Projects & Collaborations 8 foundShow per page10 10 20 50 High Performance Transmission Electron Microscope for Present and Future Nanomaterials Research Project | 9 Project MembersThe rise of nanoscience and nanotechnology would not have been happened without the impressive development of instruments that allow to resolve structure on the nanometer scale with atomic resolution. Examples are scanning-probe and electron microscopy techniques. In recent years, several major breakthroughs gave rise to an exceptional boost in the performance of today's electron microscopy (EM), both for solid-state and soft (e.g. biological) materials: 1) high-resolution through image corrections, 2) fast and highly efficient electron detectors, 3) efficient artifact-free sample fabrication (cryo-EM and FIBEM), and 4) 3D tomography and image reconstruction. This has given a leap to what can be imaged today, allowing for example to reconstruct the atomic structure of single proteins and image complex interfaces in solid-state materials with atomic scale. The University of Basel (UBAS) is nationally and internationally recognized as a leader in nanoscience and nanotechnology. It was the leading house of the National Center in Competence and Research (NCCR) on Nanoscience, which later became the Swiss Nanoscience Institute (SNI), the institution that submits the current proposal. UBAS is also co-leading the NCCR Molecular Systems Engineering and the NCCR QSIT on Quantum Science (both together with ETHZ). Nanoscience is a focus area in the research portfolio of UBAS and instrumental for the recent development of quantum science. The present proposal to the SNF R'Equip scheme has been put together by key researchers at UBAS who work on current topics in nanoscience and nanotechnology in various disciplines from quantum science, material science, polymer chemistry to molecular biology, and, who make use of EM available within the SNI. The principle investigators, who submit this proposal together, do research that relies on the availability of state-of-the-art nanoimaging tools, such as a transmission electron microscope (TEM). The proposal outlines a convincing case for the purchase of special, unique TEM that combines state-of-the-art (and fast) atomic resolution imaging with material analysis using EDX and scanning TEM (STEM). This combination is unique and crucial for the University of Basel to stay at the forefront of science. Development of novel synthetic gene transfer vectors for metabolic liver therapy Research Project | 2 Project MembersTherapeutic vectors for gene delivery remain the currently most challenging factor for human gene therapy. The translation from in vitro to in vivo applications remains a major hurdle for most nucleic acid delivery systems since there is an inherent lack of both efficient and safe carrier systems. For liver targeting of postmitotic hepatocytes, adeno-associated virus (AAV) derived vectors are thought to have the greatest potential despite concerns about a future routine clinical use. The major hurdles of AAV vectors for long-term treatment of pediatric patients are the risk of chromosomal integration and development of hepatocellular carcinoma, immune responses to viral vectors, limited loading capacity, and the difficulty to treat neonates which likely would require subsequent further injections. Consequently, the development of non-viral gene delivery systems has gained much attention due to their versatility, safety, and ease of manufacturing. During the last decades, a wide range of nanoparticle based gene delivery systems were developed and remarkable results in the field of RNA therapeutics were achieved. However, the induced pharmacological effects obtained by these siRNA or mRNA delivery strategies are short-lived and thus weekly administrations of therapeutic formulations are necessary. The use of DNA-based therapeutics would offer a favorable option for the induction of long-term therapeutic effects without need for insertion into the genome. Here, we propose an alternative approach to overcome the challenges of viral vectors or RNA-based therapeutics by developing novel nanoparticles for delivery of non-integrating, so-called minicircle (MC) vectors lacking any viral or bacterial components for liver-directed gene therapy. The successful use of MC vectors to treat genetic (metabolic) liver defects is based on the experience of one application partner with naked DNA-vectors delivered in an experimental setting by hydrodynamic pressure to either target pericentral or periportal hepatocytes to treat two classical defects in mouse models for human diseases, phenylketonuria (PKU) and ornithine transcarbamylase (OTC) deficiency, respectively. Such MC vectors exhibited persistent expression combined with basically no DNA size limitation, which made it possible to use natural promoters/enhancers in combination with introns to mimic "physiological" expression. While MC vectors bear almost ideal properties with great potential for liver gene therapy, delivery of naked DNA solely by hydrodynamic pressure is not applicable in a clinical setting. In an interdisciplinary approach, we want to develop multifunctional polymeric nanoparticles encapsulating MC vectors for non-viral gene delivery specifically to the key pathogenic cell type, i.e. hepatocytes. In order to optimize gene delivery efficiency, a novel library of polymer-peptide hybrids will be created, formulations strategies will be optimized and resulting nanoparticles will be validated in vivo using various animal models, i.e. transgenic mice, xenotransplanted mice with human liver or pig models. The combination of this novel class of polymer-peptide hybrids with a reproducible and scalable nanoparticle formulation technique (i.e. microfluidics) is expected to greatly impact further optimization of the synthetic gene delivery system for clinical applications. The overall aim of this translational project is the development of an alternative approach to AAV vectors with the potential of a breakthrough for liver gene therapy and thus a paradigm shift from potentially harmful viral vectors to safe, efficient and completely synthetic non-viral vectors. Drug Targeting to Hepatocytes: Gene Delivery using Myrcludex B Coupled Lipid Nanoparticles Research Project | 3 Project MembersHepatic disorders affect millions of people around the globe and incidence rates are further increasing. Current therapies for diseases of hepatocytes are limited and in most cases only treat symptoms. Therefore, improved therapeutic technologies are needed. Nanomedicines for the delivery of therapeutic genes have the potential to overcome the lack of satisfactory and alternative treatment options. This grant application focuses on the design of functional nanomedicines for targeted nucleic acid delivery (i.e. plasmid DNA) to liver parenchymal cells. The proposed project consists of three work-packages, which can be summarized as follows:First, specific and highly selective targeting of hepatocytes will be achieved using a targeting ligand derived from hepatitis B virus (HBV). This HBV entry inhibitor "Myrcludex B" consists of a lipid-conjugated polypeptide (i.e. the PreS1 domain of the large surface glycoprotein of HBV) and is characterized by a strong tropism for hepatocytes. Optimized Myrcludex B-derived lipopeptides will be conjugated to the surface of pegylated liposomes to mediate hepatocyte specific drug delivery. Second, in order to optimize loading and retention of DNA expression plasmids within lipid nanoparticles, a novel library of double tailed, ionizable amino-lipids will be created and screened for efficient and safe transfection activity. The combination of this new class of amino-lipids with a novel nanoparticle formulation technique (i.e. microfluidics) offers the possibility to optimize the transfection efficiency of the lipid-based delivery system. Third, in the final part of the project, both technologies will be combined to achieve targeted gene delivery to human hepatocytes; both in vitro in human liver derived cell lines as well as in vivo in different mouse models expressing the mouse or human NTCP, i.e. the entry point for HBV and at the same time our highly selective target structure on hepatocytes. With our novel targeting strategy, we have the possibility to address an unmet medical need. Non-viral gene delivery may offer well tolerated therapeutic options for diseases of the liver such as Crigler-Najjar syndrome, where a single gene defect leads to severe clinical manifestations. The proposed project will be the first step towards a future therapeutic intervention for this and other orphan liver diseases. Design of polymer nanoreactors with triggered activity Research Project | 2 Project MembersAn efficient way to overcome many of today's challenges in domains such as medicine, food science, catalysis, and environmental sciences is to design and use nanoreactors as protected reaction spaces for active compounds (enzymes, proteins, mimics) encapsulated/inserted in supramolecular polymer assemblies (micelles, spheres, vesicles) [1]. The overall aim of this PhD project is to develop triggered polymer nanoreactors by encapsulating/co-encapsulating enzymes and inserting channel proteins in polymer vesicles with sizes in the nanometer range. The triggered function is introduced by channel proteins that are modified so as to be open only in the presence of a specific chemical stimulus (protein gate). We will develop two types of nanoreactors intended to serve for: i. sensitive biosensing of pH changes, and ii. simultaneous detoxification of superoxide radicals and triggered production and release of compounds. The interactions of nanoreactors with biological systems will be tested with human liver carcinoma cell line HepG2 in terms of cellular binding, uptake and intracellular processing in order to support their further medical application [2] . [1] Palivan CG, Onaca O, Delcea M, Itel F, Meier W, Chem Soc. Rev. 2012 , 41(7), 2800 [2] Seeland S, Treiber A, Hafner M, Huwyler J, Biochim. Biophys. Acta , 2011 , 1808, 1827. Hepatotoxicity (part of SCAHT Research Projects 2013 - 2016) Research Project | 4 Project MembersThe project has three parts, which are started simultaneously since different research groups are involved in the project. Part 1 consists of the establishment of suitable cell models for performing toxicological studies with hepatocytes. We are going to add HepaRG cells to already existing HepG2 and HUH cell lines. In addition, we are going to characterize 3-dimensional human hepatocytes. We are also going to prepare hepatocytes containing genes enabling to detect ROS generation in different cellular compartments. Part 2 describes the application of these systems and analytical techniques for studying diverse known hepatotoxins. The idea is to find out mechanisms of toxicity, proof these mechanisms in cellular systems, find out potential susceptibility factors and proof such factors in genetically manipulated cells and in experimental animals. If suitable, such susceptibility factors can also be tested using a large tissue biobank and/or in cohorts of patients with drug-induced induced liver injury. In Part 3, the cellular uptake and the toxicity of nanoparticles will be tested in our established hepatocyte systems and in experimental animals. Entwicklung mikropartikel-basierter Formulierungen zur gezielten Verabreichung pharmazeutischer Wirkstoffe Research Project | 1 Project MembersZusammenarbeit mit der Firma Tillotts Pharma AG auf dem Gebiet der pharmazeutischsen Technologie. Purchase of a laser scanning microscope Research Project | 3 Project MembersThe field of microscopy applications to investigate cellular functions and disturbances that are relevant to understand disease processes and for generation of therapeutic interventions is rapidly developing. After completion of sequencing the human genome, there is a great demand to elucidate the functions of the proteins encoded by the about 40'000 genes and, furthermore, to uncover the dynamic interactions between these proteins in various pathophysiological situations. The introduction of laser scanning microscopy opened, among other applications, the possibility to determine the intracellular distribution of proteins, their colocalization with other proteins and organelle markers, their dynamic movement upon induction by stimuli, their synthesis and degradation and their relative 3D-distributions within compartments of a cell. For these applications, conventional fluorescence microscopy is not sufficient due to the lack of sensitivity. Currently, the applicants are equipped with conventional fluorescence microscopy, fluorescence 96-well plate format readers, ArrayScan High Content Imaging, and Seahorse technology. Thus, the purchase of a high sensitivity laser scanning microscope would fill the gap and allow extended analyses of cell-based studies. At the Department of Pharmaceutical Sciences of the University of Basel there is a rapidly increasing demand in the use of laser scanning microscopy for various applications. The recent decision for a more biological focus (novel Profs in Pharmaceutical Technology, J. Huwyler, and Biopharmacy, to be appointed in 2011) will further enhance the need for laser scanning microscopy. Currently, the research groups of the Department are highly restricted in the use of laser scanning microscopy and are fully dependent on external collaborations. Three Divisions of the Department (the applicants Prof. S. Krähenbühl, Prof. J. Huwyler, Prof. A. Odermatt) have ongoing research projects where laser scanning microscopy is required. Moreover, the new Prof. in Biopharmacy is likely to have a need for this technique. Thus, the research of the involved groups greatly benefits from the purchase of a highly sensitive laser scanning microscope. Mechanism of action of MDMA (Ecstasy) Research Project | 7 Project MembersIn our main project: "Mechanism of action of MDMA (Ecstasy)" we investigate how MDMA acts in the brain using both in vitro techniques and studies in humans. In the in vitro studies we assess interactions of MDMA with serotonin-, dopamine-, and norepinephrine uptake carriers in cell assays. In humans, we assess the subjective and cardiovascular effects of MDMA after the administration of substances that selectively block potential sites of action of MDMA in the brain. 1 1
High Performance Transmission Electron Microscope for Present and Future Nanomaterials Research Project | 9 Project MembersThe rise of nanoscience and nanotechnology would not have been happened without the impressive development of instruments that allow to resolve structure on the nanometer scale with atomic resolution. Examples are scanning-probe and electron microscopy techniques. In recent years, several major breakthroughs gave rise to an exceptional boost in the performance of today's electron microscopy (EM), both for solid-state and soft (e.g. biological) materials: 1) high-resolution through image corrections, 2) fast and highly efficient electron detectors, 3) efficient artifact-free sample fabrication (cryo-EM and FIBEM), and 4) 3D tomography and image reconstruction. This has given a leap to what can be imaged today, allowing for example to reconstruct the atomic structure of single proteins and image complex interfaces in solid-state materials with atomic scale. The University of Basel (UBAS) is nationally and internationally recognized as a leader in nanoscience and nanotechnology. It was the leading house of the National Center in Competence and Research (NCCR) on Nanoscience, which later became the Swiss Nanoscience Institute (SNI), the institution that submits the current proposal. UBAS is also co-leading the NCCR Molecular Systems Engineering and the NCCR QSIT on Quantum Science (both together with ETHZ). Nanoscience is a focus area in the research portfolio of UBAS and instrumental for the recent development of quantum science. The present proposal to the SNF R'Equip scheme has been put together by key researchers at UBAS who work on current topics in nanoscience and nanotechnology in various disciplines from quantum science, material science, polymer chemistry to molecular biology, and, who make use of EM available within the SNI. The principle investigators, who submit this proposal together, do research that relies on the availability of state-of-the-art nanoimaging tools, such as a transmission electron microscope (TEM). The proposal outlines a convincing case for the purchase of special, unique TEM that combines state-of-the-art (and fast) atomic resolution imaging with material analysis using EDX and scanning TEM (STEM). This combination is unique and crucial for the University of Basel to stay at the forefront of science.
Development of novel synthetic gene transfer vectors for metabolic liver therapy Research Project | 2 Project MembersTherapeutic vectors for gene delivery remain the currently most challenging factor for human gene therapy. The translation from in vitro to in vivo applications remains a major hurdle for most nucleic acid delivery systems since there is an inherent lack of both efficient and safe carrier systems. For liver targeting of postmitotic hepatocytes, adeno-associated virus (AAV) derived vectors are thought to have the greatest potential despite concerns about a future routine clinical use. The major hurdles of AAV vectors for long-term treatment of pediatric patients are the risk of chromosomal integration and development of hepatocellular carcinoma, immune responses to viral vectors, limited loading capacity, and the difficulty to treat neonates which likely would require subsequent further injections. Consequently, the development of non-viral gene delivery systems has gained much attention due to their versatility, safety, and ease of manufacturing. During the last decades, a wide range of nanoparticle based gene delivery systems were developed and remarkable results in the field of RNA therapeutics were achieved. However, the induced pharmacological effects obtained by these siRNA or mRNA delivery strategies are short-lived and thus weekly administrations of therapeutic formulations are necessary. The use of DNA-based therapeutics would offer a favorable option for the induction of long-term therapeutic effects without need for insertion into the genome. Here, we propose an alternative approach to overcome the challenges of viral vectors or RNA-based therapeutics by developing novel nanoparticles for delivery of non-integrating, so-called minicircle (MC) vectors lacking any viral or bacterial components for liver-directed gene therapy. The successful use of MC vectors to treat genetic (metabolic) liver defects is based on the experience of one application partner with naked DNA-vectors delivered in an experimental setting by hydrodynamic pressure to either target pericentral or periportal hepatocytes to treat two classical defects in mouse models for human diseases, phenylketonuria (PKU) and ornithine transcarbamylase (OTC) deficiency, respectively. Such MC vectors exhibited persistent expression combined with basically no DNA size limitation, which made it possible to use natural promoters/enhancers in combination with introns to mimic "physiological" expression. While MC vectors bear almost ideal properties with great potential for liver gene therapy, delivery of naked DNA solely by hydrodynamic pressure is not applicable in a clinical setting. In an interdisciplinary approach, we want to develop multifunctional polymeric nanoparticles encapsulating MC vectors for non-viral gene delivery specifically to the key pathogenic cell type, i.e. hepatocytes. In order to optimize gene delivery efficiency, a novel library of polymer-peptide hybrids will be created, formulations strategies will be optimized and resulting nanoparticles will be validated in vivo using various animal models, i.e. transgenic mice, xenotransplanted mice with human liver or pig models. The combination of this novel class of polymer-peptide hybrids with a reproducible and scalable nanoparticle formulation technique (i.e. microfluidics) is expected to greatly impact further optimization of the synthetic gene delivery system for clinical applications. The overall aim of this translational project is the development of an alternative approach to AAV vectors with the potential of a breakthrough for liver gene therapy and thus a paradigm shift from potentially harmful viral vectors to safe, efficient and completely synthetic non-viral vectors.
Drug Targeting to Hepatocytes: Gene Delivery using Myrcludex B Coupled Lipid Nanoparticles Research Project | 3 Project MembersHepatic disorders affect millions of people around the globe and incidence rates are further increasing. Current therapies for diseases of hepatocytes are limited and in most cases only treat symptoms. Therefore, improved therapeutic technologies are needed. Nanomedicines for the delivery of therapeutic genes have the potential to overcome the lack of satisfactory and alternative treatment options. This grant application focuses on the design of functional nanomedicines for targeted nucleic acid delivery (i.e. plasmid DNA) to liver parenchymal cells. The proposed project consists of three work-packages, which can be summarized as follows:First, specific and highly selective targeting of hepatocytes will be achieved using a targeting ligand derived from hepatitis B virus (HBV). This HBV entry inhibitor "Myrcludex B" consists of a lipid-conjugated polypeptide (i.e. the PreS1 domain of the large surface glycoprotein of HBV) and is characterized by a strong tropism for hepatocytes. Optimized Myrcludex B-derived lipopeptides will be conjugated to the surface of pegylated liposomes to mediate hepatocyte specific drug delivery. Second, in order to optimize loading and retention of DNA expression plasmids within lipid nanoparticles, a novel library of double tailed, ionizable amino-lipids will be created and screened for efficient and safe transfection activity. The combination of this new class of amino-lipids with a novel nanoparticle formulation technique (i.e. microfluidics) offers the possibility to optimize the transfection efficiency of the lipid-based delivery system. Third, in the final part of the project, both technologies will be combined to achieve targeted gene delivery to human hepatocytes; both in vitro in human liver derived cell lines as well as in vivo in different mouse models expressing the mouse or human NTCP, i.e. the entry point for HBV and at the same time our highly selective target structure on hepatocytes. With our novel targeting strategy, we have the possibility to address an unmet medical need. Non-viral gene delivery may offer well tolerated therapeutic options for diseases of the liver such as Crigler-Najjar syndrome, where a single gene defect leads to severe clinical manifestations. The proposed project will be the first step towards a future therapeutic intervention for this and other orphan liver diseases.
Design of polymer nanoreactors with triggered activity Research Project | 2 Project MembersAn efficient way to overcome many of today's challenges in domains such as medicine, food science, catalysis, and environmental sciences is to design and use nanoreactors as protected reaction spaces for active compounds (enzymes, proteins, mimics) encapsulated/inserted in supramolecular polymer assemblies (micelles, spheres, vesicles) [1]. The overall aim of this PhD project is to develop triggered polymer nanoreactors by encapsulating/co-encapsulating enzymes and inserting channel proteins in polymer vesicles with sizes in the nanometer range. The triggered function is introduced by channel proteins that are modified so as to be open only in the presence of a specific chemical stimulus (protein gate). We will develop two types of nanoreactors intended to serve for: i. sensitive biosensing of pH changes, and ii. simultaneous detoxification of superoxide radicals and triggered production and release of compounds. The interactions of nanoreactors with biological systems will be tested with human liver carcinoma cell line HepG2 in terms of cellular binding, uptake and intracellular processing in order to support their further medical application [2] . [1] Palivan CG, Onaca O, Delcea M, Itel F, Meier W, Chem Soc. Rev. 2012 , 41(7), 2800 [2] Seeland S, Treiber A, Hafner M, Huwyler J, Biochim. Biophys. Acta , 2011 , 1808, 1827.
Hepatotoxicity (part of SCAHT Research Projects 2013 - 2016) Research Project | 4 Project MembersThe project has three parts, which are started simultaneously since different research groups are involved in the project. Part 1 consists of the establishment of suitable cell models for performing toxicological studies with hepatocytes. We are going to add HepaRG cells to already existing HepG2 and HUH cell lines. In addition, we are going to characterize 3-dimensional human hepatocytes. We are also going to prepare hepatocytes containing genes enabling to detect ROS generation in different cellular compartments. Part 2 describes the application of these systems and analytical techniques for studying diverse known hepatotoxins. The idea is to find out mechanisms of toxicity, proof these mechanisms in cellular systems, find out potential susceptibility factors and proof such factors in genetically manipulated cells and in experimental animals. If suitable, such susceptibility factors can also be tested using a large tissue biobank and/or in cohorts of patients with drug-induced induced liver injury. In Part 3, the cellular uptake and the toxicity of nanoparticles will be tested in our established hepatocyte systems and in experimental animals.
Entwicklung mikropartikel-basierter Formulierungen zur gezielten Verabreichung pharmazeutischer Wirkstoffe Research Project | 1 Project MembersZusammenarbeit mit der Firma Tillotts Pharma AG auf dem Gebiet der pharmazeutischsen Technologie.
Purchase of a laser scanning microscope Research Project | 3 Project MembersThe field of microscopy applications to investigate cellular functions and disturbances that are relevant to understand disease processes and for generation of therapeutic interventions is rapidly developing. After completion of sequencing the human genome, there is a great demand to elucidate the functions of the proteins encoded by the about 40'000 genes and, furthermore, to uncover the dynamic interactions between these proteins in various pathophysiological situations. The introduction of laser scanning microscopy opened, among other applications, the possibility to determine the intracellular distribution of proteins, their colocalization with other proteins and organelle markers, their dynamic movement upon induction by stimuli, their synthesis and degradation and their relative 3D-distributions within compartments of a cell. For these applications, conventional fluorescence microscopy is not sufficient due to the lack of sensitivity. Currently, the applicants are equipped with conventional fluorescence microscopy, fluorescence 96-well plate format readers, ArrayScan High Content Imaging, and Seahorse technology. Thus, the purchase of a high sensitivity laser scanning microscope would fill the gap and allow extended analyses of cell-based studies. At the Department of Pharmaceutical Sciences of the University of Basel there is a rapidly increasing demand in the use of laser scanning microscopy for various applications. The recent decision for a more biological focus (novel Profs in Pharmaceutical Technology, J. Huwyler, and Biopharmacy, to be appointed in 2011) will further enhance the need for laser scanning microscopy. Currently, the research groups of the Department are highly restricted in the use of laser scanning microscopy and are fully dependent on external collaborations. Three Divisions of the Department (the applicants Prof. S. Krähenbühl, Prof. J. Huwyler, Prof. A. Odermatt) have ongoing research projects where laser scanning microscopy is required. Moreover, the new Prof. in Biopharmacy is likely to have a need for this technique. Thus, the research of the involved groups greatly benefits from the purchase of a highly sensitive laser scanning microscope.
Mechanism of action of MDMA (Ecstasy) Research Project | 7 Project MembersIn our main project: "Mechanism of action of MDMA (Ecstasy)" we investigate how MDMA acts in the brain using both in vitro techniques and studies in humans. In the in vitro studies we assess interactions of MDMA with serotonin-, dopamine-, and norepinephrine uptake carriers in cell assays. In humans, we assess the subjective and cardiovascular effects of MDMA after the administration of substances that selectively block potential sites of action of MDMA in the brain.
