ERC FUNDED PROJECTS

Up to one third of diabetic patients develop nephropathy, a serious complication of diabetes. Microalbuminuria is the earliest sign of the complication, which may ultimately develop to end-stage renal disease requiring dialysis or a kidney transplant. Insulin resistance and metabolic syndrome are associated with an increased risk for diabetic nephropathy. Interestingly, glomerular epithelial cells or podocytes have recently been shown to be insulin responsive. Further, nephrin, a key structural component of podocytes, is essential for insulin action in these cells. Our novel findings show that adaptor protein CD2AP, an interaction partner of nephrin, associates with regulators of insulin signaling and glucose transport in glomeruli. The results suggest that nephrin and CD2AP are involved, by association with these proteins, in the regulation of insulin signaling and glucose transport in podocytes. We hypothesize that podocytes can develop insulin resistance and that disturbances in insulin response affect podocyte function and contribute to the development of diabetic nephropathy. The aim of this project is to clarify the mechanisms leading to development of insulin resistance in podocytes and to study the association between insulin resistance and the development of diabetic nephropathy. For this we will develop transgenic zebrafish and mouse models by overexpressing/knocking down insulin signaling-associated proteins specifically in podocytes. Further, we aim to identify novel drug leads to treat insulin resistance and diabetic nephropathy by performing high-throughput small molecule library screens on the developed transgenic fish models. The ultimate goal is to find a treatment to combat the early stages of diabetic nephropathy in humans.

2 000 000 €

Start date: 2009-11-01, End date: 2014-10-31

Plants and their pathogens are in a constant process of co-evolution. Consequently, many of the known defense genes of plants against fungal pathogens are rapidly loosing effectiveness under agricultural conditions. However, there are examples for durable resistance. It is one of the main research questions in plant biology to determine the genetic basis of such naturally occurring resistance and to understand the underlying biochemical and molecular cause for durability. This durability is characterized by the apparent inability of the pathogen to adapt to the resistance mechanism. The molecular understanding of durable resistance will contribute to future attempts to develop such resistance by design. We want to use two approaches towards understanding and developing durable resistance: the first one is based on the naturally occurring durable resistance gene Lr34 against rust and mildew diseases in wheat. This gene was recently isolated in our group and it encodes a putative ABC type of transporter protein, providing a possible link between non-host and durable resistance. Its function in resistance will be studied by genetic and biochemical approaches in the crop plant wheat, as there is no Lr34-type of resistance characterized in any other plant. However, there is a close Lr34-homolog in rice and its function will be investigated in this diploid system. The second approach will be based on natural diversity found in a specific resistance gene, conferring strong, but not durable resistance. This diversity will be used for a designed improvement of durability by developing new proteins or protein combinations to which the pathogen can not adapt. We will use the 15 naturally occurring alleles of the Pm3 powdery mildew resistance genes to identify the structural basis of specific interactions. Based on this characterization, we will develop intragenic or gene combination pyramiding strategies to obtain more broad-spectrum and more durable resistance.

2 100 000 €

Start date: 2010-04-01, End date: 2015-03-31

Olfactory ensheathing cells (OECs) are a unique group of cells which were originally discovered in the olfactory bulb, the part of the brain that receives the sense of smell. These cells have now been found in the nose, and have the potential to encourage damaged nerve fibres to regenerate. When these cells are transplanted into the damaged spinal cords of rats they facilitate repair of the nerve fibres, and this results in an improved ability to climb and breath. It is now possible to obtain these cells from the noses of patients with brachial plexus avulsion (a longitudinal spinal cord injury) and to purify and multiply them for transplantation back into the same patient s damaged brachial plexus, to possibly cure injuries which were previously untreatable. However it is first necessary to find a safe and reliable way to obtain these cells from patients, develop a protocol for cleanroom manufacturing these cells under UK Good Manufacturing Practice (GMP) guidelines, and check whether human cells have the same reparative effects in the laboratory and animal studies, compared to what we already know about rat cells. The research programme consists of the following projects: 1. To develop a protocol for obtaining these cells in optimum quantities, by taking samples from volunteer patients who are undergoing nasal endoscopy for other reasons, and develop an effective culture method to maximise the yield of OECs. 2. To culture these cells under GMP conditions, using standardised reagents, and develop a protocol that ensures the maximum yield in a new culture facility. 3. To transplant autologous OECs into the site of injury in patients with complete brachial plexus avulsion, and assess the safety and efficacy of the technique. This will also allow us to obtain pilot data to allow planning of a future randomised controlled trial of OEC transplantation. 4. To study the effects of OECs derived from rat mucosa in animal models of brachial plexus avulsion

1 600 000 €

Start date: 2010-01-01, End date: 2014-12-31

Proline and related secondary amines were found to be efficient organocatalysts of aldol, alkylation, Michael addition, and other challenging bond-forming reactions. The design of even more efficient systems that would allow extending this remarkable catalytic strategy to a useful and environmentally benign synthetic methodology would be highly desirable. This proposal aims to develop a new class of biocatalysts that use the powerful proline-based enamine mechanism of organocatalysts but that take advantage of the water solubility and stereochemical control available with enzymes. As a scaffold for engineering, we will exploit the active-site template of the tautomerase superfamily proteins, the only known group of enzymes that has the unique feature of using a N-terminal proline in catalysis. By systematically screening tautomerases for promiscuous carbonyl transformation activities, we aim to demonstrate that Pro-1 residues within these tautomerases can react with carbonyl compounds to give enamine intermediates, and thus may facilitate various bond-forming reactions. The most promising enzymes will be used in directed evolution experiments to enhance the desired activities to a practical level. For this, new screening and selection methods will be developed that allow the efficient passage through protein sequence space. Furthermore, unnatural secondary amines will be introduced by total chemical synthesis. In this way, we envision to generate superior biocatalysts for carbon-carbon bond-forming reactions. The idea of using a protein scaffold in which a catalytic proline is present as nucleophile and that has a unique reactivity to form enamines would be a completely new approach to making carbon-carbon bonds. It does not copy something in Nature, but is based on the synthetic requirements of the desired bond-forming reactions. I believe that this type of approach is the future of enzyme engineering and is the key to more application of biocatalysis in industry.

2 000 000 €

Start date: 2010-05-01, End date: 2015-04-30