Retrovirus-mediated correction of the metabolic defect in cultured Farber disease cells. Hum Gene Ther 1999 May 20;10(8):1321-9
Date
06/12/1999Pubmed ID
10365663DOI
10.1089/10430349950018003Scopus ID
2-s2.0-0033587181 (requires institutional sign-in at Scopus site) 33 CitationsAbstract
Farber disease is a rare severe lysosomal storage disorder due to a deficient activity of the enzyme acid ceramidase (AC). Patients have granulomas along with lipid-laden macrophages that accumulate in a number of tissues, leading to multiple diverse clinical symptoms. There is no therapy for the disorder and most patients succumb to the disease in early childhood. The severity of the disease progression seems to correlate with the amount of the accumulated ceramide substrate. Since the cDNA for human AC has been elucidated we sought to establish if genetic transfer of this sequence would lead to enzymatic and, especially, functional correction of the catabolic defect in Farber patient cells. To do this, a novel amphotropic recombinant retrovirus was constructed that engineers transfer of the human AC cDNA. On infection of patient fibroblasts, AC enzyme activity in cell extracts was completely restored. Further, substrate-loading assays of intact living cells showed a fully normalized catabolism of lysosomal ceramide. Lastly, as reported for some other corrected enzymatic defects of lysosomes, metabolic cooperativity was seen, in that functionally corrected patient fibroblasts secreted AC that was taken up through the mannose 6-phosphate receptor and used by uncorrected fibroblasts as well as recipient Farber lymphoblastoid cells. This overall transduction and uptake scenario for Farber disease allows future treatment of this severe disorder to be envisioned using gene transfer approaches.
Author List
Medin JA, Takenaka T, Carpentier S, Garcia V, Basile JP, Segui B, Andrieu-Abadie N, Auge N, Salvayre R, Levade TAuthor
Jeffrey A. Medin PhD Professor in the Pediatrics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Acid CeramidaseAmidohydrolases
Cell Line, Transformed
Cells, Cultured
Ceramidases
DNA, Complementary
Fibroblasts
Gene Expression
Gene Transfer Techniques
Genetic Engineering
Genetic Vectors
Humans
Lysosomal Storage Diseases
Recombination, Genetic
Retroviridae
