Periodic Reporting for period 1 - DNAPC4ImunoMol (DNA modules as communicators in tumor immunomodulation)
Periodo di rendicontazione: 2023-05-01 al 2025-04-30
Certain diseases present themselves as a complex molecular scenario. In these diseases, of which the most important examples are cancer and autoimmune diseases, a single treatment, e.g. a drug, only works under strict conditions. Therefore, the disease must be characterized by so-called markers. These can be small molecules, proteins or other features that represent the biochemical state of the disease. Today, complex diseases are treated by diagnosing the disease, characterizing the conditions and then applying appropriate treatments. In the future, these three steps will be performed simultaneously by molecular computers. Molecular computers are sets of molecules that are designed to interact with each other and with target molecules so that they can be used to determine whether a molecule is present or not (e.g. a cancer marker) and then trigger a further response, the treatment, but only if the right markers are present. In other words, the diagnosis, characterization and treatment of complex diseases is done all at once. There are many types of chemical systems that can be used to build molecular computers. In this project, we explored aptamer displacement reactions. Aptamers are short DNA sequences that can bind specifically to a molecule or protein, the target. In the displacement reaction, another DNA molecule is bound to the aptamer, which is released when the target binds the aptamer. In this case, the released molecule acts as a signal that can trigger further reactions. Aptamer displacement reactions are complex, but form the basis for future drugs that can be used to diagnose and treat patients with complex diseases.
To achieve treatments that can single-handedly diagnose and cure diseases, it is necessary to develop (1) a carrier that contains the system, (2) the molecular sensor and the computer that captures the inputs and makes decisions, (3) an output that leaves the carrier when triggered and acts as a treatment.
The carrier (1) is a synthetic cell similar in size to biological cells and that perform life-like functions. We have used protocells that consist of DNA. These protocells are self-assembled structures and can host molecular systems in a spatially organized way.
The molecular computer (2) is based on aptamer displacement reactions. We used an aptamer that recognizes the protein VEGF165 as the basis for our system. VEGF165 is a protein that promotes angiogenesis and is associated with the growth of certain cancers.
The output (3) is a component that is released by a trigger. We have developed condensates consisting of dextran and lipids. These condensates, which are compatible with DNA-based systems, can be loaded with drugs or mRNA and delivered to cells. These condensates, which are stable and non-toxic to the cell, are taken up by cells and can transfect them with mRNA, leading to protein expression.
The integration of 1-3 is what will enable soft matter computers that can perform complex therapeutical treatments in vivo.
The carrier (1) is a synthetic cell similar in size to biological cells and that perform life-like functions. We have used protocells that consist of DNA. These protocells are self-assembled structures and can host molecular systems in a spatially organized way.
The molecular computer (2) is based on aptamer displacement reactions. We used an aptamer that recognizes the protein VEGF165 as the basis for our system. VEGF165 is a protein that promotes angiogenesis and is associated with the growth of certain cancers.
The output (3) is a component that is released by a trigger. We have developed condensates consisting of dextran and lipids. These condensates, which are compatible with DNA-based systems, can be loaded with drugs or mRNA and delivered to cells. These condensates, which are stable and non-toxic to the cell, are taken up by cells and can transfect them with mRNA, leading to protein expression.
The integration of 1-3 is what will enable soft matter computers that can perform complex therapeutical treatments in vivo.
The most important achievement of the project was the demonstration of the feasibility of aptamer displacement reactions as a method for integrating computers into synthetic cells. For a fully functional synthetic cell that is intelligent enough to perform relevant tasks, further research in the field of aptamers is necessary, especially with regard to the binding process and their structure.