The investigators of this proposal have developed two tools that may be of value in establishing stable mixed chimerism of patients with sickle cell anemia and beta thalassemia. The first tool is directed toward developing a minimally toxic method for producing stem cell depletion using the combination of flt-3 ligand (FL) and 5-fluorouracil (5-FU). This approach may be a useful component of transplant conditioning regimens that are directed at achieving mixed chimerism. The second tool uses chemical inducers of dimerization (CIDs) to specifically deliver a mitogenic signal to genetically modified cells. Transfer of a gene that encodes a CID-responsive protein may render normal donor stem cell responsive to CID-mediated proliferation. This approach may allow the proliferative status of normal donor stem cells in mixed chimeras to come under direct pharmacological control.
Specific Aim 1 tests whether CIDs can expand genetically modified stem and progenitor cells in vivo.
Specific Aim 2 develops a mouse model of mixed chimerism in beta thalassemia, and seeks to determine the level of normal donor stem cell engraftment needed to reverse the thalassemic phenotype.
Specific Aim 3 tests whether CID-mediated in vivo expansion of normal donor stem cells can correct the thalassemic phenotype of mice with mixed chimerism.
Specific Aim 4 evaluates whether FL can sensitize stem ells to 5-FU.
In Specific Aim 5, the FL/5-FU combination is added to an immunosuppressive condition regimen for studies in allogeneic models of mixed chimerism.
In Specific Aim 6, findings from the previous specific aims are combined to test CID-mediated in vivo expansion of normal donor stem cells in an allogeneic model of mixed chimerism.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK057525-03
Application #
6381785
Study Section
Special Emphasis Panel (ZHL1-CSR-O (M2))
Program Officer
Badman, David G
Project Start
1999-09-30
Project End
2003-08-31
Budget Start
2001-09-01
Budget End
2002-08-31
Support Year
3
Fiscal Year
2001
Total Cost
$342,301
Indirect Cost
Name
University of Washington
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195
Basha, Maureen; Labelle, Edward F; Northington, Gina M et al. (2009) Functional significance of muscarinic receptor expression within the proximal and distal rat vagina. Am J Physiol Regul Integr Comp Physiol 297:R1486-93
Richard, Robert E; Siritanaratkul, Noppadol; Jonlin, Erica et al. (2005) Collection of blood stem cells from patients with sickle cell anemia. Blood Cells Mol Dis 35:384-8
Zhao, Shengming; Weinreich, Michael A; Ihara, Kenji et al. (2004) In vivo selection of genetically modified erythroid cells using a jak2-based cell growth switch. Mol Ther 10:456-68
Richard, Robert E; De Claro, R Angelo; Yan, James et al. (2004) Differences in F36VMpl-based in vivo selection among large animal models. Mol Ther 10:730-40
Richard, Robert E; Weinreich, Michael; Chang, Kai-Hsin et al. (2004) Modulating erythrocyte chimerism in a mouse model of pyruvate kinase deficiency. Blood 103:4432-9
Richard, Robert E; Blau, C Anthony (2003) Small-molecule-directed mpl signaling can complement growth factors to selectively expand genetically modified cord blood cells. Stem Cells 21:71-8
Zhao, Shengming; Zoller, Karen; Masuko, Masayoshi et al. (2002) JAK2, complemented by a second signal from c-kit or flt-3, triggers extensive self-renewal of primary multipotential hemopoietic cells. EMBO J 21:2159-67
Neff, Tobias; Horn, Peter A; Valli, Victor E et al. (2002) Pharmacologically regulated in vivo selection in a large animal. Blood 100:2026-31
Otto, K G; Jin, L; Spencer, D M et al. (2001) Cell proliferation through forced engagement of c-Kit and Flt-3. Blood 97:3662-4
Otto, K G; Broudy, V C; Lin, N L et al. (2001) Membrane localization is not required for Mpl function in normal hematopoietic cells. Blood 98:2077-83

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