ERC FUNDED PROJECTS

Monoclonal antibodies against the EGF receptor (EGFR) provide substantive benefit to colorectal cancer (CRC) patients. However, no genetic lesions that robustly predict ‘addiction’ to the EGFR pathway have been yet identified. Further, even in tumours that regress after EGFR blockade, subsets of drug-tolerant cells often linger and foster ‘minimal residual disease’ (MRD), which portends tumour relapse. Our preliminary evidence suggests that reliance on EGFR activity, as opposed to MRD persistence, could be assisted by genetically-based variations in transcription factor partnerships and activities, gene expression outputs, and biological fates controlled by the WNT/beta-catenin pathway. On such premises, BEAT (Beta-catenin and EGFR Abrogation Therapy) will elucidate the mechanisms of EGFR dependency, and escape from it, with the goal to identify biomarkers for more efficient clinical management of CRC and develop new therapies for MRD eradication. A multidisciplinary approach will be pursued spanning from integrative gene regulation analyses to functional genomics in vitro, pharmacological experiments in vivo, and clinical investigation, to address whether: (i) specific genetic alterations of the WNT pathway affect anti-EGFR sensitivity; (ii) combined neutralisation of EGFR and WNT signals fuels MRD deterioration; (iii) data from analysis of this synergy can lead to the discovery of clinically meaningful biomarkers with predictive and prognostic significance. This proposal capitalises on a unique proprietary platform for high-content studies based on a large biobank of viable CRC samples, which ensures strong analytical power and unprecedented biological flexibility. By providing fresh insight into the mechanisms whereby WNT/beta-catenin signalling differentially sustains EGFR dependency or drug tolerance, the project is expected to put forward an innovative reinterpretation of CRC molecular bases and advance the rational application of more effective therapies.

1 793 421 €

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

In 2010, 800 billion Euros was spent on brain diseases in Europe and the cost is expected to increase due to the aging population. – Here I propose to exploit our new discovery for research to alleviate this disease burden. In work selected by Nature Medicine among the top 10 ”Notable Advances” and by Science as one of the 10 ”Breakthroughs of the year” 2015, we discovered a meningeal lymphatic vascular system that serves brain homeostasis. We want to reassess current concepts about cerebrovascular dynamics, fluid drainage and cellular trafficking in physiological conditions, in Alzheimer’s disease mouse models and in human postmortem tissues. First, we will study the development and properties of meningeal lymphatics and how they are sustained during aging. We then want to analyse the clearance of macromolecules and protein aggregates in Alzheimer’s disease in mice that lack the newly discovered meningeal lymphatic drainage system. We will study if growth factor-mediated expansion of lymphatic vessels alleviates the parenchymal accumulation of neurotoxic amyloid beta and pathogenesis of Alzheimer’s disease and brain damage after traumatic brain injury. We will further analyse the role of lymphangiogenic growth factors and lymphatic vessels in brain solute clearance, immune cell trafficking and in a mouse model of multiple sclerosis. The meningeal lymphatics could be involved in a number of neurodegenerative and neuroinflammatory diseases of considerable human and socioeconomic burden. Several of our previous concepts have already been translated to clinical development and we aim to develop proof-of-principle therapeutic concepts in this project. I feel that we are just now in a unique position to advance frontline European translational biomedical research in this suddenly emerging field, which has received great attention worldwide.

2 420 429 €

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

We aim to develop and apply a suite of new technologies in a novel cancer discovery platform that will link high-definition cancer biology, via state-of-the-art disease imaging and pathway modelling, with development of novel interrogative and therapeutic interventions to test in models of cancer that closely resemble human disease. The work will lead to a new understanding of cancer invasion, how to treat advanced disease in the metastatic niche, how to monitor therapeutic responses and the compensatory mechanisms that cause acquired resistance. Platform development will be based on combined, cross-informing technologies that will enable us to predict optimal ‘maintenance therapies’ for metastatic disease by targeting cancer evolution and spread through combination therapy. A key strand of the platform is the development of quantitative multi-modal imaging in vivo by use of optical window technology to inform detailed understanding of disease and drug mechanisms and predictive capability of pathway biomarkers. Innovative methodologies are urgently needed to address declining approval rates of novel medicines and the unmet clinical needs of treating cancer patients in the advanced disease setting, where tumour spread and survival generally continues unchecked by current therapies. This work will be largely pre-clinical, but will always be mindful of the clinical problem in managing late stage human disease through rationale design of combination therapies with companion diagnostic tests. The cancer survival statistics will be changed if we can curb continuing spread of aggressive, metastatic disease and resistance to therapy by taking smarter combined approaches that make best use of emerging technologies in an innovative way, particularly where they are more predictive of clinical efficacy.

2 499 000 €

Start date: 2012-08-01, End date: 2017-07-31

An important goal of stem cell therapy is to create “customized” cells that are genetically identical to the patient, which upon transplantation can restore damaged tissues. Such cells can be obtained by in vitro direct reprogramming of somatic cells into embryonic stem (ES)-like cells, termed induced pluripotent stem cells (iPSC). This approach also opens possibilities for modelling human diseases in vitro. However, major hurdles remain that restrain fulfilling conventional human iPSC/ESC potential, as they reside in an advanced primed pluripotent state. Such hurdles include limited differentiation capacity and functional variability. Further, in vitro iPSC based research platforms are simplistic and iPSC based “humanized” chimeric mouse models may be of great benefit. The recent isolation of distinct and new “mouse-like” naive pluripotent states in humans that correspond to earlier embryonic developmental state(s), constitutes a paradigm shift and may alleviate limitations of conventional primed iPSCs/ESCs. Thus, our proposal aims at dissecting the human naïve pluripotent state(s) and to unveil pathways that facilitate their unique identity and flexible programming. Specific goals: 1) Transcriptional and Epigenetic Design Principles of Human Naïve Pluripotency 2) Signalling Principles Governing Human Naïve Pluripotency Maintenance and Differentiation 3) Defining Functional Competence and Safety of Human Naïve Pluripotent Stem Cells in vitro 4) Novel human naïve iPSC based cross-species chimeric mice for studying human differentiation and disease modelling in vivo. These aims will be conducted by utilizing engineered human iPSC/ESC models, CRISPR/Cas9 genome-wide screening, advanced microscopy and ex-vivo whole embryo culture methods. Our goals will synergistically lead to the design of strategies that will accelerate the safe medical application of human naive pluripotent stem cells and their use in disease specific modelling and applied stem cell research.

2 000 000 €

Start date: 2017-11-01, End date: 2022-10-31

Despite decades of research, developing ways to overcome drug resistance in cancer is the most challenging bottleneck for curative therapies. This is because, in some forms of cancer, the cancer stem cells from which the diseases arise are constantly evolving, particularly under the selective pressures of drug therapies, in order to survive. The events leading to drug resistance can occur within one or more individual cancer stem cell(s) – and the features of each of these cells need to be studied in detail in order to develop drugs or drug combinations that can eradicate all of them. The BCR-ABL+ and BCR-ABL- myeloproliferative neoplasms (MPN) are a group of proliferative blood diseases that can be considered both exemplars of precision medicine and of the drug resistance bottleneck. While significant advances in the management of MPN have been made using life-long and expensive tyrosine kinase inhibitors (TKI), patients are rarely cured of their disease. This is because TKI fail to eradicate the leukaemia stem cells (LSC) from which MPN arise and which persist in patients on treatment, often leading to pervasive drug resistance, loss of response to therapy and progression to fatal forms of acute leukaemia. My goal is to change the way we study the LSC that persist in MPN patients as a means of delivering more effective precision medicine in MPN that is a “game-changer” leading to therapy-free remission (TFR) and cure. Here, I will apply an innovative strategy, ChAMPioN, to study the response of the MPN LSC to TKI in innovative pre-clinical laboratory models and directly in patients with MPN - up to the resolution of individual LSC. This work will reveal, for the first time, the molecular and clonal evolution of LSC during TKI therapies, thus enabling the development of more accurate predictions of TKI efficacy and resistance and rational approaches for curative drug therapies.

3 005 818 €

Start date: 2018-01-01, End date: 2022-12-31

Acute myeloid leukemia (AML) is a group of hematologic malignancies which, due to their molecular and clinical heterogeneity, have been traditionally difficult to classify and treat. Recently, next-generation, whole-genome sequencing has uncovered several recurrent somatic mutations that better define the landscape of AML genomics. Despite these advances in deciphering AML molecular subsets, there have been no concurrent improvements in AML therapy which still relies on the ‘antracycline+cytarabine’ scheme. Hereto, only about 40-50% of adult young patients are cured whilst most of the elderly succumb to their disease. Therefore, new therapeutic approaches which would take advantage of the new discoveries are clearly needed. In the past years, we discovered and characterized nucleophosmin (NPM1) mutations as the most frequent genetic alteration (about 30%) in AML, and today NPM1-mutated AML is a new entity in the WHO classification of myeloid neoplasms. However, mechanisms of leukemogenesis and a specific therapy for this leukemia are missing. Here, I aim to unravel the complex network of molecular interactions that take place in this distinct genetic subtype, and find their vulnerabilities to identify new targets for therapy. To address this issue, I will avail of relevant pre-clinical models developed in our laboratories and propose two complementary strategies: 1) a screening-based approach, focused either on the target, by analyzing synthetic lethal interactions through CRISPR-based genome-wide interference, or on the drug, by high-throughput chemical libraries screenings; 2) a hypothesis-driven approach, based on our recent gained novel insights on the role of specific intracellular pathways/genes in NPM1-mutated AML and on pharmacological studies with ‘old’ drugs, which we have revisited in the specific AML genetic context. I expect our discoveries will lead to find novel therapeutic approaches and make clinical trials available to patients as soon as possible.

1 883 750 €

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

Making cancer treatment more personalized and effective is one of the grand challenges in our health care system. However, many drugs have entered clinical trials but so far showed limited efficacy or induced rapid development of resistance. We critically need multi-targeted drug combinations, which shall selectively inhibit the cancer cells and block the emergence of drug resistance. This project will develop mathematical and computational tools to identify drug combinations that can be used to provide personalized and more effective therapeutic strategies that may prevent acquired resistance. Utilizing molecular profiling and pharmacological screening data from patient-derived leukaemia and ovarian cancer samples, I will develop model-based clustering methods for identification of patient subgroups that are differentially responsive to first-line chemotherapy. For patients resistant to chemotherapy, I will develop network modelling approaches to predict the most potential drug combinations by understanding the underlying drug target interactions. The drug combination prediction will be made for each patient and will be validated using a preclinical drug testing platform on patient samples. I will explore the drug combination screen data to identify significant synergy at the therapeutically relevant doses. The drug combination hits will be mapped into signalling networks to infer their mechanisms. Drug combinations with selective efficacy in individual patient samples or in sample subgroups will be further translated into in treatment options by clinical collaborators. This will lead to novel and personalized strategies to treat cancer patients.

1 500 000 €

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

Vascularization, the process in which new blood vessels assemble, is fundamental to tissue vitality. Vessel network assembly within 3D tissues can be induced in-vitro by means of multicellular culturing of endothelial cells (EC), fibroblasts and cells specific to the tissue of interest. This approach supports formation of endothelial vessels and promotes EC and tissue-specific cell interactions. Such EC-dependent tube-like openings may also form the basis for improved media penetration to the inner regions of thick 3D constructs, allowing for enhanced construct survival and for effective engineering of large complex tissues in the lab. Moreover, our own breakthrough results describe the beneficial impact of in vitro prevascularization of engineered muscle tissue on its survival and vascularization upon implantation. These studies have also demonstrated that implanted vascular networks of in vitro engineered constructs, can anastomose with host vasculature and form functional blood vessels in vivo. However, the mechanisms underlying enhanced vascularization of endothelialized engineered constructs and implant-host vessel integration remain unclear. In this proposal, our research objectives are (1) to uncover the mechanisms governing in vitro vessel network formation in engineered 3D tissues and (2) to elucidate the process of graft-host vessel network integration and implant vessel-stimulated promotion of neovascularization in vivo. In addition, the impact of construct prevascularization on implant survival and function will be explored in animal disease models. While there are still many challenges ahead, should we succeed, our research could lay the foundation for significantly enhanced tissue construct vascularization procedures and for their application in regenerative medicine. In addition, it may provide alternative models for studying the vascularization processes in embryogenesis and disease.

1 500 000 €

Start date: 2012-10-01, End date: 2017-09-30

Heart failure (HF) affects 15 million Europeans and entails higher mortality and health care costs than cancer. EPLORE addresses this issue by prospective epidemiological research in 4 European countries and by a proof-of-concept clinical trial. WP1 will for the first time at the population level document the incidence and progression of subclinical LV dysfunction and clarify whether asymptomatic LV dysfunction, as picked up by the newest echocardiographic techniques, predicts cardiovascular (CV) outcomes, including HF. WP2 will investigate the contribution of ventricular-arterial coupling disease and mechanical LV dyssynchrony to subclinical LV dysfunction. WP3 will identify a set of urinary polypeptides that signify early LV dysfunction and validate these biomarkers by showing that they predict deterioration of LV function, progression to HF and the incidence of CV complications over and beyond established risk factors. WP3 will also search for novel panels of circulating biomarkers, of which combined measurement will add information (accuracy, sensitivity and specificity) to established biomarkers (e.g., NT-proBNP) and identify genetic variants involved in the progression of LV dysfunction, either causally or as biomarker. WP4 consists of a randomised clinical trial to translate in a high-risk high-gain setting the results of WPs 1-3 into clinical practice and to identify a new treatment modality that potentially slows progression of diastolic LV dysfunction. Dissemination in WP5 will contribute to new guidelines for the prevention and treatment of HF. WP6 includes governance, monitoring research strategies and output, and protection of IPR. In conclusion, EPLORE will advance risk stratification and the early diagnosis of subclinical HF. The project will potentially result into specific treatments for diastolic LV dysfunction and inform guidelines for prevention and treatment of HF. It will benefit 20% of Europeans who currently have subclinical LV dysfunction.

2 391 440 €

Start date: 2012-07-01, End date: 2017-06-30

We are witnessing transformative results in the clinical application of both cancer immunotherapies and gene transfer technologies. Tumor vaccines are a specific modality of cancer immunotherapy. Similar to vaccination against pathogens, tumor vaccines are designed to elicit a specific immune response against cancer. They are based on the administration of inactivated cancer cells or tumor antigens, or the inoculation of antigen-presenting cells (APCs) previously exposed to tumor antigens. In spite of significant development and testing, tumor vaccines have largely delivered unsatisfactory clinical results. Indeed, while some patients show dramatic and durable cancer regressions, many do not respond, highlighting both the potential and the shortcomings of current vaccination strategies. Hence, identifying and abating the barriers to effective cancer vaccines is key to broadening their therapeutic reach. The goal of EVOLVE (EVirs to Optimize and Leverage Vaccines for cancer Eradication) is to propel the development of effective APC-based tumor vaccines using an innovative strategy that overcomes several key hurdles associated with available treatments. EVOLVE puts forward a novel APC engineering platform whereby chimeric receptors are used to both enable the specific and efficient uptake of cancer-derived extracellular vesicles (EVs) into APCs, and to promote the cross-presentation of EV-associated tumor antigens for stimulating anti-tumor immunity. EVOLVE also envisions a combination of ancillary ‘outside of the box’ interventions, primarily based on further APC engineering combined with innovative pre-conditioning of the tumor microenvironment, to facilitate the deployment of effective APC-driven, T-cellmediated anti-tumor immunity. Further to preclinical trials in mouse models of breast cancer and melanoma, our APC platform will be used to prospectively identify novel human melanoma antigens and reactive T cell clones for broader immunotherapy applications.

1 958 919 €

Start date: 2017-07-01, End date: 2022-06-30

Despite technological advances, industry struggles to develop new pharmaceuticals and therefore novel strategies for drug discovery are urgently needed. G protein-coupled receptors (GPCRs) play important roles in numerous physiological processes and are important drug targets for neurological diseases. My research focuses on modelling of GPCR-ligand interactions at the atomic level, with the goal to increase knowledge of receptor function and develop new methods for drug discovery. Breakthroughs in GPCR structural biology and access to sensitive screening assays provide opportunities to utilize fragment-based lead discovery (FBLD), a powerful approach for drug design. The objective of the project is to create a computational platform for FBLD, with a vision to transform the early drug discovery process for GPCRs. As structural information for these targets is limited, predictive models of receptor-fragment complexes will be crucial for the successful use of FBLD. In this project, computational structure-based methods for discovery of fragment ligands and further optimization of these to potent leads will be developed. These techniques will be applied to address two difficult problems in drug discovery. The first of these is to design ligands of peptide-binding GPCRs that have been challenging for existing methods. One of the promises of FBLD is to provide access to difficult targets, which will be explored by combining molecular docking and biophysical screening against peptide-GPCRs to identify novel lead candidates. A second challenge is that efficient treatment of neurological disorders often requires modulation of multiple targets, which also will be the focus of the project.

1 467 500 €

Start date: 2017-11-01, End date: 2022-10-31

This application addresses a major problem in therapy of solid tumors: poor penetration of anti-cancer drugs into tumor tissue and to infiltrating tumor cells. Recently, we have identified tumor penetrating peptides (TPP) that trigger specific penetration of co-administered un-conjugated drugs deep into tumor and increase their therapeutic index. Current TPP target angiogenic tumor vessels and may not be suitable for targeting slow-growing tumors and invasive tumor cells. TPP are composed of functional modules (tumor recruitment motif, cryptic tissue penetrating C-end Rule element, and a protease cleavage site), which can be rearranged to yield peptides of novel specificities. Our goal is to develop TPP platform for delivery of co-administered drugs to the deadliest brain tumor – glioblastoma (GBM). High-grade glioma is a target that is particularly evasive and well-suited for tissue penetrative drug delivery. We will develop glioma-specific TPP (gTPP) by combination of in vivo and ex vivo phage display of constrained peptide libraries on state-of-the-art glioma animal models. These gTPP will be able to penetrate gliomas (and potentially other tumors) independent of their angiogenic status, and to deliver drugs to infiltrating malignant cells far from the bulk glioma lesion. We will characterize, validate, and optimize the gTPP platform for enhanced glioma delivery of co-injected drugs. These studies will provide the preclinical data needed to advance the gTPP combination therapy of glioma to GLP toxicology and subsequent IND filing.

1 499 931 €

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

Targeting therapeutic genes selectively into the central nervous system (CNS) is a crucial precondition for translation of gene therapy strategies into human trials. The current multidisciplinary proposal integrates expertise identified as essential in the effective acceleration of research to overcome bottlenecks in the field including: 1) Inefficiency of therapy delivery to the CNS because of factors like the blood-brain barrier (BBB); 2) Poor understanding of disease mechanisms at the molecular and cellular levels. These problems must be overcome to develop fully effective treatments for neurological disorders. Currently the adeno-associated (AAV)-based system is one of the most refined and effective gene delivery systems for neuronal cells. In contrast to all other systems, it has been possible to engineer AAV9 to deliver genes through the BBB to the CNS by intravascular (IV) administration. However, following IV delivery, these vectors also target liver and other tissues, with significant potential for untoward effects. This has prompted us to adopt two major strategies: i) targeting of AAV9 vectors at the level of transcription by insertion of hybrid motor neuron specific promoters into the vector genome; ii) development of a CNS-targeted delivery approach based on state-of-the art nanoparticle-mediated encapsulation of AAV9 vectors. We anticipate that engineering strategies with the ability to restrict transgene expression to CNS tissue will significantly overcome various existing hurdles in CNS gene therapy development. Our objectives are to: 1) explore mechanisms leading to penetration of scAAV9 vectors through BBB since the exact mechanism of AAV9 diffusion through BBB is unknown; 2) design novel targeted strategies with enhanced tropism to CNS; 3) use CNS targeted vectors to investigate mechanisms of motor neuron death linked to mutations in RNA processing genes; 4) utilise CNS-targeted systems to test therapeutic strategies for motor neuron diseases.

2 499 959 €

Start date: 2012-03-01, End date: 2017-10-31

The liver is a vital organ for synthesis and detoxification. The most significant liver diseases are hepatitis, non alcoholic fatty liver disease (NAFLD), non-alcoholic fatty liver steatohepatitis (NASH), carcinoma and cirrhosis. An additional and important cause of liver injury is adverse drug reactions (ADRs). In particular NAFLD is the most common liver disease affecting between 20% and 44% of European adults and 43-70% of patients with type 2 diabetes, and is one prime cause for chronic and end-stage liver disease, such as cirrhosis and primary hepatocellular carcinoma. This proposal is based on recent findings in the laboratory: The development of novel 3D spheroid system with chemically defined media allowing studies of chronic drug toxicity, relevant liver disease and liver function for 5 weeks in vitro, the finding of the role of miRNA in hepatocyte dedifferentiation and that hepatocytes during spheroid formation first de-differentiate but later in spheroids re-differentiate to an in vivo relevant phenotype. This forms the basis for the main objectives: i) to study diseased liver in vitro with identification of mechanisms, biomarkers and novel drug candidates for treatment of NAFLD and fibrosis, ii) evaluate drug toxicity sensitivity and mechanisms in diseased liver systems and iii) further develop methods for hepatocyte proliferation and regeneration in vitro for transplantation purposes, including genetic editing in cases of hepatocytes obtained from patients with genetically inherited liver diseases. This work is carried out in close contact with the Hepatology unit at the Karolinska Hospital partly using resources at the Science for Life Laboratory at Karolinska. It is anticipated that the project can provide with novel mechanisms, biomarkers and new targets for treatment of liver disease as well as novel methods for clinically applicable liver regeneration without the use of stem cells or transformed cells.

2 413 449 €

Start date: 2017-09-01, End date: 2022-08-31

Human myeloid leukemias are life-threatening disorders. Although with current therapies the bulk of the tumor is readily eradicated, it is particularly the rare population of leukemic stem cells (LSCs) that is relatively quiescent and is very difficult to target. As a consequence, relapse of the disease occurs frequently resulting in poor survival rates. Nevertheless, remarkable differences do exist between subsets of patients. It is important to realize that leukemias are rather heterogeneous disorders that can be caused by a multiplicity of genetic and epigenetic changes. One of the most important challenges in the field today lies in establishing which of the differences between LSCs and normal hematopoietic stem cells (HSCs) provide attractive targets for the identification, targeting and ultimately the selective eradication of LSCs. Given the heterogeneity of the disorder, it will be important to address these aspects in detail in a leukemia subtype-specific manner. Central to this proposal is the establishment of a human xenograft mouse leukemia “clinic” in which all important human leukemia subtypes will be represented. LSCs from patients as well as from cord blood and bone marrow model systems that we have generated will be equipped with luciferase-GFP tracers in order to allow detection of leukemic development in alive immunodeficient mice, as well as with unique barcodes that will allow clonal tracking. Within this mouse clinic, in a leukemia subtype-specific manner, we will be able to: 1) identify (novel) leukemic stem cell markers and evaluate their targetability; 2) evaluate whether stem cell intrinsic versus extrinsic signaling can be used as targets in alternative approaches to eradicate LSCs, and 3) study clonal evolution from de novo to relapsed leukemia. Our studies should provide insight into the biology of leukemia and novel rational approaches to treat leukemia patients more successfully.

1 499 820 €

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

Up to one fifth of young people have had the experience of psychotic symptoms, such as hearing voices when there is no-one around, or seeing visions. We now know that young people who experience these symptoms are at increased risk of developing psychotic disorders in adulthood. We also know that these young people are at higher risk of a range of co-morbid disorders such as depression and anxiety, and particularly suicidal behaviours. On the other hand, many of these young people will remain well and, for them, the psychotic experiences were merely a transitory phenomenon. Childhood trauma is known to be associated with increased risk for psychotic symptoms and is a promising target for intervention. However we do not yet know enough about what types or timing of stressors are involved in the pathogenesis of psychotic symptoms, nor the mechanism by which early life stress may lead to changes in brain structure and function resulting in symptoms such as hallucinations. We also need to be able to identify those young people who will benefit most from intervention. This ground-breaking, multi-disciplinary programme of work sets out to address these issues by drawing together epidemiology, social science, anthropology and neuroscience to devise a comprehensive programme of work examining the relationship between early life stress and psychotic symptoms among young people. Designed as three inter-related work packages, this iHEAR programme will exploit a large population-based cohort and will capitalise on my existing unique cohort of young people, who were known to have experienced psychotic symptoms in childhood, as they enter young adulthood. This iHEAR programme will result in new information which will allow the development of innovative interventions to prevent or pre-empt severe mental illness in later life.

1 781 623 €

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

Background: Statins are essential drugs in the treatment of hypercholesterolaemia and are among the most prescribed drugs worldwide. The response to statin therapy varies widely between individuals. While most patients show good efficacy, a significant proportion of individuals show poor or even a lack of cholesterol-lowering efficacy. Moreover, a number of patients experience adverse drug reactions. These together with the lack of immediate effect on well-being likely explain the relatively poor adherence to statin therapy. Poor adherence to statins in turn increases the incidence of cardiovascular events and mortality. Aims: The objectives of this project are 1) to develop a systems pharmacology model for predicting statin efficacy and tolerability at the level of an individual patient and 2) to investigate whether selecting the statin based on the model improves treatment adherence. Methods: A systems pharmacology approach will be used to integrate data from in vitro and clinical studies. Semi-physiological pharmacokinetic-dynamic-toxicologic models will be built for each statin allowing the prediction of the pharmacokinetic and clinical outcomes for patients with different characteristics, genotypes, and concomitant medications. The ability of the systems pharmacology algorithm to enhance adherence will be investigated in a randomized clinical trial. Significance: Systems pharmacology models have been increasingly applied in drug development, for example to predict the effect of organ dysfunction on pharmacokinetics. The proposed project is the first to use systems pharmacology predictions to guide clinical drug therapy, thus going beyond the state of the art. If successful, the project will not only improve the prevention and treatment of cardiovascular disease, but it will open new horizons to individualizing drug therapies.

2 211 565 €

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

"The goal of ""InstantCount"" is to develop low-cost point-of-care blood tests based on microfluidic counting chambers and image cytometry (InstantCount tests) for use in challenging environments, e.g. remote areas in developing countries. Preliminary work on CD4 counting (measuring the concentration of helper T-cells in blood in order to diagnose the progression of an HIV infection) demonstrates the feasibility of the concept. I have assembled a prototype instrument for large area fluorescence imaging, developed the image analysis software and microfluidic counting chambers containing dried reagents. The fabrication of these chambers is simple and the used materials cheap, making affordable diagnostics even in the poorest regions appear feasible. However, a clinical evaluation of the tests outside the laboratory environment in rural areas requires improved reproducibility in the fabrication of the chambers, which would also enable the development of new assays, once the properties of the reagent layers can be tailored according to the requirements of different tests. I propose the development of printing techniques for the fabrication of InstantCount tests. Counting chambers with precisely defined thickness will be realised by printing of microbeads embedded in glue. Printing of different types of hydrogel will offer solutions like self-sealing chambers or timed release of reagents with a wide range of molecular sizes without the need for mixing other than by diffusion. With these techniques, we will realise rapid low-cost point-of-care diagnostics for primary health care in remote areas and small medical practices. Complete blood counts for general health screening, CD4 counting and the detection of parasites in red blood cells such as in malaria will be provided. The project will neither require expensive fabrication methods in clean room facilities nor high-resolution optics in the instruments. Printing can bridge the gap between prototyping and production."

1 496 400 €

Start date: 2011-09-01, End date: 2017-04-30

Chronic liver disease can progress to cirrhosis, with death due to liver failure and cancer. Cirrhosis prevalence in the EU is 0.5%-1%. However, development of therapies that prevent progression to cirrhosis is hampered by the lack of a sensitive, non-invasive method to quantify fibrosis or fibrosis progression (fibrogenesis). Liver biopsy 1) is risky, 2) shows high sampling variability, and 3) is too insensitive to assess fibrosis progression in clinical studies. Conventional radiological imaging, serum markers, and ultrasound- or MR-elastography do neither permit exact fibrosis nor any fibrogenesis measurement. We plan to develop a clinically applicable methodology to quantitate fibrosis and fibrogenesis over the whole liver using imaging agents that target and thus quantify abundant fibrillar collagen or key cells that drive fibrogenesis (activated myofibroblasts and cholangiocytes). We demonstrated the feasibility of this approach using radiolabeled conjugates of high affinity that target integrin alphaVbeta6 and PDGFbeta receptor that are cell surface molecules of activated cholangiocytes and myofibroblasts. i.v. injection of the integrin conjugate allowed quantitative imaging of alphaVbeta6 expression and correlated with whole liver fibrogenesis. We intend to optimize nonpeptide and peptide ligands for integrin alphaVbeta6, PDGF beta receptor and fibrillar collagens using novel linkers and oligomerization, using PET-radioimaging with Ga-68, Sc-44 and F-18. The targeted imaging constructs will be validated in vivo using established rodent models with defined liver fibrosis and fibrogenesis, with and without antifibrotic drug therapy. Translation to phase I and II clinical studies is planned in years 4-5 of the project. The technology will for the first time allow for 1. individual risk assessment of fibrosis progression, and 2. rapid testing of antifibrotic drugs and their combinations in small groups of individual patients.

2 454 604 €

Start date: 2012-08-01, End date: 2017-07-31

Meningitis is an inflammation of the membranes covering the brain and spinal cord (the meninges). Each year 35,000 European patients suffer from bacterial meningitis, resulting in 7000 deaths and leaving 7000 disabled. Streptococcus pneumoniae and Neisseria meningitidis are the most common causative bacteria of bacterial meningitis. These bacteria are common colonizers, but in some individuals, they are able to spread to the bloodstream, slip through the blood-brain barrier and cause meningitis, with devastating consequences. Genetics of host and pathogen are considered crucial in this host-pathogen interaction. Our objectives are to identify and characterize host genetic traits and bacterial virulence factors controlling occurrence and outcome of bacterial meningitis. In a nationwide prospective cohort study, using pathway and total exome sequencing, we will search for genetic variants in 2000 patients with community-acquired bacterial meningitis and 2000 controls. On the pathogen side, using whole genome sequencing, we will systematically search for new bacterial virulence factors in the 2000 causative bacteria. Subsequently, by analyzing clinical data, serum and cerebrospinal fluid, we will examine the potential impact and functionality of these host and pathogen factors. Next, we will validate and explore our findings in an animal model of meningitis and investigate whether treatment against these specific components can improve outcome. Finally, we will initiate an open-access European database on the genetics in bacterial meningitis. In addressing our objectives, I will combine my expertise in bacterial meningitis with groundbreaking, translational approaches using clinical data, human samples, next generation sequencing, in vitro techniques, and a mouse model. The results of this project will be translated into novel therapeutic strategies to advance patient care and will also be important for the development of disease prevention and vaccines.

1 400 000 €

Start date: 2012-02-01, End date: 2017-01-31

"I propose to develop a new generation of molecular photoacoustic imaging technologies and methods capable of detecting single deep tissue cells in preclinical studies of tissue regeneration. In order to achieve this goal, an interdisciplinary research programme involving physicists, engineers and life scientists is required to address the following objectives: 1) the development of novel photoacoustic imaging technology that provides high sensitivity and acquisition speed, 2) the development of the theoretical framework and experimental methods for quantitative imaging, 3) the development of novel genetically expressed reporters, and 4) the preclinical application in small animal models of tissue regeneration. This will result in a preclinical imaging modality has the potential to combine single cell sensitivity and microscale spatial resolution in deep (centimetre range) tissue regions with molecular, physiological, and anatomical imaging capabilities. The instrumentation and methodologies developed in this project will be applied to noninvasive, longitudinal, and quantitative studies of stem cell driven tissue regeneration, such as angiogenesis in bone fractures and muscle trauma. It will allow the detection and tracking of single stem cells and the probing of stem cell function. This will provide unprecedented opportunities for correlating cellular localization, migration, and function and with anatomical changes - knowledge that can be exploited to develop novel drugs and cell-based clinical therapies. Crucially, the technologies and methodologies developed in this project will be directly applicable to a wide range of other fields of the life sciences, such as cancer research and neurology."

1 622 736 €

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

MULTIEPIGEN seeks to solve does ancestral exposure to various stressors transmit to offspring via epigenetic mechanisms. Thus far animal models have indicated that exposure to certain stressors can lead to phenotypic changes not only in the predisposed individuals, but also in the future generations, such that individuals can acquire phenotypes caused by exposures of their ancestors. Such effects do not involve new DNA mutations, but are transmitted to offspring via epigenetic mechanisms such as the transfer of non-coding RNA molecules in the semen. In humans, intergenerational transmission has been examined extremely little because a priori designed population-based studies across several generations are lacking. To close this gap MULTIEPIGEN will expand the well-characterized Cardiovascular Risk in Young Finns Study (YFS) to the parents and offspring of the original YFS participants. During the ERC funding period, we will perform field studies involving N~9000 individuals across 3 generations and test 3 key ancestral exposures with very high plausibility causing intergenerational effects on obesity-related phenotypes, cognitive function and psychological well-being. The studied exposures are 1) tobacco smoke, 2) persistent organic pollutants, and 3) accumulation of psychosocial adversities. We will collect serum, blood and semen samples for epigenetic marker analysis to provide understanding of the mechanisms of intergenerational transmission in humans. Specifically, we will seek proof for the hypothesis that paternal stressors can lead to phenotypic changes in the offspring via non-coding RNA molecules in the semen. Multigenerational epidemiologic data showing robust links between ancestral exposures and offspring phenotypes that operate with biologically plausible epigenetic mechanism would provide a conceptual change in the developmental biology in humans and have substantial ramifications on public health.

2 498 606 €

Start date: 2017-11-01, End date: 2022-10-31

Current knowledge of neurodegenerative diseases is limited by poor understanding of how they progress through the central nervous system (CNS). It has recently been hypothesized that clinical progression in these conditions involves the systematic spreading of protein misfolding along neuronal pathways. Protein aggregates would trigger misfolding of adjacent homologue proteins in newly-affected regions, and this would propagate in a “prion-like” fashion across anatomical connections. This proposal seeks to decipher the mechanisms of network-based neurodegeneration by understanding how the complex architecture of brain networks (the connectome) shapes the evolving pathology of neurodegenerative diseases, and to develop tools for monitoring disease progression from presymptomatic to later stages of the disease. NeuroTRACK will apply emerging network science tools to longitudinal, structural and functional brain connectivity 3T magnetic resonance imaging data from patients with frontotemporal lobar degeneration (FTLD) – a devastating, relentlessly progressive, young onset, neurodegenerative disorder. The study will involve both sporadic and familial cases, including presymptomatic gene mutation carriers. The proposal addresses the following fundamental questions: i) How and where does pathological protein propagation occur in the FTLD phenotypes? ii) Can pathological spreading be predicted from brain connectome fingerprinting? iii) How do different protein abnormalities translate into large-scale network degeneration? iv) How early are brain network changes detectable in the (even presymptomatic) course of the disease? The ground-breaking nature of the experiments planned in this proposal will pave the way to the development of novel tools for understanding the biological underpinnings of other CNS proteinopathies such as Alzheimer’s disease and Parkinson’s disease, and to identifying individualized, early interventions to modify disease progression.

1 496 994 €

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

Background Osteoarthritis (OA) is one of the most prevalent disorders of the musculoskeletal system. In OA, articular cartilage degenerates and its structure and mechanical properties change, but monitoring or predicting the progression of OA has not been possible. Magnetic resonance imaging (MRI) is a potential tool for the imaging of joint tissues, estimating cartilage structure and diagnostics of OA, whereas joint loading and estimation of stresses/strains within joint tissues necessitates computational modeling. It would be a major breakthrough if one could develop a technique where, based on MRI and computational modeling, prediction and evaluation of OA progression of a patient under a certain loading condition would be possible. Objectives 1) to combine MRI with computational modeling for the estimation of stresses and possible failure points within human knee joints, and 2) to develop second generation adaptive models of articular cartilage for the prediction of altered tissue structure and composition during OA progression. For the model validation, cartilage structure, composition and biomechanical properties as well as cell responses in situ are characterized. At the end of the project these main aims will be merged 3) to estimate the effect of loading on cartilage degeneration during the progression of OA in a patient-specific manner. Significance Combining MRI information of joint tissues with computational modeling, we develop a tool to evaluate the effect of different interventions on stresses in human joints. By combining this tool with an adaptive model that can estimate the effect of loading on cartilage composition and structure, we hope to be able to predict changes in cartilage properties during OA progression in a patient-specific manner several years ahead. This would help in decision making of clinical treatments and interventions (conservative or surgical) for the prevention or further progression of OA.

1 303 056 €

Start date: 2012-02-01, End date: 2017-01-31