This Core Unit will assist the individual Research Projects by serving four general functions: 1) Analyzing expression of globin gene vectors in primary erythroid cultures and transplanted animals using both flow cytometric and molecular methods; 2) Maintaining colonies and testing globin vectors in thalassemic and sickle cell mice; 3) Preparing CD34+ cells from humans and baboons for transduction studies; 4) Producing and characterizing retrovirus vector packaging lines for titer and stability. or Project 1. 1) Analyze expression of retrovirus vectors for globin and trans- activating genes in primary mouse and human erythroid cultures and transplanted mice; 2) Provide animals and analyze expression of globin and trans-activating vectors in thalassemic and sickle cell mice; and 3) Produce retrovirus vector packaging lines for the globin, insulator, and trans- activating constructs. For Project 2: 1) Analyze expression of human foamy virus vectors for globin genes in primary mouse, baboon, and human erythroid cultures and transplanted mice and baboons; 2) provide animals and analyze expression of globin vectors in thalassemic and sickle cell mice; and 3) Prepare human and baboon CD34+ cells for transduction and transplantation studies. For the next project: 1) Analyze expression of deltaAd.AAV vectors for globin genes in primary mouse and human erythroid cultures and transplanted mice; and 2) Provide animals and analyze expression of globin vectors in thalassemic and sickle cell mice. For the next project: 1) Analyze expression of retrovirus vectors for globin genes in primary mouse erythroid cultures and transplanted mice; 2) Provide animals and analyze expression of globin vectors in transplantation studies; and 4) Produce retrovirus vector packaging lines for selection constructs. The last project: Prepare baboon CD34+ cells for transduction and transplantation studies.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
5P01HL053750-08
Application #
6501918
Study Section
Project Start
2001-09-21
Project End
2002-08-31
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
8
Fiscal Year
2001
Total Cost
$88,647
Indirect Cost
Name
University of Washington
Department
Type
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195
Constantinou, Varnavas C; Bouinta, Asimina; Karponi, Garyfalia et al. (2017) Poor stem cell harvest may not always be related to poor mobilization: lessons gained from a mobilization study in patients with ?-thalassemia major. Transfusion 57:1031-1039
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
Psatha, Nikoletta; Karponi, Garyfalia; Yannaki, Evangelia (2016) Optimizing autologous cell grafts to improve stem cell gene therapy. Exp Hematol 44:528-39
Li, Li B; Ma, Chao; Awong, Geneve et al. (2016) Silent IL2RG Gene Editing in Human Pluripotent Stem Cells. Mol Ther 24:582-91
Karponi, Garyfalia; Psatha, Nikoletta; Lederer, Carsten Werner et al. (2015) Plerixafor+G-CSF-mobilized CD34+ cells represent an optimal graft source for thalassemia gene therapy. Blood 126:616-9
Vierstra, Jeff; Reik, Andreas; Chang, Kai-Hsin et al. (2015) Functional footprinting of regulatory DNA. Nat Methods 12:927-30
Qi, Heyuan; Liu, Mingdong; Emery, David W et al. (2015) Functional validation of a constitutive autonomous silencer element. PLoS One 10:e0124588
Liu, Mingdong; Maurano, Matthew T; Wang, Hao et al. (2015) Genomic discovery of potent chromatin insulators for human gene therapy. Nat Biotechnol 33:198-203
Polak, Paz; Karli?, Rosa; Koren, Amnon et al. (2015) Cell-of-origin chromatin organization shapes the mutational landscape of cancer. Nature 518:360-364
Watts, Korashon L; Beard, Brian C; Wood, Brent L et al. (2014) No evidence of clonal dominance after transplant of HOXB4-expanded cord blood cells in a nonhuman primate model. Exp Hematol 42:497-504

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