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.
| Original language | English |
|---|---|
| Pages (from-to) | 1472-1483 |
| Number of pages | 12 |
| Journal | Molecular Therapy |
| Volume | 22 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - 2014 |
Fingerprint
ASJC Scopus subject areas
- Molecular Biology
- Molecular Medicine
- Genetics
- Drug Discovery
- Pharmacology
- Medicine(all)
Cite this
Dual-regulated lentiviral vector for gene therapy of X-linked chronic granulomatosis. / Chiriaco, Maria; Farinelli, Giada; Capo, Valentina; Zonari, Erika; Scaramuzza, Samantha; Di Matteo, Gigliola; Sergi, Lucia Sergi; Migliavacca, Maddalena; Hernandez, Raisa Jofra; Bombelli, Ferdinando; Giorda, Ezio; Kajaste-Rudnitski, Anna; Trono, Didier; Grez, Manuel; Rossi, Paolo; Finocchi, Andrea; Naldini, Luigi; Gentner, Bernhard; Aiuti, Alessandro.
In: Molecular Therapy, Vol. 22, No. 8, 2014, p. 1472-1483.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Dual-regulated lentiviral vector for gene therapy of X-linked chronic granulomatosis
AU - Chiriaco, Maria
AU - Farinelli, Giada
AU - Capo, Valentina
AU - Zonari, Erika
AU - Scaramuzza, Samantha
AU - Di Matteo, Gigliola
AU - Sergi, Lucia Sergi
AU - Migliavacca, Maddalena
AU - Hernandez, Raisa Jofra
AU - Bombelli, Ferdinando
AU - Giorda, Ezio
AU - Kajaste-Rudnitski, Anna
AU - Trono, Didier
AU - Grez, Manuel
AU - Rossi, Paolo
AU - Finocchi, Andrea
AU - Naldini, Luigi
AU - Gentner, Bernhard
AU - Aiuti, Alessandro
PY - 2014
Y1 - 2014
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84905451458&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84905451458&partnerID=8YFLogxK
U2 - 10.1038/mt.2014.87
DO - 10.1038/mt.2014.87
M3 - Article
C2 - 24869932
AN - SCOPUS:84905451458
VL - 22
SP - 1472
EP - 1483
JO - Molecular Therapy
JF - Molecular Therapy
SN - 1525-0016
IS - 8
ER -