[Beta]-globin gene transfer to human bone marrow for sickle cell disease

Autologous hematopoietic stem cell gene therapy is an approach to treating sickle cell disease (SCD) patients that may result in lower morbidity than allogeneic transplantation. We examined the potential of a lenti- viral vector (LV) (CCL-ßAS3-FB) encoding a human hemoglobin (HBB) gene engineered to...

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Published in:The Journal of clinical investigation 2013-08, Vol.123 (8), p.3317
Main Authors: Romero, Zulema, Urbinati, Fabrizia, Geiger, Sabine, Cooper, Aaron R, Wherley, Jennifer, Kaufman, Michael L, Hollis, Roger P, de Assin, Rafael Ruiz, Senadheera, Shantha, Sahagian, Arineh, Jin, Xiangyang, Gellis, Alyse, Wang, Xiaoyan, Gjertson, David, DeOliveira, Satiro, Kempert, Pamela, Shupien, Sally, Abdel-Azim, Hisham, Walters, Mark C, Meiselman, Herbert J, Wenby, Rosalinda B, Gruber, Theresa, Marder, Victor, Coates, Thomas D, Kohn, Donald B
Format: Article
Language:English
Subjects:
Biomedical research
Bone marrow
Cancer
Children & youth
Clinical trials
Flow cytometry
Gene expression
Hospitals
Mortality
Polymerization
Proteins
Sickle cell anemia
Sickle cell disease
Stem cells
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Summary:Autologous hematopoietic stem cell gene therapy is an approach to treating sickle cell disease (SCD) patients that may result in lower morbidity than allogeneic transplantation. We examined the potential of a lenti- viral vector (LV) (CCL-ßAS3-FB) encoding a human hemoglobin (HBB) gene engineered to impede sickle hemoglobin polymerization (HBBAS3) to transduce human BM CD34+ cells from SCD donors and prevent sickling of red biood cells produced by in vitro differentiation. The CCL-ßAS3-FB LV transduced BM CD34+ cells from either healthy or SCD donors at similar levels, based on quantitative PCR and colony-forming unit progenitor analysis. Consistent expression of HBBAS3 mRNA and HbAS3 protein compromised a fourth of the total ß-globin-like transcripts and hemoglobin (Hb) tetramers. Upon deoxygenation, a lower percent- age of HBBAS3-transduced red blood cells exhibited sickling compared with mock-transduced cells from sickle donors. Transduced BM CD34+ cells were transplanted into immunodeficient mice, and the human, cells recovered after 2-3 months were cultured for erythroid differentiation, which showed levels of HBBAS3 mRNA similar to those seen in the CD34+ cells that were directly differentiated in vitro. These results dem- onstrate that the CCL-ßAS3-FB LV is capable of efficient transfer and consistent expression of an effective anti-sickling ß-globin gene in human SCD BM CD34+ progenitor cells, improving physiologic parameters of the resulting red blood cells. [PUBLICATION ABSTRACT]
ISSN:0021-9738
1558-8238