Lentivirus mediated correction of artemis-deficient severe combined immunodeficiency

Divya Punwani, Misako Kawahara, Jason Yu, Ukina Sanford, Sushmita Roy, Kiran Patel, Denise A. Carbonaro, Andrea D. Karlen, Sara Khan, Kenneth Cornetta, Michael Rothe, Axel Schambach, Donald B. Kohn, Harry L. Malech, R. Scott McIvor, Jennifer M. Puck, Morton J. Cowan

Research output: Contribution to journalArticlepeer-review

17 Scopus citations


During B and T lymphocyte maturation, V(D)J recombination is initiated by creation of DNA doublestrand 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.

Original languageEnglish (US)
Pages (from-to)112-124
Number of pages13
JournalHuman gene therapy
Issue number1
StatePublished - Jan 1 2017

Bibliographical note

Funding Information:
These studies were supported by a research grant from the California Institute of Regenerative Medicine TR3-05535 and the National Institutes of Health National Institute of Allergy and Infectious Diseases, Division of Intramural Research and Warren Magnusen Clinical Center. We thank Steve Yannone, Laurence Berkeley Laboratory, for anti-Artemis antibody and Yanning Wang for technical expertise. We also thank the ART-SCID patients who volunteered to donate cells for these studies.

Publisher Copyright:
© 2017 by Mary Ann Liebert, Inc.


  • Artemis
  • Gene therapy
  • Lentivirus
  • Radiation sensitivity
  • Severe combined immunodeficiency


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