Hematopoietic stem cells (HSCs) are an important target for the gene therapy of infectious diseases, genetic disorders, and cancer. In vivo genome editing of HSCs has major advantages over currently used approaches that involve the collection of HSCs from patients, their in vitro culture/transduction, and retransplantation into myelo-conditioned patients. We have developed a new in vivo approach for HSC transduction, which is based on the GCSF/AMD3100-mediated mobilization of HSCs from the bone marrow into the peripheral blood stream and the intravenous injection of an HSC-targeting, helper-dependent adenovirus vector (HD-Ad5/35). The central goal of this proposal is to increase the percentage of stably in vivo HD-Ad5/35 - transduced HSCs while avoiding potential genotoxicity. We plan to achieve this through i) optimization of HSC mobilization and in vivo HSC transduction, ii) optimization of transgene integration through stimulation of homologous recombination, and iii) incorporation into HD- Ad5/35 vectors of systems that allow for the in vivo selection or expansion of stably transduced cells. Based on these studies, we will generate and test HD-Ad5/35 vectors for in vivo gene therapy of b-thalassemia.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL130040-04
Application #
9634982
Study Section
Therapeutic Approaches to Genetic Diseases Study Section (TAG)
Program Officer
Qasba, Pankaj
Project Start
2016-06-01
Project End
2020-01-31
Budget Start
2019-02-01
Budget End
2020-01-31
Support Year
4
Fiscal Year
2019
Total Cost
Indirect Cost
Name
University of Washington
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Wang, Hongjie; Georgakopoulou, Aphrodite; Psatha, Nikoletta et al. (2018) In vivo hematopoietic stem cell gene therapy ameliorates murine thalassemia intermedia. J Clin Invest :
Li, Chang; Psatha, Nikoletta; Gil, Sucheol et al. (2018) HDAd5/35++ Adenovirus Vector Expressing Anti-CRISPR Peptides Decreases CRISPR/Cas9 Toxicity in Human Hematopoietic Stem Cells. Mol Ther Methods Clin Dev 9:390-401
Li, Chang; Psatha, Nikoletta; Sova, Pavel et al. (2018) Reactivation of ?-globin in adult ?-YAC mice after ex vivo and in vivo hematopoietic stem cell genome editing. Blood 131:2915-2928
Wang, Hongjie; Richter, Maximilian; Psatha, Nikoletta et al. (2018) A Combined In Vivo HSC Transduction/Selection Approach Results in Efficient and Stable Gene Expression in Peripheral Blood Cells in Mice. Mol Ther Methods Clin Dev 8:52-64
Li, Chang; Psatha, Nikoletta; Wang, Hongjie et al. (2018) Integrating HDAd5/35++ Vectors as a New Platform for HSC Gene Therapy of Hemoglobinopathies. Mol Ther Methods Clin Dev 9:142-152
Richter, Maximilian; Stone, Daniel; Miao, Carol et al. (2017) In Vivo Hematopoietic Stem Cell Transduction. Hematol Oncol Clin North Am 31:771-785
Richter, Maximilian; Saydaminova, Kamola; Yumul, Roma et al. (2016) In vivo transduction of primitive mobilized hematopoietic stem cells after intravenous injection of integrating adenovirus vectors. Blood 128:2206-2217