. Br J Haematol 166:268-78
Boura, Joana S; Vance, Melisa; Yin, Weihong et al. (2014) Evaluation of gene delivery strategies to efficiently overexpress functional HLA-G on human bone marrow stromal cells. Mol Ther Methods Clin Dev 2014:
Soland, M A; Keyes, L R; Bayne, R et al. (2014) Perivascular stromal cells as a potential reservoir of human cytomegalovirus. Am J Transplant 14:820-30
Sanada, Chad; Kuo, Chung-Jung; Colletti, Evan J et al. (2013) Mesenchymal stem cells contribute to endogenous FVIII:c production. J Cell Physiol 228:1010-6
Soland, Melisa A; Bego, Mariana; Colletti, Evan et al. (2013) Mesenchymal stem cells engineered to inhibit complement-mediated damage. PLoS One 8:e60461
Almeida-Porada, Graca D (2010) Stem cell gene manipulation and delivery as systemic therapeutics. Adv Drug Deliv Rev :
Almeida-Porada, Graca; Zanjani, Esmail D; Porada, Christopher D (2010) Bone marrow stem cells and liver regeneration. Exp Hematol 38:574-80
Porada, Christopher D; Almeida-Porada, Gra├ža (2010) Mesenchymal stem cells as therapeutics and vehicles for gene and drug delivery. Adv Drug Deliv Rev 62:1156-66