The pharmaceutical sector has a huge demand for new active compounds including natural products to fill the drug pipelines and to stop the global decline in novel approved active pharmaceutical ingredients. Therefore, developing new tools to fabricate complex molecular structures in a fast and reliable way is paramount. This holds especially true for the field of antibiotics. Multidrug resistant (MDR) pathogens evolve at a terrifying rate and confer resistance to all presently available antibacterial treatments and therefore WHO has identified MDR bacteria as major threat to human health. In this ERC Advanced Grant, I propose a radically new approach to fabricate very complex molecules with minimal synthetic effort. The technology is based on nucleic acid binders (aptamers), which are evolved in a selection protocol and block several functional groups within a target molecule while allowing other functionalities not in contact with the aptamer to be selectively modified in a single reaction step. Here, we aim to establish this groundbreaking aptameric protective group (APG) method as a novel tool that gives access to compounds that would otherwise be too difficult to obtain by multistep synthesis. Toward this end, the specific objectives are: • To develop reagents and reactions that are compatible with aptamer-mediated reactions • To control the site of chemical modification within complex molecules by APGs • To establish APGs as a general paradigm in natural product derivatization to modify several kinds of substrates • To achieve site selective modification of proteins by aptamers • To synthesize novel antibiotics that kill MDR bacteria • To fabricate “image-and-activate” antibiotics by the APG technology • To employ the aptamer-target complexes for live-cell imaging of RNA The outcomes will enable future advances in drug discovery and drug design, bioimaging technologies, and the site-specific modification of therapeutic proteins.

PeriGO will pave the way for the commercialization of a unique, first-of-its-kind peripheral nerve repair strategy, as it uses an injectable, biocompatible hydrogel that provides directionality through controlled guiding elements. The properties of the smart material Anisogel, initially developed in the ERC project ANISOGEL to repair spinal cord injuries, provide a sophisticated, superior alternative for peripheral nerve regeneration. The final product can be injected inside a bridging blood vessel or artificial hollow tube after implantation to direct regenerating nerves across the injury site. PeriGO comprises 3 key actions to advance a quick translation from the laboratory to the clinic: We will 1) establish a sterilization method and a packaging/application system to perform pre-clinical tests, 2) verify the handling and efficacy of the product in a rat sciatic nerve lesion model in cooperation with the Department of Plastic Surgery at the University Hospital in Aachen, and 3) complete a market study and business plan to bring PeriGO into the clinic. PeriGO addresses essential issues associated with peripheral nerve regeneration. While the current Gold Standard of autologous nerve grafts creates a secondary defect, artificial bridging approaches for nerve gaps only reconnect short distances (< 3 cm) with marginal success. Features absent in the commercially available nerve tubes are guiding structures, controlled degradation, and a good interface with the damaged nerve stumps. The PeriGO platform will overcome these limitations as the material 1) is injected as a liquid to make a tight contact with the injured nerve and to adapt to the shape of the bridging tube before gelation, 2) provides unidirectional guiding elements, and 3) presents tunable material properties with tunable degradation kinetics adjusting to the length of the nerve gap. With PeriGO, we will thus establish the next generation of nerve repair strategies.

Today’s materials research in the field of synthetic membranes gives access to highly permeable and extremely selective membranes. However, their potential will remain ineffective as high and selective transport rates always go along with resistances emerging at the membrane fluid interface in the form diffusion limitations in the laminary boundary layers. In order to make full use of the very many new materials, also new means to control and minimize such fluid based resistances need to be developed. Yet another phenomena disturbs the full potential use of membranes: retained solutes, colloids and biological matter accumulates at the membrane interface and causes irreversible fouling and scaling. The proposed research aims to develop a rigorous translational methodology to control and improve mass transport through the fluid/membrane interface. ConFluReM will establish Strategic Tools and New Instruments to: (1) comprehend and quantify the prevalent mass transport resistances in representative membrane separation processes, (2) synthesize and fabricate new nano-, micro- and mesoscale material and device systems as instruments to control and overcome the limitations of concentration polarization and fouling, Strategic Tools are experimental and simulation methods to quantify and engineer the mass transport and hydrodynamical properties of the new membrane systems. These encompass flow imaging (flowMRI, microPIV and microfluidic transport studies) as well as computational fluidic dynamics (CFD and CFDEM). New Instruments are synthetic and fabrication means as well as process condition means to improve mixing at the membrane/fluid interface. These encompass (a) lateral patterning of chemical topology of the membrane surface by printing and stamping, (b) shaping the 3D geometry of channels using additive manufacturing techniques and (c) imposing dynamical gradients to destablize fluid side resistances.

Polymeric materials are ubiquitous in our daily lives but they have a predominantly fossil origin, with low degradability at their end-of- life. Transitioning to a circular polymer economy requires a rethinking of the entire value chain, from the raw materials, tools, and processes used to polymer design degradation and recycling. Enzymes are eco-friendly and sustainable tools that tackle many industrial applications. However, biocatalysis in the polymer field remains mostly unexplored due to i) enzymes’ high cost and low stability under reaction conditions, ii) enzymes’ inefficiency in converting bio-based monomers into cost-effective building blocks, and iii) lack of knowledge in key enzyme-polymer interactions that can control the final polymer performance and degradability features. Computational tools have shown immense power to revolutionize the field of enzyme engineering in a time and cost effective way. However, there is currently a clear lack of researchers combining computational and experimental skills, capable of determining future directions for the optimization of biocatalytic processes for the sustainable molecular design of polymers. To foster the transition to a bio-based polymer industry, COMENZE aims to develop enzymatic strategies for improving the eco-design and development of future sustainable polymers. This will be achieved by combining cutting-edge computational and experimental approaches for enzyme discovery and engineering through in-silico modeling, simulation, and translation of results into wet labs to validate enzymatic reactions. COMENZE will train the next generation of researchers by equipping 10 DCs with the skills to revolutionize the polymer circularity by delivering new optimized enzymes and bioprocesses, newly identified bio-based building blocks, and functionalized polymers with innovative bio-upcycling and biodegradation end-of-life options.

To date, light has been employed as a widespread trigger to achieve control over the activity of drugs and protein function establishing the fields of photopharmacology and optogenetics, respectively. Both techniques led to promising new therapies, the elucidation of brain function or understanding of neural disorders. However, serious limitations resulting from the low penetration depth of light into tissues are severely hampering progress in these fields. In contrast to photons, ultrasound deeply penetrates tissue and can be applied with sub-millimeter resolution and consequently has been widely established in the clinic over many decades for therapy and diagnostics. In this ERC Advanced Grant, I will develop a radically new approach to control the activity of drugs, proteins and genes by biocompatible ultrasound. Polynucleic acid carriers, which can bind a wide variety of bioactive payloads, will be designed to be sensitive to different ultrasound sources, which can be applied in clinical settings and do not harm cells or tissues. Upon ultrasound irradiation, these carriers liberate their bioactive payloads by mechanochemical principles to switch on drugs and control cellular functions. To achieve this aim, I will: investigate the effect of ultrasound (US) on nucleic acid architectures; study the loading of polynucleic acids with different payloads and their release by US; develop a technology platform to activate small molecule drugs, proteins and oligonucleotides; and showcase the huge potential of these technologies for cancer immunotherapy, diabetes research and tissue engineering. This project will boost sonopharmacology and sonogenetics. Its outcomes will enable spatiotemporal control of drug action to minimize side effects in pharmacotherapy like cancer. The remote controlled orchestration of protein and gene function by US will strongly advance medicine and the life sciences by answering fundamental questions in these fields.