RNA interference mediated silencing of α-synuclein in MN9D cells and its effects on cell viability

Liu, Dongmei; Jin, Ling; Wang, Hao; Zhao, Huanying; Zhao, Chunli; Yang, Hui

doi:10.1007/s12264-008-0096-4

Cited by 10 publications

(9 citation statements)

References 49 publications

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“…In neuronal cells, similar to that used in our experiments, it has been demonstrated that downregulation of SNCA had a specific effect on viability, but not proliferative potential (Liu et al 2008). Elevation of SNCA by inhibition of HDAC increases neuronal protection against death induced by glutamate (Leng and Chuang 2006) or 6-OHDA (Monti et al 2007).…”

Section: Discussionsupporting

confidence: 64%

Alpha-Synuclein Loss in Spinal Muscular Atrophy

Acsádi

Murphy

et al. 2010

J Mol Neurosci

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Spinal muscular atrophy, the most prevalent hereditary motor neuron disease, is caused by mutations in the survival motor neuron (SMN) 1 gene. A significant reduction in the encoded SMN protein leads to the degeneration of motor neurons. However, the molecular events leading to this process are not well understood. The present study uses a previously developed neuronal cell culture model of spinal muscular atrophy for a multiplex transcriptome analysis. Furthermore, gene expression analysis was performed on in vitro cell cultures, as well as tissue samples of spinal muscular atrophy patients and transgenic mice. RNA and subsequent Western blot protein analyses suggest that low SMN levels are associated with significantly lower alpha-synuclein expression. Examination of two genes related to vesicular transport showed a similar though less dramatic decrease in expression. The 140-amino acid protein alpha-synuclein, dominant mutations of which have previously been associated with an autosomal dominant form of Parkinson's disease, is strongly expressed in select neurons of the brain. Although not well understood, the physiologic functions of alpha-synuclein have been linked to synaptic vesicular neurotransmitter release and neuroprotection, suggesting a possible contribution to Smn-deficient motor neuron pathology. Furthermore, alpha-synuclein may be a genetic modifier or biomarker of spinal muscular atrophy.

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Section: Discussionsupporting

confidence: 64%

Alpha-Synuclein Loss in Spinal Muscular Atrophy

Acsádi

Murphy

et al. 2010

J Mol Neurosci

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“…In support of this, a recent report showed that specifically silencing endogenous rat SNCA in vivo in substantia nigra with an shRNA approach resulted in loss of DA neurons (Gorbatyuk et al, 2010). Another study showed that down-regulation of SNCA in MN9D cells decreases cell viability (Liu et al, 2008). On the other hand, it was recently reported that down regulation of endogenous SNCA in primate substantia nigra by pump delivery of siRNAs does not reduce the number and phenotype of nigral DA neurons (McCormack et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

A microRNA embedded AAV alpha-synuclein gene silencing vector for dopaminergic neurons

Han¹,

Khodr²,

Sapru³

et al. 2011

Brain Research

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Alpha-synuclein (SNCA), an abundantly expressed presynaptic protein, is implicated in Parkinson disease (PD). Since over-expression of human SNCA (hSNCA) leads to death of dopaminergic (DA) neurons in human, rodent and fly brain, hSNCA gene silencing may reduce levels of toxic forms of SNCA and ameliorate degeneration of DA neurons in PD. To begin to develop a gene therapy for PD based on hSNCA gene silencing, two AAV gene silencing vectors were designed, and tested for efficiency and specificity of silencing, as well as toxicity in vitro. The same hSNCA silencing sequence (shRNA) was used in both vectors, but in one vector, the shRNA was embedded in a microRNA backbone and driven by a pol II promoter, and in the other the shRNA was not embedded in a microRNA and was driven by a pol III promoter. Both vectors silenced hSNCA to the same extent in 293T cells transfected with hSNCA. In DA PC12 cells, neither vector decreased expression of rat SNCA, tyrosine hydroxylase (TH), dopamine transporter (DAT) or the vesicular monoamine transporter (VMAT). However, the mir30 embedded vector was significantly less toxic to both PC12 and SH-SY5Y cells. Our in vitro data suggest that this miRNA-embedded silencing vector may be ideal for chronic in vivo SNCA gene silencing in DA neurons.

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“…Of note, a-syn null mice do not develop a nigrostriatal lesion (Abeliovich et al, 2000). This plausible therapeutic approach was applied with RNAi to reduce the synthesis in vitro and in vivo (Fountaine and Wade-Martins, 2007;Liu et al, 2008;Sapru et al, 2006). To date, studies have yielded conflicting results regarding the effectiveness and tolerability of this strategy, with different groups reporting nigrostriatal degeneration (Gorbatyuk et al, 2010;Khodr et al, 2011Khodr et al, , 2014 and deleterious inflammation (Khodr et al, 2014) after depleting a-syn in the rat brain.…”

Section: Parkinson's Disease Therapeutic Approachesmentioning

confidence: 99%