Krako 2010 Abstract MiP2010
|Targeting mitochondrial amyloid β through intrabodies: new approach to study pathogenesis of Alzheimer’s disease.|
Intracellular antibodies (intrabodies), represent recombinant antibody fragments which are ectopically expressed within a cell and target specific intracellular antigens. The concept of using intrabodies, pioneered by our laboratory, is based on induction of phenotypic knockout of a relevant target protein . To exert their function, intrabodies have to be directed to the subcellular compartment where the antigen is located. Thus, unlike other knock-down approaches, intrabodies can exert subcellular-specific protein interference . Intrabody-based approaches are now being investigated for mechanistic studies and potential therapeutic treatment of diverse neurodegenerative disorders: Alzheimer’s disease, Parkinson’s disease, Huntington’s disease and prion encephalopathies. In all of these disorders the aggregation of specific misfolded proteins is linked to neurodegeneration of specific neuronal populations. Our laboratory investigates the use of conformational sensitive intrabodies against amyloid βoligomers (AβOs), which are pathologically relevant conformations of amyloid β (Aβ) peptide in Alzheimer’s disease. Functional anti-AβOs intrabodies were selected from intrabody libraries in yeast by the intracellular antibody capture technology (IACT) . The specific conformational anti-AβOs intrabodies have been used to selectively target AβOs in different subcellular compartments. Since there are growing evidences for important role of mitochondria in most common neurodegenerative diseases, we are investigating role of mitochondrial AβOs by using intrabody approach. We have prepared plasmid vectors encoding anti-AβOs intrabodies harboring mitochondrial targeting sequences in order to target the mitochondrial pool of the Aβ peptide. Functional and biochemical assays are in progress in well established cell models for production and secretion of AβOs (7PA2 cells, kindly provided by Dr D. Selkoe) in order to study functions of these intrabodies that interfere with mitochondrial Aβ accumulation and cellular toxicity.
Labels: Pathology: Alzheimer's
1-Scuola Normale Superiore, Pisa, Italy; 2-European Brain Research Inst (EBRI), Rome, Italy. - [email protected]
References and acknowledgements
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- Persic L, Righi M, Roberts A, Hoogenboom HR, Cattaneo A, Bradbury A (1997) Targeting vectors for intracellular immunization. Gene 187:1-8.
- Meli G, Visintin M, Cannistraci I, Cattaneo A (2009) Direct in vivo intracellular selection of conformation-sensitive antibody domains targeting Alzheimer’s amyloid-β oligomers. J Mol Biol 387:584-606.
This abstract was also presented at IOC57