Adult bone marrow-derived mononuclear cells expressing chondroitinase AC transplanted into CNS injury sites promote local brain chondroitin sulphate degradation.
Injury to the CNS of vertebrates leads to the formation of a glial scar and production of inhibitory molecules, including chondroitin sulphate proteoglycans. Various studies suggest that the sugar component of the proteoglycan is responsible for the inhibitory role of these compounds in axonal regeneration. By degrading chondroitin sulphate chains with specific enzymes, denominated chondroitinases, the inhibitory capacity of these proteoglycans is decreased. Chondroitinase administration involves frequent injections of the enzyme at the lesion site which constitutes a rather invasive method. We have produced a vector containing the gene for Flavobacterium heparinum chondroitinase AC for expression in adult bone marrow-derived cells which were then transplanted into an injury site in the CNS. The expression and secretion of active chondroitinase AC was observed in vitro using transfected Chinese hamster ovarian and gliosarcoma cells and in vivo by immunohistochemistry analysis which showed degraded chondroitin sulphate coinciding with the location of transfected bone marrow-derived cells. Immunolabelling of the axonal growth-associated protein GAP-43 was observed in vivo and coincided with the location of degraded chondroitin sulphate. We propose that bone marrow-derived mononuclear cells, transfected with our construct and transplanted into CNS, could be a potential tool for studying an alternative chondroitinase AC delivery method.
Coulson-Thomas
Yvette M
YM
Department of Biochemistry, UNIFESP, São Paulo 04044-020, Brazil.
Coulson-Thomas
Vivien J
VJ
Filippo
Thais R
TR
Mortara
Renato A
RA
da Silveira
Rafael B
RB
Nader
Helena B
HB
Porcionatto
Marimélia A
MA
D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism*; D016026:Bone Marrow Transplantation / Q000379:methods*; D001930:Brain Injuries / Q000378:metabolism* / Q000601:surgery*; D002460:Cell Line; D002809:Chondroitin Sulfates / Q000378:metabolism*; D002810:Chondroitinases and Chondroitin Lyases / Q000235:genetics / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D019922:GAP-43 Protein / Q000378:metabolism; D015870:Gene Expression; D018316:Gliosarcoma; D006025:Glycosaminoglycans / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D014162:Transfection / Q000379:methods
D019922:GAP-43 Protein; D006025:Glycosaminoglycans; D049452:Green Fluorescent Proteins; D002809:Chondroitin Sulfates; D002810:Chondroitinases and Chondroitin Lyases