Despite significant advances in chemotherapy, the effective treatment of malignant masses via systemically injectable agents are still limited by insufficient accumulation at the biological target (<< 10% injected dose per gram tumor) and non-specific sequestration by the reticulo-endothelial system (tumor/liver < 0.1). The goal of this proposal is to engineer Discoidal Polymeric Nanoconstructs (DPNs) to preferentially target the malignant neovasculature for the delivery of imaging agents, controlled release of therapeutic molecules and thermal energy. The central hypothesis is that the size, shape, surface properties and stiffness (4S parameters) of the DPNs can be controlled during synthesis, and that therapeutic molecules (Temozolomide), Gd(DOTA) complexes and ultra-small Super-Paramagnetic Iron Oxide nanoparticles (USPIOs) can be efficiently incorporated within the DPN polymeric matrix. This will be achieved by pursuing 3 specific aims: i) synthesis and physico-chemical characterization of poly(lactic-co-glycolic acid)/poly(ethylene glycol) DPNs with multiple 4S combinations; ii) in-silico and in vitro rational selection of DPN configurations with preferential tumor deposition, low macrophage uptake and high loading; and iii) in-vivo testing of the DPN imaging and therapeutic performance in mice bearing Glioblastoma Multiforme (GBM). The innovation stays in i) using synergistically three different targeting strategies (rational selection of the 4S parameters; magnetic guidance via external magnets acting on the USPIOs; specific ligand-receptor recognition of the tumor neovasculature); ii) combining therapeutic and imaging molecules within the same nanoconstruct; and iii) employing synergistically different therapeutic approaches (molecular and thermal ablation therapies). This would allow us to support minimally invasive screening via clinical imaging and enhance therapeutic efficacy in GBM patients.

In Naturale CG I propose transformative bioengineering approaches that will overcome severe limitations in current materials in two main areas, namely 1) Biosensing and 2) Regenerative Medicine. A key focus is on understanding and engineering the biomaterial interface using innovative designs and state of the art materials characterisation methods. Firstly I aim to transform the way that we can currently detect disease through innovations in the design and development of nanomaterials-based biosensors that could be used to detect a number of diseases with global implications, such as cancer, malaria, heart failure and tuberculosis. These innovations in biosensor design will involve both building on our existing highly successful work on plasmonic biosensors and also involve the design and development of completely new polymersome and fluorescent based biosensors. Another key aim of Naturale CG is to design first in kind biosensors for the facile detection of microRNAs. Secondly, the goal of regenerating failing organs before the body as a whole is ready to surrender, is now timelier than ever and one in which the design of new bio-inspired materials can play an important role. In Naturale CG I will build on my previous research in the design of 3-dimensional tissue engineering scaffolds and address an important new direction in the engineering of new bio-inspired conducting polymers as tissue engineering materials to promote cardiac tissue regeneration. First-in-field biomaterials-based innovations generated from this programme could enable far more effective regeneration of functional myocardial tissue which has been notoriously difficult to achieve thus far. Whilst I will lead this grant and the research within it, the proposed innovations are truly multidisciplinary in nature and will be accelerated towards clinical translation through the numerous clinical, scientific and industrial collaborations that I have established.

This project, entitled 'Hybrid polymer-peptide hydrogels for Cell Therapy' (HYCELT) will produce cost-effective neuronal stem cell (NSC) scaffolds for central nervous system regeneration using a fibrous gel that has all the necessary requisites for fast clinical translation: cell-instructive, biocompatible, injectable and biodegradable. Finding a perfect methodology to heal spinal cord damage is a challenge of current times. Peptides are used for this purpose because they are biocompatible, biodegradable and will produce suitable gelatinous environments for cell growth at extremely low concentrations. The target of HYCELT is to demonstrate this technology in vitro, with follow-on work focussed on in vivo studies in appropriate animal models and in collaboration with clinical scientists to produce a global therapy that will change the lives of thousands of people suffering from irreparable spinal cord injury. HYCELT brings together a talented young fellow with a background in polymer hydrogels with an internationally blossoming group in polymer nanotechnology and well-established UK groups in peptide/materials science and biomaterials. HYCELTs multidisciplinary solution will exploit polymer-peptide hydrogel system also be applied in the vast field of drug delivery. Moreover as it will be developed in concert with Merck Serono (Italy), the biotechnology world-leaders in the field of therapeutics, it will be immediately applicable to living system. The supervisory team has been selected from experts at Aston, Sheffield and Merck (Rome) to provide a multidisciplinary training programme for the fellow and to ensure success of the ambitious research targets of HYCELT.

The use of Acellular Technology to stimulate the body's own repair processing is an emerging direction in regenerative medicine, which may tackle one of challenges in orthopedic treatment where how to treat bone loss or bone fracture due to osteoporosis is unsolved. The main objective of this project is to biofabricate new bioinspired scaffolds enabling off-shelf, scalable and stimulating multiple cellular actions including stem cell infiltration/adhesion; differentiation of osteoblast and endothethial cells for treatment of bone defects. The proposal will develop new multifaceted 3D composite scaffolds combined the advantages of electrospun nanofibers, porous foam and active chemicals conjugation enabling to provide biological, physical and mechanical properties for mesenchymal stem cells adhesion, migration, and differentiation and angiogenesis. The proposal will also develop an in vitro dynamic culture model mimicking bone development in vivo to evaluate the efficiency of scaffolds in order to reduce the use of animal models. The project proposes to bring an outstanding young researcher, Dr Lü, who was trained in the State Key Laboratory of Bioelectronics in Southeast University, the top 20 university in China, to undertake such multidisciplinary research project with host institution in Keele University, UK. Dr Lü has worked in biofabrcation field in past 5 years, and have established excellent track record in smart nanofiber fabrication and applications. The fellow's experience and knowledge will be effectively transferred to host group and other European research groups through the delivering the targets of the project and disseminating of her previous research outcomes by seminar, joint publication and visiting other laboratories. The fellowship will also aim to build collaboration with third country through Dr Lü's research links in China after the fellowship, which will enhance European knowledge-based economy in general and healthcare in specifically.

The mechanical properties of graphene, the thinnest material in the world, will be investigated theoretically. This project will focus on the basic and advanced mechanical properties that are potentially useful for controlling: i) the strain distribution, ii) the band gap, and iii) the observation and visualization of electronic polarization in single/multilayer graphene. The theoretical

Most cancers remain 'incurable' and life-thretening. Cancer stem cells (CSCs) are the source of chemo/radioresistance and responsible for cancer recurrence which suggests the urgent requirement of CSC-targeting drugs. Drug development is a slow (15 years/drug) and costly (US$1.5bn/drug) procedure with only 5-25% of new oncology drugs in clinical development actually reaching the market mainly due to the toxicity of novel molecules. This dilemma has led to an increasing appreciation of the potential of repurposing of known drugs. We have demonstrated that Disulfiram (DS), an old anti-alcoholism drug, possesses excellent anti-CSC activity with low toxicity to normal cells. Whereas its cancer clinial indication is limited by its bio-instability (~4 min half-life in blood stream). Our pilot data demonstrated that the anticancer efficacy of DS is significantly improved when mild extending its half-life by liposome encapsulation. In this study, the Incoming Fellow, who has very strong technical knowhow in cancer research, molecular pharmacology, anticancer drug development and nano-encapsulation, will bring novel nano-biomaterials invented in China into Europe. Taking advantage of the state-of-the-art facilities, CSC models, pharmaceutical resarch and developmental expertise and scientific/technical support from the Incoming Host and the other European collaborators, we will develop a long-circulating nano-encapsulated DS. The anticancer activity of the nano-encapsulated DS will be examined in vitro and in vivo in breast and liver cancer cell lines as well as the relevant CSC models. This study will pave the path for clinical trial of DS in cancer indication. The significance of this project will be: 1. Expand and extend our FP7-IRSES (2011-16) platform to strenthen long-term collaboration between China and EU partners; 2. Develop a new cancer therapeutics for the benefic of healthcare in Europe; 3. Open a new drug developmental window to benefit European economy.

As a result of the research undertaken in the NANOFORCELLS project (ERC-StG-2011-278860), we have successfully developed instrumentation for the investigation of cell mechanics to discriminate cancer cells from healthy cells by novel physical biomarkers, placing a particular emphasis in the elasticity and viscoelasticity of the cells. In order to accelerate the market entry of the results, the present project will focus on conducting a market feasibility study of the nanomechanical flow cytometer developed, as well as clarifying the IP position strategy and making an initial approach to potential partners and investors. The nanomechanical device to be taken to proof-of-concept, provides label-free classification of cells attending to their volume, mass, density and viscoelasticity. The device is capable of enabling parallel measurements of physical properties of hundreds of individual cells per minute, opening the route for portable tests that would enable determining the health status of cells from blood samples (f.e. leukaemia) based on their mechanical properties. This device will become an ideal tool for the study of drug effects on the physical properties of the cell with research and prognostic value, once made commercially available. The expected outcomes of the PoC project will be an increased understanding of the target markets and the definition of a commercialization roadmap to exploit the device. We envision that research groups within this biomedical field would be a first target market for the product use. The work undertaken so far requires further investment in order to bring the laboratory device to a marketable device and turn the outputs of the NANOFORCELLS project into a commercial proposition.

Peptide macrocycles can bind with high affinity and selectivity to protein targets and are an attractive class of molecules for the development of therapeutics. Recently, a phage display-based strategy was developed that allowed to generate potent bicyclic peptide antagonists (Heinis, C., et al., Nat. Chem. Biol., 2009). While bicyclic peptides with nanomolar affinities to a range of protein targets could be generated, it was more difficult to obtain high-affinity binders to some proteins, particularly to those having flat surfaces and no clefts or cavities. Herein, I propose to develop rigid, tricyclic peptides that should, due to a more defined three-dimensional structure, bind to flat surfaces similar as antibodies. Two formats are envisioned for the synthesis of tricyclic peptides: in the first one, a linear peptide is anchored via four cysteine residues to a small molecule while in the second format, bicyclic peptides will be generated and their two peptide rings are connected via Huisgen cycloaddition reaction to impose an additional conformational constraint. Phage-encoded combinatorial libraries of these peptide folds will be generated and subjected to affinity selections. Tricyclic peptide binding to a variety of biological targets including (a) the well-characterized cancer-associated targets EGFR and HER2, and (b) the more challenging target of the antibiotic vancomycin, the short peptide D-Ala-D-Ala, will be developed.

Cell-to-cell communication pathways coordinate cellular functions in multicellular organisms. Cells that are nearest neighbours can communicate through specific interactions between ligand and receptor proteins present in their respective cell membranes. The objective of this research program is to address the hypothesis that the physical context of the ligand/receptor interaction contributes to defining the fundamental mechanisms of action of cell-to-cell communication pathways and their cellular outcomes. The research program relies on the development of tools that provide well-defined physical inputs to cells, not confounded by simultaneous changes in chemical inputs. Therefore, beyond state-of-the-art developments in nanotechnology are here integrated with cell biology. In particular, DNA origami technology is applied to the development of ligand nanoclusters with customized spatial organization and mechanical properties. These ligand nanoclusters are used to probe the roles of physical properties of the ligand presentation on the activation of intracellular signalling pathways. We will focus on the ephrin/Eph cell-to-cell communication pathway, which regulates embryonic development and the homeostasis of adult organs. ephrin/Eph signalling is commonly disrupted in cancer, showing tumour suppressing or tumour promoting character. The mechanisms that generate the diversity of outcomes of the ephrin/Eph pathway are largely unknown. We will use DNA origami/ephrin ligand nanoclusters to investigate whether the spatial organization and mechanical properties of ephrin ligand assemblies impact Eph receptor function and contribute to generating diversity in the pathway. Our novel approach is readily transferrable to the study of other signalling pathways. We aim to generate a knowledge foundation for the roles of mechanotransduction, the conversion of physical to biochemical signals, in cell-to-cell communication mediated by membrane-bound ligands and receptors.

Biological cells possess a chemical 'sense of smell' and a physical 'sense of touch'. Structure, dynamics, development, differentiation and even apoptosis of cells are guided by physical stimuli feeding into a regulatory network integrating biochemical and mechanical signals. Cells are equipped with both, force-generating structures, and stress sensors including force-sensitive structural proteins or mechanosensitive ion channels. Pathways from force sensing to structural and transcriptional controls are not yet understood. The goal of the proposed interdisciplinary project is to quantitatively establish such pathways, connecting the statistical physics and the mechanics to the biochemistry. We will measure and model the complex non-equilibrium mechanical structures in cells, and we will study how external and cell-generated forces activate sensory processes that (i) act (back) on the morphology of the cell structures, and (ii) lead to cell-fate decisions, such as differentiation. The most prominent stress-bearing and -generating structures in cells are actin/myosin based, and the most prominent mechanoactive and -sensitive cell types are fibroblasts in connective tissue and myocytes in muscle. We will first focus on actin/myosin bundles in fibroblasts and in sarcomeres in developing heart muscle cells. We will observe cells under the influence of exactly controlled external stresses. Forces on suspended single cells or cell clusters will be exerted by laser trapping and sensitively detected by laser interferometry. We furthermore will monitor mechanically triggered transcriptional regulation by detecting mRNA in the nucleus of mouse stem cells differentiating to cardiomyocytes. We will develop fluorescent mRNA sensors that can be imaged in cells, based on near-IR fluorescent single-walled carbon nanotubes. Understanding mechanical cell regulation has far-ranging relevance for fundamental cell biophysics, developmental biology and for human health.