Characterisation of early changes in ovine CLN5 and CLN6 Batten disease neural cultures for the rapid screening of therapeutics

Best, Hannah; Neverman, Nicole J.; Wicky, Hollie E.; Mitchell, Nadia L.; Leitch, Beulah; Hughes, Stephanie M.

doi:10.1016/j.nbd.2017.01.001

Cited by 30 publications

(55 citation statements)

References 79 publications

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“…Both lysosomal compartment and the ER were clearly enlarged in the CLN5Y392X iPSC-derived cells compared to the control cells. Accordingly, it was just recently reported that LAMP-1-positive lysosomal compartments of fetal neuronal cultures prepared from CLN5-deficient sheep have similar appearance as observed in this study in patient-specific neural lineage cells [40]. Increased lysosomal size is a typical phenomenon for lysosomal disorders, which accumulate storage material in the lysosomes.…”

Section: Discussionsupporting

confidence: 80%

Induced Pluripotent Stem Cells Derived from a CLN5 Patient Manifest Phenotypic Characteristics of Neuronal Ceroid Lipofuscinoses

Uusi-Rauva

Blom

Schantz-Fant

et al. 2017

IJMS

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Neuronal ceroid lipofuscinoses (NCLs) are autosomal recessive progressive encephalopathies caused by mutations in at least 14 different genes. Despite extensive studies performed in different NCL animal models, the molecular mechanisms underlying neurodegeneration in NCLs remain poorly understood. To model NCL in human cells, we generated induced pluripotent stem cells (iPSCs) by reprogramming skin fibroblasts from a patient with CLN5 (ceroid lipofuscinosis, neuronal, 5) disease, the late infantile variant form of NCL. These CLN5 patient-derived iPSCs (CLN5Y392X iPSCs) harbouring the most common CLN5 mutation, c.1175_1176delAT (p.Tyr392X), were further differentiated into neural lineage cells, the most affected cell type in NCLs. The CLN5Y392X iPSC-derived neural lineage cells showed accumulation of autofluorescent storage material and subunit C of the mitochondrial ATP synthase, both representing the hallmarks of many forms of NCLs, including CLN5 disease. In addition, we detected abnormalities in the intracellular organelles and aberrations in neuronal sphingolipid transportation, verifying the previous findings obtained from Cln5-deficient mouse macrophages. Therefore, patient-derived iPSCs provide a suitable model to study the mechanisms of NCL diseases.

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Section: Discussionsupporting

confidence: 80%

Induced Pluripotent Stem Cells Derived from a CLN5 Patient Manifest Phenotypic Characteristics of Neuronal Ceroid Lipofuscinoses

Uusi-Rauva

Blom

Schantz-Fant

et al. 2017

IJMS

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“…The HIV-1 derived lentiviral plasmid, pCDH-EF1-MCS-T2A-copGFP (CD521A-1, System Biosciences, Palo Alto, CA) was modified to replace EF1 with the rat neuron-specific synapsin 1 promoter (Syn) [ 21 ] to drive neuronal expression of either copGFP (LV-control) or human sAPPα [ 22 ] and copGFP, separated by a T2A cleavage signal (LV-sAPPα). Vectors were packaged in HEK293FT cells using a second-generation packaging system [ 23 ]. Viral particles were pseudotyped with either the vesicular stomatitis virus (VSVg) envelope, which has tropism for a wide variety of cells, but has limited spread from injection sites [ 24 , 25 ] or a chimeric rabies/VSVg (RabB19) envelope (Addgene #88865) containing the SADB19 (B19) extra-virion and transmembrane domains and the intra-virion domain of VSVg, which by contrast can undergo retrograde transport [ 26 ].…”

Section: Methodsmentioning

confidence: 99%

Lentivirus-mediated expression of human secreted amyloid precursor protein-alpha prevents development of memory and plasticity deficits in a mouse model of Alzheimer's disease

et al. 2018

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Alzheimer’s disease (AD) is a neurodegenerative disease driven in large part by accumulated deposits in the brain of the amyloid precursor protein (APP) cleavage product amyloid-β peptide (Aβ). However, AD is also characterised by reductions in secreted amyloid precursor protein-alpha (sAPPα), an alternative cleavage product of APP. In contrast to the neurotoxicity of accumulated Αβ, sAPPα has many neuroprotective and neurotrophic properties. Increasing sAPPα levels has the potential to serve as a therapeutic treatment that mitigates the effects of Aβ and rescue cognitive function. Here we tested the hypothesis that lentivirus-mediated expression of a human sAPPα construct in a mouse model of AD (APPswe/PS1dE9), begun before the onset of plaque pathology, could prevent later behavioural and electrophysiological deficits. Male mice were given bilateral intra-hippocampal injections at 4 months of age and tested 8–10 months later. Transgenic mice expressing sAPPα performed significantly better than untreated littermates in all aspects of the spatial water maze task. Expression of sAPPα also resulted in partial rescue of long-term potentiation (LTP), tested in vitro. These improvements occurred in the absence of changes in amyloid pathology. Supporting these findings on LTP, lentiviral-mediated expression of sAPPα for 3 months from 10 months of age, or acute sAPPα treatment in hippocampal slices from 18 to 20 months old transgenic mice, completely reversed the deficits in LTP. Together these findings suggest that sAPPα has wide potential to act as either a preventative or restorative therapeutic treatment in AD by mitigating the effects of Aβ toxicity and enhancing cognitive reserve.

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“…Viral particles were concentrated by ultra-centrifugation in a Beckman L-70 (SW28 rotor; Beckman Coulter, Carlsbad, CA, USA) at 110,000 × g for 90 min at 4°C and the viral pellet was resuspended in lactose (40 mg/mL in Dulbecco’s phosphate buffered saline) and stored at −80°C (Schoderboeck et al, 2015 ). The viral genomic titres were determined using qPCR with primers for an enhancer incorporated into the lentiviral vectors; woodchuck posttranscriptional response element (Supplementary Table S1), as previously described in Best et al ( 2017 ). The titres ranged from 7.02 × 10 9 − 3.72 × 10 10 genomes/mL.…”

Section: Methodsmentioning

confidence: 99%

RNA-Sequencing Analysis Reveals a Regulatory Role for Transcription Factor Fezf2 in the Mature Motor Cortex

Clare

Wicky

Empson

et al. 2017

Front. Mol. Neurosci.

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Forebrain embryonic zinc finger (Fezf2) encodes a transcription factor essential for the specification of layer 5 projection neurons (PNs) in the developing cerebral cortex. As with many developmental transcription factors, Fezf2 continues to be expressed into adulthood, suggesting it remains crucial to the maintenance of neuronal phenotypes. Despite the continued expression, a function has yet to be explored for Fezf2 in the PNs of the developed cortex. Here, we investigated the role of Fezf2 in mature neurons, using lentiviral-mediated delivery of a shRNA to conditionally knockdown the expression of Fezf2 in the mouse primary motor cortex (M1). RNA-sequencing analysis of Fezf2-reduced M1 revealed significant changes to the transcriptome, identifying a regulatory role for Fezf2 in the mature M1. Kyoto Encyclopedia Genes and Genomes (KEGG) pathway analyses of Fezf2-regulated genes indicated a role in neuronal signaling and plasticity, with significant enrichment of neuroactive ligand-receptor interaction, cell adhesion molecules and calcium signaling pathways. Gene Ontology analysis supported a functional role for Fezf2-regulated genes in neuronal transmission and additionally indicated an importance in the regulation of behavior. Using the mammalian phenotype ontology database, we identified a significant overrepresentation of Fezf2-regulated genes associated with specific behavior phenotypes, including associative learning, social interaction, locomotor activation and hyperactivity. These roles were distinct from that of Fezf2-regulated genes identified in development, indicating a dynamic transition in Fezf2 function. Together our findings demonstrate a regulatory role for Fezf2 in the mature brain, with Fezf2-regulated genes having functional roles in sustaining normal neuronal and behavioral phenotypes. These results support the hypothesis that developmental transcription factors are important for maintaining neuron transcriptomes and that disruption of their expression could contribute to the progression of disease phenotypes.

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Characterisation of early changes in ovine CLN5 and CLN6 Batten disease neural cultures for the rapid screening of therapeutics

Cited by 30 publications

References 79 publications

Induced Pluripotent Stem Cells Derived from a CLN5 Patient Manifest Phenotypic Characteristics of Neuronal Ceroid Lipofuscinoses

Induced Pluripotent Stem Cells Derived from a CLN5 Patient Manifest Phenotypic Characteristics of Neuronal Ceroid Lipofuscinoses

Lentivirus-mediated expression of human secreted amyloid precursor protein-alpha prevents development of memory and plasticity deficits in a mouse model of Alzheimer's disease

RNA-Sequencing Analysis Reveals a Regulatory Role for Transcription Factor Fezf2 in the Mature Motor Cortex

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