Abstract

In utero hematopoietic stem cell transplantation (IUT) offers the hope of curing a number of hematological diseases by generating a state of hematopoietic stem cells (HSCs), the levels of donor cell engraftment that can be achieved by IUT are low, limiting the use of this therapy.
Our aim i s to extend the use of IUT to the treatment of diseases such as sickle cell anemia.. To achieve the high levels of donor cell engraftment needed to treat this disease it is the goal of this proposal to engineer HSCs to have a proliferative advantage over normal HSCs. This proposal will test the hypothesis that introduction of the erythropoietin (EpoR) into HSCs will render these altered cells responsive to erythropoietin (EPO). This will result in the altered HSCs and their progeny having a proliferative advantage over normal progenitors. Truncated forms of EpoR (tEpoR) will also be tested. These tEpoR, having deletions in the negative regulatory region of their cytoplasmic domains, deliver stronger proliferative signals, than EpoR. Fetal HSCs will be used as targets since they offer proliferative advantages over adult cells and are, therefore, susceptible to transduction by retroviral vectors. Lentiviral vectors will also be tested for their capacity to modify fetal as well as postnatal sources of HSCs. The effects of introducing the EpoR genes on the proliferation and differentiation of HSCs will be determined using various in vitro culture systems. It is hypothesized that ectopic expression of either EpoR or tEpoR expression on HSCs can make these cells more competitive than their normal counterparts, modified HSCs will be tested against control HSCs in a mouse model of human fetal hematopoiesis. The in vivo model will also be used to test the effects of EPO administration on the expansion of the modified HSCs. The ability of experiments will further determine if making HSCs responsive to HPO will have detrimental effect on the long-term reconstituting- and multi- lineage potential of HSCs.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
2P01HL053762-06
Application #
6369190
Study Section
Heart, Lung, and Blood Initial Review Group (HLBP)
Project Start
1994-09-30
Project End
2005-08-31
Budget Start
Budget End
Support Year
6
Fiscal Year
2000
Total Cost
$135,852
Indirect Cost

  Institution

Name
University of California San Francisco
Department
Type
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143

  Publications

Mizunoe, Shota; Shuto, Tsuyoshi; Suzuki, Shingo et al. (2012) Synergism between interleukin (IL)-17 and Toll-like receptor 2 and 4 signals to induce IL-8 expression in cystic fibrosis airway epithelial cells. J Pharmacol Sci 118:512-520
Ye, Lin; Chang, Judy C; Lu, Ronghua et al. (2008) High oxygen environment during pregnancy rescues sickle cell anemia mice from prenatal death. Blood Cells Mol Dis 41:67-72
Han, Xiao-Dong; Lin, Chin; Chang, Judy et al. (2007) Fetal gene therapy of alpha-thalassemia in a mouse model. Proc Natl Acad Sci U S A 104:9007-11
Chang, Judy C; Ye, Lin; Kan, Yuet Wai (2006) Correction of the sickle cell mutation in embryonic stem cells. Proc Natl Acad Sci U S A 103:1036-40
Jimenez, D F; Lee, C I; O'Shea, C E et al. (2005) HIV-1-derived lentiviral vectors and fetal route of administration on transgene biodistribution and expression in rhesus monkeys. Gene Ther 12:821-30
Jimenez, Daniel F; Leapley, Alyssa C; Lee, Chang I et al. (2005) Fetal CD34+ cells in the maternal circulation and long-term microchimerism in rhesus monkeys (Macaca mulatta). Transplantation 79:142-6
Jimenez, Daniel F; Tarantal, Alice F (2003) Quantitative analysis of male fetal DNA in maternal serum of gravid rhesus monkeys (Macaca mulatta). Pediatr Res 53:18-23
Meiering, C D; Comstock, K E; Linial, M L (2000) Multiple integrations of human foamy virus in persistently infected human erythroleukemia cells. J Virol 74:1718-26
Gaensler, K M; Tu, G; Bruch, S et al. (1999) Fetal gene transfer by transuterine injection of cationic liposome-DNA complexes. Nat Biotechnol 17:1188-92
Lipshutz, G S; Sarkar, R; Flebbe-Rehwaldt, L et al. (1999) Short-term correction of factor VIII deficiency in a murine model of hemophilia A after delivery of adenovirus murine factor VIII in utero. Proc Natl Acad Sci U S A 96:13324-9

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