Abstract
α-Synuclein (α-syn) has been identified as the major component of Lewy bodies that characterize neurodegenerative synucleinopathies, including Parkinson's disease. Overexpression of α-syn, and prefibrillar α-syn oligomers, has been implicated in these pathologies; therefore, prevention of prefibril accumulation, and inhibition of other aberrant effects of overexpressed α-syn, could provide novel treatments. Here, we have selected a human single-chan Fv (scFv) antibody, D10, that binds human monomeric wild-type α-syn. We demonstrate, by retargeting assays and coimmunoprecipitation, that the D10 scFv is a specific and efficient intracellular antibody (intrabody). By transfecting the D10 scFv gene into an HEK 293 cell line that overexpresses wild-type α-syn, we show that the D10 intrabody stabilizes detergent-soluble monomeric α-syn and inhibits the formation of detergent-insoluble high-molecular-weight α-syn species. In addition, the D10 intrabody ameliorates the decreased cell adhesion that characterizes the α-syn-overexpressing cells. Given the important role of α-syn pathology, and the facility with which intrabodies can be further engineered in vitro, anti-α-syn intrabodies may represent novel molecular therapeutics for synucleinopathies, with implications for other neurodegenerative disorders caused by misfolded accumulated proteins.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1023-1031 |
| Number of pages | 9 |
| Journal | Molecular Therapy |
| Volume | 10 |
| Issue number | 6 |
| DOIs | |
| State | Published - Dec 2004 |
Keywords
- Intrabody
- Lewy body
- Neurodegeneration
- Parkinson's disease
- Synuclein
- Synucleinopathy
- scFv
ASJC Scopus subject areas
- Molecular Medicine
- Molecular Biology
- Genetics
- Pharmacology
- Drug Discovery