Periodic Reporting for period 3 - SIROCCO (Remote control of cellular signalling triggered by magnetic switching)
Okres sprawozdawczy: 2023-05-01 do 2024-10-31
It is becoming increasingly clear that successful tissue engineering rests on capitalizing on more comprehensive understanding of mechanotransduction. In fact, the study of mechanotransduction is now a burgeoning research field, helping us to understand how cells respond to mechanical stimuli and convert them into biochemical signals, which in turn sheds light on the remote control of intracellular functions and the treatment of diseases. However, external manipulation of cellular signalling constitutes an important challenge due to the lack of non-invasive techniques that can be tuned in a spatiotemporal manner at a deep-tissue level. SIROCCO aims to control different pathways related with cutaneous mechanotransduction by using magnetic nanoparticles in order to enhance wound healing. External magnetic fields will be applied to magnetic nanoparticles selectively attached to the membrane of living cells in order to create a force that can activate key intracellular pathways connected with mechanotransduction. Using genetically modified 2D and 3D in vitro models, the possibility to activate important pathways in order to enhance wound healing will be tested.
We have synthesized magnetic nanoparticles (MNPs) of different sizes and compositions. We have then produced small protein fragments and attached them to the MNPs so that they can recognize the cell membrane in a specific way. We have studied the colocalization of MNPs with the cell membrane using cells expressing fluorescent proteins. To assess if we are activating key intracellular pathways, we have generated different reporter cell lines that turn fluorescent upon the activation of the pathways. We have also developed the first prototype to deliver magnetic cues to the cells.
In order to target cell membranes, we optimized the bioconjugation of the proteins to MNPs using a rather novel alternative for orienting protein fragments on MNPs.
We expect to assess the activation of two important pathways by the end of the project.
We expect to assess the activation of two important pathways by the end of the project.