Cloning, sequencing, and expression of cDNA for human beta-glucuronidase.

Oshima, Akihiro; Kyle, John W.; Miller, Raymond D.; Hoffmann, Joseph W.; Powell, Penny P.; Grubb, Jeffrey H.; Sly, William S.; Tropak, Michael B.; Guise, K. S.; Gravel, Roy A.

doi:10.1073/pnas.84.3.685

Cited by 159 publications

(76 citation statements)

References 47 publications

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“…Following cleavage with NciI or ScrFI, the 77-bp restriction fragment present in the normal allele is distinguished on a 3% Nu Sieve agarose gel from the uncleaved 95-bp PCR fragment from the mutant allele. The presence of the human transgene was detected by PCR of tail DNA using forward primer 5Ј-GCTGGTGAATTACCAGATCTCTGT-CAA-3Ј and reverse primer 5Ј-GGAAATAGAAAGGTTTC-CCATTGATGAGG-3Ј, which amplified a 312-bp fragment spanning nucleotides 750-1062 of the human cDNA (44).…”

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confidence: 99%

Active site mutant transgene confers tolerance to human β-glucuronidase without affecting the phenotype of MPS VII mice

Sly

Vogler

Grubb

et al. 2001

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

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Mucopolysaccharidosis type VII (MPS VII; Sly syndrome) is an autosomal recessive lysosomal storage disorder due to an inherited deficiency of ␤-glucuronidase. A naturally occurring mouse model for this disease was discovered at The Jackson Laboratory and shown to be due to homozygosity for a 1-bp deletion in exon 10 of the gus gene. The murine model MPS VII (gus mps/mps ) has been very well characterized and used extensively to evaluate experimental strategies for lysosomal storage diseases, including bone marrow transplantation, enzyme replacement therapy, and gene therapy. To enhance the value of this model for enzyme and gene therapy, we produced a transgenic mouse expressing the human ␤-glucuronidase cDNA with an amino acid substitution at the active site nucleophile (E540A) and bred it onto the MPS VII (gus mps/mps ) background. We demonstrate here that the mutant mice bearing the active site mutant human transgene retain the clinical, morphological, biochemical, and histopathological characteristics of the original MPS VII (gus mps/mps ) mouse. However, they are now tolerant to immune challenge with human ␤-glucuronidase. This ''tolerant MPS VII mouse model'' should be useful for preclinical trials evaluating the effectiveness of enzyme and͞or gene therapy with the human gene products likely to be administered to human patients with MPS VII.

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mentioning

confidence: 99%

Active site mutant transgene confers tolerance to human β-glucuronidase without affecting the phenotype of MPS VII mice

Sly

Vogler

Grubb

et al. 2001

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

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“…(j) The immunogenicity of the antibody enzyme conjugate. Despite the obvious complexity of the system, a first clinical trial with a F(ab')2 fragment of a murine anti CEA MAb chemically linked to carboxypeptidase G2 (CPG2) from Pseudomonas origin combined with a para-N (mono-2-chloroethyl monomesyl) amino benzoyl glutamic acid prodrug was performed (Bagshawe, 1991 (Bosslet et al, 1988), its CDR-grafted (Jones et al, 1986;Riechmann et al, 1988) humanised version (Giissow & Seemann, 1991) and the cloned cDNA for human placental P-glucuronidase (Oshima et al, 1987), a lysomal endogenous enzyme (Brot et al, 1978). Out of these human building blocks a fusion gene was constructed .…”

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confidence: 99%

Molecular and functional characterisation of a fusion protein suited for tumour specific prodrug activation

Bosslet

Czech²,

Lorenz³

et al. 1992

Br J Cancer

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Currently, the treatment of non-disseminated solid tumours is performed by surgery and irradiation. Both methods can be considered to be relatively tumour selective, without significantly harming the rest of the body. If however the tumour has disseminated to various organ sites, the metastases can be treated by chemo-or hormone therapy only. Chemotherapy has considerable side effects and a minor influence on patients' survival due to the lack of specificity of action or the induction of resistance. Hormone therapy alone has a limited tumour spectrum.To overcome these obvious limitations of today's treatment modalities, we tailored a fusion gene consisting of the VH and CHI Exons of a humanised MAb and the human P-glucuronidase cDNA . The product encoded by the fusion gene might be suitable for performing an antibody directed enzyme prodrug therapy (ADEPT). The concept of ADEPT as developed by Philpott et al. (1973a,b;1974) and reemphasised by Bagshawe (1987) and Bagshawe et al. (1988) assumes that an antibody enzyme conjugate after selective localisation at the tumour target site, and its clearance from normal tissues, activates a nontoxic low molecular weight prodrug to a highly toxic drug in the tumour by enzymatic catalysis.The therapeutic success of this approach depends on several factors:(a) The stability of the prodrug in vivo. (j) The immunogenicity of the antibody enzyme conjugate. Despite the obvious complexity of the system, a first clinical trial with a F(ab')2 fragment of a murine anti CEA MAb chemically linked to carboxypeptidase G2 (CPG2) from

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“…This PCR also generates a 454-bp fragment from the endogenous murine GUSB gene. 20,30 When adult mice were infected with AxCAh-GUS intravenously, a clear 240-bp DNA fragment was only detected in the PCR reaction from liver DNA, suggesting that AxCAhGUS accumulated predominantly in the liver. On the other hand, clear 240-bp DNA fragments were observed in all organs, when AxCAh-GUS was intravenously administered into newborn mice, demonstrating that a significant amount of virus was distributed into multiple organs including the brain.…”

Section: Presence Of Viral Dna In C3h Mice Treated With Axcahgusmentioning

confidence: 99%

Long-term normalization in the central nervous system, ocular manifestations, and skeletal deformities by a single systemic adenovirus injection into neonatal mice with mucopolysaccharidosis VII

et al. 2003

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Systemic injection of an adenovirus vector into adult mice resulted in pathological improvements in multiple visceral organs of mice with mucopolysaccharidosis VII; however, no therapeutic efficacy was observed for mental retardation, skeletal deformities, corneal clouding, and retinal degeneration. In this study, an adenovirus vector expressing human bglucuronidase was injected into mice with mucopolysaccharidosis VII within 24 h of birth, and therapeutic efficacy was evaluated. In the brains of the mice, more than 20% of GUSB activity was maintained for at least 20 weeks after birth, and histopathological analysis showed no obvious lysosomal storage. Furthermore, no vacuolated cells were detected in corneal stroma and retinal pigment epithelium in the eyes of the mice treated in the neonatal period, while pathological improvement was not observed in adult MPSVII mice that received similar treatments. The treated mice also lacked characteristic facial skeletal deformities, and radiographic analysis demonstrated that their facial and cranial bones were morphologically normal. These results indicate that a single systemic adenovirus injection in the neonatal period could prevent the progression of mental retardation, corneal clouding, retinal degeneration, and skeletal deformities, all of which are frequently observed clinical manifestations and difficult to treat in adulthood.

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Cloning, sequencing, and expression of cDNA for human beta-glucuronidase.

Abstract: We report here the cDNA sequence for hu-

Cited by 159 publications

References 47 publications

Active site mutant transgene confers tolerance to human β-glucuronidase without affecting the phenotype of MPS VII mice

Active site mutant transgene confers tolerance to human β-glucuronidase without affecting the phenotype of MPS VII mice

Molecular and functional characterisation of a fusion protein suited for tumour specific prodrug activation

Long-term normalization in the central nervous system, ocular manifestations, and skeletal deformities by a single systemic adenovirus injection into neonatal mice with mucopolysaccharidosis VII

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