A human single-chain Fv intrabody blocks aberrant cellular effects of overexpressed α-synuclein

Chun Zhou, Sharareh Emadi, Michael Sierks, Anne Messer

Research output: Contribution to journalArticlepeer-review

101 Scopus citations

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 languageEnglish (US)
Pages (from-to)1023-1031
Number of pages9
JournalMolecular Therapy
Volume10
Issue number6
DOIs
StatePublished - Dec 1 2004

Keywords

  • Intrabody
  • Lewy body
  • Neurodegeneration
  • Parkinson's disease
  • Synuclein
  • Synucleinopathy
  • scFv

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Pharmacology
  • Drug Discovery

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