Organ transplantation is highly successful in the treatment of end-stage organ failure. Although first year graft survival rates are 90 %, this initial high success rate drastically declines to less than 50 % within 10 years. The cause for loss of graft function is chronic rejection, which, however can be prevented by the induction of transplantation tolerance, a goal that has not been achieved in clinical transplantation. One established approach to achieving tolerance is by establishing hematopoietic mixed chimerism through bone marrow transplantation of the donor into the recipient. Due to the severity of the pre-conditioning regimens required to protect bone marrow cells from rejection, this procedure is often associated with significant morbidity and economic cost, limiting its use to the treatment of life-threatening hematopoietic malignancies. In contrast, mouse embryonic stem cells (ESC) are poorly immunogenic because of their low expression of MHC class I and lack of class II molecules. By exploiting their pluripotency, we can coax ESC to form hematopoietic precursor cells (HPCs) that can safely be used to induce mixed chimerism across major histocompatibility barriers with minimal requirement for immunosuppression. Here, we will utilize the homeodomain transcription factor HOXB4 that confers self-renewal and expansion capability to hematopoietic cells allowing us to exploit existing ESC lines for therapeutic purposes. To optimize this opportunity, HOXB4 was cloned in frame with GFP, so that as the cells differentiate, they turn green. Isolation of CD45+GFP+ progenitor cells assures removal of undifferentiated cells avoiding tumor formation. The cells will be transplanted into mice to establish mixed chimerism. Once stable mixed chimerism has been established, we will transplant skin allografts in allogenic recipient mice.
Our Specific Aims will address: 1. immunogenicity of ESC-derived HPCs;2. establishment of mixed chimerism and graft tolerance and 3. cell fusion of HPCs with host cells post transplantation. Achievement of these goals will establish ESC as a new source of hematopoietic cells that can be used for tolerance induction without harsh immunosuppression of the recipient.

Public Health Relevance

Organ transplantation is a life-saving form of treatment for patients suffering from end-stage organ failure. Its success is, however severely hampered by immunological rejection. Here, we seek to develop a new approach that utilizes embryonic stem cells for the induction of mixed chimerism and tolerance.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL073015-08
Application #
8207866
Study Section
Transplantation, Tolerance, and Tumor Immunology (TTT)
Program Officer
Shah, Monica R
Project Start
2004-08-01
Project End
2013-12-31
Budget Start
2012-01-01
Budget End
2013-12-31
Support Year
8
Fiscal Year
2012
Total Cost
$315,000
Indirect Cost
$65,000
Name
University of Iowa
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
062761671
City
Iowa City
State
IA
Country
United States
Zip Code
52242
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Manzar, Gohar S; Kim, Eun-Mi; Rotti, Pavana et al. (2014) Skin deep: from dermal fibroblasts to pancreatic beta cells. Immunol Res 59:279-86
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Kim, Eun-Mi; Manzar, Gohar; Zavazava, Nicholas (2013) Mouse ES cell-derived hematopoietic progenitor cells. Methods Mol Biol 1029:109-17
Kim, Eun-Mi; Manzar, Gohar; Zavazava, Nicholas (2013) Human iPS cell-derived hematopoietic progenitor cells induce T-cell anergy in in vitro-generated alloreactive CD8(+) T cells. Blood 121:5167-75
Pilat, Sandra; Carotta, Sebastian; Klump, Hannes (2013) Development of hematopoietic stem and progenitor cells from mouse embryonic stem cells, in vitro, supported by ectopic human HOXB4 expression. Methods Mol Biol 1029:129-47
Kim, E M; Stultz, R; Bonde, S et al. (2012) Embryonic stem cell-derived T cells induce lethal graft-versus-host disease and reject allogenic skin grafts upon thymic selection. Am J Transplant 12:600-9
Fan, Rong; Bonde, Sabrina; Gao, Peng et al. (2012) Dynamic HoxB4-regulatory network during embryonic stem cell differentiation to hematopoietic cells. Blood 119:e139-47
Raikwar, Sudhanshu P; Zavazava, Nicholas (2011) Spontaneous in vivo differentiation of embryonic stem cell-derived pancreatic endoderm-like cells corrects hyperglycemia in diabetic mice. Transplantation 91:11-20

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