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

Maria Chiriaco, Giada Farinelli, Valentina Capo, Erika Zonari, Samantha Scaramuzza, Gigliola Di Matteo, Lucia Sergi Sergi, Maddalena Migliavacca, Raisa Jofra Hernandez, Ferdinando Bombelli, Ezio Giorda, Anna Kajaste-Rudnitski, Didier Trono, Manuel Grez, Paolo Rossi, Andrea Finocchi, Luigi Naldini, Bernhard Gentner, Alessandro Aiuti

Research output: Contribution to journalArticle

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 languageEnglish
Pages (from-to)1472-1483
Number of pages12
JournalMolecular Therapy
Volume22
Issue number8
DOIs
Publication statusPublished - 2014

Fingerprint

Hematopoietic Stem Cells
Genetic Therapy
Chronic Granulomatous Disease
Myeloid Cells
Transgenes
Bone Marrow
Heterologous Transplantation
Bone Development
Cell- and Tissue-Based Therapy
MicroRNAs
Monocytes
Reactive Oxygen Species
Carcinogenesis
4-ethoxymethylene-2-phenyl-2-oxazoline-5-one
Clinical Trials
Safety
Cell Line
Therapeutics

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 journalArticle

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. / Dual-regulated lentiviral vector for gene therapy of X-linked chronic granulomatosis. In: Molecular Therapy. 2014 ; Vol. 22, No. 8. pp. 1472-1483.
@article{ec5d8c03332541e8b72c0dce7689b83a,
title = "Dual-regulated lentiviral vector for gene therapy of X-linked chronic granulomatosis",
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.",
author = "Maria Chiriaco and Giada Farinelli and Valentina Capo and Erika Zonari and Samantha Scaramuzza and {Di Matteo}, Gigliola and Sergi, {Lucia Sergi} and Maddalena Migliavacca and Hernandez, {Raisa Jofra} and Ferdinando Bombelli and Ezio Giorda and Anna Kajaste-Rudnitski and Didier Trono and Manuel Grez and Paolo Rossi and Andrea Finocchi and Luigi Naldini and Bernhard Gentner and Alessandro Aiuti",
year = "2014",
doi = "10.1038/mt.2014.87",
language = "English",
volume = "22",
pages = "1472--1483",
journal = "Molecular Therapy",
issn = "1525-0016",
publisher = "Nature Publishing Group",
number = "8",

}

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 -