Regulated transgene expression may improve the safety and efficacy of hematopoietic stem cell (HSC) gene therapy. Clinical trials for X-linked chronic granulomatous disease (X-CGD) employing gammaretroviral vectors were limited by insertional oncogenesis or lack of persistent engraftment. Our novel strategy, based on regulated lentiviral vectors (LV), targets gp91(phox) expression to the differentiated myeloid compartment while sparing HSC, to reduce the risk of genotoxicity and potential perturbation of reactive oxygen species levels. Targeting was obtained by a myeloid-specific promoter (MSP) and posttranscriptional, microRNA-mediated regulation. We optimized both components in human bone marrow (BM) HSC and their differentiated progeny in vitro and in a xenotransplantation model, and generated therapeutic gp91(phox) expressing LVs for CGD gene therapy. All vectors restored gp91(phox) expression and function in human X-CGD myeloid cell lines, primary monocytes, and differentiated myeloid cells. While unregulated LVs ectopically expressed gp91(phox) in CD34(+) cells, transcriptionally and posttranscriptionally regulated LVs substantially reduced this off-target expression. X-CGD mice transplanted with transduced HSC restored gp91(phox) expression, and MSP-driven vectors maintained regulation during BM development. Combining transcriptional (SP146.gp91-driven) and posttranscriptional (miR-126-restricted) targeting, we achieved high levels of myeloid-specific transgene expression, entirely sparing the CD34(+) HSC compartment. This dual-targeted LV construct represents a promising candidate for further clinical development.

Chiriaco, M., Farinelli, G., Capo, V., Zonari, E., Scaramuzza, S., DI MATTEO, G., et al. (2014). Dual-regulated lentiviral vector for gene therapy of X-linked chronic granulomatosis. MOLECULAR THERAPY, 22(8), 1472-1483 [10.1038/mt.2014.87].

Dual-regulated lentiviral vector for gene therapy of X-linked chronic granulomatosis

DI MATTEO, GIGLIOLA;ROSSI, PAOLO;FINOCCHI, ANDREA;AIUTI, ALESSANDRO
2014-08-01

Abstract

Regulated transgene expression may improve the safety and efficacy of hematopoietic stem cell (HSC) gene therapy. Clinical trials for X-linked chronic granulomatous disease (X-CGD) employing gammaretroviral vectors were limited by insertional oncogenesis or lack of persistent engraftment. Our novel strategy, based on regulated lentiviral vectors (LV), targets gp91(phox) expression to the differentiated myeloid compartment while sparing HSC, to reduce the risk of genotoxicity and potential perturbation of reactive oxygen species levels. Targeting was obtained by a myeloid-specific promoter (MSP) and posttranscriptional, microRNA-mediated regulation. We optimized both components in human bone marrow (BM) HSC and their differentiated progeny in vitro and in a xenotransplantation model, and generated therapeutic gp91(phox) expressing LVs for CGD gene therapy. All vectors restored gp91(phox) expression and function in human X-CGD myeloid cell lines, primary monocytes, and differentiated myeloid cells. While unregulated LVs ectopically expressed gp91(phox) in CD34(+) cells, transcriptionally and posttranscriptionally regulated LVs substantially reduced this off-target expression. X-CGD mice transplanted with transduced HSC restored gp91(phox) expression, and MSP-driven vectors maintained regulation during BM development. Combining transcriptional (SP146.gp91-driven) and posttranscriptional (miR-126-restricted) targeting, we achieved high levels of myeloid-specific transgene expression, entirely sparing the CD34(+) HSC compartment. This dual-targeted LV construct represents a promising candidate for further clinical development.
ago-2014
Pubblicato
Rilevanza internazionale
Articolo
Sì, ma tipo non specificato
Settore MED/38 - PEDIATRIA GENERALE E SPECIALISTICA
Settore BIO/11 - BIOLOGIA MOLECOLARE
English
Con Impact Factor ISI
Chiriaco, M., Farinelli, G., Capo, V., Zonari, E., Scaramuzza, S., DI MATTEO, G., et al. (2014). Dual-regulated lentiviral vector for gene therapy of X-linked chronic granulomatosis. MOLECULAR THERAPY, 22(8), 1472-1483 [10.1038/mt.2014.87].
Chiriaco, M; Farinelli, G; Capo, V; Zonari, E; Scaramuzza, S; DI MATTEO, G; Sergi, L; Migliavacca, M; Hernandez, R; Bombelli, F; Giorda, E; Kajaste Ru...espandi
Articolo su rivista
File in questo prodotto:
File Dimensione Formato  
mol theraphy 2014.pdf

accesso aperto

Licenza: Creative commons
Dimensione 1.12 MB
Formato Adobe PDF
1.12 MB Adobe PDF Visualizza/Apri

Questo articolo è pubblicato sotto una Licenza Licenza Creative Commons Creative Commons

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/100773
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 57
  • ???jsp.display-item.citation.isi??? 53
social impact