Ex vivo gene therapy with lentiviral vectors rescues adenosine deaminase (ADA)–deficient mice and corrects their immune and metabolic defects

Mortellaro, Alessandra; Hernández, Raisa Jofra; Guerrini, Matteo M.; Carlucci, Filippo; Tabucchi, Antonella; Ponzoni, Maurilio; Sanvito, Francesca; Doglioni, Claudio; Serio, Clelia Di; Biasco, Luca; Follenzi, Antonia; Naldini, Luigi; Bordignon, Claudio; Roncarolo, Maria Grazia; Aiuti, Alessandro

doi:10.1182/blood-2006-05-023507

Cited by 71 publications

(61 citation statements)

References 42 publications

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“…34 Our study included a wide distribution of VCN (ranging from 0.3 to 9.8). While these VCNs are in line with previous studies from our group and others [36][37][38] it is expected that lower VCNs would be observed in the clinical application based on our previous studies in human CD34+ cells, 23 and clinical trials in other therapeutic areas. 34,39 Unfortunately, no published data is available regarding the protection from the infections in clinical trials with lentiviral vectors encoding for gp91phox.…”

Section: Discussionsupporting

confidence: 77%

Lentiviral Vector Gene Therapy Protects XCGD Mice From Acute Staphylococcus aureus Pneumonia and Inflammatory Response

Farinelli

Hernández

Rossi³

et al. 2016

Molecular Therapy

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Chronic granulomatous disease (CGD) is a primary immunodeficiency due to a deficiency in one of the subunits of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex. CGD patients are characterized by an increased susceptibility to bacterial and fungal infections, and to granuloma formation due to the excessive inflammatory responses. Several gene therapy approaches with lentiviral vectors have been proposed but there is a lack of in vivo data on the ability to control infections and inflammation. We set up a mouse model of acute infection that closely mimic the airway infection in CGD patients. It involved an intratracheal injection of a methicillin-sensitive reference strain of S. aureus. Gene therapy, with hematopoietic stem cells transduced with regulated lentiviral vectors, restored the functional activity of NADPH oxidase complex (with 20-98% of dihydrorhodamine positive granulocytes and monocytes) and saved mice from death caused by S. aureus, significantly reducing the bacterial load and lung damage, similarly to WT mice even at low vector copy number. When challenged, gene therapytreated XCGD mice showed correction of proinflammatory cytokines and chemokine imbalance at levels that were comparable to WT. Examined together, our results support the clinical development of gene therapy protocols using lentiviral vectors for the protection against infections and inflammation.

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Section: Discussionsupporting

confidence: 77%

Lentiviral Vector Gene Therapy Protects XCGD Mice From Acute Staphylococcus aureus Pneumonia and Inflammatory Response

Farinelli

Hernández

Rossi³

et al. 2016

Molecular Therapy

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“…To further analyze the LV-FL DYSF transduction efficiency we synthesized a second lentiviral vector using the same pRRL backbone subcloned with GFP transcript. Our lentivirus construction recapitulates the transduction efficiency of previously tested and published lentivirus vectors [38,39]. We performed FACS analysis to determine the expression of GFP in infected CD133+ normal blood derived cells and we showed a percentage of 60.9% of GFP positive cells demonstrating a good efficiency of transduction as previously described [40,41] (Fig.…”

Section: Patient Phenotype and Genotype Descriptionmentioning

confidence: 99%

Full‐length dysferlin expression driven by engineered human dystrophic blood derived CD133+ stem cells

et al. 2013

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The protein dysferlin is abundantly expressed in skeletal and cardiac muscles, where its main function is membrane repair. Mutations in the dysferlin gene are involved in two autosomal recessive muscular dystrophies: Miyoshi myopathy and limb‐girdle muscular dystrophy type 2B. Development of effective therapies remains a great challenge. Strategies to repair the dysferlin gene by skipping mutated exons, using antisense oligonucleotides (AONs), may be suitable only for a subset of mutations, while cell and gene therapy can be extended to all mutations. AON‐treated blood‐derived CD133+ stem cells isolated from patients with Miyoshi myopathy led to partial dysferlin reconstitution in vitro but failed to express dysferlin after intramuscular transplantation into scid/blAJ dysferlin null mice. We thus extended these experiments producing the full‐length dysferlin mediated by a lentiviral vector in blood‐derived CD133+ stem cells isolated from the same patients. Transplantation of engineered blood‐derived CD133+ stem cells into scid/blAJ mice resulted in sufficient dysferlin expression to correct functional deficits in skeletal muscle membrane repair. Our data suggest for the first time that lentivirus‐mediated delivery of full‐length dysferlin in stem cells isolated from Miyoshi myopathy patients could represent an alternative therapeutic approach for treatment of dysferlinopathies.

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“…Viral vector systems derived from lentivirus or FV are currently under investigation in small and large animal models [9,[19][20][21][22][23][24][25][26][27][28][29][30][31]. Russell and colleagues demonstrated that murine stem cells residing in 5-fluorouracil (5FU)-treated bone marrow (BM) [24] and SCID-repopulating cells derived from umbilical cord blood and G-CSF-mobilized peripheral blood could be marked with FV vectors.…”

Section: Introductionmentioning

confidence: 99%

In vivo selection of hematopoietic stem cells transduced at a low multiplicity-of-infection with a foamy viral MGMTP140K vector

Cai¹,

Ernstberger²,

Wang³

et al. 2008

Experimental Hematology

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Objective-Using a clinically relevant transduction strategy, we investigated to what extent hematopoietic stem cells in lineage-negative bone marrow (Lin neg BM) could be genetically modified with a FV vector that expresses the DNA repair protein, O 6 -methylguanine DNA methyltransferase (MGMT P140K ) and selected in vivo with submyeloablative versus myeloablative alkylator therapy.Methods-Lin neg BM was transduced at a low multiplicity-of-infection (MOI), with the FV vector, MD9-P140K, that co-expresses MGMT P140K and the enhanced green fluorescent protein, transplanted into C57BL/6 mice, and mice treated with submyeloablative or myeloablative alkylator therapy. The BM was analyzed for the presence of in vivo selected, MD9-P140K-transduced cells at 6 months post-transplantation and subsequently transplanted into secondary recipient animals.Results-Following submyeloablative therapy, 55% of the mice expressed MGMT P140K in the BM. Proviral integration was observed in ∼50% of committed BM-derived progenitors and analysis of proviral insertion sites indicated up to 2 integrations per transduced progenitor colony. Transduced BM cells selected with submyeloablative therapy reconstituted secondary recipient mice for up to 6 months post-transplantation. In contrast, following delivery of myeloablative therapy to primary recipient mice, only 25% survived. Hematopoietic stem cells were transduced since BM cells from the surviving animals reconstituted secondary recipients with MGMT P140K positive cells for 5-6 months. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. of transduced-stem cells must be present to produce sufficient numbers of genetically modified progeny to protect against the acute toxicity associated with myeloablative therapy. Conclusions-In NIH Public Access

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Ex vivo gene therapy with lentiviral vectors rescues adenosine deaminase (ADA)–deficient mice and corrects their immune and metabolic defects

Cited by 71 publications

References 42 publications

Lentiviral Vector Gene Therapy Protects XCGD Mice From Acute Staphylococcus aureus Pneumonia and Inflammatory Response

Lentiviral Vector Gene Therapy Protects XCGD Mice From Acute Staphylococcus aureus Pneumonia and Inflammatory Response

Full‐length dysferlin expression driven by engineered human dystrophic blood derived CD133+ stem cells

In vivo selection of hematopoietic stem cells transduced at a low multiplicity-of-infection with a foamy viral MGMTP140K vector

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