Project acronym ACTIVATION OF XCI
Project Molecular mechanisms controlling X chromosome inactivation
Researcher (PI) Joost Henk Gribnau
Host Institution (HI) ERASMUS UNIVERSITAIR MEDISCH CENTRUM ROTTERDAM
Call Details Starting Grant (StG), LS2, ERC-2010-StG_20091118
Summary In mammals, gene dosage of X-chromosomal genes is equalized between sexes by random inactivation of either one of the two X chromosomes in female cells. In the initial phase of X chromosome inactivation (XCI), a counting and initiation process determines the number of X chromosomes per nucleus, and elects the future inactive X chromosome (Xi). Xist is an X-encoded gene that plays a crucial role in the XCI process. At the start of XCI Xist expression is up-regulated and Xist RNA accumulates on the future Xi thereby initiating silencing in cis. Recent work performed in my laboratory indicates that the counting and initiation process is directed by a stochastic mechanism, in which each X chromosome has an independent probability to be inactivated. We also found that this probability is determined by the X:ploïdy ratio. These results indicated the presence of at least one X-linked activator of XCI. With a BAC screen we recently identified X-encoded RNF12 to be a dose-dependent activator of XCI. Expression of RNF12 correlates with Xist expression, and a heterozygous deletion of Rnf12 results in a marked loss of XCI in female cells. The presence of a small proportion of cells that still initiate XCI, in Rnf12+/- cells, also indicated that more XCI-activators are involved in XCI. Here, we propose to investigate the molecular mechanism by which RNF12 activates XCI in mouse and human, and to search for additional XCI-activators. We will also attempt to establish the role of different inhibitors of XCI, including CTCF and the pluripotency factors OCT4, SOX2 and NANOG. We anticipate that these studies will significantly advance our understanding of XCI mechanisms, which is highly relevant for a better insight in the manifestation of X-linked diseases that are affected by XCI.
Summary
In mammals, gene dosage of X-chromosomal genes is equalized between sexes by random inactivation of either one of the two X chromosomes in female cells. In the initial phase of X chromosome inactivation (XCI), a counting and initiation process determines the number of X chromosomes per nucleus, and elects the future inactive X chromosome (Xi). Xist is an X-encoded gene that plays a crucial role in the XCI process. At the start of XCI Xist expression is up-regulated and Xist RNA accumulates on the future Xi thereby initiating silencing in cis. Recent work performed in my laboratory indicates that the counting and initiation process is directed by a stochastic mechanism, in which each X chromosome has an independent probability to be inactivated. We also found that this probability is determined by the X:ploïdy ratio. These results indicated the presence of at least one X-linked activator of XCI. With a BAC screen we recently identified X-encoded RNF12 to be a dose-dependent activator of XCI. Expression of RNF12 correlates with Xist expression, and a heterozygous deletion of Rnf12 results in a marked loss of XCI in female cells. The presence of a small proportion of cells that still initiate XCI, in Rnf12+/- cells, also indicated that more XCI-activators are involved in XCI. Here, we propose to investigate the molecular mechanism by which RNF12 activates XCI in mouse and human, and to search for additional XCI-activators. We will also attempt to establish the role of different inhibitors of XCI, including CTCF and the pluripotency factors OCT4, SOX2 and NANOG. We anticipate that these studies will significantly advance our understanding of XCI mechanisms, which is highly relevant for a better insight in the manifestation of X-linked diseases that are affected by XCI.
Max ERC Funding
1 500 000 €
Duration
Start date: 2011-04-01, End date: 2016-03-31
Project acronym BioNanoPattern
Project Protein nano-patterning using DNA nanotechnology; control of surface-based immune system activation
Researcher (PI) Thomas Harry SHARP
Host Institution (HI) ACADEMISCH ZIEKENHUIS LEIDEN
Call Details Starting Grant (StG), LS9, ERC-2017-STG
Summary Protein nanopatterning concerns the geometric arrangement of individual proteins with nanometre accuracy. It is becoming apparent that protein nanopatterns are essential for cellular function, and have roles in cell signalling and protection, phagocytosis and stem cell differentiation. Recent research indicates that our immune system is activated by nanopatterned antibody platforms, which initiate the classical Complement pathway by binding to the first component of Complement, the C1 complex. DNA nanotechnology can be used to form self-assembled nanoscale structures, which are ideal for use as templates to pattern proteins with specific geometries and nanometre accuracy. I propose to use DNA to nanopattern antigens and agonistic aptamers with defined geometry to study and control Complement pathway activation by the C1 complex.
To develop and demonstrate the potential use of DNA to nanopattern proteins, the first aim of this proposal is to design DNA nanotemplates suitable for patterning antibody-binding sites. Antibodies and C1 will bind with specific geometry, and the relationship between antibody geometry and Complement activation will be assessed using novel liposome assays. Using DNA to mimic antigenic surfaces will enable high-resolution structure determination of DNA-antibody-C1 complexes, both in solution and on lipid bilayer surfaces, using phase plate cryo-electron microscopy to elucidate the structure-activation relationship of C1.
The second aim of this proposal is to evolve agonistic aptamers that directly bind to and activate C1, and incorporate these into DNA nanotemplates. These nanopatterned aptamers will enable further study of C1 activation, and allow direct targeting of Complement activation to specific cells within a population of cell types to demonstrate targeted cell killing. This may open up new and highly efficient ways to activate our immune system in vivo, with potential for targeted anti-tumour immunotherapies.
Summary
Protein nanopatterning concerns the geometric arrangement of individual proteins with nanometre accuracy. It is becoming apparent that protein nanopatterns are essential for cellular function, and have roles in cell signalling and protection, phagocytosis and stem cell differentiation. Recent research indicates that our immune system is activated by nanopatterned antibody platforms, which initiate the classical Complement pathway by binding to the first component of Complement, the C1 complex. DNA nanotechnology can be used to form self-assembled nanoscale structures, which are ideal for use as templates to pattern proteins with specific geometries and nanometre accuracy. I propose to use DNA to nanopattern antigens and agonistic aptamers with defined geometry to study and control Complement pathway activation by the C1 complex.
To develop and demonstrate the potential use of DNA to nanopattern proteins, the first aim of this proposal is to design DNA nanotemplates suitable for patterning antibody-binding sites. Antibodies and C1 will bind with specific geometry, and the relationship between antibody geometry and Complement activation will be assessed using novel liposome assays. Using DNA to mimic antigenic surfaces will enable high-resolution structure determination of DNA-antibody-C1 complexes, both in solution and on lipid bilayer surfaces, using phase plate cryo-electron microscopy to elucidate the structure-activation relationship of C1.
The second aim of this proposal is to evolve agonistic aptamers that directly bind to and activate C1, and incorporate these into DNA nanotemplates. These nanopatterned aptamers will enable further study of C1 activation, and allow direct targeting of Complement activation to specific cells within a population of cell types to demonstrate targeted cell killing. This may open up new and highly efficient ways to activate our immune system in vivo, with potential for targeted anti-tumour immunotherapies.
Max ERC Funding
1 499 850 €
Duration
Start date: 2018-01-01, End date: 2022-12-31
Project acronym BURSTREG
Project Single-molecule visualization of transcription dynamics to understand regulatory mechanisms of transcriptional bursting and its effects on cellular fitness
Researcher (PI) Tineke LENSTRA
Host Institution (HI) STICHTING HET NEDERLANDS KANKER INSTITUUT-ANTONI VAN LEEUWENHOEK ZIEKENHUIS
Call Details Starting Grant (StG), LS1, ERC-2017-STG
Summary Transcription in single cells is a stochastic process that arises from the random collision of molecules, resulting in heterogeneity in gene expression in cell populations. This heterogeneity in gene expression influences cell fate decisions and disease progression. Interestingly, gene expression variability is not the same for every gene: noise can vary by several orders of magnitude across transcriptomes. The reason for this transcript-specific behavior is that genes are not transcribed in a continuous fashion, but can show transcriptional bursting, with periods of gene activity followed by periods of inactivity. The noisiness of a gene can be tuned by changing the duration and the rate of switching between periods of activity and inactivity. Even though transcriptional bursting is conserved from bacteria to yeast to human cells, the origin and regulators of bursting remain largely unknown. Here, I will use cutting-edge single-molecule RNA imaging techniques to directly observe and measure transcriptional bursting in living yeast cells. First, bursting properties will be quantified at different endogenous and mutated genes to evaluate the contribution of cis-regulatory promoter elements on bursting. Second, the role of trans-regulatory complexes will be characterized by dynamic depletion or gene-specific targeting of transcription regulatory proteins and observing changes in RNA synthesis in real-time. Third, I will develop a new technology to visualize the binding dynamics of single transcription factor molecules at the transcription site, so that the stability of upstream regulatory factors and the RNA output can directly be compared in the same cell. Finally, I will examine the phenotypic effect of different bursting patterns on organismal fitness. Overall, these approaches will reveal how bursting is regulated at the molecular level and how different bursting patterns affect the heterogeneity and fitness of the organism.
Summary
Transcription in single cells is a stochastic process that arises from the random collision of molecules, resulting in heterogeneity in gene expression in cell populations. This heterogeneity in gene expression influences cell fate decisions and disease progression. Interestingly, gene expression variability is not the same for every gene: noise can vary by several orders of magnitude across transcriptomes. The reason for this transcript-specific behavior is that genes are not transcribed in a continuous fashion, but can show transcriptional bursting, with periods of gene activity followed by periods of inactivity. The noisiness of a gene can be tuned by changing the duration and the rate of switching between periods of activity and inactivity. Even though transcriptional bursting is conserved from bacteria to yeast to human cells, the origin and regulators of bursting remain largely unknown. Here, I will use cutting-edge single-molecule RNA imaging techniques to directly observe and measure transcriptional bursting in living yeast cells. First, bursting properties will be quantified at different endogenous and mutated genes to evaluate the contribution of cis-regulatory promoter elements on bursting. Second, the role of trans-regulatory complexes will be characterized by dynamic depletion or gene-specific targeting of transcription regulatory proteins and observing changes in RNA synthesis in real-time. Third, I will develop a new technology to visualize the binding dynamics of single transcription factor molecules at the transcription site, so that the stability of upstream regulatory factors and the RNA output can directly be compared in the same cell. Finally, I will examine the phenotypic effect of different bursting patterns on organismal fitness. Overall, these approaches will reveal how bursting is regulated at the molecular level and how different bursting patterns affect the heterogeneity and fitness of the organism.
Max ERC Funding
1 950 775 €
Duration
Start date: 2018-01-01, End date: 2022-12-31
Project acronym deFIBER
Project Dissecting the cellular and molecular dynamics of bone marrow fibrosis for improved diagnostics and treatment
Researcher (PI) Rebekka SCHNEIDER-KRAMANN
Host Institution (HI) ERASMUS UNIVERSITAIR MEDISCH CENTRUM ROTTERDAM
Call Details Starting Grant (StG), LS4, ERC-2017-STG
Summary Bone marrow (BM) fibrosis is the continuous replacement of blood forming cells in the bone marrow by scar tissue, ultimately leading to failure of the body to produce blood cells. Primary myelofibrosis (PMF), an incurable blood cancer, is the prototypic example of the step-wise development of BM fibrosis. The specific mechanisms that cause BM fibrosis are not understood, in particular as the cells driving fibrosis have remained obscure.
My recent findings demonstrate that Gli1+ cells are fibrosis-driving cells in PMF, that their frequency correlates with fibrosis severity in patients, and that their ablation ameliorates BM fibrosis. These results indicate that Gli1+ cells are the primary effector cells in BM fibrosis and that they represent a highly attractive therapeutic target. This puts me in a unique position to vastly expand our knowledge of the BM fibrosis pathogenesis, improve diagnostics, and discover new therapeutic strategies for this fatal disease. I will do this by: 1) dissecting the molecular and cellular mechanisms of the fibrotic transformation, 2) defining the stepwise disease evolution by genetic fate tracing and analysis of the previously unknown critical effector cells of BM fibrosis , 3) understanding early forms of BM fibrosis for improved diagnostics in patients, all with the ultimate aim to identify novel therapeutic targets to directly block the cellular and molecular changes occuring in BM fibrosis.
I will apply state-of-the-art techniques, including genetic fate tracing experiments, conditional genetic knockout mouse models, tissue engineering of the bone marrow niche and in vivo and in vitro CRISPR/Cas9 gene editing, to unravel the complex molecular and cellular interaction between fibrosis-causing cells and the malignant hematopoietic cells. I will translate these findings into patient samples with the aim to improve the early diagnosis of the disease and to ultimately develop novel targeted therapies with curative intentions.
Summary
Bone marrow (BM) fibrosis is the continuous replacement of blood forming cells in the bone marrow by scar tissue, ultimately leading to failure of the body to produce blood cells. Primary myelofibrosis (PMF), an incurable blood cancer, is the prototypic example of the step-wise development of BM fibrosis. The specific mechanisms that cause BM fibrosis are not understood, in particular as the cells driving fibrosis have remained obscure.
My recent findings demonstrate that Gli1+ cells are fibrosis-driving cells in PMF, that their frequency correlates with fibrosis severity in patients, and that their ablation ameliorates BM fibrosis. These results indicate that Gli1+ cells are the primary effector cells in BM fibrosis and that they represent a highly attractive therapeutic target. This puts me in a unique position to vastly expand our knowledge of the BM fibrosis pathogenesis, improve diagnostics, and discover new therapeutic strategies for this fatal disease. I will do this by: 1) dissecting the molecular and cellular mechanisms of the fibrotic transformation, 2) defining the stepwise disease evolution by genetic fate tracing and analysis of the previously unknown critical effector cells of BM fibrosis , 3) understanding early forms of BM fibrosis for improved diagnostics in patients, all with the ultimate aim to identify novel therapeutic targets to directly block the cellular and molecular changes occuring in BM fibrosis.
I will apply state-of-the-art techniques, including genetic fate tracing experiments, conditional genetic knockout mouse models, tissue engineering of the bone marrow niche and in vivo and in vitro CRISPR/Cas9 gene editing, to unravel the complex molecular and cellular interaction between fibrosis-causing cells and the malignant hematopoietic cells. I will translate these findings into patient samples with the aim to improve the early diagnosis of the disease and to ultimately develop novel targeted therapies with curative intentions.
Max ERC Funding
1 498 544 €
Duration
Start date: 2018-01-01, End date: 2022-12-31
Project acronym ENABLE
Project Advancing cell based therapies by supporting implant survival
Researcher (PI) Jeroen Leijten
Host Institution (HI) UNIVERSITEIT TWENTE
Call Details Starting Grant (StG), LS7, ERC-2017-STG
Summary Tissue engineering aims at the creation of living implants to replace, repair, or regenerate damaged, diseased, or aged tissues, which holds tremendous possibilities to both extend our lives and improve our quality of life. During the last decades, our ability to create small tissues to heal small animals e.g. mice and rats has taken a breath taking leap. However, we have relentlessly struggled to create viable tissues of human-relevant sizes. Creating solid large tissues imposes lethal nutrient diffusion limitations, which causes the living implant to suffer from starvation, loss of function, and inevitable failure.
I hypothesize that this key challenge can be tackled by recruiting and developing advanced enabling nano- and micro-technologies. The ENABLE project begins with the design and development of a widely applicable platform that will enable large solid engineered tissues to survive and function by actively sustaining the implants metabolic needs. This platform is based on a unique two pronged strategy that rely on distinct technologies: oxygen releasing micromaterials, fabricated using a next-generation droplet generator, to enable short term survival of the implant, while embedded bioprinting will endow implants with a complex 3D vascular network to enable their long term survival. As proof of principle, the effects of ENABLE’s platform will be investigated using a critical bone defect in which I analyse the survival and function of the created living implants.
The anticipated outcomes of this proposal are three fold: first, I will develop a next-generation engineered tissue that will overcome the current size restrictions via the use of enabling technologies; second, I will reveal new knowledge on the role of the oxygen tension on vascularization and tissue formation by enabling control over the in vivo oxygen tension; and third, I will develop a novel strategy that enables the treatment of critical bone defects.
Summary
Tissue engineering aims at the creation of living implants to replace, repair, or regenerate damaged, diseased, or aged tissues, which holds tremendous possibilities to both extend our lives and improve our quality of life. During the last decades, our ability to create small tissues to heal small animals e.g. mice and rats has taken a breath taking leap. However, we have relentlessly struggled to create viable tissues of human-relevant sizes. Creating solid large tissues imposes lethal nutrient diffusion limitations, which causes the living implant to suffer from starvation, loss of function, and inevitable failure.
I hypothesize that this key challenge can be tackled by recruiting and developing advanced enabling nano- and micro-technologies. The ENABLE project begins with the design and development of a widely applicable platform that will enable large solid engineered tissues to survive and function by actively sustaining the implants metabolic needs. This platform is based on a unique two pronged strategy that rely on distinct technologies: oxygen releasing micromaterials, fabricated using a next-generation droplet generator, to enable short term survival of the implant, while embedded bioprinting will endow implants with a complex 3D vascular network to enable their long term survival. As proof of principle, the effects of ENABLE’s platform will be investigated using a critical bone defect in which I analyse the survival and function of the created living implants.
The anticipated outcomes of this proposal are three fold: first, I will develop a next-generation engineered tissue that will overcome the current size restrictions via the use of enabling technologies; second, I will reveal new knowledge on the role of the oxygen tension on vascularization and tissue formation by enabling control over the in vivo oxygen tension; and third, I will develop a novel strategy that enables the treatment of critical bone defects.
Max ERC Funding
1 500 000 €
Duration
Start date: 2018-01-01, End date: 2022-12-31
Project acronym ENCODING IN AXONS
Project Identifying mechanisms of information encoding in myelinated single axons
Researcher (PI) Maarten Kole
Host Institution (HI) KONINKLIJKE NEDERLANDSE AKADEMIE VAN WETENSCHAPPEN - KNAW
Call Details Starting Grant (StG), LS5, ERC-2010-StG_20091118
Summary A major challenge in neuroscience is to understand how information is stored and coded within single nerve cells (neurons) and across neuron populations in the brain. Nerve cell fibres (axons) are thought to provide the wiring to connect neurons and conduct the electrical nerve impulse (action potential; AP). Recent discoveries, however, show that the initial part of axons actively participates in modulating APs and providing a means to enhance the computational repertoire of neurons in the central nervous system. To decrease the temporal delay in information transmission over long distances most axons are myelinated. Here, we will test the hypothesis that the degree of myelination of single axons directly and indirectly influences the mechanisms of AP generation and neural coding. We will use a novel approach of patch-clamp recording combined with immunohistochemical and ultrastructural identification to develop a detailed model of single myelinated neocortical axons. We also will investigate the neuron-glia interactions responsible for the myelination process and measure whether their development follows an activity-dependent process. Finally, we will elucidate the physiological and molecular similarities and discrepancies between myelinated and experimentally demyelinated single neocortical axons. These studies will provide a novel methodological framework to study central nervous system axons and yield basic insights into myelin physiology and pathophysiology.
Summary
A major challenge in neuroscience is to understand how information is stored and coded within single nerve cells (neurons) and across neuron populations in the brain. Nerve cell fibres (axons) are thought to provide the wiring to connect neurons and conduct the electrical nerve impulse (action potential; AP). Recent discoveries, however, show that the initial part of axons actively participates in modulating APs and providing a means to enhance the computational repertoire of neurons in the central nervous system. To decrease the temporal delay in information transmission over long distances most axons are myelinated. Here, we will test the hypothesis that the degree of myelination of single axons directly and indirectly influences the mechanisms of AP generation and neural coding. We will use a novel approach of patch-clamp recording combined with immunohistochemical and ultrastructural identification to develop a detailed model of single myelinated neocortical axons. We also will investigate the neuron-glia interactions responsible for the myelination process and measure whether their development follows an activity-dependent process. Finally, we will elucidate the physiological and molecular similarities and discrepancies between myelinated and experimentally demyelinated single neocortical axons. These studies will provide a novel methodological framework to study central nervous system axons and yield basic insights into myelin physiology and pathophysiology.
Max ERC Funding
1 994 640 €
Duration
Start date: 2011-04-01, End date: 2016-03-31
Project acronym EvoCellBio
Project A combined in vitro and in vivo approach to dissect biochemical network evolution.
Researcher (PI) Liedewij LAAN
Host Institution (HI) TECHNISCHE UNIVERSITEIT DELFT
Call Details Starting Grant (StG), LS3, ERC-2017-STG
Summary How do organisms evolve? I propose to study how biochemical networks reorganize during evolution without compromising fitness. This is a complex problem: firstly, it is hard to know if a mutation increased fitness because this depends on the environment it arose in, which is typically unknown. Secondly, it is hard to find out how adaptive mutations improve fitness, because in cells, all biochemical networks are connected. I will reduce the complexity by two approaches, focused on symmetry-breaking in budding yeast, a functionally conserved process, which is the first step for polarity establishment and essential for proliferation.
First, I will study how adaptive mutations improve fitness in yeast cells, which are evolved after the deletion of an important symmetry-breaking gene. I will use fluorescent live-cell microscopy of polarisation markers to measure fitness, defined as the rate of symmetry breaking. I will combine my data with a kinetic mathematical model to determine how specific network structures facilitate evolutionary network reorganisation.
Second, to test predicted network structures, I will build minimal evolvable networks for symmetry breaking in vitro. In my definition of such a network, all of the components are essential for either fitness or evolvability. I will encapsulate the necessary proteins in emulsion droplets to form a functional evolvable network and use fluorescence microscopy to measure its fitness (the rate of a single protein-spot formation on a droplet membrane) and evolvability (the number of accessible neutral or adaptive mutations in the one-step mutational landscape of the network). Next, I will study how increasing the number of components affects the network’s evolvability and fitness.
This research will explain how proteins essential in one species have been lost in closely related species. My expertise with in vitro systems, modelling, biophysics and evolution makes me uniquely qualified for this ambitious project.
Summary
How do organisms evolve? I propose to study how biochemical networks reorganize during evolution without compromising fitness. This is a complex problem: firstly, it is hard to know if a mutation increased fitness because this depends on the environment it arose in, which is typically unknown. Secondly, it is hard to find out how adaptive mutations improve fitness, because in cells, all biochemical networks are connected. I will reduce the complexity by two approaches, focused on symmetry-breaking in budding yeast, a functionally conserved process, which is the first step for polarity establishment and essential for proliferation.
First, I will study how adaptive mutations improve fitness in yeast cells, which are evolved after the deletion of an important symmetry-breaking gene. I will use fluorescent live-cell microscopy of polarisation markers to measure fitness, defined as the rate of symmetry breaking. I will combine my data with a kinetic mathematical model to determine how specific network structures facilitate evolutionary network reorganisation.
Second, to test predicted network structures, I will build minimal evolvable networks for symmetry breaking in vitro. In my definition of such a network, all of the components are essential for either fitness or evolvability. I will encapsulate the necessary proteins in emulsion droplets to form a functional evolvable network and use fluorescence microscopy to measure its fitness (the rate of a single protein-spot formation on a droplet membrane) and evolvability (the number of accessible neutral or adaptive mutations in the one-step mutational landscape of the network). Next, I will study how increasing the number of components affects the network’s evolvability and fitness.
This research will explain how proteins essential in one species have been lost in closely related species. My expertise with in vitro systems, modelling, biophysics and evolution makes me uniquely qualified for this ambitious project.
Max ERC Funding
1 500 000 €
Duration
Start date: 2018-02-01, End date: 2023-01-31
Project acronym GlycoEdit
Project New Chemical Tools for Precision Glycotherapy
Researcher (PI) Thomas BOLTJE
Host Institution (HI) STICHTING KATHOLIEKE UNIVERSITEIT
Call Details Starting Grant (StG), PE5, ERC-2017-STG
Summary Glycosylation, the expression of carbohydrate structures on proteins and lipids, is found in all the domains of life. The collection of all glycans found on a cell is called the “glycome” which is information rich and a key player in a plethora of physiological and pathological processes. The information that the glycome holds can be written, read and erased by glycosyltransferases, lectins and glycosidases, respectively. The immense structural complexity and the fact that glycan biosynthesis is not under direct genetic control makes it very difficult to study the glycome.
The glycosylation pattern of cancer cells is very different from that of healthy cells. It is still unclear whether aberrant glycosylation of cancer cells is a cause or consequence of tumorigenesis but it is associated with aggressive and invasive forms of cancer and hence poor prognosis. Malignant glycans are directly involved in a number of mechanisms that suppress the immune response, increase migration and extravasation (metastasis), block apoptosis and increase resistance to chemotherapy.
The aim of this proposal is develop new glycomimetics that can be used to edit the glycome of cancer cells to target such evasive mechanisms. Using combinations of new glycan based inhibitors, a coordinated attack on the cancer glycome can be carried out which is expected to severely cripple the cancers ability to grow and metastasize. This will make the tumor more susceptible to immune mediated killing which may be further enhanced in combination with other anti-cancer strategies.
To minimize systemic side effects, new methods for the local delivery/activation of glycan inhibitors will be developed. The developed methods are expected to have a much broader than just cancer alone since the studied mechanisms are also associated with autoimmune and neurodegenerative disease.
Summary
Glycosylation, the expression of carbohydrate structures on proteins and lipids, is found in all the domains of life. The collection of all glycans found on a cell is called the “glycome” which is information rich and a key player in a plethora of physiological and pathological processes. The information that the glycome holds can be written, read and erased by glycosyltransferases, lectins and glycosidases, respectively. The immense structural complexity and the fact that glycan biosynthesis is not under direct genetic control makes it very difficult to study the glycome.
The glycosylation pattern of cancer cells is very different from that of healthy cells. It is still unclear whether aberrant glycosylation of cancer cells is a cause or consequence of tumorigenesis but it is associated with aggressive and invasive forms of cancer and hence poor prognosis. Malignant glycans are directly involved in a number of mechanisms that suppress the immune response, increase migration and extravasation (metastasis), block apoptosis and increase resistance to chemotherapy.
The aim of this proposal is develop new glycomimetics that can be used to edit the glycome of cancer cells to target such evasive mechanisms. Using combinations of new glycan based inhibitors, a coordinated attack on the cancer glycome can be carried out which is expected to severely cripple the cancers ability to grow and metastasize. This will make the tumor more susceptible to immune mediated killing which may be further enhanced in combination with other anti-cancer strategies.
To minimize systemic side effects, new methods for the local delivery/activation of glycan inhibitors will be developed. The developed methods are expected to have a much broader than just cancer alone since the studied mechanisms are also associated with autoimmune and neurodegenerative disease.
Max ERC Funding
1 500 000 €
Duration
Start date: 2017-11-01, End date: 2022-10-31
Project acronym MINDS
Project Microvesicle-inspired drug delivery systems
Researcher (PI) Raymond Michel Schiffelers
Host Institution (HI) UNIVERSITAIR MEDISCH CENTRUM UTRECHT
Call Details Starting Grant (StG), LS7, ERC-2010-StG_20091118
Summary Current state-of-the-art drug carrier systems deliver new ‘biological drugs’ (like proteins and nucleic acids) poorly to the target site. This is something I daily experience in my research on delivery of small interfering RNA. The in vivo challenges are threefold:
• Biological drugs are fragile molecules (subject to degradation and denaturation)
• They need to gain access to the target site
• They need delivery to a specific cell type and even to a specific subcellular location to be active.
Recent research points out an endogenous communication system transporting proteins and nucleic acids between cells, outperforming current synthetic drug delivery systems. These carriers, known as microvesicles, appear Nature’s choice for delivery of biologicals and have created excitement in the research community. Microvesicles encompass a variety of submicron vesicular structures that include exosomes, shedding vesicles, and microparticles. The lipids, proteins, mRNA and microRNA delivered by these microvesicles change the phenotype of the receiving cells. Microvesicles appear to play an important role in many disease processes, most notably inflammation and cancer, where their efficient functional delivery of biological cargo contributes to the disease progress. Up to now, most research addresses the role of microvesicles in cell biology. At the same time, surprisingly little is known about their in vivo kinetics, targeting behavior and tissue distribution from a drug carrier standpoint.
The aim of my proposal is to design and develop microvesicle-inspired drug delivery systems to improve targeting and delivery of biological drugs.
The work plan is divided into two approaches:
1-A synthetic approach based on liposomes or isolated microvesicle constituents
2-A biological approach based on biotechnologically-engineered and cell-produced microvesicles.
The results of this research are expected to improve insights into in vivo behavior of microvesicles and the critical molecules that trigger their delivery and targeting success. It should also be clear which of the two approaches is best suited for the production of pharmaceutically acceptable microvesicle-mimics. Finally, the research should result in a prototype microvesicle-inspired carrier. These results can form the basis for an attractive new generation of microvesicle mimicking drug delivery systems.
Summary
Current state-of-the-art drug carrier systems deliver new ‘biological drugs’ (like proteins and nucleic acids) poorly to the target site. This is something I daily experience in my research on delivery of small interfering RNA. The in vivo challenges are threefold:
• Biological drugs are fragile molecules (subject to degradation and denaturation)
• They need to gain access to the target site
• They need delivery to a specific cell type and even to a specific subcellular location to be active.
Recent research points out an endogenous communication system transporting proteins and nucleic acids between cells, outperforming current synthetic drug delivery systems. These carriers, known as microvesicles, appear Nature’s choice for delivery of biologicals and have created excitement in the research community. Microvesicles encompass a variety of submicron vesicular structures that include exosomes, shedding vesicles, and microparticles. The lipids, proteins, mRNA and microRNA delivered by these microvesicles change the phenotype of the receiving cells. Microvesicles appear to play an important role in many disease processes, most notably inflammation and cancer, where their efficient functional delivery of biological cargo contributes to the disease progress. Up to now, most research addresses the role of microvesicles in cell biology. At the same time, surprisingly little is known about their in vivo kinetics, targeting behavior and tissue distribution from a drug carrier standpoint.
The aim of my proposal is to design and develop microvesicle-inspired drug delivery systems to improve targeting and delivery of biological drugs.
The work plan is divided into two approaches:
1-A synthetic approach based on liposomes or isolated microvesicle constituents
2-A biological approach based on biotechnologically-engineered and cell-produced microvesicles.
The results of this research are expected to improve insights into in vivo behavior of microvesicles and the critical molecules that trigger their delivery and targeting success. It should also be clear which of the two approaches is best suited for the production of pharmaceutically acceptable microvesicle-mimics. Finally, the research should result in a prototype microvesicle-inspired carrier. These results can form the basis for an attractive new generation of microvesicle mimicking drug delivery systems.
Max ERC Funding
1 338 000 €
Duration
Start date: 2010-12-01, End date: 2015-11-30
Project acronym MRS in diabetes
Project Novel methods in Magnetic Resonance Spectroscopy to investigate mechanisms underlying metabolic disease
Researcher (PI) Vera SCHRAUWEN-HINDERLING
Host Institution (HI) UNIVERSITEIT MAASTRICHT
Call Details Starting Grant (StG), LS4, ERC-2017-STG
Summary The high prevalence of obesity-related metabolic disease such as diabetes and cardiovascular disease urgently requires earlier interventions in the pathogenesis of these diseases and the identification of new therapeutic targets that work in humans. To show the human relevance of mechanistic information gained from rodent and cell studies on the pathogenesis of insulin resistance and diabetes, human translational research is needed. Non-invasive techniques are key in human translational research. Magnetic Resonance Spectroscopy (MRS) is used in metabolic research, e.g. to determine ectopic lipids, but its potential is far from fully explored, and novel, dedicated MRS sequences can be developed to target new metabolites in vivo. As an example, I recently showed that the metabolite acetylcarnitine can be quantified by a modified MRS protocol, which allowed me to demonstrate in humans that acetylcarnitine concentrations in muscle strongly associate with insulin sensitivity. Furthermore, it lead to pilot data that show that the capacity to form acetylcarnitine can be hampered by low carnitine availability and that this is a determinant of metabolic flexibility, which can be succesfully targeted by carnitine supplementation. In the current proposal I aim to develop novel MRS methodology to detect levels of NAD+, another metabolite that is emerging from animal research as a central regulator of metabolic health. I will develop a non-invasive method to reliably quantify NAD+ in skeletal muscle by MRS. After careful validation of the method, I will determine the physiological relevance in a human cross-sectional study. I hypothesize that NAD+ and formation of acetylcarnitine act synergistically in determining protein acetylation and thereby affect metabolic flexibility. I will test this original hypothesis by increasing NAD+ and free carnitine using human interventional experiments and will investigate if this is a novel strategy to improve metabolic health.
Summary
The high prevalence of obesity-related metabolic disease such as diabetes and cardiovascular disease urgently requires earlier interventions in the pathogenesis of these diseases and the identification of new therapeutic targets that work in humans. To show the human relevance of mechanistic information gained from rodent and cell studies on the pathogenesis of insulin resistance and diabetes, human translational research is needed. Non-invasive techniques are key in human translational research. Magnetic Resonance Spectroscopy (MRS) is used in metabolic research, e.g. to determine ectopic lipids, but its potential is far from fully explored, and novel, dedicated MRS sequences can be developed to target new metabolites in vivo. As an example, I recently showed that the metabolite acetylcarnitine can be quantified by a modified MRS protocol, which allowed me to demonstrate in humans that acetylcarnitine concentrations in muscle strongly associate with insulin sensitivity. Furthermore, it lead to pilot data that show that the capacity to form acetylcarnitine can be hampered by low carnitine availability and that this is a determinant of metabolic flexibility, which can be succesfully targeted by carnitine supplementation. In the current proposal I aim to develop novel MRS methodology to detect levels of NAD+, another metabolite that is emerging from animal research as a central regulator of metabolic health. I will develop a non-invasive method to reliably quantify NAD+ in skeletal muscle by MRS. After careful validation of the method, I will determine the physiological relevance in a human cross-sectional study. I hypothesize that NAD+ and formation of acetylcarnitine act synergistically in determining protein acetylation and thereby affect metabolic flexibility. I will test this original hypothesis by increasing NAD+ and free carnitine using human interventional experiments and will investigate if this is a novel strategy to improve metabolic health.
Max ERC Funding
1 500 000 €
Duration
Start date: 2018-03-01, End date: 2023-02-28
