Hematopoietic stem cells reside within the bone marrow post-natally, providing all hematopoietic lineages for the life of the host, and their extensive characterization over the last several decades has led to clinical application including bone marrow transplantation for benign and malignant disorders affecting the hematopoietic compartment. Isolation of bone marrow cells based upon expression of the CD34 antigen enriches for the primitive compartment, and techniques to isolate the CD34-positive population are commonly used in clinical practice. Assays for true hematopoietic stem cells require transplantation and repopulation, and thus studies querying such potential among human cells are difficult. We have thus developed a xenograft model with robust human hematopoietic stem cell engraftment, and have demonstrated engraftment of all cell lineages. This assay will allow us to test globin vectors for the first time among engrafted human cells in a relevant model. Furthermore, ex vivo differentiation of hematopoietic stem cells along the erythroid lineage can be utilized to assay viral vector directed beta-globin expression after transfer of the human beta globin gene. These systems have proven useful in developing viral vector systems for clinical application including a forward oriented globin vector which has been developed by our group and demonstrates 10 fold higher vector titer as well as 10 fold higher transduction efficiency for long term repopulating cells.

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Budget End
Support Year
8
Fiscal Year
2015
Total Cost
Indirect Cost
Name
U.S. National Heart Lung and Blood Inst
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Leonard, Alexis; Tisdale, John F (2018) Stem cell transplantation in sickle cell disease: therapeutic potential and challenges faced. Expert Rev Hematol 11:547-565
Uchida, Naoya; Haro-Mora, Juan J; Fujita, Atsushi et al. (2017) Efficient Generation of ?-Globin-Expressing Erythroid Cells Using Stromal Cell-Derived Induced Pluripotent Stem Cells from Patients with Sickle Cell Disease. Stem Cells 35:586-596
Uchida, Naoya; Fujita, Atsushi; Hsieh, Matthew M et al. (2017) Bone Marrow as a Hematopoietic Stem Cell Source for Gene Therapy in Sickle Cell Disease: Evidence from Rhesus and SCD Patients. Hum Gene Ther Clin Dev :
Fujita, Atsushi; Uchida, Naoya; Haro-Mora, Juan J et al. (2016) ?-Globin-Expressing Definitive Erythroid Progenitor Cells Generated from Embryonic and Induced Pluripotent Stem Cell-Derived Sacs. Stem Cells 34:1541-52
Uchida, Naoya; Green, Rashidah; Ballantine, Josiah et al. (2016) Kinetics of lentiviral vector transduction in human CD34(+) cells. Exp Hematol 44:106-15
Evans, Molly E; Kumkhaek, Chutima; Hsieh, Matthew M et al. (2014) TRIM5? variations influence transduction efficiency with lentiviral vectors in both human and rhesus CD34(+) cells in vitro and in vivo. Mol Ther 22:348-58
Kovtunovych, Gennadiy; Ghosh, Manik C; Ollivierre, Wade et al. (2014) Wild-type macrophages reverse disease in heme oxygenase 1-deficient mice. Blood 124:1522-30
Herman, S E M; Sun, X; McAuley, E M et al. (2013) Modeling tumor-host interactions of chronic lymphocytic leukemia in xenografted mice to study tumor biology and evaluate targeted therapy. Leukemia 27:2311-21
Kumkhaek, Chutima; Aerbajinai, Wulin; Liu, Wenli et al. (2013) MASL1 induces erythroid differentiation in human erythropoietin-dependent CD34+ cells through the Raf/MEK/ERK pathway. Blood 121:3216-27
Perl, Shira; Perlman, Jordan; Weitzel, R P et al. (2013) Addition of rapamycin to anti-CD3 antibody improves long-term glycaemia control in diabetic NOD mice. PLoS One 8:e67189

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