Nanomagnetic formulations for integrated nucleic acid delivery, cell labelling and imaging with a focus on mRNA transfection for cell reprogramming
Final Report Abstract
During the funding period, a collection of optimised magnetic nanoparticles was tailor-made for highly efficient non-viral, lentiviral and adenoviral nucleic acid delivery. Optimised magnetic adenoviral formulations were used to boost the oncolytic potential of replicating adenovirus up to 50-fold. We achieved high-dose magnetic cell labelling with low toxicity and with particles of high transverse r2* relaxivity for magnetic resonance imaging, allowing the detection of labelled cells with high sensitivity. Scalable magnetic nanoparticle synthesis was established in a system with automated process control. Furthermore, magnetic microbubbles with high magnetic and acoustic responsiveness were developed and successfully used for magnetically targeted and ultrasound-triggered non-viral and viral nucleic acid delivery in vitro and in vivo, and this approach shows promising therapeutic results in animal models. Integrated magnetic cell separation and magnetofection, termed “magselectofection”, has been established as an efficient procedure to genetically modify cell lines and primary cells, including hematopoietic and mesenchymal stem cells. Specific magnetic cell labelling (magnetic activation) was shown to enhance efficacy of the nucleic acid delivery in an optimized magnetofection protocol. Well-characterised nanomagnetic formulations were developed for delivery of the siRNA and messenger RNA into cultured and the primary cells of interest. Then, these methods was translated for use with functional chemically modified messenger RNAs (cmRNAs), which involved a detailed analysis of timing and repeated dosing to obtain the optimal yield, viability and functionality of the derived cell populations. The work program was carried out in close collaboration with Research Unit Partners 1-6 and resulted in robust, high-yield protocols of nucleic acid delivery for research and possible therapeutic applications. This project continuously provided the Research Unit Partners with comprehensively characterised magnetic nanoparticles (MNPs) for viral and nonviral nucleic acid delivery, magnetic cell labelling and manipulation, magnetic resonance imaging, relaxometry, magnetic measurments and the generation of magnetic microbubbles for magnetically targeted and ultrasound-activated nucleic acid delivery. In this context, our focus was on understanding the nano-interactions (including the magnetophoretic mobility and relaxivity of the magnetic delivery entities) involved in these processes to optimise the design of nanomagnetic composites. These achievements have resulted in 38 published project-related papers.
Publications
- (2010). Magnetic and Acoustically Active Lipospheres for Magnetically Targeted Nucleic Acid Delivery. Advanced Functional Materials 20, 3881–3894
Vlaskou, D., Mykhaylyk, O., Krötz, F., Hellwig, N., Renner, R., Schillinger, U., Gleich, B., Heidsieck, A., Schmitz, G., Hensel, K., Plank, C.
(See online at https://doi.org/10.1002/adfm.200902388) - (2011). Magselectofection: an integrated method of nanomagnetic separation and genetic modification of target cells. Blood 117, E171-E181
Sanchez-Antequera, Y., Mykhaylyk, O., van Til, N.P., Cengizeroglu, A., de Jong, J.H., Huston, M.W., Anton, M., Johnston, I.C.D., Pojda, Z., Wagemaker, G., Plank, C.
(See online at https://doi.org/10.1182/blood-2010-08-302646) - (2012). Effects of nanoparticle coatings on the activity of oncolytic adenovirus-magnetic nanoparticle complexes. Biomaterials 33, 256-269
Tresilwised, N., Pithayanukul, P., Holm, P.S., Schillinger, U., Plank, C., and Mykhaylyk, O.
(See online at https://doi.org/10.1016/j.biomaterials.2011.09.028) - (2012). Silica-iron oxide magnetic nanoparticles modified for gene delivery: a search for optimum and quantitative criteria. Pharm Res 29, 1344-1365
Mykhaylyk, O., Sobisch, T., Almstatter, I., Sanchez-Antequera, Y., Brandt, S., Anton, M., Doblinger, M., Eberbeck, D., Settles, M., Braren, R., Plank, C.
(See online at https://doi.org/10.1007/s11095-011-0661-9) - (2015). Characterization of Magnetic Viral Complexes for Targeted Delivery in Oncology. Theranostics 5, 667-685
Almstätter, I., Mykhaylyk, O., Settles, M., Altomonte, J., Aichler, M., Walch, A., Rummeny, E., Ebert, O., Plank, C., and Braren, R.
(See online at https://dx.doi.org/10.7150%2Fthno.10438) - (2015). Nanomagnetic Activation as a Way to Control the Efficacy of Nucleic Acid Delivery. Pharmaceutical Research 32, 103-121
Grzeskowiak, B.F., Sanchez-Antequera, Y., Hammerschmid, E., Doblinger, M., Eberbeck, D., Wozniak, A., Slomski, R., Plank, C., and Mykhaylyk, O.
(See online at https://doi.org/10.1007/s11095-014-1448-6) - (2016). Chemically modified RNA induces osteogenesis of stem cells and human tissue explants as well as accelerates bone healing in rats. Biomaterials 87, 131-146
Balmayor, E.R., Geiger, J.P., Aneja, M.K., Berezhanskyy, T., Utzinger, M., Mykhaylyk, O., Rudolph, C., and Plank, C.
(See online at https://doi.org/10.1016/j.biomaterials.2016.02.018)
