Molecular evidence of genome editing in a mouse model of immunodeficiency

• Verfasst von: Abdul-Razak, Hayder [VerfasserIn]
• Verfasst von: Gabriel, Richard [VerfasserIn]
• Verfasst von: Bartholomä, Cynthia C. [VerfasserIn]
• Verfasst von: Kalle, Christof von [VerfasserIn]
• Verfasst von: Schmidt, Michael [VerfasserIn]
• Titel: Molecular evidence of genome editing in a mouse model of immunodeficiency
• Verf.angabe: H.H. Abdul-Razak, C.J. Rocca, S.J. Howe, M.E. Alonso-Ferrero, J. Wang, R. Gabriel, C.C. Bartholomae, C.H.V. Gan, M.I. Garín, A. Roberts, M.P. Blundell, V. Prakash, F.J. Molina-Estevez, J. Pantoglou, G. Guenechea, M.C. Holmes, P.D. Gregory, C. Kinnon, C. von Kalle, M. Schmidt, J.A. Bueren, A.J. Thrasher & R.J. Yáñez-Muñoz
• E-Jahr: 2018
• Jahr: 29 May 2018
• Umfang: 13 S.
• Fussnoten: Gesehen am 02.04.2019
• Titel Quelle: Enthalten in: Scientific reports
• Ort Quelle: [London] : Macmillan Publishers Limited, part of Springer Nature, 2011
• Jahr Quelle: 2018
• Band/Heft Quelle: 8(2018) Artikel-Nummer 8214, 13 Seiten
• ISSN Quelle: 2045-2322
• DOI: doi:10.1038/s41598-018-26439-9
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 1662708122

Abstract

Genome editing is the introduction of directed modifications in the genome, a process boosted to therapeutic levels by designer nucleases. Building on the experience of ex vivo gene therapy for severe combined immunodeficiencies, it is likely that genome editing of haematopoietic stem/progenitor cells (HSPC) for correction of inherited blood diseases will be an early clinical application. We show molecular evidence of gene correction in a mouse model of primary immunodeficiency. In vitro experiments in DNA-dependent protein kinase catalytic subunit severe combined immunodeficiency (Prkdc scid) fibroblasts using designed zinc finger nucleases (ZFN) and a repair template demonstrated molecular and functional correction of the defect. Following transplantation of ex vivo gene-edited Prkdc scid HSPC, some of the recipient animals carried the expected genomic signature of ZFN-driven gene correction. In some primary and secondary transplant recipients we detected double-positive CD4/CD8 T-cells in thymus and single-positive T-cells in blood, but no other evidence of immune reconstitution. However, the leakiness of this model is a confounding factor for the interpretation of the possible T-cell reconstitution. Our results provide support for the feasibility of rescuing inherited blood disease by ex vivo genome editing followed by transplantation, and highlight some of the challenges.