Assessing the Impact of Cyclosporin A on Lentiviral Transduction and Preservation of Human Hematopoietic Stem Cells in Clinically Relevant Ex Vivo Gene Therapy Settings

Carolina Petrillo, Andrea Calabria, Francesco Piras, Alessia Capotondo, Giulio Spinozzi, Ivan Cuccovillo, Fabrizio Benedicenti, Luigi Naldini, Eugenio Montini, Alessandra Biffi, Bernhard Gentner, Anna Kajaste-Rudnitski

Research output: Contribution to journalArticle


Improving hematopoietic stem and progenitor cell (HSPC) permissiveness to lentiviral vector (LV) transduction without compromising their biological properties remains critical for broad-range implementation of gene therapy as a treatment option for several inherited diseases. This study demonstrates that the use of one-hit ex vivo LV transduction protocols based on either cyclosporin A (CsA) or rapamycin enable as efficient gene transfer as the current two-hit clinical standard into bone marrow-derived CD34+ cells while better preserving their engraftment capacity in vivo. CsA was additive with another enhancer of transduction, prostaglandin E2, suggesting that tailored enhancer combinations may be applied to overcome multiple blocks to transduction simultaneously in HSPC. Interestingly, besides enhancing LV transduction, CsA also significantly reduced HSPC proliferation, preserving the quiescent G0 fraction and the more primitive multipotent progenitors, thereby yielding the highest engraftment levels in vivo. Importantly, no alterations in the vector integration profiles could be detected between CsA and control transduced HSPC. Overall, the present findings contribute to the development of more efficient and sustainable LV gene therapy protocols, underscoring the benefits of scaling down required vector doses, as well as shortening the HSPC ex vivo culture time.

Original languageEnglish
Pages (from-to)1133-1146
Number of pages14
JournalHuman Gene Therapy
Issue number9
Publication statusPublished - Sep 2019



  • engraftment and stemness
  • human hematopoietic stem cells
  • lentiviral transduction
  • transduction enhancers

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this