Lentivirus Mediated Correction of Artemis-Deficient Severe Combined Immunodeficiency.
During B and T lymphocyte maturation, V(D)J recombination is initiated by creation of DNA double-strand breaks. Artemis is an exonuclease essential for their subsequent repair by nonhomologous end-joining. Mutations in DCLRE1C, the gene encoding Artemis, cause T-B-NK+severe combined immunodeficiency (ART-SCID) and also confer heightened sensitivity to ionizing radiation and alkylating chemotherapy. Although allogeneic hematopoietic cell transplantation can treat ART-SCID, conditioning regimens are poorly tolerated, leading to early mortality and/or late complications, including short stature, endocrinopathies, and dental aplasia. However, without alkylating chemotherapy as preconditioning, patients usually have graft rejection or limited T cell and no B cell recovery. Thus, addition of normal DCLRE1C cDNA to autologous hematopoietic stem cells is an attractive strategy to treat ART-SCID. We designed a self-inactivating lentivirus vector containing human Artemis cDNA under transcriptional regulation of the human endogenous Artemis promoter (AProArt). Fibroblasts from ART-SCID patients transduced with AProArt lentivirus showed correction of radiosensitivity. Mobilized peripheral blood CD34+cells from an ART-SCID patient as well as hematopoietic stem cells from Artemis-deficient mice demonstrated restored T and B cell development following AProArt transduction. Murine hematopoietic cells transduced with AProArt exhibited no increase in replating potential in an in vitro immortalization assay, and analysis of AProArt lentivirus insertions showed no predilection for sites that could activate oncogenes. These efficacy and safety findings support institution of a clinical trial of gene addition therapy for ART-SCID.
1 Department of Pediatrics, University of California School of Medicine and University of California San Francisco Benioff Children's Hospital , San Francisco, San Francisco, California.
Divya
D
Punwani
1 Department of Pediatrics, University of California School of Medicine and University of California San Francisco Benioff Children's Hospital , San Francisco, San Francisco, California.
Misako
M
Kawahara
1 Department of Pediatrics, University of California School of Medicine and University of California San Francisco Benioff Children's Hospital , San Francisco, San Francisco, California.
Jason
J
Yu
1 Department of Pediatrics, University of California School of Medicine and University of California San Francisco Benioff Children's Hospital , San Francisco, San Francisco, California.
Ukina
U
Sanford
1 Department of Pediatrics, University of California School of Medicine and University of California San Francisco Benioff Children's Hospital , San Francisco, San Francisco, California.
Sushmita
S
Roy
1 Department of Pediatrics, University of California School of Medicine and University of California San Francisco Benioff Children's Hospital , San Francisco, San Francisco, California.
Kiran
K
Patel
2 Departments of Microbiology, Immunology and Molecular Genetics and Pediatrics, University of California Los Angeles , Los Angeles, California.
Denise A
DA
Carbonaro
3 Department of Genetics, Cell Biology and Development, University of Minnesota , Minneapolis, Minnesota.
Andrea D
AD
Karlen
1 Department of Pediatrics, University of California School of Medicine and University of California San Francisco Benioff Children's Hospital , San Francisco, San Francisco, California.
Sara
S
Khan
4 Department of Medical and Molecular Genetics, Indiana University, and the Indiana University Viral Production Facility, Indianapolis, Indiana.
Kenneth
K
Cornetta
5 Institute for Experimental Hematology, Hannover Medical School , Hannover, Germany.
Michael
M
Rothe
5 Institute for Experimental Hematology, Hannover Medical School , Hannover, Germany.
Axel
A
Schambach
2 Departments of Microbiology, Immunology and Molecular Genetics and Pediatrics, University of California Los Angeles , Los Angeles, California.
Donald B
DB
Kohn
6 Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, Division of Intramural Research, National Institutes of Health , Bethesda, Maryland.
Harry L
HL
Malech
3 Department of Genetics, Cell Biology and Development, University of Minnesota , Minneapolis, Minnesota.
R Scott
RS
McIvor
1 Department of Pediatrics, University of California School of Medicine and University of California San Francisco Benioff Children's Hospital , San Francisco, San Francisco, California.
Jennifer M
JM
Puck
1 Department of Pediatrics, University of California School of Medicine and University of California San Francisco Benioff Children's Hospital , San Francisco, San Francisco, California.
Morton J
MJ
Cowan
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000378:metabolism / Q000528:radiation effects; D002478:Cells, Cultured; D003131:Combined Modality Therapy; D004260:DNA Repair / Q000528:radiation effects; D004268:DNA-Binding Proteins; D004195:Disease Models, Animal; D004720:Endonucleases / Q000172:deficiency / Q000235:genetics*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000528:radiation effects; D005720:Gamma Rays; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage*; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism / Q000528:radiation effects; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D018345:Mice, Knockout; D016513:Mice, SCID; D009687:Nuclear Proteins / Q000172:deficiency / Q000235:genetics*; D011836:Radiation Tolerance / Q000235:genetics; D016511:Severe Combined Immunodeficiency / Q000235:genetics / Q000628:therapy*; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism / Q000528:radiation effects
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C426344:DCLRE1C protein, human; D004720:Endonucleases; C473337:Dclre1c protein, mouse
Artemis; gene therapy; lentivirus; radiation sensitivity; severe combined immunodeficiency