ERC FUNDED PROJECTS

Amine containing compounds are ubiquitous in everyday life and find applications ranging from polymers to pharmaceuticals. The vast majority of amines are synthetic and manufactured on large scale which creates waste as well as requiring high temperatures and pressures. The increasing availability of biocatalysts, together with an understanding of how they can be used in organic synthesis (biocatalytic retrosynthesis), has stimulated chemists to consider new ways of making target molecules. In this context, the iterative construction of C-N bonds via biocatalytic hydrogen borrowing represents a powerful and unexplored way to synthesise a wide range of target amine molecules in an efficient manner. Hydrogen borrowing involves telescoping redox neutral reactions together using only catalytic amounts of hydrogen. In this project we will engineer the three key target biocatalysts (reductive aminase, amine dehydrogenase, alcohol dehydrogenase) required for biocatalytic hydrogen borrowing such that they possess the required regio-, chemo- and stereo-selectivity for practical application. Recently discovered reductive aminases (RedAms) and amine dehydrogenases (AmDHs) will be engineered for enantioselective coupling of alcohols (1o, 2o) with ammonia/amines (1o, 2o, 3o) under redox neutral conditions. Alcohol dehydrogenases will be engineered for low enantioselectivity. Hydrogen borrowing requires mutually compatible cofactors shared by two enzymes and in some cases will require redesign of cofactor specificity. Thereafter we shall develop conditions for the combined use of these biocatalysts under hydrogen borrowing conditions (catalytic NADH, NADPH), to enable the conversion of simple and sustainable feedstocks (alcohols) into amines using ammonia as the nitrogen source. The main deliverables of BIO-H-BORROW will be a set of novel engineered biocatalysts together with redox neutral cascades for the synthesis of amine products from inexpensive and renewable precursors.

2 337 548 €

Start date: 2017-06-01, End date: 2022-05-31

Hydrogels cross-linked through supramolecular interactions are highly dependant on the dynamic charac- teristics of the physical cross-links. Few fundamental studies have been undertaken to quantitatively de- scribe structure-property relationships for these types of systems. Hydrogels formed from CB[8]-mediated supramolecular physical cross-linking mechanisms have gained significant interest on account of their excel- lent physical and mechanical properties such as self-healing and shear-thinning. This supramolecular motif has been further exploited to introduce and compatibilise a wide variety of different materials into hydrogel networks without phase separation, forming hybrid composite hydrogels attributed with unique and emergent properties. This proposal aims to pioneer the combination of several state-of-the-art characterisation tech- niques into an unique experimental setup (CAM-RIG), which will combine super-resolution and confocal microscopy imaging modalities with simultaneous strain-controlled rheological measurements to investigate fundamental structure-property relationships of these systems. For the first time it will be possible to decon- volute the molecular-level dynamics of the supramolecular physical cross-links from chain entanglement of the polymeric networks and understand their relative contributions on the resultant properties of the hydrogels. Using the fundamental insight gained, a set of key parameters will be determined to maximise the potential of supramolecular biocompatible hydrogels, driving paradigm shifts in sustainable science and biomaterial applications through the precise tuning of physical properties.

2 038 120 €

Start date: 2017-05-01, End date: 2022-04-30

Cancer genome sequencing has led to a paradigm shift in our understanding of oncogenesis. It has identified thousands of genetic alterations that segregate into two groups, a small number of frequently mutated genes and a much larger number of infrequently mutated genes. The causative role of frequently mutated genes is often clear and are the focus of concerted therapeutic development efforts. The role of those infrequently mutated is often unclear and can be difficult to separate from ‘mutational noise’. Determining the relevance of low frequency mutations is important for providing a full understanding of processes driving tumourigenesis and if functionally relevant may have broader implications on the applicability of targeted therapies. This project aims to begin addressing this by defining the function of all genes mutated in colorectal cancer (CRC) in the earliest stages of tumour formation. I have performed a whole genome screen in a 3D organoid CRC initiation model identifying several potentially important mediators of this process. Crucially, some of these genes are mutated in CRC at low frequency but not described as cancer driver genes. Thus, I hypothesize that rather than ‘mutational noise’ infrequently mutated genes contribute to CRC initiation. I will test this by addressing two aims: 1) Determine the role of genes mutated in CRC during tumour initiation 2) Validate and determine the function of a subset of identified genes potentially defining novel cancer mechanisms I will use a combination of CRISPR genetic disruption in state-of-the-art 3D mouse and human organoid cultures and advanced mouse models to address these aims. This comprehensive approach will provide a foundation for understanding the importance of the entire spectrum of mutations in CRC and open new avenues of research into the function of these genes. More broadly, it has the potential to make a profound impact on how we think about tumourigenic mechanisms and cancer therapeutics.

1 498 618 €

Start date: 2017-08-01, End date: 2022-07-31

Obesity and related co-morbidities give rise to severe health and socioeconomic problems. Surgical treatment for obesity (bariatric surgery) is remarkably effective in the control of morbid obesity and rapid resolution of Type 2 Diabetes, and the number of such procedures is increasing rapidly in many obesity-prevalent countries. We, and others, have demonstrated that surgical interventions such as Roux-en-Y Gastric Bypass (RYGB) modulates gut hormone levels, induces systemic metabolic changes and results in the shift of the microbiome from Firmicutes to the Proteobacteria phylum. Although the gut microbiota have been implicated in the reduction of adiposity post-surgery, the long-term effect of altered gut microbiota on patients who have undergone RYGB, remains to be studied. Our recent data suggested that microbial activities are highly associated with inflammation and cancer. My research programme aims to investigate the RYGB-specific gut microbiota impacts on host physiology and colon cancer risk. To achieve this goal, I will employ a multidisciplinary approach that combines systems biology techniques with a bottom-up approach. This work will deliver phenotypic and mechanistic characterisation of the interplay between the host and the gut microbiota. The research findings will significantly contribute towards the understanding of fundamental molecular and cellular processes that are key in host and gut microbiota interactions. This will provide knowledge-based evidence of the gut microbial impact on human physiology, and has the potential to unravel novel prevention targets and promote a more thorough healthcare strategy for bariatric patients.

1 499 091 €

Start date: 2017-08-01, End date: 2022-07-31

This proposal aims to understand and control glycosylation reactions. In a glycosylation reaction a “donor” glycoside and an “acceptor” (the nucleophile) are united to form an oligosaccharide. Although it is the central reaction in carbohydrate chemistry, our understanding of this reaction, in terms of stereoselectivity and productivity is still limited. The structural variation in the building blocks leads to a complex continuum of SN2-SN1 mechanisms that operates and it is currently impossible to predict where in the continuum the reaction exactly takes place. This proposal provides fundamental insight into the outcome of glycosylations by studying both the activated donor glycoside and the acceptor nucleophile. Activation of a donor glycoside leads to different reactive intermediates, covalent anomeric species (most often triflates) and oxocarbenium ion-like species. The relative reactivity of these species is quantified to generate novel reactivity charts. The covalent species are studied by innovative competition experiments, kinetic studies and NMR spectroscopy. The (fleeting) oxocarbenium ion-like intermediates are probed by a computational approach and by “super-acid NMR” studies in which stable glycosyl cations are generated and studied in super-acid media. The reactivity of glycosyl acceptors is systematically studied in a set of SN2 or SN1-type glycosylations. Using kinetic studies and competition reactions charts of acceptor nucleophilicity are compiled. The reactivity of the donors and acceptors is matched using a family of tailor made “reactivity modulators”, spanning a broad reactivity window bridging the reactivity gap between the building blocks leading to predictable glycosylations. The developed methodology is employed in automated solid phase syntheses of libraries of oligosaccharides featuring multiple cis-glycosidic linkages. The proposal is a major step forward in the development of a general glycosylation procedure.

2 000 000 €

Start date: 2017-05-01, End date: 2022-04-30

Molecular chirality is a central theme in chemistry; in 2015 approximately 13% of publications in J. Am. Chem. Soc. and 12% in Angew. Chem. concerned chirality. All previously studied forms of molecular asymmetry (central, axial, planar and helical chirality) have found applications throughout the sub-disciplines of chemistry including as catalysts, materials and sensors. Mechanically chiral rotaxanes are molecules in which the mechanical bond between a macrocycle and dumbbell-shaped component is the source of asymmetry rather than the covalent structure of the components themselves. These unusual molecules represent a novel and unexplored chiral supramolecular environment as the lack of a scalable synthetic approach for their isolation in enantiopure form has prevented all but the most cursory investigation of their properties. Thus, mechanical chirality remains an unexplored frontier of molecular asymmetry with the potential to deliver novel functions and impact across a range of chemical disciplines from materials chemistry to the synthesis of pharmaceutically active compounds. The Goldup Group has recently demonstrated the first practical method for the synthesis of enantiopure mechanically chiral rotaxanes using a flexible active template methodology and thus the stage is finally set for the study and exploitation of this novel form of supramolecular asymmetry. Within the period of this ERC Consolidator Grant the PI will lead a team to investigate the synthesis, properties and applications of these intriguing mechanically chiral molecules.

1 998 928 €

Start date: 2017-04-01, End date: 2022-03-31

We will study the adsorption, binding, transport and reactivity of N-containing substrates within the nanopores of functionalized and doped metal-organic framework (MOF) materials. A range of important target molecules (e.g., NH3, N2H4, N2O, NO, NO2 and N2O4) will be studied in this project, which aims to re-define the molecular chemistry for these energetic N-compounds in confined space, and to develop selective catalytic reduction (SCR) of NOx in the presence of NH3, urea and hydrocarbons. The PI has extensive experience in the field of coordination chemistry and hybrid materials, and seeks to change research direction to develop new gas phase catalysts and to gain fundamental understanding of molecular interactions, properties and function of specific energy, environmentally-related substrates within MOFs. Research objectives include the: • Design and synthesis of new porous MOFs with emphasis on the decoration of their pore environment and improvement of their structural stability and function; • Characterisation of host and substrate-loaded materials by state-of-the-art in situ structural, dynamic and spectroscopic methods for the construction of structure-function relationships, supported by computational analysis and modelling; • Adsorption, binding, release and separation of NH3, N2H4, N2O, NO, NO2 and N2O4 via both static and dynamic experiments; • Tests of degradation and selective catalytic reduction (SCR) of captured NOx molecules with NH3, urea and hydrocarbons under mild conditions using nanoporous MOFs as host catalysts; • Assembly of a MOF-based (i) catalytic deNOx reactor and (ii) NH3 storage system for potential portable applications. This project will deliver new functional materials as high capacity portable NH3 stores, efficient capture medium for NOx, and new catalysts for reduction and mitigation of NOx to deliver significant impacts to academia, industry and society of direct relevance to clean energy and sustainable environment.

2 498 645 €

Start date: 2017-06-01, End date: 2022-05-31

Type 2 diabetes mellitus, one of the major healthcare challenges of our time, is characterized by failure of beta cells to functionally adapt to increased peripheral insulin resistance. The resulting chronic elevations in blood glucose concentration are associated with heart, kidney, liver, nerve and retinal disease, as well as cancer. Here, by combining novel optogenetic, photopharmacological and innovative imaging approaches, we aim to unravel the complexity underlying the multicellular regulation of insulin secretion from islets of Langerhans during health and disease. In particular, we will examine a role for privileged pacemakers/hubs in orchestrating population responses to stimuli, identify what makes these specialized cells unique at the RNA/protein level, and understand how they contribute to islet development and failure. Furthermore, we will address whether the intraislet regulation of insulin secretion operates in vivo to determine glucose homeostasis, focusing on the neural-endocrine interface. Lastly, the mechanisms underlying islet cross-talk will be investigated directly in situ within the pancreas of living mice, paying close attention to the roles of the vasculature and secreted factors. As such, these studies should unveil a new route for restoration of insulin secretion in man, as well as provide the foundation for the de novo construction of islets for transplantation.

1 681 468 €

Start date: 2017-06-01, End date: 2022-05-31

Cancer development is characterized by uncontrolled proliferation, cell survival and metabolic reprogramming. Tumour cells are surrounded by a fluid-mosaic membrane that contains tetraspanins (Tspans) which are evolutionary conserved proteins important in the formation of multiprotein complexes at the cell surface (‘tetraspanin web’). Increasing evidence indicates that Tspans are involved in cancer, still the architecture of the Tspan web in native tumour membranes and its (patho)physiological functions have not been resolved. Based on my preliminary data, I hypothesize that tumour cells contain a disrupted Tspan web in which Tspan interactions are modified leading to aberrant metabolic signalling and tumour development. This is supported by my discovery that loss of Tspan CD37 leads to spontaneous lymphomagenesis due to activation of the Akt survival pathway. The overall aim of Secret Surface is to unravel the composition, physiological functions and molecular mechanisms of the Tspan web on tumour development and clinical outcome. To achieve this, I will focus on studying lymphomas using a multidisciplinary approach: I. Detailed analyses of Tspan web composition in lymphoma to select clinically relevant Tspans (high-throughput tissue microarray technology, multispectral imaging). II. Resolve the endogenous Tspan web on lymphoma cells (super-resolution microscopy), and generation and analysis of lymphoma cells that have a complete deficiency of multiple Tspans (CRISPR/Cas9 technology). III. Decipher molecular mechanisms underlying Tspan web function in lymphoma cells (membrane organization, membrane-proximal signalling, metabolic reprogramming). With my unique toolbox of Tspan knock-outs coupled to advanced microscopy and metabolic studies, I expect that Secret Surface will lead to a new concept in cellular physiology in which cell surface organization by the Tspan web drives tumour development, which may open new horizons for the generation of new cancer therapies.

2 000 000 €

Start date: 2017-10-01, End date: 2022-09-30

Metabolism generates vast quantities of acid, which exerts broad-spectrum biological effects because protein protonation is a powerful post-translational modification. Regulation of intracellular pH (pHi) is therefore a homeostatic priority, but carefully orchestrated proton dynamics are a versatile signal. Extracellular acidity is an established chemical signature of tumours and has recently been proposed to convey a signal that shapes the phenotypic landscape of cancer. Cancer’s genetic instability yields diversity in acid handling and signalling, forming a substrate for selection under acid-stress. This is a plausible mechanism for disease progression and an analogy can be drawn to experimentally-verified hypoxic selection. Current models of acid handling in cancer are, however, based on population-averages of observations made at the cell level. This fails to appreciate diversity and the complexity inherent in tissues. We will produce a more complete understanding of acid handling that accounts for diffusive transport across tissue compartments and the role of the tumour stroma. A systems-approach of characterising pH-regulatory processes cell-by-cell will identify which components are liable to vary, and thus are a substrate for acid-driven somatic evolution. The long-term effects of proton signals on gene expression have not been tested, despite evidence for proton-sensing transcription factors. To address the mechanism for adaptation to acid-stress, proton-sensing transcription factors will be characterised from studies of gene expression under chemically and optogenetically operated pH stimuli. The definition of a cell’s fitness to survive at a particular microenvironment pH and its relationship with stemness remain unclear. Phenotyping pHi-gated subpopulations in terms of growth, stemness and tumourigenicity will define pH-fitness and its role in aggressiveness. In evolving to survive metabolism, cancer cells may acquire the ability to thrive in new niches.

1 922 575 €

Start date: 2017-06-01, End date: 2022-05-31