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

• Verfasst von: Bogert, Nicolai [VerfasserIn]
• Verfasst von: Furkel, Jennifer [VerfasserIn]
• Verfasst von: Din, Shabana [VerfasserIn]
• Verfasst von: Braren, I. [VerfasserIn]
• Verfasst von: Eckstein, Volker [VerfasserIn]
• Verfasst von: Müller, J. A. [VerfasserIn]
• Verfasst von: Uhlmann, Lorenz [VerfasserIn]
• Verfasst von: Katus, Hugo [VerfasserIn]
• Verfasst von: Konstandin, Mathias [VerfasserIn]
• Titel: A novel approach to genetic engineering of T-cell subsets by hematopoietic stem cell infection with a bicistronic lentivirus
• Verf.angabe: N.V. Bogert, J. Furkel, S. Din, I. Braren, V. Eckstein, J.A. Müller, L. Uhlmann, H.A. Katus & M.H. Konstandin
• E-Jahr: 2020
• Jahr: 13 August 2020
• Umfang: 10 S.
• Fussnoten: Gesehen am 12.10.2020
• Titel Quelle: Enthalten in: Scientific reports
• Ort Quelle: [London] : Springer Nature, 2011
• Jahr Quelle: 2020
• Band/Heft Quelle: 10(2020) Artikel-Nummer 13740, 10 Seiten
• ISSN Quelle: 2045-2322
• DOI: doi:10.1038/s41598-020-70793-6
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 1735364835
• K10plus-PPN:
  Universitätsbibliothek
• Lokale URL UB: Zum Volltext

Abstract

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.