Unknown

Dataset Information

0

Human single-chain Fv intrabodies counteract in situ huntingtin aggregation in cellular models of Huntington's disease.


ABSTRACT: This investigation was pursued to test the use of intracellular antibodies (intrabodies) as a means of blocking the pathogenesis of Huntington's disease (HD). HD is characterized by abnormally elongated polyglutamine near the N terminus of the huntingtin protein, which induces pathological protein-protein interactions and aggregate formation by huntingtin or its exon 1-containing fragments. Selection from a large human phage display library yielded a single-chain Fv (sFv) antibody specific for the 17 N-terminal residues of huntingtin, adjacent to the polyglutamine in HD exon 1. This anti-huntingtin sFv intrabody was tested in a cellular model of the disease in which huntingtin exon 1 had been fused to green fluorescent protein (GFP). Expression of expanded repeat HD-polyQ-GFP in transfected cells shows perinuclear aggregation similar to human HD pathology, which worsens with increasing polyglutamine length; the number of aggregates in these transfected cells provided a quantifiable model of HD for this study. Coexpression of anti-huntingtin sFv intrabodies with the abnormal huntingtin-GFP fusion protein dramatically reduced the number of aggregates, compared with controls lacking the intrabody. Anti-huntingtin sFv fused with a nuclear localization signal retargeted huntingtin analogues to cell nuclei, providing further evidence of the anti-huntingtin sFv specificity and of its capacity to redirect the subcellular localization of exon 1. This study suggests that intrabody-mediated modulation of abnormal neuronal proteins may contribute to the treatment of neurodegenerative diseases such as HD, Alzheimer's, Parkinson's, prion disease, and the spinocerebellar ataxias.

SUBMITTER: Lecerf JM 

PROVIDER: S-EPMC31908 | biostudies-literature | 2001 Apr

REPOSITORIES: biostudies-literature

altmetric image

Publications

Human single-chain Fv intrabodies counteract in situ huntingtin aggregation in cellular models of Huntington's disease.

Lecerf J M JM   Shirley T L TL   Zhu Q Q   Kazantsev A A   Amersdorfer P P   Housman D E DE   Messer A A   Huston J S JS  

Proceedings of the National Academy of Sciences of the United States of America 20010401 8


This investigation was pursued to test the use of intracellular antibodies (intrabodies) as a means of blocking the pathogenesis of Huntington's disease (HD). HD is characterized by abnormally elongated polyglutamine near the N terminus of the huntingtin protein, which induces pathological protein-protein interactions and aggregate formation by huntingtin or its exon 1-containing fragments. Selection from a large human phage display library yielded a single-chain Fv (sFv) antibody specific for t  ...[more]

Similar Datasets

| S-EPMC8934500 | biostudies-literature
| S-EPMC7459410 | biostudies-literature
| S-EPMC7511939 | biostudies-literature
| S-EPMC4718563 | biostudies-literature
| S-EPMC1183604 | biostudies-literature
| S-EPMC8664476 | biostudies-literature
| S-EPMC5505970 | biostudies-literature
| S-EPMC6681014 | biostudies-literature
| S-EPMC6064984 | biostudies-literature
| S-EPMC7746729 | biostudies-literature