Gene silencing of EXTL2 and EXTL3 as a substrate deprivation therapy for heparan sulphate storing mucopolysaccharidoses

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dc.contributor.authorKaidonis, X.
dc.contributor.authorLiaw, W.
dc.contributor.authorDerrick Roberts, A.
dc.contributor.authorLy, M.
dc.contributor.authorAnson, D.
dc.contributor.authorByers, S.
dc.date.issued2010
dc.description.abstractNeurological pathology is characteristic of the mucopolysaccharidoses (MPSs) that store heparan sulphate (HS) glycosaminoglycan (gag) and has been proven to be refractory to systemic therapies. Substrate deprivation therapy (SDT) using general inhibitors of gag synthesis improves neurological function in mouse models of MPS, but is not specific to an MPS type. We have investigated RNA interference (RNAi) as a method of targeting SDT to the HS synthesising enzymes, EXTL2 and EXTL3. Multiple shRNA molecules specific to EXTL2 or EXTL3 were designed and validated in a reporter gene assay, with four out of six shRNA constructs reducing expression by over 90%. The three EXTL2-specific shRNA constructs reduced endogenous target gene expression by 68, 32 and 65%, and decreased gag synthesis by 46, 50 and 27%. One EXTL3-specific shRNA construct reduced endogenous target gene expression by 14% and gag synthesis by 39%. Lysosomal gag levels in MPS IIIA and MPS I fibroblasts were also reduced by EXTL2 and EXTL3-specific shRNA. Incorporation of shRNAs into a lentiviral expression system reduced gene expression, and one EXTL2-specific shRNA reduced gag synthesis. These results indicate that deprivation therapy through shRNA-mediated RNAi has potential as a therapy for HS-storing MPSs.
dc.description.statementofresponsibilityXenia Kaidonis, Wan Chin Liaw, Ainslie Derrick Roberts, Marleesa Ly, Donald Anson and Sharon Byers
dc.identifier.citationEuropean Journal of Human Genetics, 2010; 18(2):194-199
dc.identifier.doi10.1038/ejhg.2009.143
dc.identifier.issn1018-4813
dc.identifier.issn1476-5438
dc.identifier.orcidByers, S. [0000-0001-5576-3636]
dc.identifier.urihttp://hdl.handle.net/2440/59835
dc.language.isoen
dc.publisherNature Publishing Group
dc.rightsCopyright © 2010, Nature Publishing Group
dc.source.urihttps://doi.org/10.1038/ejhg.2009.143
dc.subjectT-Lymphocytes
dc.subjectLysosomes
dc.subjectHumans
dc.subjectMucopolysaccharidoses
dc.subjectN-Acetylglucosaminyltransferases
dc.subjectHeparitin Sulfate
dc.subjectMembrane Proteins
dc.subjectGene Products, gag
dc.subjectRNA
dc.subjectDNA Primers
dc.subjectTransfection
dc.subjectPolymerase Chain Reaction
dc.subjectGene Expression Regulation
dc.subjectGene Silencing
dc.subjectGenes, Reporter
dc.subjectChild
dc.titleGene silencing of EXTL2 and EXTL3 as a substrate deprivation therapy for heparan sulphate storing mucopolysaccharidoses
dc.typeJournal article
pubs.publication-statusPublished

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