Amyloid-beta oligomers (AbetaOs) are well recognized neurotoxic species in Alzheimer's Disease (AD) but they are still difficult targets for therapy due to their polymorphic and complex assembly state and to the lack of selective targeting tools. Conformational sensitive antibodies are unique tools to study complex amyloid assembly states and dynamics and to establish new Abeta targeting approaches. We recently generated and exploited recombinant antibody fragments as intracellular antibodies (intrabodies) for a subcellularlocalized interference to block or modulate the function of AbetaOs in living cells. In this way, we established a new experimental paradigm of subcellularlocalized and conformationalselective interference (CSI) (Meli et al., Nature Comm 2014). Our approach selectively controls both levels and toxic conformations of biologically active AbetaO in living cells, by targeting an intrabody in the Endoplasmic Reticulum (ER). In this way, we also demostrated that pools of endogenous Abeta oligomerize into pathological forms starting from the ER. We reported the functional relevance of CSI on modulating pathways of intracellular homeostasis and synaptic functions. Now we are investigating the functional effects mediated by the ERlocalized CSI on some subcellular alterations and mitochondrial disfunctions, describing an altered link between ER and mitochondria as a relevant mechanism of AD pathogenesis. We are also targeting the intrabody through lentiviral systems in primary neuronal stem cells (NSC) derived from neurogenic niches of the adult brain of AD mouse model Tg2576 and in primary human fibroblasts from AD patients. In conclusion, the intrabodybased CSI is a promising strategy for in vivo therapeutic applications to selectively target AbetaOs at subcellular level and its use to dissect new cellular mechanisms of AbetaO generation, trafficking and actions will uncover new targets for drugs development.
Intrabodies targeting Amyloid-beta oligomers in the Endoplasmic Reticulum: preclinical evidences for new twist in immunotherapy
R Scardigli;
2016
Abstract
Amyloid-beta oligomers (AbetaOs) are well recognized neurotoxic species in Alzheimer's Disease (AD) but they are still difficult targets for therapy due to their polymorphic and complex assembly state and to the lack of selective targeting tools. Conformational sensitive antibodies are unique tools to study complex amyloid assembly states and dynamics and to establish new Abeta targeting approaches. We recently generated and exploited recombinant antibody fragments as intracellular antibodies (intrabodies) for a subcellularlocalized interference to block or modulate the function of AbetaOs in living cells. In this way, we established a new experimental paradigm of subcellularlocalized and conformationalselective interference (CSI) (Meli et al., Nature Comm 2014). Our approach selectively controls both levels and toxic conformations of biologically active AbetaO in living cells, by targeting an intrabody in the Endoplasmic Reticulum (ER). In this way, we also demostrated that pools of endogenous Abeta oligomerize into pathological forms starting from the ER. We reported the functional relevance of CSI on modulating pathways of intracellular homeostasis and synaptic functions. Now we are investigating the functional effects mediated by the ERlocalized CSI on some subcellular alterations and mitochondrial disfunctions, describing an altered link between ER and mitochondria as a relevant mechanism of AD pathogenesis. We are also targeting the intrabody through lentiviral systems in primary neuronal stem cells (NSC) derived from neurogenic niches of the adult brain of AD mouse model Tg2576 and in primary human fibroblasts from AD patients. In conclusion, the intrabodybased CSI is a promising strategy for in vivo therapeutic applications to selectively target AbetaOs at subcellular level and its use to dissect new cellular mechanisms of AbetaO generation, trafficking and actions will uncover new targets for drugs development.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
