Mitochondria are cell organelles that provide the energetic requirements of the body. The majority of cellular ATP is produced by the membrane protein ATP synthase through a proton gradient across the mitochondrial inner membrane. Alterations in ATP synthase biogenesis can result in severe mitochondrial diseases affecting tissues with high energy requirements as brain and muscles. Mitochondrial diseases affect approximately 20 million people in the EU, causing 35 % of deaths during the first year of life of newborns. However, the available therapeutic approaches, are still extremely limited and there is no specific treatment for ATP synthase deficiencies. To improve the treatments currently available for mitochondrial diseases, the project will focus on the realization of artificial mitochondria (AM). Based on artificial lipid vesicles, AM will be fabricated by means of microfluidics methods, a powerful tool able to produce identical replicas of a given bio-inspired membrane-object. ATP synthase will be expressed and assembled within the lipid bilayer by encapsulating cell-free protein expression systems. To test the ability of AM as in-situ energy fabrication systems, targeting-AM will be endocytosed inside cultured cells and ATP synthesis will be triggered by taking advantage of the proton gradient provided by endosomes. Finally, by enclosing other plasmids encoding for diverse proteins, AM can be used as energy-factoring pockets to elicit protein expression just when internalized within cells. This novel approach may constitute an advanced new concept in gene therapy to more effectivelycreate breakthroughs in improving human health.
MITOCHON. Artificial mitochondria for health
Mid-Term Report Summary – MITOCHON (Artificial Mitochondria for Health)
mitochon quests to build a biomimetic system able to produce the chemical energy in form of ATP inside the eukaryotic cell. To achieve this goal we have been working on three parallel research lines: the collection of the biological building blocks, the targeting and internalization of the biomimetic system into cultured cells and the release and tracing of the newly formed ATP inside the cell. Up to date, proteins, lipids and peptides have been successfully assembled into a functional artificial system. In addition, the vectorization of non-functional prototypes into target cells are routinely performed. Ongoing research is now optimizing the best strategy to supply and to accurately trace the molecular cargo within living cells. With the tools in hand, now the logical consequence and progression is to converge these research lines and to assemble the puzzle that will lead to the fabrication of artificial mitochondria.