Role of transgene regulation in ex vivo lentiviral correction of artemis deficiency.

Abstract

Artemis is a single-stranded endonuclease, deficiency of which results in a radiation-sensitive form of severe combined immunodeficiency (SCID-A) most effectively treated by allogeneic hematopoietic stem cell (HSC) transplantation and potentially treatable by administration of genetically corrected autologous HSCs. We previously reported cytotoxicity associated with Artemis overexpression and subsequently characterized the human Artemis promoter with the intention to provide Artemis expression that is nontoxic yet sufficient to support immunodevelopment. Here we compare the human Artemis promoter (APro) with the moderate-strength human phosphoglycerate kinase (PGK) promoter and the strong human elongation factor-1α (EF1α) promoter to regulate expression of Artemis after ex vivo lentiviral transduction of HSCs in a murine model of SCID-A. Recipient animals treated with the PGK-Artemis vector exhibited moderate repopulation of their immune compartment, yet demonstrated a defective proliferative T lymphocyte response to in vitro antigen stimulation. Animals treated with the EF1α-Artemis vector displayed high levels of T lymphocytes but an absence of B lymphocytes and deficient lymphocyte function. In contrast, ex vivo transduction with the APro-Artemis vector supported effective immune reconstitution to wild-type levels, resulting in fully functional T and B lymphocyte responses. These results demonstrate the importance of regulated Artemis expression in immune reconstitution of Artemis-deficient SCID.

• Authors: Megan M Multhaup, Kelly M Podetz-Pedersen, Andrea D Karlen, Erik R Olson, Roland Gunther, Nikunj V Somia, Bruce R Blazar, Morton J Cowan, R Scott McIvor
• Journal: Human gene therapy (Hum Gene Ther) Vol. 26 Issue 4 Pg. 232-43 (Apr 2015) ISSN: 1557-7422 [Electronic] United States
• PMID: 25738323 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Nuclear Proteins
• Endonucleases
• Dclre1c protein, mouse

Topics

• Animals
• Endonucleases (biosynthesis, deficiency, genetics)
• Genetic Therapy
• HEK293 Cells
• Hematopoietic Stem Cell Transplantation
• Humans
• Lentivirus (genetics)
• Lymphocytes (immunology)
• Mice
• Mice, Inbred C57BL
• Mice, SCID
• NIH 3T3 Cells
• Nuclear Proteins (biosynthesis, deficiency, genetics)
• Severe Combined Immunodeficiency (immunology, therapy)
• Transcriptional Activation
• Transduction, Genetic
• Transgenes