NR AHCV
AU Lecerf,J.M.; Shirley,T.L.; Zhu,Q.; Kazantsev,A.; Amersdorfer,P.; Housman,D.E.; Messer,A.; Huston,J.S.
TI Human single-chain Fv intrabodies counteract in situ huntingtin aggregation in cellular models of Huntington's disease
QU Proceedings of the National Academy of Sciences of the United States of America 2001 Apr 10; 98(8): 4764-9
PT journal article
AB 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.
MH Amino Acid Sequence; Animal; COS Cells; Human; Huntington Disease/*pathology; Immunoglobulin Variable Region/*immunology; Luminescent Proteins/metabolism; Molecular Sequence Data; Nerve Tissue Proteins/immunology/*metabolism; Nuclear Proteins/immunology/*metabolism; Protein Binding; Support, U.S. Gov't, P.H.S.
AD IntraImmune Therapies, Inc., Lexington, MA 02215, USA
SP englisch
PO USA