Final Report Summary - FORCEMAP (Intramolecular force mapping of enzymes in action: the role of strain in motor mechanisms)
Technology for sustained local control of molecular effects within organs, cells or subcellular regions is currently unavailable, even though such a technology would be pivotal for unveiling the molecular actions underlying collective mechanisms of neuronal networks, signaling systems, complex machineries and organism development. Although existing uncaging and photoswitch methods allow in-cell perturbation of molecular processes, in these methods the drug effects are exerted for a very short time due to millisecond-range diffusion from the targeted area.
We have developed a novel optopharmacological technology, we termed Molecular tattooing, which combines photoaffinity labeling with two-photon microscopy. Molecular tattooing covalently attaches a photoreactive bioactive compound to its target by two-photon irradiation without any systemic effects outside the targeted area, thereby achieving subfemtoliter, long-term confinement of target-specific effects in vivo. As we demonstrated in melanoma cells and zebrafish embryos, Molecular tattooing is suitable for dissecting collective activities by the separation of autonomous and non-autonomous molecular processes in vivo ranging from subcellular to organism level. Molecular tattooing is the first technology that establishes a broad application potential for optopharmacology by confining drug effects to a few cells of a developing organ in a live animal, or even a subcellular region of a single cell. Molecular tattoo will revolutionize local targeting techniques of bioactive ligands because it is able to dissect specific interactions of complex collective processes. It will provide answers to the following questions: What is the specific role of an enzyme located in a certain cell of a developing organ? How does the subcellular location of an enzyme modulate different cellular processes such as cell movement, blebbing, cytokinesis or signalling? What is the function of a single synapse within a multi-synaptic neuronal connection? The key advantage of Molecular tattooing is that it can induce local and permanent confinement of a drug effect without significant systemic and side effects outside the targeted area. Importantly, a series of photoreactive bioactive ligands are already available for a wide variety of targets and further photoreactive compounds can be synthesized in a single step from existing drug derivatives. Thus the wide variety of compounds will facilitate the rapid, widespread propagation of Molecular tattooing. This technology provides an immediate opportunity for technology transfer both in research and health industries by developing novel photochemical drugs and scanning microscopy instruments optimized for Molecular tattooing. Realizing this opportunity, we established a SME company called Optopharma Ltd. which was supported by ERC PoC scheme.
We have developed a novel optopharmacological technology, we termed Molecular tattooing, which combines photoaffinity labeling with two-photon microscopy. Molecular tattooing covalently attaches a photoreactive bioactive compound to its target by two-photon irradiation without any systemic effects outside the targeted area, thereby achieving subfemtoliter, long-term confinement of target-specific effects in vivo. As we demonstrated in melanoma cells and zebrafish embryos, Molecular tattooing is suitable for dissecting collective activities by the separation of autonomous and non-autonomous molecular processes in vivo ranging from subcellular to organism level. Molecular tattooing is the first technology that establishes a broad application potential for optopharmacology by confining drug effects to a few cells of a developing organ in a live animal, or even a subcellular region of a single cell. Molecular tattoo will revolutionize local targeting techniques of bioactive ligands because it is able to dissect specific interactions of complex collective processes. It will provide answers to the following questions: What is the specific role of an enzyme located in a certain cell of a developing organ? How does the subcellular location of an enzyme modulate different cellular processes such as cell movement, blebbing, cytokinesis or signalling? What is the function of a single synapse within a multi-synaptic neuronal connection? The key advantage of Molecular tattooing is that it can induce local and permanent confinement of a drug effect without significant systemic and side effects outside the targeted area. Importantly, a series of photoreactive bioactive ligands are already available for a wide variety of targets and further photoreactive compounds can be synthesized in a single step from existing drug derivatives. Thus the wide variety of compounds will facilitate the rapid, widespread propagation of Molecular tattooing. This technology provides an immediate opportunity for technology transfer both in research and health industries by developing novel photochemical drugs and scanning microscopy instruments optimized for Molecular tattooing. Realizing this opportunity, we established a SME company called Optopharma Ltd. which was supported by ERC PoC scheme.