Stable and functional lymphoid reconstitution in artemis-deficient mice following lentiviral artemis gene transfer into hematopoietic stem cells.
Patients with mutations in the Artemis gene display a complete absence of T- and B lymphocytes, together with increased cellular radiosensitivity; this leads to a radiosensitive severe combined immunodeficiency (RS-SCID). Allogenic hematopoietic stem-cell (HSC) transplantation is only partially successful in the absence of an human leukocyte antigen-genoidentical donor, and this has prompted a search for alternative therapeutic approaches such as gene therapy. In this study, a self-inactivated lentiviral vector expressing Artemis was used to complement the Artemis knockout mouse (Art(-/-)). Transplantation of Artemis-transduced HSCs into irradiated Art(-/-) mice restored a stable (over a 15-month period of follow-up) and functional T- and cell repertoire that was comparable to that of control mice. The success of secondary transplantations demonstrated that the HSCs had been transduced. One of thirteen mice developed a thymoma 6 months after gene therapy. Although thymic cells were seen to be carrying two lentiviral integration sites, there was no evidence of lentivirus-driven oncogene activation. The Art(-/-) mice were found to be prone to develop T-cell lymphomas, either spontaneously or after irradiation. These data indicate that the observed lymphoproliferation was probably the consequence of the chromosomal instability associated with the Artemis-deficient background. As a whole, our work provides a basis for supporting the gene therapy approach in Artemis-deficient SCID.
Institut National de la Santé et de la Recherche (INSERM), U768, Université Paris Descartes, Hôpital Necker-Enfants Malades, Paris, France.
Fatine
F
Benjelloun
Alexandrine
A
Garrigue
Corinne
C
Demerens-de Chappedelaine
Pauline
P
Soulas-Sprauel
Michele
M
Malassis-Séris
Daniel
D
Stockholm
Julia
J
Hauer
Johanna
J
Blondeau
Julie
J
Rivière
Annick
A
Lim
Marc
M
Le Lorc'h
Serge
S
Romana
Nicole
N
Brousse
Frederique
F
Pâques
Anne
A
Galy
Pierre
P
Charneau
Alain
A
Fischer
Jean-Pierre
JP
de Villartay
Marina
M
Cavazzana-Calvo
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D017252:CD3 Complex / Q000032:analysis; D015704:CD4 Antigens / Q000032:analysis; D016827:CD8 Antigens / Q000032:analysis; D002478:Cells, Cultured; D004720:Endonucleases; D005260:Female; D005434:Flow Cytometry; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D018960:Hyaluronan Receptors / Q000032:analysis; D053645:Interleukin-2 Receptor alpha Subunit / Q000032:analysis; D016086:Lentivirus / Q000235:genetics*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009687:Nuclear Proteins / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D016511:Severe Combined Immunodeficiency / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D013601:T-Lymphocytes / Q000276:immunology; D013950:Thymus Gland / Q000276:immunology / Q000473:pathology; D014161:Transduction, Genetic
D017252:CD3 Complex; D015704:CD4 Antigens; D016827:CD8 Antigens; D018960:Hyaluronan Receptors; D053645:Interleukin-2 Receptor alpha Subunit; D009687:Nuclear Proteins; D004720:Endonucleases; C473337:Dclre1c protein, mouse