- We successfully engineered mechano-responsive cells by expressing
mechanosensitive ion channels MscS and P3:FAStm:TRPC1 in mammalian cells.
- Localization of mechanosensitive channel TRPC1 on the plasma membrane was
demonstrated and improved by fusing it with segments of FAS receptor, including
the transmembrane domain.
- Addition of gas-filled lipid microbubbles increased the sensitivity of mammalian
cells to ultrasound.
- We demonstrated for the first time that gas vesicle-forming proteins are
expressed in a human cell line, are not toxic and improve the sensitivity of
cells to mechanical stimuli.
- A custom-made ultrasound stimulation device (Moduson), suitable for use in
different experimental setups that require ultrasound stimulation of cells, was
developed.
- New graphical analysis software that enables fast analysis of fluorescent
microscopy data was also developed to quantify the response to ultrasound
stimulation.
- A new split calcium sensing/reporting system was designed that is able to report,
by emitting light, the increase of the cytosolic calcium ions induced by
mechanoreceptor stimulation.
Cells interact with other cells and the environment in various ways in order to
appropriately respond to microenvironment changes. An important extracellular physical
signal is represented
by mechanical forces and adaptation upon mechanical stimuli is crucial for regulating
the cell volume, signalization, growth, cell to cell interactions and overall
survival.
Mechanical forces such as changes in osmolality, fluid flow, gravity or direct pressure
result in changes in tension of the phospholipid bilayer, arrangement of the
cytoskeleton
and opening of cation-permeable channels.
This mechanism serves as a force-sensing system
Haswell2011, Zheng2013
. Furthermore, it has already been shown that living organisms can detect
and respond to mechanical stress generated by ultrasound, which represents an external
stimulus with many potential applications
Ibsen2015
. Ultrasound
offers remarkable advantages over other external stimuli used for targeted cell
stimulation. In our project we aimed to explore the potential of mechanosensors and to
improve the sensitivity of cells to mechanical stimulation with the idea of designing
ultrasound-responsive devices.
