scholarly journals Restoration of arylsulphatase B activity in human mucopolysaccharidosis-type-VI fibroblasts by retroviral-vector-mediated gene transfer

1991 ◽  
Vol 276 (2) ◽  
pp. 499-504 ◽  
Author(s):  
C Peters ◽  
W Rommerskirch ◽  
S Modaressi ◽  
K von Figura
Keyword(s):  
Gene Transfer ◽  
Retroviral Vector ◽  
Mucopolysaccharidosis Type ◽  
Lysosomal Storage ◽  
Packaging Cell Line ◽  
Retroviral Gene Transfer ◽  
Gene Replication ◽  
Packaging Cell ◽  
Mucopolysaccharidosis Type Vi ◽  
Mps Vi

The Maroteaux-Lamy syndrome (mucopolysaccharidosis type VI; MPS VI) is a lysosomal storage disease caused by deficiency of the enzyme arylsulphatase B (ASB). A human ASB cDNA has been subcloned into the retroviral vector pXT1 containing the bacterial neomycin-resistance gene and an internal thymidine kinase promoter for transcription of the inserted gene. Replication defective retrovirus was generated by transfecting the construct into the amphotropic packaging cell line PA317. Human MPS VI fibroblasts infected with recombinant retrovirus integrated the provirus into their genome and expressed retrovirus-encoded ASB mRNAs. In infected fibroblasts the level of ASB was up to 36-fold higher than in normal fibroblasts. Biosynthesis and processing of ASB in infected MPS VI fibroblasts was accomplished as in normal fibroblasts, and mature, enzymically active, ASB accumulated in dense lysosomes, indicating that the ASB deficiency in MPS VI fibroblasts was corrected by the retroviral gene transfer.

scholarly journals Early Diagnosis and Results of Enzyme Replacement Therapy in the Patient with Mucopolysaccharidosis Type VI: Clinical Case

2021 ◽  
Author(s):  
Dmitry V. Ivanov ◽  
Anna I. Ostrun ◽  
Vladimir M. Kenis ◽  
Tatiana V. Markova ◽  
Ekaterina Yu. Zakharova
Keyword(s):  
Enzyme Replacement Therapy ◽  
Replacement Therapy ◽  
Clinical Case ◽  
Mucopolysaccharidosis Type ◽  
Lysosomal Storage ◽  
Enzyme Replacement ◽  
Mucopolysaccharidosis Type Vi ◽  
Mps Vi ◽  
Arylsulfatase B ◽  
Arsb Gene

Background. Mucopolysaccharidosis type VI (MPS VI, Maroteaux–Lamy syndrome) is rare autosomal-recessive multisystem disease, one of the group of lysosomal storage diseases. The MPS VI pathogenesis is determined by arylsulfatase B enzyme deficiency caused by mutations in the ARSB gene. There are only few published clinical examples of this disease that covers the results of early enzyme replacement therapy (ERT) onset.Clinical case description. The child was suspected to have lysosomal storage disease at the age of 1.5 months, it was based on microscopic analysis of blood smears: Alder abnormality was revealed (granulations and red-violet inclusions in neutrophils, monocytes, lymphocytes cytoplasm). The diagnosis was confirmed at the age of 3 months: increased glycosaminoglycans (GAGs) concentration in the urine, arylsulfatase B activity decrease in dried blood spots, and pathogenic variant c.943C>T (p. R315X) in the ARSB gene in homozygous state were revealed. ERT with galsulfase was started at the age of 7 months. There was decrease in excretion of GAGs in urine to normal level after 9 and 15 months of therapy. Normal growth and body proportions for the patient’s age were determined 3 years after continuous ERT. However, there was progression of multiple dysostosis and joint stiffness, as well as eyes lesion.Conclusion. Early ERT onset cannot completely stop MPS VI progression but it allows to reduce the severity of several symptoms and improves patient’s quality of life.

scholarly journals Restoration of arylsulphatase A activity in human-metachromatic-leucodystrophy fibroblasts via retroviral-vector-mediated gene transfer

1991 ◽  
Vol 280 (2) ◽  
pp. 459-461 ◽  
Author(s):  
W Rommerskirch ◽  
A L Fluharty ◽  
C Peters ◽  
K von Figura ◽  
V Gieselmann
Keyword(s):  
Storage Disease ◽  
Metachromatic Leukodystrophy ◽  
Normal Individual ◽  
Human Fibroblasts ◽  
Retroviral Vector ◽  
Lysosomal Storage ◽  
Packaging Cell Line ◽  
Packaging Cell ◽  
Metachromatic Leucodystrophy ◽  
Arylsulphatase A

Metachromatic leukodystrophy is a lysosomal storage disease caused by the deficiency of arylsulphatase A (ASA). A human ASA cDNA was subcloned into the retroviral vector pXT1. Replication-defective retrovirus was generated by transfection of the vector into the amphotropic packaging cell line PA317. Human fibroblasts from a patient suffering from metachromatic leucodystrophy was infected with the recombinant retrovirus. Infected fibroblasts expressed ten times more ASA compared with control fibroblasts from a normal individual. The ASA encoded by the integrated provirus was shown to be correctly transported into the lysosomes and to normalize the impaired degradation of cerebroside sulphate.

scholarly journals Successful Reconstitution of Human Hematopoiesis in the SCID-hu Mouse by Genetically Modified, Highly Enriched Progenitors Isolated From Fetal Liver

Blood ◽  
1997 ◽  
Vol 90 (9) ◽  
pp. 3496-3506 ◽  
Author(s):  
Laurent Humeau ◽  
Christian Chabannon ◽  
Meri T. Firpo ◽  
Patrice Mannoni ◽  
Claude Bagnis ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fetal Liver ◽  
Genetically Modified ◽  
Retroviral Vector ◽  
Hematopoietic Progenitors ◽  
Packaging Cell Line ◽  
Retroviral Gene Transfer ◽  
Packaging Cell ◽  
Hematopoietic Reconstitution ◽  
B Lineage

Abstract Highly purified CD34++CD38−Lin− hematopoietic progenitors isolated from human fetal liver were infected with the murine retroviral vector, MFG nls-LacZ, which encodes a modified version of the Escherichia coli β-galactosidase gene. Progenitors that were cocultured with the packaging cell line could reconstitute human bone marrow or thymus implanted in SCID-hu mice. Expression of the β-galactosidase gene was observed in primitive and committed clonogenic progenitors, mature myeloid, B-lineage cells, and T-lineage cells for up to 4 months after injection into SCID-hu mice. Furthermore, hematopoietic reconstitution by genetically modified progenitor cells could be achieved by the injection of the cells generated from as few as 500 CD34++CD38−Lin− cells, suggesting efficient retroviral gene transfer into fetal liver progenitors.

Thrombocytopenia associated with galsulfase treatment

2010 ◽  
Vol 30 (7) ◽  
pp. 768-771 ◽  
Author(s):  
Murat Doğan ◽  
Yasar Cesur ◽  
Erdal Peker ◽  
Ahmet F Oner ◽  
Sekibe Zehra Dogan
Keyword(s):  
Enzyme Replacement Therapy ◽  
Replacement Therapy ◽  
Lysosomal Storage Disorder ◽  
Mucopolysaccharidosis Type ◽  
Lysosomal Storage ◽  
Cultured Fibroblasts ◽  
Enzyme Replacement ◽  
Mucopolysaccharidosis Type Vi ◽  
Mps Vi ◽  
Arylsulfatase B

Mucopolysaccharidosis type VI (MPS VI), or Maroteaux-Lamy syndrome, is a lysosomal storage disorder that results from a deficiency of the enzyme N-acetylgalactosamine-4-sulfatase or arylsulfatase B (ASB). It is a relatively rare disorder, with an estimated incidence of 1 in 248,000 to 1 in 300,000. The diagnosis is made on the basis of findings of elevated urine glycosaminoglycans and a deficiency of ASB activity in leukocytes or cultured fibroblasts. In treatment of MPS VI, enzyme replacement therapy (galsulfase; human recombinant ASB enzyme) became available. Infusions of galsulfase were generally well tolerated. But in some patients, infusion-associated reactions including rash, urticaria, headache, hypotension, nausea, and vomiting were documented and were managed successfully by interrupting or slowing the rate of infusion and/or by the administration of antihistamines, antipyretics, corticosteroids, or oxygen. Here, we report a case with MPS VI who developed thrombocytopenia after third dose of therapy. To the best of our knowledge, this is the first report about thrombocytopenia associated with galsulfase therapy in the literature. Additionally, with this report, we want to share our approach for this case.

Efficient transduction and selection of human T-lymphocytes with bicistronic Thy1/HSV1-TK retroviral vector produced by a human packaging cell line

2004 ◽  
Vol 6 (4) ◽  
pp. 374-386 ◽  
Author(s):  
François M. Lemoine ◽  
Mariana Mesel-Lemoine ◽  
Mustapha Cherai ◽  
Géraldine Gallot ◽  
Henri Vié ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Cell Line ◽  
Retroviral Vector ◽  
Packaging Cell Line ◽  
Packaging Cell ◽  
Human T Lymphocytes ◽  
Selection Of

scholarly journals Overexpression of N-acetylgalactosamine-4-sulphatase induces a multiple sulphatase deficiency in mucopolysaccharidosis-type-VI fibroblasts

1993 ◽  
Vol 294 (3) ◽  
pp. 657-662 ◽  
Author(s):  
D S Anson ◽  
V Muller ◽  
J Bielicki ◽  
G S Harper ◽  
J J Hopwood
Keyword(s):  
Practical Importance ◽  
High Titre ◽  
Skin Fibroblasts ◽  
Mucopolysaccharidosis Type ◽  
Subcellular Compartment ◽  
Mucopolysaccharidosis Type Vi ◽  
Steroid Sulphatase ◽  
Mps Vi ◽  
Genetically Determined ◽  
Processing Mechanism

High-titre stocks of an amphotropic retrovirus, constructed so as to express a full-length cDNA encoding the human lysosomal enzyme N-acetylgalactosamine-4-sulphatase (4-sulphatase) from the cytomegalovirus immediate early promoter, were used to infect skin fibroblasts from a clinically severe mucopolysaccharidosis type VI (MPS VI) patient. The infected MPS VI cells showed correction of the enzymic defect with the enzyme being expressed at high levels and in the correct subcellular compartment. Surprisingly this did not result in correction of glycosaminoglycan turnover as measured by accumulation of 35S in metabolically labelled cells. We demonstrate that this is apparently caused by an induced reduction of the activities of other lysosomal sulphatases, presumably due to competition for a sulphatase-specific processing mechanism by the over-expressed 4-sulphatase. The level of steroid sulphatase, which is a microsomal sulphatase, was also reduced. Infection of skin fibroblasts from a second, clinically mildly affected, MPS VI patient with the same virus also resulted in no significant change in the level of glycosaminoglycan storage. However, in this case the cause of the observed phenomenon was less clear. These results are of obvious practical importance when considering gene therapy for a sulphatase deficiency such as MPS VI and also provide possible new avenues for exploration of the processes involved in sulphatase synthesis and genetically determined multiple sulphatase deficiency.

Highly efficient gene transfer into baboon marrow repopulating cells using GALV-pseudotype oncoretroviral vectors produced by human packaging cells

Blood ◽  
2002 ◽  
Vol 100 (12) ◽  
pp. 3960-3967 ◽  
Author(s):  
Peter A. Horn ◽  
Max S. Topp ◽  
Julia C. Morris ◽  
Stanley R. Riddell ◽  
Hans-Peter Kiem
Keyword(s):  
Stem Cells ◽  
Gene Transfer ◽  
Cell Line ◽  
Flow Cytometric Analysis ◽  
Lentiviral Vectors ◽  
Transduction Efficiency ◽  
Packaging Cell Line ◽  
Hematopoietic Stem ◽  
Packaging Cell ◽  
Cd34 Cells

Vector-containing medium harvested from murine packaging cell lines has been shown to contain factors that can negatively influence the transduction and maintenance of hematopoietic stem cells. Thus, we generated a human packaging cell line with a gibbon ape leukemia virus pseudotype (Phoenix-GALV), and we evaluated vectors produced by Phoenix-GALV for their ability to transduce hematopoietic progenitor/stem cells. In 3 baboons, we used a competitive repopulation assay to directly compare GALV-pseudotype retrovirus vectors produced by either Phoenix-GALV or by the NIH 3T3–derived packaging cell line, PG13. In 3 additional baboons we compared Phoenix-GALV–derived vectors to more recently developed lentiviral vectors. Gene transfer efficiency into hematopoietic repopulating cells was assessed by evaluating the number of genetically modified peripheral blood and marrow cells using flow cytometry and real-time polymerase chain reaction. Transduction efficiency of hematopoietic repopulating cells was significantly higher using the Phoenix-GALV–derived vector as compared with the PG13-derived vectors or lentiviral vectors, with stable transduction levels up to 25%. We followed 2 animals for more than one year. Flow cytometric analysis of hematopoietic subpopulations in these animals revealed transgene expression in CD13+ granulocytes, CD20+ B lymphocytes, CD3+ T lymphocytes, CD61+ platelets, as well as red blood cells, indicating multilineage engraftment of cells transduced by Phoenix-GALV–pseudotype vectors. In addition, transduction of human CD34+ cells was significantly more efficient than transduction of baboon CD34+ cells, suggesting that Phoenix-GALV–derived oncoretroviral vectors may be even more efficient in human stem cell gene therapy applications.

Sign in / Sign up

Export Citation Format

Share Document