The long-term goal of this research project is to develop strategies for establishing safe, durable engraftment of normal donor stem cells in human fetuses diagnosed with immunodeficiency diseases, hemoglobinopathies or inborn errors of metabolism. In the mouse model of in utero transplantation (IUT), the majority of recipients engraft but at very low levels, insufficient to induce tolerance. This is comparable to what has been seen in monkeys and humans following IUT. We have found that marrow derived dendritic cell progenitors (pDC) added to the marrow inoculum result in virtually full donor hematopoietic chimerism in some animals but at the expense of significant graft versus host disease (GvHD). We believe this is due to the rapid maturation of pDC which then interact with donor T cells. The immediate goal of this research proposal is to understand the mechanisms by which tolerance to allogeneic mismatched hematopoietic stem cells (HSC) can be induced in utero in the fetal mouse.
The specific aims of this proposal are: 1) to understand why tolerance is not reliably induced in utero and the role of DC in its induction and 2) to characterize the role of a growth advantage for donor HSC by selectively destroying host hematopoiesis in utero or postnatally. We will utilize mature DC from CD80 and CD86 single and double knockout mice and maturation resistant DC to study the mechanisms for tolerance induction in utero. We will also evaluate the role of """"""""space"""""""" by using donor NK cells or sensitized donor CD8+ cells to prepare the recipient for IUT. Finally, we will study the mechanisms of both central and peripheral tolerance in tolerant animals in order to define those that are critical during the early phases of engraftment. The results of these experiments will lead to the development of therapeutic strategies for the induction of tolerance and optimization of engraftment of donor cells pre and postnatally and could have an impact on other areas of transplantation, specifically, induction and maintenance of tolerance to solid organs, and engraftment of HLA-mismatched stem cells post non-myeloablative conditioning.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL058842-06
Application #
6726098
Study Section
Special Emphasis Panel (ZRG1-ET-1 (06))
Program Officer
Thomas, John
Project Start
1998-01-01
Project End
2006-03-31
Budget Start
2004-04-01
Budget End
2005-03-31
Support Year
6
Fiscal Year
2004
Total Cost
$265,125
Indirect Cost
Name
University of California San Francisco
Department
Pediatrics
Type
Schools of Medicine
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Xiao, Tony Z; Singh, Kanal; Dunn, Elizabeth et al. (2012) T cell and B Cell immunity can be reconstituted with mismatched hematopoietic stem cell transplantation without alkylator therapy in artemis-deficient mice using anti-natural killer cell antibody and photochemically treated sensitized donor T cells. Biol Blood Marrow Transplant 18:200-9
Xiao, Zheng; Yannone, Steven M; Dunn, Elizabeth et al. (2009) A novel missense RAG-1 mutation results in T-B-NK+ SCID in Athabascan-speaking Dine Indians from the Canadian Northwest Territories. Eur J Hum Genet 17:205-12
Xiao, Zheng; Dunn, Elizabeth; Singh, Kanal et al. (2009) A non-leaky Artemis-deficient mouse that accurately models the human severe combined immune deficiency phenotype, including resistance to hematopoietic stem cell transplantation. Biol Blood Marrow Transplant 15:1-11
Povirk, Lawrence F; Zhou, Tong; Zhou, Ruizhe et al. (2007) Processing of 3'-phosphoglycolate-terminated DNA double strand breaks by Artemis nuclease. J Biol Chem 282:3547-58
Bhattacharyya, Swati; Cowan, Morton J (2005) B7.2-/- mature dendritic cells generate T-helper 2 and regulatory T donor cells in fetal mice after in utero allogeneic bone marrow transplantation. Biol Blood Marrow Transplant 11:657-71
Bhattacharyya, Swati; Chawla, Anjulika; Smith, Kristofer et al. (2002) Multilineage engraftment with minimal graft-versus-host disease following in utero transplantation of S-59 psoralen/ultraviolet a light-treated, sensitized T cells and adult T cell-depleted bone marrow in fetal mice. J Immunol 169:6133-40
Cowan, M J; Chou, S H; Tarantal, A F (2001) Tolerance induction post in utero stem cell transplantation. Ernst Schering Res Found Workshop :145-71
Chou, S H; Chawla, A; Lee, T H et al. (2001) Increased engraftment and GVHD after in utero transplantation of MHC-mismatched bone marrow cells and CD80low, CD86(-) dendritic cells in a fetal mouse model. Transplantation 72:1768-76