Allogeneic hematopoietic stem and progenitor cell (HSPC) transplantation has the potential to cure hematologic disease. However, many patients do not have an HLA matched donor, and graft-versus-host disease is a significant problem. Autologous patient HSPCs can be genetically corrected to cure the disease, but low yields of autologous HSPCs and ex vivo manipulation cause a loss of """"""""stemness"""""""" leading to reduced engraftment. Thus, HSPC production from patient-specific induced pluripotent stem cells (iPSCs) would solve these problems and represent an unlimited cell source. To advance clinical translation of iPSC therapeutics, we propose a novel strategy to expand iPSC-derived HSPCs for hematopoietic transplantation. Specifically, we propose to engineer endothelial cells (ECs) for generation, expansion, and engraftment of putative HSPCs from pigtail macaque (Mn)iPSCs in the clinically relevant nonhuman primate model. In a promising collaboration with Dr. Shahin Rafii, we developed an effective, novel platform to expand macaque CD34+ LT- HSCs up to 25-fold by co-culture with Akt-activated human endothelial cells (E4+ECs). We recently found that iPSC-HSPCs expanded on E4+ECs have high levels of engraftment in NSG mice (up to 50% CD45+ cells). This evidence substantiates our novel approach to alter iPSC-HSPC biology through direct contact culture with angiocrine/hematopoietic signals unique to E4+ECs. The proposed studies will translate these findings to nonhuman primates and thus provide a major step toward producing iPSC-HSPCs with the capacity for hematopoietic reconstitution and correction of genetic diseases.

Public Health Relevance

The only cure for many genetic blood diseases affecting bone marrow and/or blood cells is a blood stem cell transplant. Only about 25% of patients will have a matched sibling donor. While alternative donors can be used, the risk of side effects from graft-versus-host disease and infectious complications will be substantially increased. Thus, the goal of this project is to develop an alternative stem cell therapy approach using vascular cells to direct a patient's own cells to become blood stem cells. Using the nonhuman primate as a model, reprogrammed nonhuman primate cells are converted into blood stem cells with help of a unique vascular platform. We will also add genetic elements that provide quality control of the blood stem cells and help them take up residence in the bone marrow of the original nonhuman primate, where they will produce all types of blood cells for the lifespan of the donor. Our primary goal is to enable the use of the patients'own cells to safely treat their underlying blood or marrow genetic disease.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL115128-01A1
Application #
8579473
Study Section
Special Emphasis Panel (ZRG1-VH-A (80))
Program Officer
Zou, Shimian
Project Start
2013-08-01
Project End
2017-04-30
Budget Start
2013-08-01
Budget End
2014-04-30
Support Year
1
Fiscal Year
2013
Total Cost
$823,092
Indirect Cost
$272,697
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
078200995
City
Seattle
State
WA
Country
United States
Zip Code
98109
Humbert, Olivier; Peterson, Christopher W; Norgaard, Zachary K et al. (2018) A Nonhuman Primate Transplantation Model to Evaluate Hematopoietic Stem Cell Gene Editing Strategies for ?-Hemoglobinopathies. Mol Ther Methods Clin Dev 8:75-86
Itkin, Tomer; Gómez-Salinero, Jesús María; Rafii, Shahin (2017) Open the gates: vascular neurocrine signaling mobilizes hematopoietic stem and progenitor cells. J Clin Invest 127:4231-4234
Guo, Peipei; Poulos, Michael G; Palikuqi, Brisa et al. (2017) Endothelial jagged-2 sustains hematopoietic stem and progenitor reconstitution after myelosuppression. J Clin Invest 127:4242-4256
Lis, Raphael; Karrasch, Charles C; Poulos, Michael G et al. (2017) Conversion of adult endothelium to immunocompetent haematopoietic stem cells. Nature 545:439-445
Schachterle, William; Badwe, Chaitanya R; Palikuqi, Brisa et al. (2017) Sox17 drives functional engraftment of endothelium converted from non-vascular cells. Nat Commun 8:13963
Haworth, Kevin G; Ironside, Christina; Norgaard, Zachary K et al. (2017) In Vivo Murine-Matured Human CD3+ Cells as a Preclinical Model for T Cell-Based Immunotherapies. Mol Ther Methods Clin Dev 6:17-30
Chiarelli, Peter A; Revia, Richard A; Stephen, Zachary R et al. (2017) Nanoparticle Biokinetics in Mice and Nonhuman Primates. ACS Nano 11:9514-9524
Gori, Jennifer L; Butler, Jason M; Kunar, Balvir et al. (2017) Endothelial Cells Promote Expansion of Long-Term Engrafting Marrow Hematopoietic Stem and Progenitor Cells in Primates. Stem Cells Transl Med 6:864-876
Rafii, Shahin; Butler, Jason M; Ding, Bi-Sen (2016) Angiocrine functions of organ-specific endothelial cells. Nature 529:316-25
Radtke, Stefan; Haworth, Kevin G; Kiem, Hans-Peter (2016) The frequency of multipotent CD133(+)CD45RA(-)CD34(+) hematopoietic stem cells is not increased in fetal liver compared with adult stem cell sources. Exp Hematol 44:502-7

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