Efficient transcriptional targeting of human hematopoietic stem cells and blood cell lineages by lentiviral vectors containing the regulatory element of the Wiskott-Aldrich syndrome gene

Valeria Leuci, Loretta Gammaitoni, Sonia Capellero, Dario Sangiolo, Maria Mesuraca, Heather M. Bond, Giorgia Migliardi, Cristina Cammarata, Massimo Aglietta, Giovanni Morrone, Wanda Piacibello

Research output: Contribution to journalArticle

Abstract

The ability to effectively transduce human hematopoietic stem cells (HSCs) and to ensure adequate but "physiological" levels of transgene expression in different hematopoietic lineages represents some primary features of a gene-transfer vector. The ability to carry, integrate, and efficiently sustain transgene expression in HSCs strongly depends on the vector. We have constructed lentiviral vectors (LV) containing fragments of different lengths of the hematopoietic-specific regulatory element of the Wiskott-Aldrich syndrome (WAS) gene - spanning approximately 1,600 and 170 bp - that direct enhanced green fluorescent protein (EGFP) expression. The performance of vectors carrying the 1,600 and 170 bp fragments of the WAS gene promoter was compared with that of a vector carrying the UbiquitinC promoter in human cord blood CD34 + cells and their differentiated progeny both in vitro and in vivo in non-obese diabetic mice with severe combined immunodeficiency. All vectors displayed a similar transduction efficiency in CD34+ cells and promoted long-term EGFP expression in different hematopoietic lineages, with an efficiency comparable to, and in some instances (for example, the 170-bp promoter) superior to, that of the UbiquitinC promoter. Our results clearly demonstrate that LV containing fragments of the WAS gene promoter/enhancer region can promote long-term transgene expression in different hematopoietic lineages in vitro and in vivo and represent suitable and highly efficient vectors for gene transfer in gene-therapy applications for different hematological diseases and for research purposes. In particular, the 170-bp carrying vector, for its reduced size, could significantly improve the transduction/expression of large-size genes.

Original languageEnglish
Pages (from-to)2815-2823
Number of pages9
JournalStem Cells
Volume27
Issue number11
DOIs
Publication statusPublished - Nov 2009

Fingerprint

Wiskott-Aldrich Syndrome
Cell Lineage
Hematopoietic Stem Cells
Blood Cells
Transgenes
Genes
Severe Combined Immunodeficiency
Inbred NOD Mouse
Hematologic Diseases
Fetal Blood
Genetic Promoter Regions
Genetic Therapy
Research

Keywords

  • Gene therapy
  • Hematopoietic stem cells
  • Lentiviral vectors
  • Promoters

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Molecular Medicine

Cite this

Efficient transcriptional targeting of human hematopoietic stem cells and blood cell lineages by lentiviral vectors containing the regulatory element of the Wiskott-Aldrich syndrome gene. / Leuci, Valeria; Gammaitoni, Loretta; Capellero, Sonia; Sangiolo, Dario; Mesuraca, Maria; Bond, Heather M.; Migliardi, Giorgia; Cammarata, Cristina; Aglietta, Massimo; Morrone, Giovanni; Piacibello, Wanda.

In: Stem Cells, Vol. 27, No. 11, 11.2009, p. 2815-2823.

Research output: Contribution to journalArticle

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