This project will investigate how we can use proteins derived from a parasitic worm to either amplify or suppress "IL-33", a protein used for communication within the immune system. IL-33 is released on damage to various barrier sites (lung, skin, gut), and can lead to either allergic or inflammatory responses, depending on the context. These responses can be beneficial, e.g. they can efficiently clear bacterial infections, or they can be detrimental, e.g. they can lead to the development of allergic or inflammatory damage such as in asthma or acute respiratory distress syndrome. Therefore being able to effectively "tune" the IL-33 pathway up or down would be a powerful technique for treating a wide range of diseases including asthma, eczema, fungal or bacterial infections, and acute respiratory distress syndrome. We have identified two proteins derived from a single parasitic worm (Heligmosomoides polygyrus) which act to suppress IL-33 responses; we have named these proteins HpARI and HpBARI. Suppression of IL-33 responses is advantageous to parasites, as it allows them to avoid triggering immune responses which could lead to their ejection or damage to their host. When we produced a mutant form of one of these IL-33-suppressive proteins, we found the mutant form had the surprising effect of amplifying (rather than suppressing) IL-33 responses due to stabilisation of the IL-33 molecule. Therefore we can use these proteins and mutants to either increase or decrease IL-33 responses and potentially treat a long list of diseases in which IL-33 has a causative, or curative, role. This project will investigate the use of these proteins and their derivatives in mouse systems where IL-33 drives allergic responses (such as in asthma), damaging inflammatory responses (such as in acute lung injury or acute respiratory distress syndrome) or beneficial anti-bacterial responses (such as in pneumonia). We will translate findings from the mouse towards human responses by using genetically-engineered mice which express the human form of the IL-33 molecule, and stimulating human blood cells with human IL-33 in the lab, and testing whether our proteins affect the responses of these cells. Furthermore, we will engineer hybrid molecules, taking the active regions of our proteins, and combining them with proteins normally present in our blood. This will have the advantage that the resulting proteins will be largely ignored by the immune system as they look like one of the body's own proteins. This avoids a common problem (known as "immunogenicity") of using foreign proteins as medicines, where the immune system rejects the protein, preventing it from carrying out its function. These engineered proteins will be further assessed for activity against IL-33 responses and for immunogenicity (the level of recognition by the immune system) in mouse and human tests, as described above. In summary, this project will investigate and characterise new parasite-derived proteins which can suppress or amplify the immune response, with the potential to be used as new medicines or tools for research in a range of allergic, inflammatory and infectious diseases.

Doctoral Training Partnerships: a range of postgraduate training is funded by the Research Councils. For information on current funding routes, see the common terminology at https://www.ukri.org/apply-for-funding/how-we-fund-studentships/. Training grants may be to one organisation or to a consortia of research organisations. This portal will show the lead organisation only.

This project will investigate how we can use proteins derived from a parasitic worm to either amplify or suppress "IL-33", a protein used for communication within the immune system. IL-33 is released on damage to various barrier sites (lung, skin, gut), and can lead to either allergic or inflammatory responses, depending on the context. These responses can be beneficial, e.g. they can efficiently clear bacterial infections, or they can be detrimental, e.g. they can lead to the development of allergic or inflammatory damage such as in asthma or acute respiratory distress syndrome. Therefore being able to effectively "tune" the IL-33 pathway up or down would be a powerful technique for treating a wide range of diseases including asthma, eczema, fungal or bacterial infections, and acute respiratory distress syndrome. We have identified two proteins derived from a single parasitic worm (Heligmosomoides polygyrus) which act to suppress IL-33 responses; we have named these proteins HpARI and HpBARI. Suppression of IL-33 responses is advantageous to parasites, as it allows them to avoid triggering immune responses which could lead to their ejection or damage to their host. When we produced a mutant form of one of these IL-33-suppressive proteins, we found the mutant form had the surprising effect of amplifying (rather than suppressing) IL-33 responses due to stabilisation of the IL-33 molecule. Therefore we can use these proteins and mutants to either increase or decrease IL-33 responses and potentially treat a long list of diseases in which IL-33 has a causative, or curative, role. This project will investigate the use of these proteins and their derivatives in mouse systems where IL-33 drives allergic responses (such as in asthma), damaging inflammatory responses (such as in acute lung injury or acute respiratory distress syndrome) or beneficial anti-bacterial responses (such as in pneumonia). We will translate findings from the mouse towards human responses by using genetically-engineered mice which express the human form of the IL-33 molecule, and stimulating human blood cells with human IL-33 in the lab, and testing whether our proteins affect the responses of these cells. Furthermore, we will engineer hybrid molecules, taking the active regions of our proteins, and combining them with proteins normally present in our blood. This will have the advantage that the resulting proteins will be largely ignored by the immune system as they look like one of the body's own proteins. This avoids a common problem (known as "immunogenicity") of using foreign proteins as medicines, where the immune system rejects the protein, preventing it from carrying out its function. These engineered proteins will be further assessed for activity against IL-33 responses and for immunogenicity (the level of recognition by the immune system) in mouse and human tests, as described above. In summary, this project will investigate and characterise new parasite-derived proteins which can suppress or amplify the immune response, with the potential to be used as new medicines or tools for research in a range of allergic, inflammatory and infectious diseases.

Doctoral Training Partnerships: a range of postgraduate training is funded by the Research Councils. For information on current funding routes, see the common terminology at https://www.ukri.org/apply-for-funding/how-we-fund-studentships/. Training grants may be to one organisation or to a consortia of research organisations. This portal will show the lead organisation only.

Many aspects of biology rely on molecules coming together in response to specific signals. An important example of this is gene regulation. The vast majority of cells in our body contain the same DNA, but they turn on specific genes in response to the signals that they receive. When genes are switched on or off inappropriately the behaviour of cells can change, potentially giving rise to diseases such as cancer or diabetes. Systems that enable us to turn genes on and off artificially would be powerful tools that could be used in a wide range of medical and biotechnological applications. Genes are regulated by proteins called transcription factors (TFs), which bind to specific sites on DNA, and either turn genes on by recruiting other proteins, or turn genes off by blocking the binding of such proteins. Many of these TFs are switches that respond to small molecules. For example, a bacterial TF called Lac repressor controls a gene involved in breaking down sugar. When the sugar is present a small molecule binds to Lac repressor, changes the shape of the protein and turns the gene on. Proteins like the Lac repressor are used as tools to control gene expression in engineered cells, such as bacteria that have been altered to produce human proteins. However, our ability and desire to engineer cells with increasingly complicated biochemistry means that there is now a pressing need for new switchable TFs. Ideally, these would not interfere with normal cell biology, referred to as being orthogonal; they should have predictable and tuneable properties, leading to a reliable set of biological parts that can be adapted for different uses; and they should be controlled by small non-toxic molecules that can enter cells, and could therefore potentially be used as drugs. We have developed rules that allow proteins called coiled coils to be designed and synthesised in the lab. Coiled coils can be designed to assemble in different ways; e.g., to bring together 2, 3, 4 or more proteins, which can bind to each other tightly or weakly. We have shown that these designed coiled coils can be used inside cells to bring together the proteins needed to control a gene. By altering the strength of the interaction between the components of the coiled coil we can control how much the targeted gene is turned on or off. These new TFs have predictable and tuneable properties, but they cannot currently be controlled by small molecules to act as a switch. In the proposed work we aim to build on our findings to produce coiled coils that assemble only in the presence of a small molecule. To do this, we will "hollow out" the centre of a coiled coil (i) to weaken the interactions that normally hold it together, and (ii) to generate a space for small-molecule drugs to bind. Under the right conditions, the small molecule will tightly and specifically fill the gap created, and will act as the missing piece of the jigsaw to allow the coiled coil to form and bring together the proteins needed for gene regulation. To find such conditions, we will test a large and diverse range of coiled coils that have hollowed cores of different shapes, sizes and chemistries. We will also test a diverse sample of potentially complementary small molecules. For this reason, the work is being done in collaboration with AstraZeneca, who have large libraries of small molecules, and the expertise to guide us towards the most promising of these. The outcome of this project will be a series of compact, well-understood coiled coils whose assembly inside or outside of cells can be controlled by adding a small molecule or drug. The work will also develop the knowledge and procedures needed to make new switches in future. We will use these systems to produce TFs that can turn genes on and off in both bacteria and in human cells, and with Medimmune we will apply these tools to current problems in the production of biological molecules of medical interest.