Aspartylglucosaminidase (AGA) is efficiently produced and endocytosed by glial cells: implication for the therapy of a lysosomal storage disorder

Abstract

These data indicate the importance of glial cells in the expression and transport of AGA. Subsequently, new approaches can be developed for therapeutic intervention.

Use of nonviral promoters in adenovirus‐mediated gene therapy: reduction of lysosomal storage in the aspartylglucosaminuria mouse

“…Interestingly, the AGA promoter was shown to be a very powerful promoter in glia [20]. In the in vitro studies the glial cells showed much higher expression, exocytosing and endocytosing capacity of AGA than neurons.…”

“…The sections were fixed in 4% PFA for 15 min, washed three times with PBS, blocked and permeabilized in 0.5% bovine serum albumin (BSA) and 0.2% saponin buffer. Double stainings were performed using a rabbit anti-human AGA at 1 : 200 dilution [2,10,16] and rat Lamp-1 at 1 : 50 dilution (ID4B), mouse NeuN at 1 : 100 dilution or pGFAP at a 1 : 200 dilution as primary antibodies [20]. Corresponding secondary antibodies were goat anti-rabbit rhodamine (TRITC) and goat anti-rat/mouse/rabbit fluorescein (FITC) (Jackson Immunoresearch).…”

“…Four different adenoviruses mediating AGA expression were constructed as previously described [20]. Each adenovirus contains the 1041 bp AGA gene-coding region under the control of one of the following promoters: neuron-specific enolase (NSE) promoter, astrocyte-specific glial fibrillary acidic protein (GFAP) promoter and two endogenous human AGA promoters.…”

“…This is crucial in the case of the AGA enzyme, based on our previous in vitro study showing the importance of non-neuronal cells both in the production and endocytosis of AGA. Especially in glia, AGA is efficiently exocytosed and endocytosed and we have recently demonstrated that the endogenous AGA promoter is very powerful in cultured astrocytes [20].…”

Bone marrow transplantation in young aspartylglucosaminuria mice: improved clearance of lysosomal storage in brain by using wild type as compared to heterozygote donors

“…Interestingly, human AGA promoter was found to be a very powerful glia promoter. 3 We have also used adenovirus for overexpression of AGA in vivo in AGU mouse brain resulting in partial correction of brain pathology. 4 The AGU mouse model presents tissue pathology similar to humans including lysosomal storage in all tissues.…”