PrPTSE detection

MicroRNA profiling

MicroRNA profiling in blood as a basis for novel diagnostics and early treatment of transmissible spongiform encephalopathies

2012: 1 year / Grant: 20 000 euros

Detection of Exosomal PrPTSE in blood

An exosomal misfolded prion protein (PrPTSE) in blood as an early diagnostic marker of transmissible spongiform encephalopathies (TSEs)

2013: 1 year / Grant: 32 800 euros

Projects: The aim of the project “MicroRNA profiling” was to search for and identify microRNA biomarkers and PrPTSE in blood exosomesfrom TSE-infected mice using small RNA sequencing and PMCA, respectively. PrPTSE detection was further pursued in a second project named “Detection of Exosomal PrPTSE in blood”, in which perfectioned exosome isolation from plasma was combined with PMCA technique for the detection of PrPTSE in exosomesfrom blood ofvCJD-infected squirrel monkeys and mice. Exosomes were also characterized by various available techniques.

Main results and related published data: Scientists performed profiling bysequencing of miRNA isolated from exosomal preparations from either conditioned media or blood plasma required significant optimization. Results of miRNA profiling from conditioned cell culture media revealed no differences between TSE-infected and non-infected cells. Work is still in progress to resolve outstanding issues. Addressing the second project, scientists were able to demonstrate PrPTSE by PMCA in exosomal fractions isolated from TSE-infected cell culture media and from plasma of infected mice and squirrel monkeys. They also showed the presence of PrPTSE in exosomal preparations from plasma obtained during the incubation period and at clinical phase in mouse models of vCJD. The presence of exosome in used preparation was addressed by various methods.
The final proof whether PrPTSE is carried by exosomes still remains unresolved.