In vivo gene therapy of metachromatic leukodystrophy by lentiviral vectors: correction of neuropathology and protection against learning impairments in affected mice

Consiglio, Antonella; Quattrini, Angelo; Martino, Sabata; Bensadoun, J.C.; Dolcetta, Diego; Trojani, A.; Benaglia, Giuliana; Marchesini, Sergio; Cestari, Vincenzo; Oliverio, Alberto; Bordignon, Claudio; Naldini, Luigi

doi:10.1038/85454

Cited by 198 publications

(109 citation statements)

References 41 publications

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“…In addition, we observed tissue damage after expression of Cre for several weeks in the brain. With use of lentiviral vectors that carry other transgenes, we and others have observed long-term expression of the transgenes without obvious indications of tissue damage (18,24,35). Cre-induced cytotoxicity might be an explanation for the reduced number of Cre-expressing, ␤-gal ϩ cells that we observed in vitro and after prolonged (3-5 weeks) Cre expression in the brain.…”

Section: Discussionmentioning

confidence: 61%

Delivery of the Cre recombinase by a self-deleting lentiviral vector: Efficient gene targeting in vivo

Pfeifer

Brandon²,

Kootstra³

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

221

201

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The Cre recombinase (Cre) from bacteriophage P1 is an important tool for genetic engineering in mammalian cells. We constructed lentiviral vectors that efficiently deliver Cre in vitro and in vivo. Surprisingly, we found a significant reduction in proliferation and an accumulation in the G2͞M phase of Cre-expressing cells. To minimize the toxic effect of Cre, we designed a lentiviral vector that integrates into the host genome, expresses Cre in the target cell, and is subsequently deleted from the genome in a Credependent manner. Thus, the activity of Cre terminates its own expression (self-deleting). We showed efficient modification of target genes in vitro and in the brain after transduction with the self-deleting vectors. In contrast to sustained Cre expression, transient expression of Cre from the self-deleting vector induced significantly less cytotoxicity. Such a self-deleting Cre vector is a promising tool for the induction of conditional gene modifications with minimal Cre toxicity in vivo.gene transfer ͉ regulated gene expression ͉ Cre toxicity T he site-specific recombinase of the bacteriophage P1, Cre, catalyzes the recombination of two 34-bp sequences called loxP, which consist of two 13-bp palindromic sequences flanking an 8-bp core sequence (1, 2). Because of the high efficiency of the Cre͞loxP system in mammalian cells (3, 4), this system is now widely used for genetic engineering in vitro and in vivo (5, 6). Transfer of Cre in vivo can be achieved by the use of transgenic mice or by viral delivery to the target organ(s). Thus far, the transfer of Cre by using nonviral delivery vehicles such as liposomes has only been described for use in vitro (7).We have generated lentiviral vectors that carry the Cre recombinase and have analyzed their ability to mediate loxPspecific recombination. Lentiviruses are enveloped RNA viruses that belong to the family of complex retroviruses (for review see refs. 8 and 9). In contrast to prototypic retroviruses, such as murine leukemia virus, lentiviruses are able to transduce both dividing and nondividing cells. Lentiviral vectors pseudotyped with the G protein of the vesicular stomatitis virus can transduce a wide range of cells and have great potential for human gene therapy (10-13).The Cre lentiviral vectors efficiently transferred Cre to the target cells and induced Cre-mediated recombination of loxP sites in vitro and in vivo. The Cre-expressing cells unexpectedly exhibited a reduction in proliferation and accumulated in the G 2 ͞M phase of the cell cycle. To circumvent this toxic effect of Cre, we designed a Cre lentivirus vector that is itself subject to Cre-mediated excision (self-deleting or LV-Cre-SD). In this way, Cre expression is limited to only the time necessary for recombination. Our studies show that this approach works efficiently in vitro and in vivo. Materials and MethodsAnimals. Homozygous R26R mice (12-20 weeks of age) were anesthetized by i.p. injection of a ketamine (150 mg/kg)͞xylazine (15 mg/kg) mixture. Virus (3 l) was injected either ster...

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Section: Discussionmentioning

confidence: 61%

Delivery of the Cre recombinase by a self-deleting lentiviral vector: Efficient gene targeting in vivo

Pfeifer

Brandon²,

Kootstra³

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

221

201

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“…Many invasive strategies have been used or proposed for circumventing the blood-brain barrier, including intraparenchymal injections of therapeutic proteins, intraparenchymal gene therapy, chemical or physical agents to open the blood-brain barrier (such as mannitol), and hematopoietic stem cell transplantation [1][2][3][4][5][6][7][8][9]. Treatment via the cerebrospinal fluid has not been successful in the past due to the inability of the proteins to traverse the ependymal layer and diffuse through brain tissue, even though this route would be clinically easier for application to patients.…”

Section: Introductionmentioning

confidence: 99%

Intrathecal enzyme replacement therapy: Successful treatment of brain disease via the cerebrospinal fluid

Dickson

McEntee

Vogler

et al. 2007

Molecular Genetics and Metabolism

161

138

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Treatment of brain disease with recombinant proteins is difficult due to the blood-brain barrier. As an alternative to direct injections into the brain, we studied whether application of high concentrations of therapeutic enzymes via intrathecal (IT) injections could successfully drive uptake across the ependyma to treat brain disease. We studied IT enzyme replacement therapy with recombinant human iduronidase (rhIDU) in canine mucopolysaccharidosis I (MPS I, Hurler syndrome), a lysosomal storage disorder with brain and meningeal involvement. Monthly or quarterly IT treatment regimens with rhIDU achieved supranormal iduronidase enzyme levels in the brain, spinal cord, and spinal meninges. All regimens normalized total brain glycosaminoglycan (GAG) storage and reduced spinal meningeal GAG storage by 58-70%. The improvement in GAG storage levels persisted three months after the final IT dose. The successful use of enzyme therapy via the CSF represents a potentially useful approach for lysosomal storage disorders.

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“…2 The direct injection of viral vectors into the brain parenchyma allows only a restricted diffusion-few centimeters away from the injection site-of viral particles and large molecules (except for proteins actively transported by neurons). 3,4 Systemic as well as intraparenchymal delivery of viral vectors usually induces humoral and cell-mediated immune responses against the vectors and/or transgene products, leading to inflammation, short-term persistence of gene expression and elimination of transduced cells. [5][6][7] We, and others, have previously shown that injection of viral vectors into the cisterna magna, or trough lumbar puncture, might represent an efficient CNS delivery system in both small (mice) and large (non-human primates) animals.…”

Section: Introductionmentioning

confidence: 99%

IL4 gene delivery to the CNS recruits regulatory T cells and induces clinical recovery in mouse models of multiple sclerosis

et al. 2008

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Central nervous system (CNS) delivery of anti-inflammatory cytokines, such as interleukin 4 (IL4), holds promise as treatment for multiple sclerosis (MS). We have previously shown that short-term herpes simplex virus type 1-mediated IL4 gene therapy is able to inhibit experimental autoimmune encephalomyelitis (EAE), an animal model of MS, in mice and non-human primates. Here, we show that a single administration of an IL4-expressing helper-dependent adenoviral vector (HD-Ad) into the cerebrospinal fluid (CSF) circulation of immunocompetent mice allows persistent transduction of neuroepithelial cells and long-term (up to 5 months) CNS transgene expression without toxicity.Mice affected by chronic and relapsing EAE display clinical and neurophysiological recovery from the disease once injected with the IL4-expressing HD-Ad vector. The therapeutic effect is due to the ability of IL4 to increase, in inflamed CNS areas, chemokines (CCL1, CCL17 and CCL22) capable of recruiting regulatory T cells (CD4 + CD69 À CD25 + Foxp3 + ) with suppressant functions. CSF delivery of HD-Ad vectors expressing anti-inflammatory molecules might represent a valuable therapeutic option for CNS inflammatory disorders.

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In vivo gene therapy of metachromatic leukodystrophy by lentiviral vectors: correction of neuropathology and protection against learning impairments in affected mice

Cited by 198 publications

References 41 publications

Delivery of the Cre recombinase by a self-deleting lentiviral vector: Efficient gene targeting in vivo

Delivery of the Cre recombinase by a self-deleting lentiviral vector: Efficient gene targeting in vivo

Intrathecal enzyme replacement therapy: Successful treatment of brain disease via the cerebrospinal fluid

IL4 gene delivery to the CNS recruits regulatory T cells and induces clinical recovery in mouse models of multiple sclerosis

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