α-Synuclein accumulates in huntingtin inclusions but forms independent filaments and its deficiency attenuates early phenotype in a mouse model of Huntington's disease

Tomás-Zapico, Cristina; Diez-Zaera, María; Ferrer, Isidre; Gómez‐Ramos, Pilar; Morán, María A.; Dı́az-Hernández, Miguel; Lucas, José J.

doi:10.1093/hmg/ddr507

Cited by 36 publications

(60 citation statements)

References 62 publications

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“…Proteasome targeting of intrabodies was initially developed for HD by Butler et al [13] using the ST14 cell model transiently transfected the mhttex1-72Q-GFP plus the huntingtin specific scFv intrabody anti-htt N17 C4-PEST to demonstrate reduction of cellular toxicity in a flow cytometry assay. Recently, in the models using transfected neurons and in vivo in HD mice, Tomas-Zapico [31] identified colocalization of microaggregates of α-Syn and N-terminal mutant htt (N-mut-htt) in the brains of R6/1 and HD94 inducible mice expressing α-Syn. Expression of N-mut-htt increased the number of α-Syn filaments, while knocking out α-Syn decreased the number of N-mut-htt aggregates.…”

Section: Discussionmentioning

confidence: 99%

“…The number of RFP positive htt foci in the presence of mutant A53T-Syn was significantly reduced by VH14PEST (S2 Fig). It is possible that polyQ expression may overwhelm the cellular folding machinery and α-Syn may add to the stress [31]. It is also possible that these two proteins may be competing for the lysosomal degrading machinery (Ravikumar [30, 43–45].…”

Section: Discussionmentioning

confidence: 99%

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Bifunctional Anti-Non-Amyloid Component α-Synuclein Nanobodies Are Protective In Situ

et al. 2016

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Misfolding, abnormal accumulation, and secretion of α-Synuclein (α-Syn) are closely associated with synucleinopathies, including Parkinson’s disease (PD). VH14 is a human single domain intrabody selected against the non-amyloid component (NAC) hydrophobic interaction region of α-Syn, which is critical for initial aggregation. Using neuronal cell lines, we show that as a bifunctional nanobody fused to a proteasome targeting signal, VH14PEST can counteract heterologous proteostatic effects of mutant α-Syn on mutant huntingtin Exon1 and protect against α-Syn toxicity using propidium iodide or Annexin V readouts. We compared this anti-NAC candidate to NbSyn87, which binds to the C-terminus of α-Syn. NbSyn87PEST degrades α-Syn as well or better than VH14PEST. However, while both candidates reduced toxicity, VH14PEST appears more effective in both proteostatic stress and toxicity assays. These results show that the approach of reducing intracellular monomeric targets with novel antibody engineering technology should allow in vivo modulation of proteostatic pathologies.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Bifunctional Anti-Non-Amyloid Component α-Synuclein Nanobodies Are Protective In Situ

et al. 2016

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“…Aggregates of HTT have also been shown to stain for α-Syn [91]. It appears that this is not owing to heterologous oligomers, but to smaller aggregates of mHTT and oligomeric (or fibrillar?)…”

Section: Interactions Between Misfolded Proteinsmentioning

confidence: 99%

“…In vivo, there are clear effects on the onset and progression of HD with manipulations of α-Syn levels. HTT transgenic mice show an earlier onset and death when crossed to α-syn overexpression transgenic mice, while expressing the HTT transgene in mice that lack the α-Syn gene slows the HTT phenotype [91,93]. Although there is still much work to be done with the genetic epidemiology to determine if known α-Syn promoter polymorphisms are modifiers for the human HD phenotype, and if the size of the CAG repeat in HTT and additional trinucleotide repeat proteins can affect PD risk, these emerging data suggest that there may be an option for using therapies directed against one misfolding protein to modify the disease course in another.…”

Section: Interactions Between Misfolded Proteinsmentioning

confidence: 99%

Intrabodies as Neuroprotective Therapeutics

Messer

Joshi

2013

Neurotherapeutics

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The process of misfolding of proteins that can trigger a pathogenic cascade leading to neurodegenerative diseases largely originates intracellularly. It is possible to harness the specificity and affinity of antibodies to counteract either protein misfolding itself, or the aberrant interactions and excess stressors immediately downstream of the primary insult. This review covers the emerging field of engineering intracellular antibody fragments, intrabodies and nanobodies, in neurodegeneration. Huntington's disease has provided the clearest proof of concept for this approach. The model systems and readouts for this disorder power the studies, and the potential to intervene therapeutically at early stages in known carriers with projected ages of onset increases the chances of meaningful clinical trials. Both single-chain Fv and single-domain nanobodies have been identified against specific targets; data have allowed feedback for rational design of bifunctional constructs, as well as target validation. Intrabodies that can modulate the primary accumulating protein in Parkinson's disease, alphasynuclein, are also reviewed, covering a range of domains and conformers. Recombinant antibody technology has become a major player in the therapeutic pipeline for cancer, infectious diseases, and autoimmunity. There is also tremendous potential for applying this powerful biotechnology to neurological diseases.Keywords Intrabodies . Huntington's disease . Immunotherapy . Intrabody-PEST fusions . α-synuclein . Parkinson's disease . Polyglutamine Biological Antibody Therapies for Misfolding ProteinsThe problem of misfolding proteins appears to underlie a significant fraction of neurodegenerative diseases. Protein homeostasis, often referred to as proteostasis, is an especially critical process in postmitotic neurons. The balance of pathways for protein folding, interactions, intracellular localizations, and degradation is a complex one, and can be dysregulated by combinations of mutations, environmental stressors, and aging. We have taken the neurotherapeutic approach of normalizing or manipulating proteostasis using engineered intracellular antibody fragments-intrabodies-that bind with high specificity to selected targets. These intrabodies can be selected and manipulated as genes, allowing the full spectrum of genetic engineering to produce multifunctional constructs that can alter the folding, interactions, intracellular localization, and turnover kinetics/ levels of the target protein. Intrabodies are also valuable molecular tools, which can be used to dissect the functional and pathogenic motifs in these proteins and that could be useful for the development of alternative therapeutics. In this review, we will cover the development of this approach for Huntington's disease (HD), which has a well-described target, evaluating effects in several cell culture and in vivo systems with strong readouts for therapeutic efficacy, and advances in engineering anti-HD intrabodies. We also cover a small, but growing, literature...

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“…For example, it was found that polyQ inclusions in HD and other polyQ disorders are immunopositive for α -syn; after this observation it was speculated that α -syn might be recruited as an additional mediator of polyQ toxicity; the hypothesis was confirmed because the accumulation of α -syn in polyQ inclusions was found in HD postmortem brains and in the R6/1 mouse model of HD. It was also found that N-terminal mutant huntingtin (N-mutHtt) and α -syn form independent filamentous microaggregates in R6/1 mouse brain as well as in the inducible HD94 mouse model and that N-mutHtt expression increases the load of α -syn filaments [88]. In the same direction, however, using a bimolecular fluorescence complementation assay, it was found that the initial steps of the coaggregation of Htt and α -syn it was found that HTT (exon 1) oligomerized with α -syn and sequestered it in the cytosol.…”

Section: Cross Talk Between α-Synuclein and Huntingtinmentioning

confidence: 99%

The Aggregation of Huntingtin and α-Synuclein

Chánez‐Cárdenas

Vázquez‐Contreras

2012

Journal of Biophysics

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Huntington's and Parkinson's diseases are neurodegenerative disorders associated with unusual protein interactions. Although the origin and evolution of these diseases are completely different, characteristic deposits of protein aggregates (huntingtin and α-synuclein resp.), are a common feature in both diseases. After these observations, many studies are performed with both proteins. Some of them try to understand the nature and driving forces of the aggregation process; others try to find a correlation between the genetic and failure in protein function. Finally with the combination of both approaches, it was proposed that possible strategies deal with pathologic aggregation. Unfortunately, if protein aggregation is a cause or a consequence of the neurodegeneration observed in these pathologies, it is still debatable. This paper describes the process of aggregation of two proteins: huntingtin and α synuclein. The characteristics of the aggregation reaction of these proteins have been followed with novel methods both in vivo and in vitro; these studies include both the combination with other proteins and the presence of various chemical compounds. The ultimate goal of this study was to summarize recent findings on protein aggregation and its possible role as a therapeutic target in neurodegenerative diseases and their role in biomaterial science.

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α-Synuclein accumulates in huntingtin inclusions but forms independent filaments and its deficiency attenuates early phenotype in a mouse model of Huntington's disease

Cited by 36 publications

References 62 publications

Bifunctional Anti-Non-Amyloid Component α-Synuclein Nanobodies Are Protective In Situ

Bifunctional Anti-Non-Amyloid Component α-Synuclein Nanobodies Are Protective In Situ

Intrabodies as Neuroprotective Therapeutics

The Aggregation of Huntingtin and α-Synuclein

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