Z7-2059 — Final report
1.
NLRP3 inflammasome activation by prion protein fibrils

Transmissible spongiform encephalopathies (prion diseases) are characterized by accumulation of abnormal form of prion protein, spongiform change, gliosis and progressive neuronal cell loss; however, the underlying cause of neurodegeneration is not known. Previous studies have indicated the role of innate immunity, particularly of NLRP3 inflammasome in other amyloid diseases - Alzheimer's disease and type 2 diabetes. Production of proinflammatory cytokine IL-1b has also been observed in brain in several types of prion disease and IL-1R deficiency significantly prolonged the onset of the disease. The aim of our study was to investigate whether prion protein assemblies can instigate NLRP3 inflammasome. We prepared several types of assemblies of prion protein and biophysically characterized them. We show that PrP fibrils, converted from alpha- to predominantly beta-type conformation trigger clevage of pro-IL-1b, which requires the inflammasome components NLRP3, ASC and caspase-1, whereas the native cellular form of prion protein does not induce activation of IL-1b. Several of the previously proposed NLRP3 inflammasome activation pathways may contribute to prion fibril induced inflammasome activation, since it can be blocked by ROS inhibitors, inhibition of K+ efflux and inhibition of phagocytosis. Proteinase K resistant PrP fibrils, but not monomers, induce release of neurotoxic species by microglial cells and can thus contribute to neuronal cell death. This work was presented as a poster at international conference and as a short talk at conference with international participation. Scientific paper was submitted (in attachment).

B.03 Paper at an international scientific conference

COBISS.SI-ID: 4737562
2.
Polypeptide material with adjustable pore properties

We prepared a polypeptide material with adaptable pore properties, which self-assembles from fusion proteins composed of at least two protein domains. This material can be used for separation of molecules on the basis of their properties and for chemical or enzyme catalysis.

F.33 Slovenian patent

COBISS.SI-ID: 4382234
3.
Self-assembled structures composed of coiled-coil elements

In innovative way we prepared self-assembling nanostructures, composed of polypeptides, which are able to form coiled-coils. In this way we are able to prepare nano-cages, nano-nets or nano- tubes.

F.33 Slovenian patent

COBISS.SI-ID: 4381210
4.
DNA-based nanomaterials

DNA origami technique is able to produce a variety of different shapes by constraining a long single stranded DNA molecule with a large number of short oligonucleotides. By designing and observing DNA origamis of Slovenia, we demonstrated that DNA origami technique can be used for construction of irregular asymmetric shapes. High school student research project including these results was awarded gold prize at the national competition of young researchers. With assembly of NanoSlovenia we proved the feasibility of this technique. Taking part in international student competition Biomod 2011, where we also received several prizes, we showed it is possible to enrich DNA origami at specific positions with different protein molecules. Currently we are exploring further applications in material science and medicine.

D.10 Educational activities

COBISS.SI-ID: 4630298
5.
Polypeptide material composed of elastin-like segments and coiled coil segments

We describe the design and preparation of a polypeptide material composed of elastin-like segments, coiled-coil-forming segments and optionally also functional polypeptide domains. Possible applications of these materials are in enhancement of the cell, tissue and organ growth, for differentiation of cells, inhibition of reproduction of pathogens, treatment of living human or animal tissues and as a medical and pharmaceutical material which will be applicable for treatment of living tissue.

F.33 Slovenian patent

COBISS.SI-ID: 4381978