In vitro treatment of congenital disorder of glycosylation type Ia using PLGA nanoparticles loaded with GDPMan.
Int J Mol Med
; 44(1): 262-272, 2019 Jul.
Article
em En
| MEDLINE
| ID: mdl-31115488
ABSTRACT
Congenital disorder of glycosylation (CDG) type Ia is a multisystem disorder that occurs due to mutations in the phosphomannomutase 2 (PMM2) gene, which encodes for an enzyme involved in the Nglycosylation pathway. Mutated PMM2 leads to the reduced conversion of mannose6P to mannose1P, which results in low concentration levels of guanosine 5'diphosphoDmannose (GDPMan), a nucleotideactivated sugar essential for the construction of protein oligosaccharide chains. In the present study, an in vitro therapeutic approach was used, based on GDPManloaded poly (D,Llactidecoglycolide) (PLGA) nanoparticles (NPs), which were used to treat CDGIa fibroblast cultures, thus bypassing the glycosylation pathway reaction catalysed by PMM2. To assess the degree of hypoglycosylation in vitro, the present study examined the activities of αmannosidase, ßglucoronidase and ßgalactosidase in defective and normal fibroblasts. GDPMan (30 µg/ml GDPMan PLGA NPs) was incubated for 48 h with the cells and the specific activities of αmannosidase and ßgalactosidase were estimated at 69 and 92% compared with healthy controls. The residual activity of ßglucoronidase increased from 6.5 to 32.5% and was significantly higher compared with that noted in the untreated CDGIa fibroblasts. The glycosylation process of fibroblasts was also analysed by twodimensional electrophoresis. The results demonstrated that treatment caused the reappearance of several glycosylated proteins. The data in vitro showed that GDPMan PLGA NPs have desirable efficacy and warrant further evaluation in a preclinical validation animal model.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Portadores de Fármacos
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Fosfotransferases (Fosfomutases)
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Defeitos Congênitos da Glicosilação
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Nanopartículas
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Copolímero de Ácido Poliláctico e Ácido Poliglicólico
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Guanosina Difosfato Manose
Limite:
Humans
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article