Prion Diagnosis: Application of Real-Time Quaking-Induced Conversion

Kang, Hae‐Eun; Mo, Youngwon; Rahim, Raihah Abd; Lee, Hyemi; Ryou, Chongsuk

doi:10.1155/2017/5413936

Cited by 26 publications

(20 citation statements)

References 75 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…We found that gentle shaking of the reaction mix was required in order for PrP to form aggregates. This is consistent with conditions used for QuIC experiments 25 and was possibly due to a fraction of the aggregates adsorbed on the microcentrifuge tube surface. Interestingly, we also observed fast disappearance of PrP aggregates after the reaction reached a plateau.…”

Section: Resultssupporting

confidence: 86%

Direct Observation of Murine Prion Protein Replication in Vitro

et al. 2018

View full text Add to dashboard Cite

Prions are believed to propagate when an assembly of prion protein (PrP) enters a cell and replicates to produce two or more fibrils, leading to an exponential increase in PrP aggregate number with time. However, the molecular basis of this process has not yet been established in detail. Here, we use single-aggregate imaging to study fibril fragmentation and elongation of individual murine PrP aggregates from seeded aggregation in vitro. We found that PrP elongation occurs via a structural conversion from a PK-sensitive to PK-resistant conformer. Fibril fragmentation was found to be length-dependent and resulted in the formation of PK-sensitive fragments. Measurement of the rate constants for these processes also allowed us to predict a simple spreading model for aggregate propagation through the brain, assuming that doubling of the aggregate number is rate-limiting. In contrast, while α-synuclein aggregated by the same mechanism, it showed significantly slower elongation and fragmentation rate constants than PrP, leading to much slower replication rate. Overall, our study shows that fibril elongation with fragmentation are key molecular processes in PrP and α-synuclein aggregate replication, an important concept in prion biology, and also establishes a simple framework to start to determine the main factors that control the rate of prion and prion-like spreading in animals.

show abstract

Section: Resultssupporting

confidence: 86%

Direct Observation of Murine Prion Protein Replication in Vitro

et al. 2018

View full text Add to dashboard Cite

show abstract

“…However, RT-QuIC does offer us a means to rapidly assess large numbers of amino acid substitutions between and within species associated with CWD to determine whether. In this study we did not observe changes in the key parameters to differentiate RT-QuIC (lag time, amyloid formation, or ThT fluorescence signal intensity) [38] based on https://doi.org/10.1371/journal.pone.0227487.g009 the genotypes of CWD or with different sources of CWD. Since the RT-QuIC assay monitors the fibril formation in real-time via binding of the fluorescence marker ThT to the amyloid fibrils, different lag times indicate different seeding rate of reactions, which is often used to differentiate the reaction.…”

Section: Discussioncontrasting

confidence: 61%

Role of donor genotype in RT-QuIC seeding activity of chronic wasting disease prions using human and bank vole substrates

2020

View full text Add to dashboard Cite

Chronic wasting disease is a transmissible spongiform encephalopathy of cervids. This fatal neurodegenerative disease is caused by misfolding of the cellular prion protein (PrP C) to pathogenic conformers (PrP Sc), and the pathogenic forms accumulate in the brain and other tissues. Real-time Quaking Induced Conversion (RT-QuIC) can be used for the detection of prions and for prion strain discrimination in a variety of biological tissues from humans and animals. In this study, we evaluated how either PrP Sc from cervids of different genotypes or PrP Sc from different sources of CWD influence the fibril formation of recombinant bank vole (BV) or human prion proteins using RT-QuIC. We found that reaction mixtures seeded with PrP Sc from different genotypes of white-tailed deer or reindeer brains have similar conversion efficiency with both substrates. Also, we observed similar results when assays were seeded with different sources of CWD. Thus, we conclude that the genotypes of all sources of CWD used in this study do not influence the level of conversion of PrP C to PrP Sc .

show abstract

“…RT-QuIC has been successfully used to detect small amounts of PrP Sc in brain tissue, cerebrospinal fluid, blood, saliva, and nasal fluid from human, cervid, ovine, bovine, and rodent species. PrP amyloid resulting from the conversion of rPrP to its aggregated form is indicated by increased ThT fluorescence, which directly indicates beta-sheet structure formation in protein aggregates [44].…”

Section: Discussionmentioning

confidence: 99%

BMD42-2910, a Novel Benzoxazole Derivative, Shows a Potent Anti-prion Activity and Prolongs the Mean Survival in an Animal Model of Prion Disease

et al. 2020

View full text Add to dashboard Cite

Prion diseases are a group of neurodegenerative and fatal central nervous system disorders. The pathogenic mechanism involves the conversion of cellular prion protein (PrP C) to an altered scrapie isoform (PrP Sc), which accumulates in amyloid deposits in the brain. However, no therapeutic drugs have demonstrated efficacy in clinical trials. We previously reported that BMD42-29, a synthetic compound discovered in silico, is a novel anti-prion compound that inhibits the conversion of PrP C to protease K (PK)-resistant PrP Sc fragments (PrP res). In the present study, 14 derivatives of BMD42-29 were obtained from BMD42-29 by modifying in the side chain by in silico feedback, with the aim to determine whether they improve anti-prion activity. These derivatives were assessed in a PrP Sc-infected cell model and some derivatives were further tested using real timequaking induced conversion (RT-QuIC). Among them, BMD42-2910 showed high anti-prion activity at low concentrations in vitro and also no toxic effects in a mouse model. Interestingly, abundant PrP res was reduced in brains of mice infected with prion strain when treated with BMD42-2910, and the mice survived longer than control mice and even that treated with BMD42-29. Finally, high binding affinity was predicted in the virtual binding sites (Asn159, Gln 160, Lys194, and Glu196) when PrP C was combined with BMD-42-2910. Our findings showed that BMD42-2910 sufficiently reduces PrP res generation in vitro and in vivo and may be a promising novel anti-prion compound.

show abstract

Prion Diagnosis: Application of Real-Time Quaking-Induced Conversion

Cited by 26 publications

References 75 publications

Direct Observation of Murine Prion Protein Replication in Vitro

Direct Observation of Murine Prion Protein Replication in Vitro

Role of donor genotype in RT-QuIC seeding activity of chronic wasting disease prions using human and bank vole substrates

BMD42-2910, a Novel Benzoxazole Derivative, Shows a Potent Anti-prion Activity and Prolongs the Mean Survival in an Animal Model of Prion Disease

Contact Info

Product

Resources

About