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

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

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)232-243
Number of pages12
JournalHuman gene therapy
Volume26
Issue number4
DOIs
StatePublished - Apr 1 2015

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Transgenes
Phosphoglycerate Kinase
Peptide Elongation Factor 1
T-Lymphocytes
B-Lymphocytes
Severe Combined Immunodeficiency
Endonucleases
Hematopoietic Stem Cell Transplantation
Lymphocytes
Radiation
Antigens

Cite this

Role of transgene regulation in ex vivo lentiviral correction of artemis deficiency. / Multhaup, Megan M.; Podetz-Pedersen, Kelly M.; Karlen, Andrea D.; Olson, Erik R.; Gunther, Roland; Somia, Nikunj V.; Blazar, Bruce R.; Cowan, Morton J.; McIvor, R. Scott.

In: Human gene therapy, Vol. 26, No. 4, 01.04.2015, p. 232-243.

Research output: Contribution to journalArticle

Multhaup, MM, Podetz-Pedersen, KM, Karlen, AD, Olson, ER, Gunther, R, Somia, NV, Blazar, BR, Cowan, MJ & McIvor, RS 2015, 'Role of transgene regulation in ex vivo lentiviral correction of artemis deficiency', Human gene therapy, vol. 26, no. 4, pp. 232-243. https://doi.org/10.1089/hum.2014.062
Multhaup MM, Podetz-Pedersen KM, Karlen AD, Olson ER, Gunther R, Somia NV et al. Role of transgene regulation in ex vivo lentiviral correction of artemis deficiency. Human gene therapy. 2015 Apr 1;26(4):232-243. https://doi.org/10.1089/hum.2014.062
Multhaup, Megan M. ; Podetz-Pedersen, Kelly M. ; Karlen, Andrea D. ; Olson, Erik R. ; Gunther, Roland ; Somia, Nikunj V. ; Blazar, Bruce R. ; Cowan, Morton J. ; McIvor, R. Scott. / Role of transgene regulation in ex vivo lentiviral correction of artemis deficiency. In: Human gene therapy. 2015 ; Vol. 26, No. 4. pp. 232-243.
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