Mehrije Ferizi

The focus of my work will be the preparation of "SNIM RNA" (stabilized, non-immunogenic messenger RNA) as a novel therapeutic entity for transcript therapies. In brief, chemically modified ribonucleotides are used to prepare "SNIM RNA". Further these molecules are going to be optimized in terms of stability, non-immunogenicity and in terms of protein production level when transfected into target cells. The optimization is based on a biological selection procedure; establishing optimized nanotechnological delivery systems, such as magnetofection, for the molecules in cell culture. Magnetofection is defined as nucleic acid delivery under the influence of a magnetic field acting on nucleic acid vectors that are associated with magnetic nanoparticles.

First preliminary transfections studies of SNIM RNA in two different cell types using magnetofection were already performed (see fig. 1 and 2). Until now the results are showing different expression profiles. On the one hand transfection studies with the magnetic lipoplexes using SM4-31 as transfection reagent demonstrated significant dose-dependent improvement of the target protein expression after magnetofection. On the other hand transfections studies with NIH 3T3 cells using Dmrie-C instead of SM4-31 showed an improvement after 96h using the same nanotechnological delivery system. For that purpose further studies regarding formulation, binding affinity and internalization has to be investigated.

In the next step the best molecules in terms of long-term expression and in acting less immunogenic are going to be selected. Afterwards mesenchymal stem cells are transfected with these molecules by magnetofection to induce osteogenic differentiation. Finally the differentiated stem cells are loaded into different biomaterials first and then tested in the animal model we select.