Periodic Reporting for period 1 - MMC (Membrane Micro-Compartments)
Reporting period: 2024-02-01 to 2025-07-31
More than two-thirds of all described drug targets are membrane proteins, but structure analysis and protein production for these hydrophobic proteins is inherently challenging. Consequently, investigations of their mechanisms of function and drug interactions are often only derived from purified truncated parts of these proteins, taken out of the native biological context. Without the effects of the surrounding membrane components, conclusions on drug mechanisms are incomplete. Recent advances of in situ structure determination techniques, i.e. analysis directly within the cellular environment, have the power to overcome the limitations of classical reconstituted approaches. Complementary, we have developed a system that facilitates structural analysis of membrane protein targets and has large potential for the production of native membrane proteins - an essential prerequisite for studying membrane protein function and drug interactions.
Within the scope of our proposal, we have further developed our system and transferred it into a pharmaceutically-relevant human cell system. Furthermore, we achieved proof-of-concept and assessed our system with clinically-relevant membrane proteins for its suitability for membrane protein production, as well as suitability to solve 3D structures of membrane protein complexes by cryo-electron tomography (cryo-ET) directly in cells. Our approach can have a transformative impact on how membrane protein interactions can be studied, and could in the future be exploited for drug development by the pharmaceutical industry.
Within the scope of our proposal, we have further developed our system and transferred it into a pharmaceutically-relevant human cell system. Furthermore, we achieved proof-of-concept and assessed our system with clinically-relevant membrane proteins for its suitability for membrane protein production, as well as suitability to solve 3D structures of membrane protein complexes by cryo-electron tomography (cryo-ET) directly in cells. Our approach can have a transformative impact on how membrane protein interactions can be studied, and could in the future be exploited for drug development by the pharmaceutical industry.
One of our main objectives of the PoC was to transfer our system for structural analysis of membrane proteins developed in yeast cells into human cells. We developed a mammalian version of our system and demonstrated conservation from yeast to human for several candidate membrane proteins. Our system was further validated and characterized by different microscopy techniques, such as fluorescence microscopy and cryo-electron microscopy. The establishment of a human system (WP1 - milestone 1) laid the foundation for further scientific work packages.
We next assessed the potential of our system for in situ structural analysis by cryo-ET. For this, we established cryo-ET procedures using our yeast system and transferred this strategy to human membrane proteins using cryo-ET and subtomogram averaging. With promising results for both the yeast and human system, the structural work is still ongoing. Another main objective of the project was to test the application of the system for purifying and isolating membrane proteins. We successfully developed a protocol for isolating membranes that are enriched for two different classes of proteins. Next steps involve more in-depth characterization to further determine the economic potential of the technology.
We next assessed the potential of our system for in situ structural analysis by cryo-ET. For this, we established cryo-ET procedures using our yeast system and transferred this strategy to human membrane proteins using cryo-ET and subtomogram averaging. With promising results for both the yeast and human system, the structural work is still ongoing. Another main objective of the project was to test the application of the system for purifying and isolating membrane proteins. We successfully developed a protocol for isolating membranes that are enriched for two different classes of proteins. Next steps involve more in-depth characterization to further determine the economic potential of the technology.
We have filed a European patent application of our inducible synthetic membrane organelle system in May 2025. In addition to our IPR strategy, we plan to make the research available to the scientific community through a peer-reviewed publication.
Over the course of the project, we have carried out market analysis by ourselves and by a professional company. This included interviews with representatives from biotechnology and pharma companies to evaluate the commercialization and development potential our technology.
Following the overall positive outlook and potential impact of our technology for the production of proteins, we plan as a next step to scale and further characterize the production process to determine the commercial potential for protein production. Based on our results, we have conceptualized a research and development plan and a business plan to expand our synthetic membrane organelle system and are reaching out to companies for collaborations on pilot studies to generate a first use case of our system in drug development. Based on this plan, we are currently applying to non-dilutive funding programs to further develop our technology into commercial applications for the pharmaceutical industry.
Over the course of the project, we have carried out market analysis by ourselves and by a professional company. This included interviews with representatives from biotechnology and pharma companies to evaluate the commercialization and development potential our technology.
Following the overall positive outlook and potential impact of our technology for the production of proteins, we plan as a next step to scale and further characterize the production process to determine the commercial potential for protein production. Based on our results, we have conceptualized a research and development plan and a business plan to expand our synthetic membrane organelle system and are reaching out to companies for collaborations on pilot studies to generate a first use case of our system in drug development. Based on this plan, we are currently applying to non-dilutive funding programs to further develop our technology into commercial applications for the pharmaceutical industry.