ERC FUNDED PROJECTS

A key synthetic challenge of widespread interest in chemical science involves the creation of well-defined 2D functional materials that exist on a length-scale of nanometers to microns. In this ambitious 5 year proposal we aim to tackle this issue by exploiting the unique opportunities made possible by recent developments with the living crystallization-driven self-assembly (CDSA) platform. Using this solution processing approach, amphiphilic block copolymers (BCPs) with crystallizable blocks, related amphiphiles, and polymers with charged end groups will be used to predictably construct monodisperse samples of tailored, functional soft matter-based 2D nanostructures with controlled shape, size, and spatially-defined chemistries. Many of the resulting nanostructures will also offer unprecedented opportunities as precursors to materials with hierarchical structures through further solution-based “bottom-up” assembly methods. In addition to fundamental studies, the proposed work also aims to make important impact in the cutting-edge fields of liquid crystals, interface stabilization, catalysis, supramolecular polymers, and hierarchical materials.

2 499 597 €

Start date: 2018-05-01, End date: 2023-04-30

A persistent challenge in unravelling mechanisms that regulate memory function is how to bridge the gap between inter-molecular dynamics of single proteins, activity of individual synapses and emerging properties of neuronal circuits. The prototype condition of disintegrating neuronal circuits is Alzheimer’s Disease (AD). Since the early time of Alois Alzheimer at the turn of the 20th century, scientists have been searching for a molecular entity that is in the roots of the cognitive deficits. Although diverse lines of evidence suggest that the amyloid-beta peptide (Abeta) plays a central role in synaptic dysfunctions of AD, several key questions remain unresolved. First, endogenous Abeta peptides are secreted by neurons throughout life, but their physiological functions are largely unknown. Second, experience-dependent physiological mechanisms that initiate the changes in Abeta composition in sporadic, the most frequent form of AD, are unidentified. And finally, molecular mechanisms that trigger Abeta-induced synaptic failure and memory decline remain elusive. To target these questions, I propose to develop an integrative approach to correlate structure and function at the level of single synapses in hippocampal circuits. State-of-the-art techniques will enable the simultaneous real-time visualization of inter-molecular dynamics within signalling complexes and functional synaptic modifications. Utilizing FRET spectroscopy, high-resolution optical imaging, electrophysiology, molecular biology and biochemistry we will determine the casual relationship between ongoing neuronal activity, temporo-spatial dynamics and molecular composition of Abeta, structural rearrangements within the Abeta signalling complexes and plasticity of single synapses and whole networks. The proposed research will elucidate fundamental principles of neuronal circuits function and identify critical steps that initiate primary synaptic dysfunctions at the very early stages of sporadic AD.

2 000 000 €

Start date: 2011-12-01, End date: 2017-09-30

Elucidating how neural circuits coordinate behaviour, and how molecules underpin the properties of individual neurons are major goals of neuroscience. Optogenetics and neural imaging combined with the powerful genetics and well-described nervous system of C. elegans offer special opportunities to address these questions. Previously, we identified a series of sensory neurons that modulate aggregation of C. elegans. These include neurons that respond to O2, CO2, noxious cues, satiety state, and pheromones. We propose to take our analysis to the next level by dissecting how, in mechanistic molecular terms, these distributed inputs modify the activity of populations of interneurons and motoneurons to coordinate group formation. Our strategy is to develop new, highly parallel approaches to replace the traditional piecemeal analysis. We propose to: 1) Harness next generation sequencing (NGS) to forward genetics, rapidly to identify a molecular ¿parts list¿ for aggregation. Much of the genetics has been done: we have identified almost 200 mutations that inhibit or enhance aggregation but otherwise show no overt phenotype. A pilot study of 50 of these mutations suggests they identify dozens of genes not previously implicated in aggregation. NGS will allow us to molecularly identify these genes in a few months, providing multiple entry points to study molecular and circuitry mechanisms for behaviour. 2) Develop new methods to image the activity of populations of neurons in immobilized and freely moving animals, using genetically encoded indicators such as the calcium sensor cameleon and the voltage indicator mermaid. This will be the first time a complex behaviour has been dissected in this way. We expect to identify novel conserved molecular and circuitry mechanisms.

2 439 996 €

Start date: 2011-04-01, End date: 2017-03-31

A significant advance in the field of development has been the appreciation that radial glial cells are progenitors and give birth to neurons in the brain. In order to advance this exciting area of biology, we need approaches that combine structural and functional studies of these cells. This is reflected by the emerging realisation that dynamic interactions involving radial glia may be critical for the regulation of their proliferative behaviour. It has been observed that radial glia experience transient elevations in intracellular Ca2+ but the nature of these signals, and the information that they convey, is not known. The inability to observe these cells in vivo and over the course of their development has also meant that basic questions remain unexplored. For instance, how does the behaviour of a radial glial cell at one point in development, influence the final identity of its progeny? I propose to build a research team that will capitalise upon methods we have developed for observing individual radial glia and their progeny in an intact vertebrate nervous system. The visual system of Xenopus Laevis tadpoles offers non-invasive optical access to the brain, making time-lapse imaging of single cells feasible over minutes and weeks. The system s anatomy lends itself to techniques that measure the activity of the cells in a functional sensory network. We will use this to examine signalling mechanisms in radial glia and how a radial glial cell s experience influences its proliferative behaviour and the types of neuron it generates. We will also examine the interactions that continue between a radial glial cell and its daughter neurons. Finally, we will explore the relationships that exist within neuronal progeny derived from a single radial glial cell.

1 284 808 €

Start date: 2010-02-01, End date: 2015-01-31

Microporous materials are an important class of solid; the two main members of this family are zeolites and metal-organic frameworks (MOFs). Zeolites are industrial solids whose applications range from catalysis, through ion exchange and adsorption technologies to medicine. MOFs are some of the most exciting new materials to have been developed over the last two decades, and they are just beginning to be applied commercially. Over recent years the applicant’s group has developed new synthetic strategies to prepare microporous materials, called the Assembly-Disassembly-Organisation-Reassembly (ADOR) process. In significant preliminary work the ADOR process has shown to be an extremely important new synthetic methodology that differs fundamentally from traditional solvothermal methods. In this project I will look to overturn the conventional thinking in materials science by developing methodologies that can target both zeolites and MOF materials that are difficult to prepare using traditional methods – the so-called ‘unfeasible’ materials. The importance of such a new methodology is that it will open up routes to materials that have different properties (both chemical and topological) to those we currently have. Since zeolites and MOFs have so many actual and potential uses, the preparation of materials with different properties has a high chance of leading to new technologies in the medium/long term. To complete the major objective I will look to complete four closely linked activities covering the development of design strategies for zeolites and MOFs (activities 1 & 2), mechanistic studies to understand the process at the molecular level using in situ characterisation techniques (activity 3) and an exploration of potential applied science for the prepared materials (activity 4).

2 489 220 €

Start date: 2018-10-01, End date: 2023-09-30

The aim of the ALIGN project is to understand, predict, and optimize the photovoltaic energy conversion in third-generation solar cells, starting from an atomic-scale quantum-mechanical modelling of the photovoltaic interface. The quest for photovoltaic materials suitable for low-cost synthesis, large-area production, and functional architecture has driven substantial research efforts towards third-generation photovoltaic devices such as plastic solar cells, organic-inorganic cells, and photo-electrochemical cells. The physical and chemical processes involved in the harvesting of sunlight, the transport of electrical charge, and the build-up of the photo-voltage in these devices are fundamentally different from those encountered in traditional semiconductor heterojunction solar cells. A detailed atomic-scale quantum-mechanical description of such processes will lay down the basis for a rational approach to the modelling, optimization, and design of new photovoltaic materials. The short name of the proposal hints at one of the key materials parameters in the area of photovoltaic interfaces: the alignment of the quantum energy levels between the light-absorbing material and the electron acceptor. The level alignment drives the separation of the electron-hole pairs formed upon absorption of sunlight, and determines the open circuit voltage of the solar cell. The energy level alignment not only represents a key parameter for the design of photovoltaic devices, but also constitutes one of the grand challenges of modern computational materials science. Within this project we will develop and apply new ground-breaking computational methods to understand, predict, and optimize the energy level alignment and other design parameters of third-generation photovoltaic devices.

1 000 000 €

Start date: 2010-03-01, End date: 2016-02-29

"I propose a structured multidisciplinary research programme that seeks to combine advanced materials, such as metal oxides and organics, with novel fabrication methods to develop devices for application in: (1) large area electronics, (2) integrated nanoelectronics and (3) sensors. At the heart of this programme lies the development of novel oxide semiconductors. These will be synthesised from solution using precursors. Chemical doping via physical blending will be explored for the tuning of the electronic properties of these compounds. This simple approach will enable the rapid development of a library of materials far beyond those accessible by traditional methods. Oxides will then be combined with inorganic/organic dielectrics to demonstrate low power transistors. Ultimate target for application area (1) is the development of transistors with hole/electron mobilities exceeding 20/200 cm^2/Vs respectively. For application area (2) I will combine the precursor formulations with advanced scanning thermochemical nanolithography. A heated atomic force microscope tip will be used for the local chemical conversion of the precursor to oxide with sub-50 nm resolution. This will enable patterning of nanostructures with desirable shape and size. Sequential patterning of semi/conductive layers combined with SAM dielectrics would enable fabrication of nano-sized devices and circuits. For application area (3), research effort will focus on novel hybrid phototransistors. Use of different light absorbing organic dyes functionalised onto the oxide channel will be explored as a mean for developing high sensitivity phototransistors and full colour sensing arrays. Organic dyes will also be combined with nano-sized transistors to demonstrate integrated nano-scale optoelectronics. The unique combination of bottom-up and top-down strategies adopted in this project will lead to the development of novel high performance devices with a host of existing and new applications."

1 497 798 €

Start date: 2012-01-01, End date: 2016-12-31

Within a 12 month period, 20% of adults will meet criteria for one or more clinical anxiety disorders (ADs). These disorders are hugely disruptive, placing an emotional burden on individuals and their families. While both cognitive behavioural therapy and pharmacological treatment are widely viewed as effective strategies for managing ADs, systematic review of the literature reveals that only 30–45% of patients demonstrate a marked response to treatment (anxiety levels being reduced into the nonaffected range). In addition, a significant proportion of initial responders relapse after treatment is discontinued. There is hence a real and marked need to improve upon current approaches to AD treatment. One possible avenue for improving response rates is through optimizing initial treatment selection. Specifically, it is possible that certain individuals might respond better to cognitive interventions while others might respond better to pharmacological treatment. Recently it has been suggested that there may be two or more distinct biological pathways disrupted in anxiety. If this is the case, then specification of these pathways may be an important step in predicting which individuals are likely to respond to which treatment. Few studies have focused upon this issue and, in particular, upon identifying neural markers that might predict response to cognitive (as opposed to pharmacological) intervention. The proposed research aims to address this. Specifically, it tests the hypothesis that there are at least two mechanisms disrupted in ADs, one entailing amygdala hyper-responsivity to cues that signal threat, the other impoverished recruitment of frontal regions that support cognitive and emotional regulation. Two series of functional magnetic resonance imaging experiments will be conducted. These will investigate differences in amygdala and frontal function during (a) attentional processing and (b) fear conditioning. Initial clinical experiments will investigate whether Generalised Anxiety Disorder and Specific Phobia involve differing degrees of disruption to frontal versus amygdala function during these tasks. This work will feed into training studies, the goal being to characterize AD patient subgroups that benefit from cognitive training.

1 708 407 €

Start date: 2011-04-01, End date: 2016-08-31

Supramolecular polymers are of major interest in the field of self assembly with a promising outlook in areas of viscosity modification, compartmentalized architectures, bio-conjugates and drug-delivery applications. They are dynamic macromolecular materials prepared by simple mixing of relatively small components bearing complementary or self-complementary recognition motifs. A major limitation in the field, however, has been access to synthetic systems capable of undergoing self assembly in an aqueous environment. This research proposal develops well-defined, self-organizing macromolecular structures that will overcome this limitation by focusing on systems that rely on several non-covalent interactions occurring in concert rather than on single interactions alone. The envisioned supramolecular polymers and bio-conjugates are designed as dynamic water-soluble smart materials, whose architectures can be controlled and exhibit reversibility upon exposure to external stimuli such as electrochemical, temperature or pH changes. Molecular recognition events occurring between functional handles on both synthetic and bio-polymers will be investigated in order to control the formation of desired functional architectures through stoichiometrically controlled complexation. Preparation of synthetic core motifs to assemble discrete peptide aggregates such as the dimeric through hexameric oligomers of amyloid-beta(40/42) will lead to structural elucidation and insight into several peptide misfolding pathologies like Alzheimer's or Parkinson's disease.

1 700 000 €

Start date: 2009-11-01, End date: 2015-10-31

ATM is the protein kinase that is mutated in the hereditary autosomal recessive disease ataxia telangiectasia (A-T). A-T patients display immune deficiencies, cancer predisposition and radiosensitivity. The molecular role of ATM is to respond to DNA damage by phosphorylating its substrates, thereby promoting repair of damage or arresting the cell cycle. Following the induction of double-strand breaks (DSBs), the NBS1 protein is required for activation of ATM. But ATM can also be activated in the absence of DNA damage. Treatment of cultured cells with hypotonic stress leads to the activation of ATM, presumably due to changes in chromatin structure. We have recently described a second ATM cofactor, ATMIN (ATM INteractor). ATMIN is dispensable for DSBs-induced ATM signalling, but ATM activation following hypotonic stress is mediated by ATMIN. While the biological role of ATM activation by DSBs and NBS1 is well established, the significance, if any, of ATM activation by ATMIN and changes in chromatin was up to now completely enigmatic. ATM is required for class switch recombination (CSR) and the suppression of translocations in B cells. In order to determine whether ATMIN is required for any of the physiological functions of ATM, we generated a conditional knock-out mouse model for ATMIN. ATM signaling was dramatically reduced following osmotic stress in ATMIN-mutant B cells. ATMIN deficiency led to impaired CSR, and consequently ATMIN-mutant mice developed B cell lymphomas. Thus ablation of ATMIN resulted in a severe defect in ATM function. Our data strongly argue for the existence of a second NBS1-independent mode of ATM activation that is physiologically relevant. While a large amount of scientific effort has gone into characterising ATM signaling triggered by DSBs, essentially nothing is known about NBS1-independent ATM signaling. The experiments outlined in this proposal have the aim to identify and understand the molecular pathway of ATMIN-dependent ATM signaling.

1 499 881 €

Start date: 2012-02-01, End date: 2018-01-31

"The origin of life is not well understood, and is one of the great remaining questions in science. Autocatalytic chemical reactions have been extensively studied with the aim of providing insight into the principles underlying living systems. In biology, organisms can be thought of as imperfect self-replicators, which produce closely related species, allowing for selection and evolution. Autocatalysis is also an important part of many other biological processes. This project aims to develop new autocatalytic reactions where two simple chemical building blocks come together to give a more complex product, and then the product aggregates to give primitive cell-like structures or "protocells" such as micelles or vesicles. The protocells allow the starting materials to mix more efficiently, speeding up the reaction in time and giving rise to complex behaviour of the protocells. These reactions will serve as models that I hope will contribute to understanding how cell-like systems can emerge from simpler chemicals and be relevant to how life started on earth. This project will give the opportunity to study chemical systems that may be able to evolve in time, allow development of useful chemical models of important biological processes, and provide ‘bottom-up’ approaches to synthetic biology. This research will potential allow the study evolution in a new ways, develop technology useful to a number of scientific fields, and potentially shed light on the processes that allowed chemistry to become biology on the primitive Earth."

2 278 073 €

Start date: 2016-05-01, End date: 2021-04-30

Axon growth potential declines during development, contributing to the lack of effective regeneration in the adult central nervous system. What determines the intrinsic growth potential of neurites, and how such growth is regulated during development, disease and following injury is a fundamental question in neuroscience. Although multiple lines of evidence indicate that intrinsic growth capability is genetically encoded, its nature remains poorly defined. Neuronal remodeling of the Drosophila mushroom body offers a unique opportunity to study the mechanisms of various types of axon degeneration and growth. We have recently demonstrated that regrowth of axons following developmental pruning is not only distinct from initial outgrowth but also shares molecular similarities with regeneration following injury. In this proposal we combine state of the art tools from genomics, functional genetics and microscopy to perform a comprehensive study of the mechanisms underlying axon growth during development and following injury. First, we will combine genetic, biochemical and genomic studies to gain a mechanistic understanding of the developmental regrowth program. Next, we will perform extensive transcriptomic analyses and comparisons aimed at defining the genetic programs involved in initial axon growth, developmental regrowth, and regeneration following injury. Finally, we will harness the genetic power of Drosophila to perform a comprehensive functional analysis of genes and pathways, those previously known and new ones that we will discover, in various neurite growth paradigms. Importantly, these functional assays will be performed in the same organism, allowing us to use identical genetic mutations across our analyses. To this end, our identification of a new genetic program regulating developmental axon regrowth, together with emerging tools in genomics, places us in a unique position to gain a broad understanding of axon growth during development and following injury.

2 000 000 €

Start date: 2014-03-01, End date: 2019-02-28

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

Despite that C–H functionalization represents a paradigm shift from the standard logic of organic synthesis, the selective activation of non-functionalized alkanes has puzzled chemists for centuries and is always referred to one of the remaining major challenges in chemical sciences. Alkanes are inert compounds representing the major constituents of natural gas and petroleum. Converting these cheap and widely available hydrocarbon feedstocks into added-value intermediates would tremendously affect the field of chemistry. For long saturated hydrocarbons, one must distinguish between non-equivalent but chemically very similar alkane substrate C−H bonds, and for functionalization at the terminus position, one must favor activation of the stronger, primary C−H bonds at the expense of weaker and numerous secondary C-H bonds. The goal of this work is to develop a general principle in organic synthesis for the preparation of a wide variety of more complex molecular architectures from saturated hydrocarbons. In our approach, the alkane will first be transformed into an alkene that will subsequently be engaged in a metal-catalyzed hydrometalation/migration sequence. The first step of the sequence, ideally represented by the removal of two hydrogen atoms, will be performed by the use of a mutated strain of Rhodococcus. The position and geometry of the formed double bond has no effect on the second step of the reaction as the metal-catalyzed hydrometalation/migration will isomerize the double bond along the carbon skeleton to selectively produce the primary organometallic species. Trapping the resulting organometallic derivatives with a large variety of electrophiles will provide the desired functionalized alkane. This work will lead to the invention of new, selective and efficient processes for the utilization of simple hydrocarbons and valorize the synthetic potential of raw hydrocarbon feedstock for the environmentally benign production of new compounds and new materials.

2 499 375 €

Start date: 2018-11-01, End date: 2023-10-31

"In the course of biomineralization, organisms produce a large variety of functional biogenic crystals that exhibit fascinating mechanical, optical, magnetic and other characteristics. More specifically, when living organisms grow crystals they can effectively control polymorph selection as well as the crystal morphology, shape, and even atomic structure. Materials existing in nature have extraordinary and specific functions, yet the materials employed in nature are quite different from those engineers would select. I propose to emulate specific strategies used by organisms in forming structural biogenic crystals, and to apply these strategies biomimetically so as to form new structural materials with new properties and characteristics. This bio-inspired approach will involve the adoption of three specific biological strategies. We believe that this procedure will open up new ways to control the structure and properties of smart materials. The three bio-inspired strategies that we will utilize are: (i) to control the short-range order of amorphous materials, making it possible to predetermine the polymorph obtained when they transform from the amorphous to the succeeding crystalline phase; (ii) to control the morphology of single crystals of various functional materials so that they can have intricate and curved surfaces and yet maintain their single-crystal nature; (iii) to entrap organic molecules into single crystals of functional materials so as to tailor and manipulate their electronic structure. The proposed research has significant potential for opening up new routes for the formation of novel functional materials. Specifically, it will make it possible for us (1) to produce single, intricately shaped crystals without the need to etch, drill or polish; (2) to control the short-range order of amorphous materials and hence the polymorph of the successive crystalline phase; (3) to tune the band gap of semiconductors via incorporation of tailored bio-molecules."

1 500 000 €

Start date: 2013-09-01, End date: 2018-08-31

Tight regulation of RNA metabolism is essential for normal brain function. This includes co and post-transcriptional regulation, which are extremely prevalent in neurons. Recently, circular RNAs (circRNAs), a highly abundant new type of regulatory non-coding RNA have been found across the animal kingdom. Two of these RNAs have been shown to act as miRNA sponges but no function is known for the thousands of other circRNAs, indicating the existence of a widespread layer of previously unknown gene regulation. The present proposal aims to comprehensively determine the role and mode of actions of circRNAs in gene expression and RNA metabolism in the fly brain. We will do so by studying their biogenesis, transport, and mechanism of action, as well as by determining the roles of circRNAs in neuronal function and behaviour. Briefly, we will: 1) identify factors involved in the biogenesis, localization, and stabilization of circRNAs; 2) determine neuro-developmental, molecular, neural and behavioural phenotypes associated with down or up regulation of specific circRNAs; 3) study the molecular mechanisms of action of circRNAs: identify circRNAs that work as miRNA sponges and determine whether circRNAs can encode proteins or act as signalling molecules and 4) perform mechanistic studies in order to determine cause-effect relationships between circRNA function and brain physiology and behaviour. The present proposal will reveal the key pathways by which circRNAs control gene expression and influence neuronal function and behaviour. Therefore it will be one of the pioneer works in the study of this new and important area of research, which we predict will fundamentally transform the study of gene expression regulation in the brain

1 971 750 €

Start date: 2016-02-01, End date: 2021-01-31

Aggregates of proteins such as amyloid-beta and alpha-synuclein circulate the extracellular space of the brain (ECS) and are thought to be key players in the development of neurodegenerative diseases. The clearance of these aggregates (among other toxic metabolites) is a fundamental physiological feature of the brain which is poorly understood due to the lack of techniques to study the nanoscale organisation of the ECS. Exciting advances in this field have recently shown that clearance is enhanced during sleep due to a major volume change in the ECS, facilitating the flow of the interstitial fluid. However, this process has only been characterised at a low spatial resolution while the physiological changes occur at the nanoscale. The recently proposed “glymphatic” pathway still remains controversial, as there are no techniques capable of distinguishing between diffusion and bulk flow in the ECS of living animals. Understanding these processes at a higher spatial resolution requires the development of single-molecule imaging techniques that can study the brain in living animals. Taking advantage of the strategies I have recently developed to target single-molecules in the brain in vivo with nanoparticles, we will do “nanoscopy” in living animals. Our proposal will test the glymphatic pathway at the spatial scale in which events happen, and explore how sleep and wake cycles alter the ECS and the diffusion of receptors in neuronal plasma membrane. Overall, BrainNanoFlow aims to understand how nanoscale changes in the ECS facilitate clearance of protein aggregates. We will also provide new insights to the pathological consequences of impaired clearance, focusing on the interactions between these aggregates and their putative receptors. Being able to perform single-molecule studies in vivo in the brain will be a major breakthrough in neurobiology, making possible the study of physiological and pathological processes that cannot be studied in simpler brain preparations.

1 552 948 €

Start date: 2018-12-01, End date: 2023-11-30

We are currently witnessing a paradigm shift in our understanding of human brain function, moving towards a clearer description of cortical processing. Sensory systems are no longer considered as 'passively recording' but rather as dynamically anticipating and adapting to the rapidly changing environment. These new ideas are encompassed in the predictive coding framework, and indeed in a unifying theory of the brain (Friston, 2010). In terms of brain computation, a predictive model is created in higher cortical areas and communicated to lower sensory areas through feedback connections. Based on my pioneering research I propose experiments that are capable of ‘brain-reading’ cortical feedback– which would contribute invaluable data to theoretical frameworks. The proposed research project will advance our understanding of ongoing brain activity, contextual processing, and cortical feedback - contributing to what is known about general cortical functions. By providing new insights as to the information content of cortical feedback, the proposal will fill one of the most important gaps in today’s knowledge about brain function. Friston’s unifying theory of the brain (Friston, 2010) and contemporary models of the predictive-coding framework (Hawkins and Blakeslee, 2004;Mumford, 1992;Rao and Ballard, 1999) assign feedback processing an essential role in cortical processing. Compared to feedforward information processing, our knowledge about feedback processing is in its infancy. The proposal introduces parametric and explorative brain reading designs to investigate this feedback processing. The chief goal of my proposal will be precision measures of cortical feedback, and a more ambitious objective is to read mental images and inner thoughts.

1 494 714 €

Start date: 2012-12-01, End date: 2017-11-30

An expanded GGGGCC repeat in a non-coding region of the C9orf72 gene is the most common known cause of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). The repeat RNA is transcribed and accumulates in neuronal RNA aggregates, implicating RNA toxicity as a key pathogenic mechanism. However, the pathways that lead to neurodegeneration are unknown. My lab has made pioneering contributions to the understanding of C9orf72 FTD/ALS, and reported the first structure of the repeat RNA, and the first description of both sense and antisense RNA aggregates in patient brain. We have now developed new disease models that allow, for the first time, the dissection of RNA toxicity both in vivo and in sophisticated neuronal culture models. We have also used our knowledge of the repeat structure to identify novel small molecules that show very strong binding to the repeats. We will utilise our innovative disease models in a multidisciplinary approach to fully dissect the cellular pathways underlying C9orf72 repeat RNA toxicity in vivo, on a genome-wide scale. Altered RNA metabolism has been implicated in a wide range of neurodegenerative diseases, indicating that our findings will provide profound new insight into fundamental mechanisms of neuronal maintenance and survival. This research programme will also deliver a step change in our understanding of C9orf72 FTD/ALS pathogenesis and provide essential insight for the identification of small molecules with genuine therapeutic potential. RNA-mediated mechanisms are now known to be a common theme in neurodegeneration, suggesting these findings will have broad significance.

1 985 699 €

Start date: 2015-10-01, End date: 2020-09-30

Medicinal chemistry requires more efficient and diverse methods for the asymmetric synthesis of chiral scaffolds. Over 60% of the world’s top selling small molecule drug compounds are chiral and, of these, approximately 80% are marketed as single enantiomers. There is a compelling correlation between drug candidate “chiral complexity” and the likelihood of progression to the marketplace. Surprisingly, and despite the tremendous advances made in catalysis over the past several decades, the “chiral complexity” of drug discovery libraries has actually decreased, while, at the same time, for the reasons mentioned above, the “chiral complexity” of marketed drugs has increased. Since the mid-1990s, there has been a notable acceleration of this “complexity divergence”. Consequently, there is now an urgent need to provide efficient processes that directly access privileged chiral scaffolds. It is our philosophy that catalysis holds the key here and new processes should be based upon platforms that can exert control over both absolute and relative stereochemistry. In this proposal we outline the development of a range of N-heteroannulation processes based upon the catalytic generation and trapping of unique or unusual classes of organometallic intermediate derived from transition metal insertion into C-C and C-N sigma-bonds. We will provide a variety of enabling methodologies and demonstrate applicability in flexible total syntheses of important natural product scaffolds. The processes proposed are synthetically flexible, operationally simple and amenable to asymmetric catalysis. Likely starting points, based upon preliminary results, will set the stage for the realisation of aspirational and transformative goals. Through the study of the organometallic intermediates involved here, there is potential to generalise these new catalytic manifolds, such that this research will transcend N heterocyclic chemistry to provide enabling methods for organic chemistry as a whole.

1 548 738 €

Start date: 2015-04-01, End date: 2020-03-31

Humans are endowed with a number of advanced cognitive abilities not seen in other species. So what allows the human brain to stand out from the rest in these capabilities? In general, the brains of primates, including humans, have more neurons per unit volume than other mammals. But humans are also in the fortunate position of having the largest of the primate brains, making the number of neurons in the human cerebral cortex greatly expanded. Thus, the difference seems to be a matter of quantity, not quality. My laboratory is interested in understanding how neuron number, and thus brain size, is determined in human brain development. The research proposed here is aimed at taking an evolutionary approach to this question and comparing brain development in an in vitro 3D model system, cerebral organoids. This method, which relies on self-organization from differentiating pluripotent stem cells, recapitulates remarkably well the endogenous developmental program of the human brain. Having previously established the brain organoid approach, and more recently improved upon it with the application of bioengineering, my laboratory is in a unique position to carry out functional studies of human brain development. I propose to use this approach to compare developing human brain tissue to that of other hominid species and tease apart unique features of human neural stem cells and progenitors that allow them to generate more neurons and therefore a greater cerebral cortical size. Furthermore, we will perform transcriptomic and functional screening to identify factors underlying this expansion, followed by careful genetic substitution to test the contributions of putative evolutionary changes. In this way, we will functionally test putative human evolutionary changes in a manner not previously possible.

1 444 911 €

Start date: 2018-07-01, End date: 2023-06-30

This research proposal’s goal is to investigate the role of cortico-hippocampal interactions during the encoding and consolidation of a memory. Current memory consolidation models postulate that memory storage in our brains occurs by a dynamic process- a recent episodic experience is initially encoded in the hippocampus, and during off-line states such as sleep, the encoded memory is gradually transferred to neocortex for long-term storage. One potential neural mechanism by which this could occur is replay, a phenomenon where neural activity patterns in the hippocampus evoked by a previous experience reactivate spontaneously during non-REM sleep, leading to coordinated cortical reactivation. While previous work suggests that hippocampal replay is important for encoding new memories, how memory consolidation is accomplished through cortico-hippocampal interactions is not well understood. This research project has three major aims- 1) examine how cortical feedback influences which spatial trajectory is replayed by the hippocampus, 2) investigate how the hippocampal replay of a behavioural episode modifies cortical circuits, 3) measure the causal role of cortico-hippocampal interactions in consolidating memories. We will record ensemble activity from freely moving rats during an auditory-spatial association task and during post-behavioural sleep sessions. We will focus our ensemble recordings on two brain regions: 1) the dorsal CA1 region of the hippocampus, where the phenomenon of sleep replay has been most extensively examined, and 2) auditory cortex, a region of the brain critical for both auditory perception and long-term memory storage. This work will use behavioral and molecular-genetic techniques in combination with large-scale electrophysiological recordings, to help elucidate the role of cortico-hippocampal interactions in memory encoding and consolidation.

1 500 000 €

Start date: 2015-04-01, End date: 2021-03-31

Following their synthesis during DNA replication, sister chromatids remain paired by the cohesin complex, which forms the basis for their faithful segregation during cell division. Cohesin is a large ring-shaped protein complex, incorporating an ABC-type ATPase module. Despite its importance for genome stability, the molecular mechanism of cohesin action remains as intriguing as it remains poorly understood. How is cohesin topologically loaded onto chromatin? How is it unloaded again? What happens to cohesin during DNA replication in S-phase, so that it establishes cohesion between newly synthesized sister chromatids? We propose to capitalise on our recent success in the biochemical reconstitution of topological cohesin loading onto DNA. This lays the foundation for a work programme encompassing a combination of biochemical, single molecule, structural and genetic approaches to address the above questions. Five work packages will investigate cohesin’s molecular behaviour during its life-cycle on chromosomes, including the ATP binding and hydrolysis-dependent conformational changes that make this molecular machine work. It will be complemented by mechanistic analyses of the cofactors that help cohesin to load onto chromosomes and establish sister chromatid cohesion. The insight gained will not only advance our molecular knowledge of sister chromatid cohesion. It will more generally advance our understanding of the ubiquitous family of chromosomal SMC ATPases, of which cohesin is a member, and their activity of shaping and segregating genomes.

2 120 100 €

Start date: 2015-10-01, End date: 2020-09-30

DNA replication represents a dangerous moment in the life of the cell as endogenous and exogenous events challenge genome integrity by interfering with the progression, stability and restart of the replication fork. Failure to protect stalled forks or to process the replication fork appropriately contribute to the pathological mechanisms giving rise to cancer, therefore an understanding of the intricate mechanisms that ensure fork integrity can provide targets for new chemotherapeutic assays. Smc5-Smc6 is a multi-subunit complex with a poorly understood function in DNA replication and repair. One of its subunits, Nse2, is able to promote the addition of a small ubiquitin-like protein modifier (SUMO) to specific target proteins. Recent work has revealed that the Smc5-Smc6 complex is required for the progression of replication forks through damaged DNA and is recruited de novo to forks that undergo collapse. In addition, Smc5-Smc6 mediate repair of DNA breaks by homologous recombination between sister-chromatids. Thus, Smc5-Smc6 is anticipated to promote recombinational repair at stalled/collapsed replication forks. My laboratory proposes to develop molecular techniques to study replication at the level of single replication forks. We will employ these assays to identify and dissect the function of factors involved in replication fork stability and repair. We will place an emphasis on the study of the Smc5-Smc6 complex in these processes because of its potential roles in recombination-dependent fork repair and restart. We also propose to identify novel Nse2 substrates involved in DNA repair using yeast model systems. Specifically, we will address the following points: (1) Development of assays for analysis of factors involved in stabilisation, collapse and re-start of single-forks, (2) Analysis of the roles of Smc5-Smc6 in fork biology using developed techniques, (3) Isolation and functional analysis of novel Nse2 substrates.

893 396 €

Start date: 2008-09-01, End date: 2013-08-31

The overarching objective of this interdisciplinary project is to elucidate mechanisms through which billions of individual clocks in the body communicate with each other and tick in harmony. The mammalian circadian timing system consists of a master clock in the brain and subsidiary oscillators in almost every cell of the body. Since these clocks anticipate environmental changes and function together to orchestrate daily physiology and behavior their temporal synchronization is critical. Our recent finding that oxygen serves as a resetting cue for circadian clocks points towards the unprecedented involvement of blood gases as time signals. We will apply cutting edge continuous physiological measurements in freely moving animals, alongside biochemical/molecular biology approaches and advanced cell culture setup to determine the molecular role of oxygen, carbon dioxide and pH in circadian clock communication and function. The intricate nature of the mammalian circadian system demands the presence of communication mechanisms between clocks throughout the body at multiple levels. While previous studies primarily addressed the role of the master clock in resetting peripheral clocks, our knowledge regarding the communication among clocks between and within peripheral organs is rudimentary. We will reconstruct the mammalian circadian system from the bottom up, sequentially restoring clocks in peripheral tissues of a non-rhythmic animal to (i) obtain a system-view of the peripheral circadian communication network; and (ii) study novel tissue-derived circadian communication mechanisms. This integrative proposal addresses fundamental aspects of circadian biology. It is expected to unravel the circadian communication network and shed light on how billions of clocks in the body function in unison. Its impact extends beyond circadian rhythms and bears great potential for research on communication between cells/tissues in various fields of biology.

1 999 945 €

Start date: 2018-03-01, End date: 2023-02-28

An important question of modern neurobiology is how neurons regulate synaptic function in response to excitation. In particular, the roles of alternative pre-mRNA splicing and mRNA translation regulation in this response are poorly understood. We will study the RNA-binding proteins (RBPs) that control these post-transcriptional changes using a UV crosslinking-based purification method (CLIP) and ultra-high throughput sequencing. Computational analysis of the resulting data will define the sequence and structural features of RNA motifs recognized by each RBP. Splicing microarrays and translation reporter assays will then allow us to examine the regulatory functions of RBPs and RNA motifs. By integrating the biochemical and functional datasets, we will relate the position of RNA motifs to the activity of bound RBPs, and predict the interactions that act as central nodes in the regulatory network. The physiological role of these core RBP-RNA interactions will then be tested using antisense RNAs. Together, these projects will provide insights to the regulatory mechanisms underlying neuronal activity-dependent changes, and provide new opportunities for future treatments of neurodegenerative disorders.

900 000 €

Start date: 2008-09-01, End date: 2013-08-31

In this project, the basal ganglia are defined as actor-critic reinforcement learning networks that aim at an optimal tradeoff between the maximization of future cumulative rewards and the minimization of the cost (the reinforcement driven multi objective optimization RDMOO model). This computational model will be tested by multiple neuron recordings in the major basal ganglia structures of monkeys engaged in a similar behavioral task. We will further validate the RMDOO computational model of the basal ganglia by extending our previous studies of neural activity in the MPTP primate model of Parkinson's disease to a primate model of central serotonin depletion and emotional dysregulation disorders. The findings in the primate model of emotional dysregulation will then be compared to electrophysiological recordings carried out in human patients with treatment-resistant major depression and obsessive compulsive disorder during deep brain stimulation (DBS) procedures. I aim to find neural signatures (e.g., synchronous gamma oscillations in the actor part of the basal ganglia as predicted by the RMDOO model) characterizing these emotional disorders and to use them as triggers for closed loop adaptive DBS. Our working hypothesis holds that, as for the MPTP model of Parkinson's disease, closed loop DBS will lead to greater amelioration of the emotional deficits in serotonin depleted monkeys. This project incorporates extensive collaborations with a team of neurosurgeons, neurologists, psychiatrists, and computer science/ neural network researchers. If successful, the findings will provide a firm understanding of the computational physiology of the basal ganglia networks and their disorders. Importantly, they will pave the way to better treatment of human patients with severe mental disorders.

2 476 922 €

Start date: 2013-12-01, End date: 2018-11-30

Targeted drug delivery across the blood brain barrier (BBB) to the central nervous system is a large challenge for the treatment of neurological disorders. This 4 year ERC program is aimed towards the evaluating the BBB penetration capacity and toxicological potential of novel carbon nanotube (CNT) carriers using an integrated multidisciplinary approach. State-of-art characterisation techniques developed by the PI will be applied and further developed to detect the interaction of carbon nanotubes with in vitro BBB model and neuronal cells. Specific aims: 1. Identify the mechanisms of translocation of CNT across the endothelial cells which comprise the BBB, as well as uptake by neuronal cells in vitro. 2. To investigate the effect of length, diameter and surface charge of CNTs on the BBB and neuronal cells penetration capacity in vitro. 3. To investigate the toxicological profile of CNT on the BBB and the various neuronal cell types (immortalised and primary neuronal cultures). 4. Develop protocols to assess whether the CNTs degrade inside the cell. The ERC Grant will consolidate the new Research Group in nanomaterials-cell interfaces, and allow them to perform stimulating investigator-initiated frontier research in nanotoxicology and nanomedicine. To this end, a multi-disciplinary laboratory will be realized within the framework of this 4-year the ERC Programme. This will permit the group around the PI, to expand activities, push limits, create new boundaries, and develop new protocols for studying nanoparticle-cell interactions in close collaboration with ICL s Department of medicine and chemistry. Within the proposed program there is an underlying ambition both to gain a fundamental understanding for which parameters of CNTs determine their penetration capacity through the BBB and also to assess their toxicological potential at the BBB two highlighted themes by the ERC.

1 229 998 €

Start date: 2011-02-01, End date: 2017-01-31

The idea of this research project is to take advantage of molecular self-assembly to create a new generation of periodically-organized porous organic materials that, acting as specific molecular hosts, can structurally control the positioning of multiple functional guests on surfaces, opening new horizons toward the understanding and development of rationale protocols for the patterning of unprecedented materials. Taking advantage of a supramolecular approach to engineer extended mono- and two-dimensional organic networks, the ultimate aim of COLORLANDS is to create novel hosting frameworks accommodating in a predetermined fashion organic chromophores and/or fluorophores. For instance, these can be oligophenylenes as blue emitters, cumarines/oligophenylethylenes as green emitters, or perylenebisimides conjugates as red emitters. Depending on their spatial organization, such materials will be the springboard for further technological development in the fields of electroluminescent devices or artificial leafs mimicking natural light harvesting antenna systems. The self-assembly of selected rigid molecular modules alternatively functionalized with complementary connectors (PNA strands) will yield, under equilibrium conditions, one exclusive structural pattern. This will feature controllable (in shape, size and chemical nature) periodic receptor sites, each programmed to selectively accommodate a specific molecular chromophore and/or fluorophore. Particular attention will be given to the design and fundamental understanding of specific orthogonal interactions between the self-assembled receptor sites and the functional molecular guests. This will be achieved through the lateral organic functionalization of the PNA strands with novel orthogonal H-bonding-based recognition motifs. Depending on the ratio between the different receptors, one can tailor the desired emission or absorption colour, virtually enabling unlimited surfing through the color coordinate diagram.

1 295 400 €

Start date: 2012-01-01, End date: 2016-12-31

The fundamental objective of the research described in this proposal is to lay the foundations for understanding how structural complexity can give rise to materials properties inaccessible to structurally-simple states. The long-term vision is a paradigm shift in the way we as chemists design materials—the “Complexity Revolution”—where we move to thinking beyond the unit cell and harness unconventional order to generate emergent states with entirely novel behaviour. The key methodologies of the project are (i) exploitation of the rich structural information accessible using 3D-PDF / diffuse scattering techniques, (ii) exploration of the phase behaviour of unconventional ordered states using computational methods, and (iii) experimental/computational studies of a broad range of materials in which complexity arises from a large variety of different phenemona. In this way, the project will establish how we might controllably introduce complexity into materials by varying chemical composition and synthesis, how we might then characterise these complex states, and how we might exploit this complexity when designing next-generation materials with unprecedented electronic, catalytic, photonic, information storage, dielectric, topological, and magnetic properties.

3 362 635 €

Start date: 2018-10-01, End date: 2023-09-30

The urgent need to reduce carbon emissions in order to mitigate climate change requires the development of clean, renewable energy sources. Solar power offers a virtually unlimited supply of energy, providing it can be harnessed efficiently. Traditional silicon solar cells demonstrate high performance (~20%) but their required method of manufacture prohibits large area production rendering them too expensive to be used on a global scale. Organic solar cells (made from conjugated polymers and fullerenes) have the potential to be fabricated by low cost printing methods allowing for large scale modules to be produced cheaply. Conventional organic solar cells function by generating charge from a singlet excited state. In order to achieve optimum performance the precise morphology of polymer and fullerene must be controlled which can be extremely challenging. These devices however, have attained good efficiencies (10%) but are hampered by severe loss mechanisms which generally involve the formation of a lower energy triplet excited state. We propose to develop novel materials for organic solar cells which will instead utilise this triplet excited state to generate charges. This will enable us to not only eliminate this loss mechanism but due to the unique properties of the triplet excited state will allow for numerous benefits. Firstly, the long lifetime of the triplet excited state will be exploited to allow for a simpler organic solar cell where precise morphological control is not required. Secondly, the proposed new materials will allow for the utilisation of near-IR light which is typically wasted in ALL current solar cell devices. Thirdly, exploiting a unique photophysical process we will produce materials capable of delivering efficiencies in excess of the theoretical limit available to conventional solar cells. Thus we propose that utilisation of triplet excitons is the required step-change to allow for organic solar cells to achieve their ultimate efficiencies

1 499 223 €

Start date: 2016-04-01, End date: 2021-03-31

Neurons in primary visual cortex (area V1) receive feedforward inputs from thalamic afferents and lateral inputs from other cortical neurons. Little is known about how these components interact to determine the responses of a V1 neuron. One camp ascribes most responses to feedforward mechanisms. The other camp ascribes them mostly to lateral interactions. We propose that these two apparently opposed views can be simply reconciled in a single framework. We hypothesize that area V1 can operate both in a feedforward regime and in a lateral interaction regime, depending on the nature of the stimulus and on the cognitive task at hand, and that the transition from one regime to the other is governed by synaptic inhibition. We will test these hypotheses by recording from individual V1 neurons while monitoring the activity of nearby populations of cortical neurons via multiprobe electrodes. In Aim 1 we will relate the activity of V1 neurons to that of nearby populations. We will use simple measures of correlation and nonlinear models that predict individual spikes to measure how responses depend on a feedforward contribution (the receptive field ) and on a lateral contribution (the connection field ). We will test our first hypothesis, concerning the role of the stimulus in changing this dependence. In Aim 2 we will extend these results to a behaving animal. We will record from V1 of mice performing a 2-alternative forced-choice psychophysical task, and we will test our second hypothesis, concerning the role of the cognitive task in determining the operating regime of the cortex. In Aim 3 we will seek a biophysical interpretation of the functional mechanisms and effective connectivity revealed by the previous Aims. We will test our third hypothesis, concerning the role of synaptic inhibition. The tools involved will include intracellular recordings and optical stimulation in transgenic mice whose cortical neurons are sensitive to light.

2 499 921 €

Start date: 2009-04-01, End date: 2014-03-31

I propose to bridge the gap between simple in vitro measurements of biological processes, and the complexities of the cellular environment. This requires reduced in vitro systems that are sufficiently complex to reproduce the subtleties of the in vivo biological phenomenon, but sufficiently controllable to test how quantitative changes in a particular property affects function. The challenge is to step beyond the most simple and straightforward in vitro mimics of the cell membrane, and create model systems that more closely reproduce the conditions in vivo. I propose to tackle two specific, but interrelated membrane phenomena, that are currently not captured in artificial bilayers and create new complex mimics of the cell membrane capable of tackling these systems; namely (1) protein crowding and the cytoskeleton, and (2) lateral forces and membrane curvature. Testing our synthetic mimics with models that we understand in vivo is vital. This benchmarking will ensure that the mimics we create are relevant and will help ensure the more ambitious later goals of the this proposal are successful.We will then take these tools to go on and aim to create a synthetic mimic of the bacterial membrane. However we are not limited to creating purely natural duplicates, and we can exploit a much wider range of building material than nature. In addition to creating complex mimics, we will also create totally new synthetic systems inspired by the properties of the cell membrane, but possessing unique properties.

1 498 523 €

Start date: 2013-02-01, End date: 2018-10-31

The human genome is highly transcribed, with over 90% of sequences contributing to the production of RNA. The function of the vast majority of these RNAs is unknown. Evidence over many years has revealed that transcription factors and chromatin regulators are associated with a variety of non-coding (nc)RNAs, but their function remains largely unknown. There are a few cases where a role has been ascribed for ncRNAs in transcription, but no clear mechanistic insight has been defined yet. We predict that many of the newly identified ncRNAs emanating from the genome will play a role in transcriptional processes. We intend to identify and characterise such ncRNAs. This will take place in two phases. In the first phase we will use biochemical approaches to identify ncRNAs involved in the regulation of chromatin and transcription. Our investigations will focus on proteins leading to the induction of pluripotency and oncogenesis. ncRNAs associated with such proteins will be identified using targeted screens. In the second phase, the importance of these RNAs in determining pluripotency and oncogenesis will be analysed. In addition, a variety of molecular approaches will be used to investigate the mechanism by which these ncRNAs regulate the function of the proteins or complexes they associate with. One particular hypothesis we will explore is that such ncRNAs play a role in guiding proteins to DNA sequences, via the formation of RNA/DNA triplexes. This concerted and focused analysis will provide mechanistic insights into the functions of ncRNAs in transcriptional regulation and validate their role in key biological processes. The identification of such new ncRNA-regulated pathways may open up new avenues for therapeutic intervention.

2 141 470 €

Start date: 2011-05-01, End date: 2017-04-30

In eukaryotic cells, DNA replication is tightly regulated to ensure that the genome is duplicated only once per cell cycle. Errors in the control mechanisms that regulate chromosome ploidy cause genomic instability, which is linked to the development of cellular abnormalities, genetic disease and the onset of cancer. Recent reconstitution experiments performed with purified proteins revealed that initiation of eukaryotic genome duplication requires three distinct steps. First, DNA replication start sites are identified and targeted for the loading of an inactive MCM helicase motor, which encircles the double helix. Second, MCM activators are recruited, causing duplex-DNA untwisting. Third, upon interaction with a firing factor, the MCM ring opens to eject one DNA strand, leading to unwinding of the replication fork and duplication by dedicated replicative polymerases. These three events are not understood at a molecular level. Structural investigations so far aimed at imaging artificially isolated replication steps and used simplified templates, such as linear duplex DNA to study helicase loading or pre-formed forks to understand unwinding. However, the natural substrate of the eukaryotic replication machinery is not DNA but rather chromatin, formed of nucleosome arrays that compact the genome. Chromatin plays important regulatory roles in all steps of DNA replication, by dictating origin start-site selection and stimulating replication fork progression. Only by studying chromatin replication, we argue, will we understand the molecular basis of genome propagation. To this end, we have developed new protocols to perform visual biochemistry experiments under the cryo-electron microscope, to image chromatin duplication at high resolution, frozen as it is being catalysed. Using these strategies we want to generate a molecular movie of the entire replication reaction. Our achievements will change the way we think about genome stability in eukaryotic cells.

2 000 000 €

Start date: 2019-03-01, End date: 2024-02-29

The prime objective for every life form is to deliver its genetic material, intact, to the next generation. Each human cell receives tens-of-thousands of DNA lesions per day. These lesions can block genome replication and transcription, and if not repaired or repaired incorrectly, they lead to mutations or wider genome aberrations that threaten cell viability. To counter such threats, life has evolved the DNA-damage response (DDR), to detect DNA damage, signal its presence and mediate its repair. DDR events impact on many cellular processes and, crucially, prevent diverse human diseases that include cancer, neurodegenerative diseases, immune-deficiencies and premature ageing. While much progress has been made in identifying DDR proteins, much remains to be learned about the molecular and cellular functions that they control. Furthermore, the frequent reporting of new DDR proteins in the literature suggests that many others await identification. The main goals for the proposed research are to: identify important new DDR-proteins and DDR-modulators, particularly those responding to DNA double-strand breaks (DSBs); provide mechanistic insights into how these proteins function; and determine how DDR events are affected by chromatin structure, by molecular chaperones and components of the Ubiquitin and Sumo systems. To achieve these ends, we will use molecular biology, biochemical, cell-biology and molecular genetics approaches, including synthetic-lethal and phenotypic-suppression screening methods in human cells and in the nematode worm. This work will not only be of academic importance, but will also indicate how DDR dysfunction can cause human disease and how such diseases might be better diagnosed and treated.

2 482 492 €

Start date: 2011-05-01, End date: 2016-04-30

Bacterial dehalorespiration is a microbial respiratory process in which halogenated hydrocarbons, from natural or anthropogenic origin, act as terminal electron acceptors. This leads to effective dehalogenation of these compounds, and as such their degradation and detoxification. The bacterial species, their enzymes and other components responsible for this unusual metabolism have only recently been identified. Unlocking the full potential of this process for bioremediation of persistent organohalides, such as polychlorinated biphenyls (PCBs) and tetrachloroethene, requires detailed understanding of the underpinning biochemistry. However, the regulation, mechanism and structure of the reductive dehalogenase (the enzyme responsible for delivering electrons to the halogenated substrates) are poorly understood. This ambitious proposal seeks to study representatives of the distinct reductive dehalogenase classes as well as key elements of the associated regulatory systems. Our group has been at the forefront of studying the biochemistry underpinning transcriptional regulation of dehalorespiration, providing detailed insights in the protein CprK at the atomic level. However, it is now apparent that only a subset of dehalogenases are regulated by CprK homologues with little known about the other regulators. In addition, studies on the reductive dehalogenases have been hampered by the inability to purify sufficient quantities. Using an interdisciplinary, biophysical approach focused around X-ray crystallography, enzymology and molecular biology, combined with novel reductive dehalogenase production methods, we aim to provide a detailed understanding and identification of the structural elements crucial to reductive dehalogenase mechanism and regulation. At the same time, we aim to apply the knowledge gathered and study the feasibility of generating improved dehalorespiratory components for biosensing or bioremediation applications through laboratory assisted evolution.

1 148 522 €

Start date: 2008-09-01, End date: 2013-08-31

This proposal aims to address a simple question: what is the fundamental unit of computation in the brain? Answering this question is crucial not only for understanding how the brain works, but also if we are to build accurate models of brain function, which require abstraction based on identification of the essential elements for carrying out computations relevant to behaviour. In this proposal, we will build on recent work demonstrating that dendrites are highly electrically excitable to test the possibility that single dendritic branches may act as individual computational units during behaviour, challenging the classical view that the neuron is the fundamental unit of computation. We will address this question using a combination of electrophysiolgical, anatomical, imaging, molecular, and modeling approaches to probe dendritic integration in pyramidal cells and Purkinje cells in mouse cortex and cerebellum. We will first define the computational rules for integration of synaptic input in single and multiple dendrites by examining the somatic and dendritic responses to different spatiotemporal patterns of excitatory and inhibitory inputs in brain slices. Next, we will determine how these rules are engaged by patterns of sensory stimulation in vivo, by using various strategies to map the spatiotemporal patterns of synaptic inputs onto single dendrites. To understand how physiological patterns of activity in the circuit engage these dendritic computations, we will use anatomical approaches to map the wiring diagram of synaptic inputs to individual dendrites. Finally, we will perturb the dendritic computational rules by manipulating dendritic function using molecular and optogenetic tools, in order to provide causal links between specific dendritic computations and sensory processing relevant to behaviour. These experiments will provide us with deeper insights into how single neurons act as computing devices.

2 495 563 €

Start date: 2016-07-01, End date: 2021-06-30

This proposal will address how a multimodal cognitive system, the neural representation of space in the hippocampus, emerges during development. There is a long tradition in neuroscience of studying the development of primary sensory systems, but fewer studies have concentrated on the development of brain networks supporting higher-order cognitive representations. Our recent findings (Wills, Cacucci et al. Science, 2010) provide a starting point to fill this gap, charting the emergence of spatial responses of hippocampal formation neurons, using in vivo recording in awake, behaving rats. The hippocampal formation supports neural representations of the environment ('cognitive maps') by means of which an animal can locate itself and navigate to a goal location. It contains three classes of spatially-tuned cells: place cells, which code for location, head direction cells, which code for directional orientation and grid cells, which may code for distance travelled. The key aim of this proposal is to delineate the developmental processes that create this neural representation of space, focusing on the representations of place and direction. We will delineate which sensory information is capable of driving spatial firing, and whether early hippocampal coding is truly spatial in the sense of representing configurations of stimuli and not single cues. How are abstract spatial constructs (place and head direction) built from raw sensory information during development? We will test whether boundary sensitive neurons and angular velocity tuned neurons are the elemental 'building blocks' making up place and directional signals, as suggested by many theoretical models. We will also investigate the role of experience in the construction of spatial representations. Do the network architectures underlying spatial firing emerge through experience-dependent learning mechanisms, or are they the result of self-organizing processes which take place independently of experience?

1 491 930 €

Start date: 2012-01-01, End date: 2017-12-31

MRI is indispensable in the diagnosis of neurodegenerative diseases. These are poorly understood while their prevalence and socio-economic burden continue to rise. Structural and functional MRI can provide biomarkers for early diagnosis and potential therapeutic intervention. My research vision is to develop novel MRI methods for structural and functional mapping of tissue magnetic susceptibility and electrical conductivity as these show great promise for neuroimaging in diseases such as Alzheimer’s (AD). Susceptibility mapping (SM), which I pioneered, is uniquely sensitive to tissue composition including iron content affected in AD while conductivity mapping (CM) probably reflects cellular disruption in AD. Resting-state functional MRI (rsfMRI) reveals how AD affects brain networks without any tasks or stimulation equipment. However, each technique currently needs a separate time-consuming MRI scan. I will develop an integrated scan for simultaneous structural SM and CM, and rsfMRI functional connectivity characterisation. This efficient scan, ideal for AD patients, will reveal totally new resting-state networks based on electromagnetic properties: resting-state functional SM and resting-state functional CM for the first time. As changes in blood susceptibility underlie fMRI, rsfSM should measure functional connectivity more directly. This also makes it sensitive to physiological noise so I will develop noise removal methods building on fMRI techniques I established. Initial fSM studies have been at 7 Tesla but I will target the more widespread 3T field to maximise applicability. As a leader in both SM and rsfMRI physiological noise removal I have the ideal background to integrate SM and CM with fMRI and extend them for ground-breaking functional electromagnetic connectivity. This research will yield a rich set of novel, multimodal MRI contrasts to allow development of new combined structural and functional biomarkers for early diagnosis of AD and other diseases.

1 721 726 €

Start date: 2019-04-01, End date: 2024-03-31

For cells and organisms to survive and propagate, they must accurately pass on their genetic information to the next generation. Errors in this process may arise from spontaneous mistakes in normal cellular metabolism, or from exposure to external agents, such as chemical mutagens and radiation. To protect themselves from the consequences of DNA damage, cells have evolved a vast array of pathways DNA repair mechanisms, each optimised for the resolution of a particular problem. One method of DNA repair, called homologous recombination (HR), involves using intact undamaged DNA sequences as a template to repair the damaged copy. HR is used extensively in meiotic cells to repair DNA breaks that are purposely created by the cell. In this context, HR is not just a repair mechanism, but also a method to drive interaction and genetic exchange between maternally and paternally inherited chromosomes, creating haploid genomes which are chimeras of the parental genetic information. Thus, the study of DNA repair and recombination informs our understanding of mechanisms that maintain genome stability, but which also generate genetic diversity, topics that are as critical to the survival of an individual cell as they are for the evolution and survival of an entire ecosystem. In recent decades a great deal has been learned of the genetic and biochemical control of the DNA repair and recombination mechanism. In general we infer gene function from what happens (or doesn’t happen) when we mutate a pathway of interest, and use biochemistry to test function using surrogate, simplified in vitro assays. Here, to bridge the divide between these classic approaches, I propose to develop biochemical methods using intact chromatin prepared from living cells. I believe that integrating chromatin biochemistry, with cell biology and genome-wide analysis will enable a new mode of scientific investigation, detailing how molecular reactions occur on biologically-relevant chromosomal substrates.

1 747 823 €

Start date: 2013-01-01, End date: 2018-12-31

Our genetic material is continually subjected to damage, either from endogenous sources such as reactive oxygen species produced as by-products of oxidative metabolism, from the breakdown of replication forks during cell growth, or by agents in the environment such as ionising radiation or carcinogenic chemicals. To cope with DNA damage, cells employ elaborate and effective repair processes that specifically recognise a wide variety of lesions in DNA. These repair systems are essential for the maintenance of genome integrity. Unfortunately, some individuals are genetically predisposed to crippling diseases or cancers that are the direct result of mutations in genes involved in the DNA damage response. For several years our work has been at the forefront of basic biological research in the area of DNA repair, and in particular we have made significant contributions to the understanding of inheritable diseases such as breast cancer, Fanconi anemia, and the neurodegenerative disease Ataxia with Oculomotor Apraxia-1 (AOA-1). The focus of this ERC proposal is: (i) to define the phenotypic interplay between three inheritable cancer predisposition syndromes, Fanconi anemia, Bloom s syndrome and breast cancers caused by mutation of BRCA2, (ii) to determine the biological role of the newly discovered GEN1 Holliday junction resolvase in homologous recombination and repair, and (iii) to understand the actions of Aprataxin and Senataxin in relation to the inheritable neurodegenerative diseases AOA-1 and AOA-2, respectively. Our studies will provide an improved understanding of basic mechanisms of DNA repair and thereby underpin future therapeutic developments that will help individuals afflicted with these diseases.

2 449 091 €

Start date: 2010-06-01, End date: 2015-05-31

The unique properties of nucleic acids have made them the material of choice for complex nanofabrication. High fidelity formation of duplexes via non-covalent interactions between complementary sequences provides a straightforward approach to molecular programming of multicomponent self-assembly processes. The structure of the nucleic acid backbone and bases can be changed without destroying these properties, suggesting that there are all kinds of unexplored polymeric structures that will also show sequence selective duplex formation. This proposal investigates this rich new area at the interface of supramolecular, biological and polymer chemistry. The appeal of nucleic acids is that we can dial up any desired sequence via chemical solid phase synthesis or via biological template synthesis. With recent advances in polymerisation processes, which proceed under mild conditions compatible with non-covalent chemistry, we are now in a position to develop comparable processes for synthetic polymers. This proposal explores a ground-breaking approach to the synthesis of polymeric systems equipped with defined sequences of recognition sites. The aim is to establish protocols for routine solid phase synthesis of one class of oligomer, which can be used to template the synthesis of different classes of oligomer. This template chemistry will provide tools for polymerisation of conventional monomers using templates to determine the sequence of recognition sites and hence incorporate the selective recognition properties of nucleic acids into bulk polymers like polystyrene. The ability to program polymers with recognition information will open the way to new materials of unprecedented complexity and functionality with applications in all areas of nanotechnology where precise control over macromolecular structure and supramolecular organisation will be used to program mechanical, photochemical and electronic properties into sophisticated assemblies that rival biology.

2 457 947 €

Start date: 2013-03-01, End date: 2019-02-28

This project will take inspiration from biomineralisation to achieve exceptional, dynamic control over crystallisation processes. Understanding the fundamental mechanisms which govern crystallisation promises the ability to inhibit or promote crystallisation as desired, and to tailor the properties of crystalline materials towards a huge range of applications. Biomineralisation provides a perfect precedent for this approach, where organisms achieve control currently unparalleled in synthetic systems. This is achieved because mineralisation occurs within controlled environments in which an organism can interact with the nascent mineral. Thanks to recent advances in microfabrication techniques and analytical methods we finally have the tools required to bring such control to the laboratory. DYNAMIN will exploit microfluidic and confined systems to study and interact with crystallisation processes with outstanding spatial and temporal resolution. Flowing droplet devices will be coupled to synchrotron techniques to investigate and control nucleation, using soluble additives and nucleating particles to direct the crystallisation pathway. Static chambers will be used to interact with crystallisation processes over longer length and time scales to achieve spatio-temporal control to rival that in biomineralisation, while a unique confined system – titania nanotubes – will enable the study and control of organic-mediated mineralisation, using fresh reagents and proteinases to interact with the process. Finally, a key biogenic strategy will provide the inspiration to develop a simple and potentially general method to trigger and control the transformation of amorphous precursor phases to single crystal products. This will generate a new framework for studying and controlling crystallisation processes, where these new skills will find applications in sectors ranging from the Chemical Industry, to Healthcare, Advanced Materials, Formulated Products and the Environment.

2 632 375 €

Start date: 2018-09-01, End date: 2023-08-31

The project addresses the long-term vision of man-made materials with chemical selectivity and functional efficiency produced by dynamic structural flexibility. These materials are not intended as protein mimics; they are however inspired by nature’s use of flexible rather than rigid systems, with their ability to dynamically restructure around guests and thus perform highly specific chemistry. Such materials would transform chemical processes through their precision, for example by reorganising to accelerate each step of a cascade reaction without reagent or product inhibition. The road to this vision is blocked as we do not have the methodology and understanding to control such materials. The aim is to develop synergic, multidisciplinary experimental and computational capability to harness the dynamics of flexible crystalline porous solids for function, demonstrated in separation and catalysis. This will enable design and synthesis of materials that controllably adopt distinct structures according to their chemical environment to optimise performance. We will create a new workflow that integrates understanding of the structure-composition-dynamics-property relationship into the materials design and discovery process. This workflow builds on proof-of-concept in (i) chemical control of dynamical restructuring in flexible crystalline porous materials and in the use of dynamics to (ii) enhance function and (iii) guide synthesis. Crystalline flexible porous materials are selected because crystallinity maximises the atomic-scale understanding generated, which is transferable to other materials classes, whilst porosity permits sorption and organisation of guests that controls function. This inorganic materials chemistry project develops integrated capability in chemical synthesis (new metal-organic frameworks and linkers), computation (prediction and evaluation of structure and dynamical guest response), characterisation (e.g. by diffraction) and measurement of function.

2 493 425 €

Start date: 2016-10-01, End date: 2021-09-30

This high-impact, challenging CoG Proposal integrates multiple novel ideas in boron and zinc chemistry into an overarching project to open up new horizons across synthesis and catalysis. The Applicant’s successful ERC StG has opened up new avenues of pioneering research in main group element mediated transformations that were not conceivable before the work was done. Components of this proposal extend out from the StG into new, exciting research areas that are completely different. Developing low toxicity earth abundant catalysts for important transformations is vital to the EU with the focus herein being on; (i) the Suzuki-Miyaura (S-M) cross coupling reaction which is ubiquitous in industry and academia, and (ii) the formation of organoboranes that are essential synthetic intermediates. Both of these are currently dominated by toxic, expensive and low abundance precious metal catalysts (e.g. Pd, Ir). This project will deliver innovation through utilising combinations of main group Lewis acids and nucleophilic anions that do not react with each other, i.e. are frustrated pairs. This “frustration” enables the two species to concertedly transform substrates to achieve: (i) Precious metal-free S-M cross coupling reactions of sp3C electrophiles catalysed by zinc and boron compounds, including stereospecific couplings and one pot two step cross electrophile couplings. (ii) Trans-elementoboration of alkynes, including the unprecedented fluoroboration of alkynes. Other new approaches will be developed to access novel (hetero)arylboronic acid derivatives using only simple boranes and without requiring noble metal catalysts, specifically: (i) boron directed C-H borylation and (ii) directed ortho borylation to enable subsequent meta selective SEAr C-H functionalisation. This CoG will afford the freedom and impetus via consolidated funding to undertake fundamental research to deliver high impact results, including developing a new area of cross coupling catalysis research.

2 070 093 €

Start date: 2018-05-01, End date: 2023-04-30

Structure determination of G protein-coupled receptors (GPCRs) has been exceedingly successful over the last 5 years due to the development of complimentary generic methodologies that will now allow the structure determination of virtually any GPCR. However, these technologies address only two aspects of the process, namely the stability of the receptors during purification and the ability to form well-diffracting crystals. The strategies also apply only to GPCRs and not transporters or ion channels. The recent successes have been of GPCRs that are expressed in either yeasts or in insect cells using the baculovirus expression system, but many membrane proteins are expressed poorly in these systems or may be expressed in a misfolded non-functional form. A second issue with the future structure determination of GPCRs is the lack of generic technologies to allow the crystallisation of arrestin-GPCR and G protein-GPCR complexes. Although one G protein GPCR complex has been crystallised this was exceedingly diffciult and resulted in poor resolution of the GPCR component of the complex. We believe that it is possible to thermostabilise both arrestin and heterotrimeric G proteins, which will allow a simplified strategy for the crystallisation and structure determination of GPCR complexes. This is based on the development of the strategy of conformational thermostabilisation of GPCRs developed in our lab that has resulted in the structure determination of 3 different GPCRs bound to either antagonists, partial agonists, full agonists and/or biased agonists. The aims are: 1. The development of generic methodology for the production of eukaryotic membrane proteins in mammalian cells. 2. The development of a thermostable functional arrestin mutant 3. Structures of β1-adrenoceptor, adenosine A2A receptor and angiotensin receptor bound to a G protein and arrestin 4. Understanding the role of each amino acid residue in the activation process of GPCRs through saturation mutagenes

2 378 162 €

Start date: 2014-02-01, End date: 2019-01-31

ENBION will engineer a platform to direct the differentiation of stem cells by developing principles for the rational design of the biointerface of nanowires. It is increasingly evident that efficient tissue regeneration can only ensue from combining the regenerative potential of stem cells with regulatory stimuli from gene therapy and niche engineering. Yet, despite significant advances towards integrating these technologies, the necessary degree of control over cell fate remains elusive. Vertical arrays of high aspect ratio nanostructures (nanowires) are rapidly emerging as promising tools to direct cell fate. Thanks to their unique biointerface, nanowires enable gene delivery, intracellular sensing, and direct stimulation of signalling pathways, achieving dynamic manipulation of cells and their environment. This broad manipulation potential highlights the importance and timeliness of engineering nanowires for regenerative medicine. However, developing a nanowire platform to direct stem cell fate requires design principles based on the largely unknown biological processes governing their interaction with cells. Enabling localized, vector-free gene therapy through efficient transfection relies on understanding the still debated mechanisms by which nanowires induce membrane permeability. Directing cell reprogramming requires understanding the largely unexplored mechanosensory processes and the resulting epigenetic effects arising from the direct interaction of nanowires with multiple organelles within the cell. Engineering the cell microenvironment requires yet undeveloped strategies to localize signalling and transfection with a resolution comparable to the lengthscale of cells. ENBION will develop this critical knowledge and integrate it into guidelines for dynamic manipulation of cells. Beyond the nanowire platform, the principles highlighted by this unique interface can guide the development of nanomaterials with improved control over cellular processes.

1 495 430 €

Start date: 2018-02-01, End date: 2023-01-31

The remarkable way that Nature prepares complex natural products has always been a source of inspiration to scientists, stimulating the development of new synthetic methods and strategies, as elegantly demonstrated by biomimetic approaches to total synthesis. Similarly, the performance and specificity of enzymes, perfected though evolution, offer ideals of selectivity and specificity that synthetic chemists aspire to. This proposal aims to develop an internationally leading research programme inspired by Nature’s ability to selectively generate diverse products from simple materials with exquisite levels of regio- and enantiocontrol. We aspire to synthetically emulate the elegant behaviour of Nature’s building blocks, such as co-enzyme A, in their ability to generate synthetic diversity (such as polyketides and alkaloids) from a simple and common starting material. Using this blueprint, we intend to selectively control the synthesis of a diverse range of bespoke stereodefined carbo- and heterocycles from readily available starting materials using simple man-made catalysts. We specifically aim to develop new strategies within the field of organic catalysis, focused upon the development of methods for the in situ catalytic generation of chiral ammonium enolates from carboxylic acids and their employment in catalysis. We also propose to develop a comprehensive mechanistic understanding of these processes. In preliminary work we have delineated a simple and efficient approach to this problem by employing chiral isothioureas as asymmetric catalysts, and we aim to build on the insight provided by these studies to develop this powerful concept into a generally applicable synthetic strategy.

1 497 005 €

Start date: 2011-10-01, End date: 2017-06-30