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A novel approach to genetic engineering of T-cell subsets by hematopoietic stem cell infection with a bicistronic lentivirus

Lentiviral modification of hematopoietic stem cells (HSCs) paved the way for in vivo experimentation and therapeutic approaches in patients with genetic disease. A disadvantage of this method is the use of a ubiquitous promoter leads not only to genetic modification of the leukocyte subset of intere...

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Bibliographic Details
Main Authors: Bogert, Nicolai (Author) , Furkel, Jennifer (Author) , Din, Shabana (Author) , Braren, I. (Author) , Eckstein, Volker (Author) , Müller, J. A. (Author) , Uhlmann, Lorenz (Author) , Katus, Hugo (Author) , Konstandin, Mathias (Author)
Format: Article (Journal)
Language:English
Published: 13 August 2020
In: Scientific reports
Year: 2020, Volume: 10
ISSN:2045-2322
DOI:10.1038/s41598-020-70793-6
Online Access:Verlag, kostenfrei, Volltext: https://doi.org/10.1038/s41598-020-70793-6
Verlag, kostenfrei, Volltext: https://www.nature.com/articles/s41598-020-70793-6
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Author Notes:N.V. Bogert, J. Furkel, S. Din, I. Braren, V. Eckstein, J.A. Müller, L. Uhlmann, H.A. Katus & M.H. Konstandin
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Summary:Lentiviral modification of hematopoietic stem cells (HSCs) paved the way for in vivo experimentation and therapeutic approaches in patients with genetic disease. A disadvantage of this method is the use of a ubiquitous promoter leads not only to genetic modification of the leukocyte subset of interest e.g. T-cells, but also all other subsequent leukocyte progeny of the parent HSCs. To overcome this limitation we tested a bicistronic lentivirus, enabling subset specific modifications. Designed novel lentiviral constructs harbor a global promoter (mPGK) regulating mCherry for HSCs selection and a T-cell specific promoter upstream of eGFP. Two T-cell specific promoters were assessed: the distal Lck—(dLck) and the CD3δ-promoter. Transduced HSCs were FACS sorted by mCherry expression and transferred into sublethally irradiated C57/BL6 mice. Successful transplantation and T-cell specific expression of eGFP was monitored by peripheral blood assessment. Furthermore, recruitment response of lentiviral engineered leukocytes to the site of inflammation was tested in a peritonitis model without functional impairment. Our constructed lentivirus enables fast generation of subset specific leukocyte transgenesis as shown in T-cells in vivo and opens new opportunities to modify other HSCs derived subsets in the future.
Item Description:Gesehen am 12.10.2020
Physical Description:Online Resource
ISSN:2045-2322
DOI:10.1038/s41598-020-70793-6

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