Optimization of recombinant adeno-associated viral vectors for human β-globin gene transfer and transgene expression

Njeri Maina, Li Zhong, Xiaomiao Li, Weihong Zhao, Zongchao Han, Daniela Bischof, George Aslanidi, Sergei Zolotukhin, Kirsten A. Weigel-Van Aken, Angela E. Rivers, William B. Slayton, Mervin Yoder, Arun Srivastava

Research output: Contribution to journalArticle

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Abstract

Therapeutic levels of expression of the β-globin gene have been difficult to achieve with conventional retroviral vectors without the inclusion of DNase I-hypersensitive site (HS2, HS3, and HS4) enhancer elements. We generated recombinant adeno-associated viral (AAV) vectors carrying an antisickling human β-globin gene under the control of either the β-globin gene promoter/enhancer or the erythroid cell-specific human parvovirus B19 promoter at map unit 6 (B19p6) without any enhancer, and tested their efficacy in a human erythroid cell line (K-562) and in primary murine hematopoietic progenitor cells (c-kit+lin-). We report here that (1) self-complementary AAV serotype 2 (scAAV2)-β-globin vectors containing only the HS2 enhancer are more efficient than single-stranded AAV (ssAAV2)-β-globin vectors containing the HS2+HS3+HS4 enhancers; (2) scAAV2-β-globin vectors recombine with scAAV2-HS2+HS3+HS4 vectors after dual-vector transduction, leading to transgene expression; (3) scAAV2-β-globin as well as scAAV1-β-globin vectors containing the B19p6 promoter without the HS2 enhancer element are more efficient than their counterparts containing the HS2 enhancer/β-globin promoter; and (4) scAAV2-B19p6-β-globin vectors in K-562 cells, and scAAV1-B19p6-β- globin vectors in murine c-kit+lin- cells, yield efficient expression of the β-globin protein. Thus, the combined use of scAAV vectors and the parvovirus B19 promoter may lead to expression of therapeutic levels the β-globin gene in human erythroid cells, which has implications in the use of these vectors in gene therapy of β-thalassemia and sickle cell disease.

Original languageEnglish
Pages (from-to)365-375
Number of pages11
JournalHuman Gene Therapy
Volume19
Issue number4
DOIs
StatePublished - Apr 1 2008

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Globins
Transgenes
Genes
Erythroid Cells
Human Parvovirus B19
Parvovirus
Thalassemia
Deoxyribonuclease I
Sickle Cell Anemia
Hematopoietic Stem Cells
Genetic Therapy
Self Report

ASJC Scopus subject areas

  • Genetics

Cite this

Optimization of recombinant adeno-associated viral vectors for human β-globin gene transfer and transgene expression. / Maina, Njeri; Zhong, Li; Li, Xiaomiao; Zhao, Weihong; Han, Zongchao; Bischof, Daniela; Aslanidi, George; Zolotukhin, Sergei; Weigel-Van Aken, Kirsten A.; Rivers, Angela E.; Slayton, William B.; Yoder, Mervin; Srivastava, Arun.

In: Human Gene Therapy, Vol. 19, No. 4, 01.04.2008, p. 365-375.

Research output: Contribution to journalArticle

Maina, N, Zhong, L, Li, X, Zhao, W, Han, Z, Bischof, D, Aslanidi, G, Zolotukhin, S, Weigel-Van Aken, KA, Rivers, AE, Slayton, WB, Yoder, M & Srivastava, A 2008, 'Optimization of recombinant adeno-associated viral vectors for human β-globin gene transfer and transgene expression', Human Gene Therapy, vol. 19, no. 4, pp. 365-375. https://doi.org/10.1089/hum.2007.173
Maina, Njeri ; Zhong, Li ; Li, Xiaomiao ; Zhao, Weihong ; Han, Zongchao ; Bischof, Daniela ; Aslanidi, George ; Zolotukhin, Sergei ; Weigel-Van Aken, Kirsten A. ; Rivers, Angela E. ; Slayton, William B. ; Yoder, Mervin ; Srivastava, Arun. / Optimization of recombinant adeno-associated viral vectors for human β-globin gene transfer and transgene expression. In: Human Gene Therapy. 2008 ; Vol. 19, No. 4. pp. 365-375.
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AU - Han, Zongchao

AU - Bischof, Daniela

AU - Aslanidi, George

AU - Zolotukhin, Sergei

AU - Weigel-Van Aken, Kirsten A.

AU - Rivers, Angela E.

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