Correction in trans for Fabry disease: expression, secretion and uptake of alpha-galactosidase A in patient-derived cells driven by a high-titer recombinant retroviral vector. Proc Natl Acad Sci U S A 1996 Jul 23;93(15):7917-22
Date
07/23/1996Pubmed ID
8755577Pubmed Central ID
PMC38849DOI
10.1073/pnas.93.15.7917Scopus ID
2-s2.0-0029836764 (requires institutional sign-in at Scopus site) 72 CitationsAbstract
Fabry disease is an X-linked metabolic disorder due to a deficiency of alpha-galactosidase A (alpha-gal A; EC 3.2.1.22). Patients accumulate glycosphingolipids with terminal alpha-galactosyl residues that come from intracellular synthesis, circulating metabolites, or from the biodegradation Of senescent cells. Patients eventually succumb to renal, cardio-, or cerebrovascular disease. No specific therapy exists. One possible approach to ameliorating this disorder is to target corrective gene transfer therapy to circulating hematopoietic cells. Toward this end, an amphotropic virus-producer cell line has been developed that produces a high titer (>10(6) i.p. per ml) recombinant retrovirus constructed to transduce and correct target cells. Virus-producer cells also demonstrate expression of large amounts of both intracellular and secreted alpha-gal A. To examine the utility of this therapeutic vector, skin fibroblasts from Fabry patients were corrected for the metabolic defect by infection with this recombinant virus and secreted enzyme was observed. Furthermore, the secreted enzyme was found to be taken up by uncorrected cells in a mannose-6-phosphate receptor-dependent manner. In related experiments, immortalized B cell lines from Fabry patients, created as a hematologic delivery test system, were transduced. As with the fibroblasts, transduced patient B cell lines demonstrated both endogenous enzyme correction and a small amount of secretion together with uptake by uncorrected cells. These studies demonstrate that endogenous metabolic correction in transduced cells, combined with secretion, may provide a continuous source of corrective material in trans to unmodified patient bystander cells (metabolic cooperativity).
Author List
Medin JA, Tudor M, Simovitch R, Quirk JM, Jacobson S, Murray GJ, Brady ROAuthor
Jeffrey A. Medin PhD Professor in the Pediatrics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
3T3 CellsAnimals
B-Lymphocytes
Base Sequence
Cell Line
Cells, Cultured
DNA Primers
Fabry Disease
Fibroblasts
Genetic Therapy
Genetic Vectors
Humans
Leukocytes
Mice
Molecular Sequence Data
Polymerase Chain Reaction
Retroviridae
Skin
Transfection
X Chromosome
alpha-Galactosidase
