Project acronym BIOMEMOS
Project Higher order structure and function of biomembranes
Researcher (PI) Poul Nissen
Host Institution (HI) AARHUS UNIVERSITET
Call Details Advanced Grant (AdG), LS1, ERC-2009-AdG
Summary The biomembrane is a prerequisite of life. It enables the cell to maintain a controlled environment and to establish electrochemical gradients as rapidly accessible energy stores. Biomembranes also provide scaffold for organisation and spatial definition of signal transmission in the cell. Crystal structures of membrane proteins are determined with an increasing pace. Along with functional studies integral studies of individual membrane proteins are now widely implemented. The BIOMEMOS proposal goes a step further and approaches the function of the biomembrane at the higher level of membrane protein complexes. Through a combination of X-ray crystallography, electrophysiology, general biochemistry, biophysics and bioinformatics and including also the application of single-particle cryo-EM and small-angle X-ray scattering, the structure and function of membrane protein complexes of key importance in life will be investigated. The specific targets for investigation in this proposal include: 1) higher-order complexes of P-type ATPase pumps such as signalling complexes of Na+,K+-ATPase, and 2) development of methods for structural studies of membrane protein complexes Based on my unique track record in structural studies of large, difficult structures (ribosomes and membrane proteins) in the setting of a thriving research community in structural biology and biomembrane research in Aarhus provides a critical momentum for a long-term activity. The activity will take advantage of the new possibilities offered by synchrotron sources in Europe. Furthermore, a single-particle cryo-EM research group formed on my initiative in Aarhus, and a well-established small-angle X-ray scattering community provides for an optimal setting through multiple cues in structural biology and functional studies
Summary
The biomembrane is a prerequisite of life. It enables the cell to maintain a controlled environment and to establish electrochemical gradients as rapidly accessible energy stores. Biomembranes also provide scaffold for organisation and spatial definition of signal transmission in the cell. Crystal structures of membrane proteins are determined with an increasing pace. Along with functional studies integral studies of individual membrane proteins are now widely implemented. The BIOMEMOS proposal goes a step further and approaches the function of the biomembrane at the higher level of membrane protein complexes. Through a combination of X-ray crystallography, electrophysiology, general biochemistry, biophysics and bioinformatics and including also the application of single-particle cryo-EM and small-angle X-ray scattering, the structure and function of membrane protein complexes of key importance in life will be investigated. The specific targets for investigation in this proposal include: 1) higher-order complexes of P-type ATPase pumps such as signalling complexes of Na+,K+-ATPase, and 2) development of methods for structural studies of membrane protein complexes Based on my unique track record in structural studies of large, difficult structures (ribosomes and membrane proteins) in the setting of a thriving research community in structural biology and biomembrane research in Aarhus provides a critical momentum for a long-term activity. The activity will take advantage of the new possibilities offered by synchrotron sources in Europe. Furthermore, a single-particle cryo-EM research group formed on my initiative in Aarhus, and a well-established small-angle X-ray scattering community provides for an optimal setting through multiple cues in structural biology and functional studies
Max ERC Funding
2 444 180 €
Duration
Start date: 2010-04-01, End date: 2015-03-31
Project acronym CHILIC
Project Child health intervention interactions in low-income countries
Researcher (PI) Christine Benn
Host Institution (HI) STATENS SERUM INSTITUT
Call Details Starting Grant (StG), LS7, ERC-2009-StG
Summary Vitamin A supplementation (VAS) and vaccines are the most powerful tools to reduce child mortality in low-income countries. However, we may not use these interventions optimally because we disregard that the interventions may have immunomodulatory effects which differ for boys and girls and which may interact with the effects of other interventions. I have proposed the hypothesis that VAS and vaccines interact. This hypothesis is supported by randomised and observational studies showing that the combination of VAS and DTP may be harmful. I have furthermore proposed that VAS has sex-differential effects. VAS seems beneficial for boys but may not carry any benefits for girls. These findings challenge the current understanding that VAS and vaccines have only targeted effects and can be given together without considering interactions. This is of outmost importance for policy makers. The global trend is to combine health interventions for logistic reasons. My research suggests that this may not always be a good idea. Furthermore, the concept of sex-differential response to our common health interventions opens up for a completely new understanding of the immunology of the two sexes and may imply that we need to treat the two sexes differently in order to treat them optimally possibly also in high-income countries. In the present proposal I outline a series of inter-disciplinary epidemiological and immunological studies, which will serve to determine the overall and sex-differential effects of VAS and vaccines, the mechanisms behind these effects, and the basis for the immunological difference between boys and girls. If my hypotheses are true we can use the existing tools in a more optimal way to reduce child mortality without increasing costs. Thus, the results could lead to shifts in policy as well as paradigms.
Summary
Vitamin A supplementation (VAS) and vaccines are the most powerful tools to reduce child mortality in low-income countries. However, we may not use these interventions optimally because we disregard that the interventions may have immunomodulatory effects which differ for boys and girls and which may interact with the effects of other interventions. I have proposed the hypothesis that VAS and vaccines interact. This hypothesis is supported by randomised and observational studies showing that the combination of VAS and DTP may be harmful. I have furthermore proposed that VAS has sex-differential effects. VAS seems beneficial for boys but may not carry any benefits for girls. These findings challenge the current understanding that VAS and vaccines have only targeted effects and can be given together without considering interactions. This is of outmost importance for policy makers. The global trend is to combine health interventions for logistic reasons. My research suggests that this may not always be a good idea. Furthermore, the concept of sex-differential response to our common health interventions opens up for a completely new understanding of the immunology of the two sexes and may imply that we need to treat the two sexes differently in order to treat them optimally possibly also in high-income countries. In the present proposal I outline a series of inter-disciplinary epidemiological and immunological studies, which will serve to determine the overall and sex-differential effects of VAS and vaccines, the mechanisms behind these effects, and the basis for the immunological difference between boys and girls. If my hypotheses are true we can use the existing tools in a more optimal way to reduce child mortality without increasing costs. Thus, the results could lead to shifts in policy as well as paradigms.
Max ERC Funding
1 686 043 €
Duration
Start date: 2010-01-01, End date: 2014-12-31
Project acronym DEFACT
Project DNA repair factories how cells do biochemistry
Researcher (PI) Michael Lisby
Host Institution (HI) KOBENHAVNS UNIVERSITET
Call Details Starting Grant (StG), LS1, ERC-2009-StG
Summary The integrity of a cell's genome is constantly challenged by DNA lesions such as base modifications and DNA strand breaks. A single double-strand break is lethal if unrepaired and may lead to loss-of-heterozygosity, mutations, deletions, genomic rearrangements and chromosome loss if repaired improperly. Such genetic alterations are the main cause of cancer and other genetic diseases. Homologous recombination is an error-free pathway for repairing DNA lesions such as single- and double-strand breaks, and for the restart of collapsed replication forks. This pathway is catalyzed by giga-Dalton protein complexes consisting of dozens of different proteins. These DNA repair factories are able to catalyze complex, multi-step biochemical processes, which have so far failed reconstitution in vitro. The aim of this project is to establish an understanding of how cells catalyze complex biochemical processes such as homologous recombination in vivo. To reach this goal, we will seek to define the complete set of RNA and protein components of DNA repair factories using a combination of genetic, cell biological and biochemical approaches in the yeast Saccharomyces cerevisiae. Further, we will characterize the molecular architecture of DNA repair factories using fluorescence resonance energy transfer (FRET) and by applying systematic hybrid loss-of-heterozygosity (LOH) to physical interactions among DNA repair proteins. Key findings will be extended to metazoans using the chicken DT40 model system. My aim is to determine the fundamental molecular principles that govern protein factories in living cells. As such, our results are likely to be directly relevant to other protein factories such as DNA replication factories, PML bodies, nuclear pore complexes and transcription clusters.
Summary
The integrity of a cell's genome is constantly challenged by DNA lesions such as base modifications and DNA strand breaks. A single double-strand break is lethal if unrepaired and may lead to loss-of-heterozygosity, mutations, deletions, genomic rearrangements and chromosome loss if repaired improperly. Such genetic alterations are the main cause of cancer and other genetic diseases. Homologous recombination is an error-free pathway for repairing DNA lesions such as single- and double-strand breaks, and for the restart of collapsed replication forks. This pathway is catalyzed by giga-Dalton protein complexes consisting of dozens of different proteins. These DNA repair factories are able to catalyze complex, multi-step biochemical processes, which have so far failed reconstitution in vitro. The aim of this project is to establish an understanding of how cells catalyze complex biochemical processes such as homologous recombination in vivo. To reach this goal, we will seek to define the complete set of RNA and protein components of DNA repair factories using a combination of genetic, cell biological and biochemical approaches in the yeast Saccharomyces cerevisiae. Further, we will characterize the molecular architecture of DNA repair factories using fluorescence resonance energy transfer (FRET) and by applying systematic hybrid loss-of-heterozygosity (LOH) to physical interactions among DNA repair proteins. Key findings will be extended to metazoans using the chicken DT40 model system. My aim is to determine the fundamental molecular principles that govern protein factories in living cells. As such, our results are likely to be directly relevant to other protein factories such as DNA replication factories, PML bodies, nuclear pore complexes and transcription clusters.
Max ERC Funding
1 700 030 €
Duration
Start date: 2009-12-01, End date: 2014-11-30
Project acronym DIADRUG
Project Insulin resistance and diabetic nephropathy - development of novel in vivo models for drug discovery
Researcher (PI) Sanna Lehtonen
Host Institution (HI) HELSINGIN YLIOPISTO
Call Details Starting Grant (StG), LS9, ERC-2009-StG
Summary 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.
Summary
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.
Max ERC Funding
2 000 000 €
Duration
Start date: 2009-11-01, End date: 2014-10-31
Project acronym FUTUREGENES
Project Gene transfer techniques in the treatment of cardiovascular diseases and malignant glioma
Researcher (PI) Seppo Yla-Herttuala
Host Institution (HI) ITA-SUOMEN YLIOPISTO
Call Details Advanced Grant (AdG), LS7, ERC-2009-AdG
Summary Background: Poor angiogenesis and collateral vessel formation lead to coronary heart disease, claudication, infarctions and amputations while malignant glioma is one of the most aggressive proangiogenic tumors leading to death in a few months. For these diseases either stimulation or blocking, respectively, of angiogenesis may provide novel treatment options. Advancing State-of-the-Art: Our hypothesis is that in ischemia it will be possible to support natural growth of blood vessels with Therapeutic angiogenesis and lymphangiogenesis by using local gene transfer of the new members of vascular endothelial growth factor (VEGF) family and their receptors. New co-receptors, designer mutants and PCR suffling products of VEGFs will be used. New vector technology will be used to achieve long-lasting effects of VEGFs. We aim to develop novel site-specifically integrating, targeted, regulated vectors to precisely express the new VEGFs, their soluble decoy receptors and single-chain therapeutic antibodies (scFv) for pro- and anti-angiogenic purposes. As novel approaches, we have developed metabolically biotinylated lenti- and adenoviruses suitable for targeting and Epigenetherapy where siRNA/miRNAs and short nuclear RNAs regulate endogenous gene expression at the VEGF promoter level via modification of histone code. scFv library for endothelial cells and lentivirus-siRNA library directed to all human and mouse kinases will be screened to identify new mediators of angiogenesis in order to develop next generation pro- and antiangiogenic therapies. Based on our strong track record in Clinical applications, the best new pro- and antiangiogenic approaches will be taken to phase I clinical studies in myocardial ischemia and malignant glioma. Significance: This work should lead to significant advances and new therapies for severe ischemia and malignant glioma. Epigenetherapy and new site-specifically integrating, regulated vectors should be widely applicable in medicine.
Summary
Background: Poor angiogenesis and collateral vessel formation lead to coronary heart disease, claudication, infarctions and amputations while malignant glioma is one of the most aggressive proangiogenic tumors leading to death in a few months. For these diseases either stimulation or blocking, respectively, of angiogenesis may provide novel treatment options. Advancing State-of-the-Art: Our hypothesis is that in ischemia it will be possible to support natural growth of blood vessels with Therapeutic angiogenesis and lymphangiogenesis by using local gene transfer of the new members of vascular endothelial growth factor (VEGF) family and their receptors. New co-receptors, designer mutants and PCR suffling products of VEGFs will be used. New vector technology will be used to achieve long-lasting effects of VEGFs. We aim to develop novel site-specifically integrating, targeted, regulated vectors to precisely express the new VEGFs, their soluble decoy receptors and single-chain therapeutic antibodies (scFv) for pro- and anti-angiogenic purposes. As novel approaches, we have developed metabolically biotinylated lenti- and adenoviruses suitable for targeting and Epigenetherapy where siRNA/miRNAs and short nuclear RNAs regulate endogenous gene expression at the VEGF promoter level via modification of histone code. scFv library for endothelial cells and lentivirus-siRNA library directed to all human and mouse kinases will be screened to identify new mediators of angiogenesis in order to develop next generation pro- and antiangiogenic therapies. Based on our strong track record in Clinical applications, the best new pro- and antiangiogenic approaches will be taken to phase I clinical studies in myocardial ischemia and malignant glioma. Significance: This work should lead to significant advances and new therapies for severe ischemia and malignant glioma. Epigenetherapy and new site-specifically integrating, regulated vectors should be widely applicable in medicine.
Max ERC Funding
2 200 000 €
Duration
Start date: 2010-06-01, End date: 2015-05-31
