Prion diseases are a group of lethal neurodegenerative diseases in humans and animals. The central event of prion diseases pathogenesis is the conversion of the normal prion protein (PrPC) into pathogenic prion protein (PrPSc). To diagnose human prion diseases (Creutzfeldt-Jakob disease, CJD), CSF 1...
Prion diseases are a group of lethal neurodegenerative diseases in humans and animals. The central event of prion diseases pathogenesis is the conversion of the normal prion protein (PrPC) into pathogenic prion protein (PrPSc). To diagnose human prion diseases (Creutzfeldt-Jakob disease, CJD), CSF 14-3-3 protein has been used as a indirect marker, but the specificity and variety of different methods are controversial. Previous studies have investigated pharmacotherapy approaches to treat prion diseases and most potential anti-prion compounds have been validated using PrPSc-infected cell models. However, there is no differentiation between the abilities of the anti-prion compounds to inhibit prion conversion and to degrade PrPSc. Also, the potential compounds have been shown to be ineffective in PrPSc-infected mouse model. This study attempts to improve the diagnostic accuracy and evaluate both a novel anti-prion screening method and compounds obtained from in silico simulation. Potential diagnostic activity of tau protein as an alternative marker was examined in 62 sporadic CJD (sCJD) and 89 non-CJD CSF samples. Total tau (t-tau) and phosphorylated tau (p-tau) levels were measured and the p-tau/t-tau ratio (p/t ratio) was calculated. The results show that combined use of 14-3-3 protein and p/t ratio improved the specificity of the diagnosis compared with 14-3-3 protein alone. In addition, the p/t ratio allowed discrimination between true positive and negative from ambiguous weak positive 14-3-3. Recently, the small quantity of PrPSc in CSF has been detected using in vitro amplification assays, called real time-quaking induced conversion (RT-QuIC). RT-QuIC enables controlling the experimental conversion of PrPC into PrPSc, and was applied to the anti-prion compound screening, to evaluate its usefulness as a screening method. Inhibitory activities of acridine, dextran sulfate sodium (DSS), and tannic acid were assessed using RT-QuIC and compared with those from a PrPSc-infected cell assay. The results show that acridine and tannic acid inhibited prion conversion and reduced PrPSc, whereas DSS only inhibited the conversion in RT-QuIC. This difference was not observed in the PrPSc-infected cell screening. Thus, RT-QuIC enabled a more specific classification of compounds according to their action, as well as the evaluation of their activity. No suitable anti-prion compounds have been identified to date in in vivo models. The novel and potential anti-prion compounds were therefore selected by in silico simulation to predict the in vivo as well as in vitro activities in advance. A total of 37 potential compounds were screened in vitro using a multimer detection system (MDS), cell-based assay and surface plasmon resonance (SPR) assay. Among these, BMD42-29 and -35 were the most effective, and derivatives were produced for both compounds. Eventually, BMD42-2910 was selected by an additional in vitro screening. BMD42-29 and -2910 were tested in a PrPSc-infected mouse model. Finally, BMD42-2910 showed the highest anti-prion activity in the brains of in vivo disease model. Taken together, this study investigated a more accurate diagnosis method using tau protein as well as a novel screening method for the anti-prion compounds and a potential compound from in silico to in vivo. In management and assessment of human prion disease, the findings of this study provide essential data that will help the accurate diagnosis and effective treatment in the future.
Prion diseases are a group of lethal neurodegenerative diseases in humans and animals. The central event of prion diseases pathogenesis is the conversion of the normal prion protein (PrPC) into pathogenic prion protein (PrPSc). To diagnose human prion diseases (Creutzfeldt-Jakob disease, CJD), CSF 14-3-3 protein has been used as a indirect marker, but the specificity and variety of different methods are controversial. Previous studies have investigated pharmacotherapy approaches to treat prion diseases and most potential anti-prion compounds have been validated using PrPSc-infected cell models. However, there is no differentiation between the abilities of the anti-prion compounds to inhibit prion conversion and to degrade PrPSc. Also, the potential compounds have been shown to be ineffective in PrPSc-infected mouse model. This study attempts to improve the diagnostic accuracy and evaluate both a novel anti-prion screening method and compounds obtained from in silico simulation. Potential diagnostic activity of tau protein as an alternative marker was examined in 62 sporadic CJD (sCJD) and 89 non-CJD CSF samples. Total tau (t-tau) and phosphorylated tau (p-tau) levels were measured and the p-tau/t-tau ratio (p/t ratio) was calculated. The results show that combined use of 14-3-3 protein and p/t ratio improved the specificity of the diagnosis compared with 14-3-3 protein alone. In addition, the p/t ratio allowed discrimination between true positive and negative from ambiguous weak positive 14-3-3. Recently, the small quantity of PrPSc in CSF has been detected using in vitro amplification assays, called real time-quaking induced conversion (RT-QuIC). RT-QuIC enables controlling the experimental conversion of PrPC into PrPSc, and was applied to the anti-prion compound screening, to evaluate its usefulness as a screening method. Inhibitory activities of acridine, dextran sulfate sodium (DSS), and tannic acid were assessed using RT-QuIC and compared with those from a PrPSc-infected cell assay. The results show that acridine and tannic acid inhibited prion conversion and reduced PrPSc, whereas DSS only inhibited the conversion in RT-QuIC. This difference was not observed in the PrPSc-infected cell screening. Thus, RT-QuIC enabled a more specific classification of compounds according to their action, as well as the evaluation of their activity. No suitable anti-prion compounds have been identified to date in in vivo models. The novel and potential anti-prion compounds were therefore selected by in silico simulation to predict the in vivo as well as in vitro activities in advance. A total of 37 potential compounds were screened in vitro using a multimer detection system (MDS), cell-based assay and surface plasmon resonance (SPR) assay. Among these, BMD42-29 and -35 were the most effective, and derivatives were produced for both compounds. Eventually, BMD42-2910 was selected by an additional in vitro screening. BMD42-29 and -2910 were tested in a PrPSc-infected mouse model. Finally, BMD42-2910 showed the highest anti-prion activity in the brains of in vivo disease model. Taken together, this study investigated a more accurate diagnosis method using tau protein as well as a novel screening method for the anti-prion compounds and a potential compound from in silico to in vivo. In management and assessment of human prion disease, the findings of this study provide essential data that will help the accurate diagnosis and effective treatment in the future.
주제어
#RT-QuIC ScN2a Anti-prion Acridine Dextran sulfate sodium Tannic acid Multimer detection system Prion Creutzfeldt-Jakob disease Diagnosis Tau
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