Project acronym Agglomerates
Project Infinite Protein Self-Assembly in Health and Disease
Researcher (PI) Emmanuel Doram LEVY
Host Institution (HI) WEIZMANN INSTITUTE OF SCIENCE
Call Details Consolidator Grant (CoG), LS2, ERC-2018-COG
Summary Understanding how proteins respond to mutations is of paramount importance to biology and disease. While protein stability and misfolding have been instrumental in rationalizing the impact of mutations, we recently discovered that an alternative route is also frequent, where mutations at the surface of symmetric proteins trigger novel self-interactions that lead to infinite self-assembly. This mechanism can be involved in disease, as in sickle-cell anemia, but may also serve in adaptation. Importantly, it differs fundamentally from aggregation, because misfolding does not drive it. Thus, we term it “agglomeration”. The ease with which agglomeration can occur, even by single point mutations, shifts the paradigm of how quickly new protein assemblies can emerge, both in health and disease. This prompts us to determine the basic principles of protein agglomeration and explore its implications in cell physiology and human disease.
We propose an interdisciplinary research program bridging atomic and cellular scales to explore agglomeration in three aims: (i) Map the landscape of protein agglomeration in response to mutation in endogenous yeast proteins; (ii) Characterize how yeast physiology impacts agglomeration by changes in gene expression or cell state, and, conversely, how protein agglomerates impact yeast fitness. (iii) Analyze agglomeration in relation to human disease via two approaches. First, by predicting single nucleotide polymorphisms that trigger agglomeration, prioritizing them using knowledge from Aims 1 & 2, and characterizing them experimentally. Second, by providing a proof-of-concept that agglomeration can be exploited in drug design, whereby drugs induce its formation, like mutations can do.
Overall, through this research, we aim to establish agglomeration as a paradigm for protein assembly, with implications for our understanding of evolution, physiology, and disease.
Summary
Understanding how proteins respond to mutations is of paramount importance to biology and disease. While protein stability and misfolding have been instrumental in rationalizing the impact of mutations, we recently discovered that an alternative route is also frequent, where mutations at the surface of symmetric proteins trigger novel self-interactions that lead to infinite self-assembly. This mechanism can be involved in disease, as in sickle-cell anemia, but may also serve in adaptation. Importantly, it differs fundamentally from aggregation, because misfolding does not drive it. Thus, we term it “agglomeration”. The ease with which agglomeration can occur, even by single point mutations, shifts the paradigm of how quickly new protein assemblies can emerge, both in health and disease. This prompts us to determine the basic principles of protein agglomeration and explore its implications in cell physiology and human disease.
We propose an interdisciplinary research program bridging atomic and cellular scales to explore agglomeration in three aims: (i) Map the landscape of protein agglomeration in response to mutation in endogenous yeast proteins; (ii) Characterize how yeast physiology impacts agglomeration by changes in gene expression or cell state, and, conversely, how protein agglomerates impact yeast fitness. (iii) Analyze agglomeration in relation to human disease via two approaches. First, by predicting single nucleotide polymorphisms that trigger agglomeration, prioritizing them using knowledge from Aims 1 & 2, and characterizing them experimentally. Second, by providing a proof-of-concept that agglomeration can be exploited in drug design, whereby drugs induce its formation, like mutations can do.
Overall, through this research, we aim to establish agglomeration as a paradigm for protein assembly, with implications for our understanding of evolution, physiology, and disease.
Max ERC Funding
2 574 819 €
Duration
Start date: 2019-04-01, End date: 2024-03-31
Project acronym ANTHROPOID
Project Great ape organoids to reconstruct uniquely human development
Researcher (PI) Jarrett CAMP
Host Institution (HI) INSTITUT FUR MOLEKULARE UND KLINISCHE OPHTHALMOLOGIE BASEL
Call Details Starting Grant (StG), LS2, ERC-2018-STG
Summary Humans diverged from our closest living relatives, chimpanzees and other great apes, 6-10 million years ago. Since this divergence, our ancestors acquired genetic changes that enhanced cognition, altered metabolism, and endowed our species with an adaptive capacity to colonize the entire planet and reshape the biosphere. Through genome comparisons between modern humans, Neandertals, chimpanzees and other apes we have identified genetic changes that likely contribute to innovations in human metabolic and cognitive physiology. However, it has been difficult to assess the functional effects of these genetic changes due to the lack of cell culture systems that recapitulate great ape organ complexity. Human and chimpanzee pluripotent stem cells (PSCs) can self-organize into three-dimensional (3D) tissues that recapitulate the morphology, function, and genetic programs controlling organ development. Our vision is to use organoids to study the changes that set modern humans apart from our closest evolutionary relatives as well as all other organisms on the planet. In ANTHROPOID we will generate a great ape developmental cell atlas using cortex, liver, and small intestine organoids. We will use single-cell transcriptomics and chromatin accessibility to identify cell type-specific features of transcriptome divergence at cellular resolution. We will dissect enhancer evolution using single-cell genomic screens and ancestralize human cells to resurrect pre-human cellular phenotypes. ANTHROPOID utilizes quantitative and state-of-the-art methods to explore exciting high-risk questions at multiple branches of the modern human lineage. This project is a ground breaking starting point to replay evolution and tackle the ancient question of what makes us uniquely human?
Summary
Humans diverged from our closest living relatives, chimpanzees and other great apes, 6-10 million years ago. Since this divergence, our ancestors acquired genetic changes that enhanced cognition, altered metabolism, and endowed our species with an adaptive capacity to colonize the entire planet and reshape the biosphere. Through genome comparisons between modern humans, Neandertals, chimpanzees and other apes we have identified genetic changes that likely contribute to innovations in human metabolic and cognitive physiology. However, it has been difficult to assess the functional effects of these genetic changes due to the lack of cell culture systems that recapitulate great ape organ complexity. Human and chimpanzee pluripotent stem cells (PSCs) can self-organize into three-dimensional (3D) tissues that recapitulate the morphology, function, and genetic programs controlling organ development. Our vision is to use organoids to study the changes that set modern humans apart from our closest evolutionary relatives as well as all other organisms on the planet. In ANTHROPOID we will generate a great ape developmental cell atlas using cortex, liver, and small intestine organoids. We will use single-cell transcriptomics and chromatin accessibility to identify cell type-specific features of transcriptome divergence at cellular resolution. We will dissect enhancer evolution using single-cell genomic screens and ancestralize human cells to resurrect pre-human cellular phenotypes. ANTHROPOID utilizes quantitative and state-of-the-art methods to explore exciting high-risk questions at multiple branches of the modern human lineage. This project is a ground breaking starting point to replay evolution and tackle the ancient question of what makes us uniquely human?
Max ERC Funding
1 500 000 €
Duration
Start date: 2019-06-01, End date: 2024-05-31
Project acronym BactRNA
Project Bacterial small RNAs networks unravelling novel features of transcription and translation
Researcher (PI) Maude Audrey Guillier
Host Institution (HI) CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS
Call Details Consolidator Grant (CoG), LS2, ERC-2018-COG
Summary Regulation of gene expression plays a key role in the ability of bacteria to rapidly adapt to changing environments and to colonize extremely diverse habitats. The relatively recent discovery of a plethora of small regulatory RNAs and the beginning of their characterization has unravelled new aspects of bacterial gene expression. First, the expression of many bacterial genes responds to a complex network of both transcriptional and post-transcriptional regulators. However, the properties of the resulting regulatory circuits on the dynamics of gene expression and in the bacterial adaptive response have been poorly addressed so far. In a first part of this project, we will tackle this question by characterizing the circuits that are formed between two widespread classes of bacterial regulators, the sRNAs and the two-component systems, which act at the post-transcriptional and the transcriptional level, respectively. The study of sRNAs also led to major breakthroughs regarding the basic mechanisms of gene expression. In particular, we recently showed that repressor sRNAs can target activating stem-loop structures located within the coding region of mRNAs that promote translation initiation, in striking contrast with the previously recognized inhibitory role of mRNA structures in translation. The second objective of this project is thus to draw an unprecedented map of non-canonical translation initiation events and their regulation by sRNAs.
Overall, this project will greatly improve our understanding of how bacteria can so rapidly and successfully adapt to many different environments, and in the long term, provide clues towards the development of anti-bacterial strategies.
Summary
Regulation of gene expression plays a key role in the ability of bacteria to rapidly adapt to changing environments and to colonize extremely diverse habitats. The relatively recent discovery of a plethora of small regulatory RNAs and the beginning of their characterization has unravelled new aspects of bacterial gene expression. First, the expression of many bacterial genes responds to a complex network of both transcriptional and post-transcriptional regulators. However, the properties of the resulting regulatory circuits on the dynamics of gene expression and in the bacterial adaptive response have been poorly addressed so far. In a first part of this project, we will tackle this question by characterizing the circuits that are formed between two widespread classes of bacterial regulators, the sRNAs and the two-component systems, which act at the post-transcriptional and the transcriptional level, respectively. The study of sRNAs also led to major breakthroughs regarding the basic mechanisms of gene expression. In particular, we recently showed that repressor sRNAs can target activating stem-loop structures located within the coding region of mRNAs that promote translation initiation, in striking contrast with the previously recognized inhibitory role of mRNA structures in translation. The second objective of this project is thus to draw an unprecedented map of non-canonical translation initiation events and their regulation by sRNAs.
Overall, this project will greatly improve our understanding of how bacteria can so rapidly and successfully adapt to many different environments, and in the long term, provide clues towards the development of anti-bacterial strategies.
Max ERC Funding
1 999 754 €
Duration
Start date: 2019-09-01, End date: 2024-08-31
Project acronym BRAIN-MATCH
Project Matching CNS Lineage Maps with Molecular Brain Tumor Portraits for Translational Exploitation
Researcher (PI) Stefan PFISTER
Host Institution (HI) DEUTSCHES KREBSFORSCHUNGSZENTRUM HEIDELBERG
Call Details Consolidator Grant (CoG), LS2, ERC-2018-COG
Summary Brain tumors represent an extremely heterogeneous group of more than 100 different molecularly distinct diseases, many of which are still almost uniformly lethal despite five decades of clinical trials. In contrast to hematologic malignancies and carcinomas, the cell-of-origin for the vast majority of these entities is unknown. This knowledge gap currently precludes a comprehensive understanding of tumor biology and also limits translational exploitation (e.g., utilizing lineage targets for novel therapies and circulating brain tumor cells for liquid biopsies).
The BRAIN-MATCH project represents an ambitious program to address this challenge and unmet medical need by taking an approach that (i) extensively utilizes existing molecular profiles of more than 30,000 brain tumor samples covering more than 100 different entities, publicly available single-cell sequencing data of normal brain regions, and bulk normal tissue data at different times of development across different species; (ii) generates unprecedented maps of normal human CNS development by using state-of-the art novel technologies; (iii) matches these molecular portraits of normal cell types with tumor datasets in order to identify specific cell-of-origin populations for individual tumor entities; and (iv) validates the most promising cell-of-origin populations and tumor-specific lineage and/or surface markers in vivo.
The expected outputs of BRAIN-MATCH are four-fold: (i) delivery of an unprecedented atlas of human normal CNS development, which will also be of great relevance for diverse fields other than cancer; (ii) functional validation of at least three lineage targets; (iii) isolation and molecular characterization of circulating brain tumor cells from patients´ blood for at least five tumor entities; and (iv) generation of at least three novel mouse models of brain tumor entities for which currently no faithful models exist.
Summary
Brain tumors represent an extremely heterogeneous group of more than 100 different molecularly distinct diseases, many of which are still almost uniformly lethal despite five decades of clinical trials. In contrast to hematologic malignancies and carcinomas, the cell-of-origin for the vast majority of these entities is unknown. This knowledge gap currently precludes a comprehensive understanding of tumor biology and also limits translational exploitation (e.g., utilizing lineage targets for novel therapies and circulating brain tumor cells for liquid biopsies).
The BRAIN-MATCH project represents an ambitious program to address this challenge and unmet medical need by taking an approach that (i) extensively utilizes existing molecular profiles of more than 30,000 brain tumor samples covering more than 100 different entities, publicly available single-cell sequencing data of normal brain regions, and bulk normal tissue data at different times of development across different species; (ii) generates unprecedented maps of normal human CNS development by using state-of-the art novel technologies; (iii) matches these molecular portraits of normal cell types with tumor datasets in order to identify specific cell-of-origin populations for individual tumor entities; and (iv) validates the most promising cell-of-origin populations and tumor-specific lineage and/or surface markers in vivo.
The expected outputs of BRAIN-MATCH are four-fold: (i) delivery of an unprecedented atlas of human normal CNS development, which will also be of great relevance for diverse fields other than cancer; (ii) functional validation of at least three lineage targets; (iii) isolation and molecular characterization of circulating brain tumor cells from patients´ blood for at least five tumor entities; and (iv) generation of at least three novel mouse models of brain tumor entities for which currently no faithful models exist.
Max ERC Funding
1 999 875 €
Duration
Start date: 2019-05-01, End date: 2024-04-30
Project acronym CRISPRsition
Project Developing CRISPR adaptation platforms for basic and applied research
Researcher (PI) Ehud Itzhak Qimron
Host Institution (HI) TEL AVIV UNIVERSITY
Call Details Consolidator Grant (CoG), LS2, ERC-2018-COG
Summary The CRISPR-Cas system has been extensively studied for its ability to cleave DNA. In contrast, studies of the ability of the system to acquire and integrate new DNA from invaders as a form of prokaryotic adaptive immunity, have lagged behind. This delay reflects the extreme enthusiasm surrounding the potential of using the system’s cleavage capabilities as a genome editing tool. However, the enormous potential of the adaptation process can and should arouse a similar degree of enthusiasm. My lab has pioneered studies on the CRISPR adaptation process by establishing new methodologies, and applying them to demonstrate the essential role of the proteins and DNA elements, as well as the molecular mechanisms, operating in this process. In this project, I will establish novel platforms for studying adaptation and develop them into biotechnological applications and research tools. These tools will allow me to identify the first natural and synthetic inhibitors of the adaptation process. This, in turn, will provide genetic tools to control adaptation, as well as advance the understanding of the arms race between bacteria and their invaders. I will also harness the adaptation process as a platform for diversifying genetic elements for phage display, and for extending phage recognition of a wide range of hosts. Lastly, I will provide the first evidence for an association between the CRISPR adaptation system and gene repression. This linkage will form the basis of a molecular scanner and recorder platform that I will develop and that can be used to identify crucial genetic elements in phage genomes as well as novel regulatory circuits in the bacterial genome. Together, my findings will represent a considerable leap in the understanding of CRISPR adaptation with respect to the process, potential applications, and the intriguing evolutionary significance.
Summary
The CRISPR-Cas system has been extensively studied for its ability to cleave DNA. In contrast, studies of the ability of the system to acquire and integrate new DNA from invaders as a form of prokaryotic adaptive immunity, have lagged behind. This delay reflects the extreme enthusiasm surrounding the potential of using the system’s cleavage capabilities as a genome editing tool. However, the enormous potential of the adaptation process can and should arouse a similar degree of enthusiasm. My lab has pioneered studies on the CRISPR adaptation process by establishing new methodologies, and applying them to demonstrate the essential role of the proteins and DNA elements, as well as the molecular mechanisms, operating in this process. In this project, I will establish novel platforms for studying adaptation and develop them into biotechnological applications and research tools. These tools will allow me to identify the first natural and synthetic inhibitors of the adaptation process. This, in turn, will provide genetic tools to control adaptation, as well as advance the understanding of the arms race between bacteria and their invaders. I will also harness the adaptation process as a platform for diversifying genetic elements for phage display, and for extending phage recognition of a wide range of hosts. Lastly, I will provide the first evidence for an association between the CRISPR adaptation system and gene repression. This linkage will form the basis of a molecular scanner and recorder platform that I will develop and that can be used to identify crucial genetic elements in phage genomes as well as novel regulatory circuits in the bacterial genome. Together, my findings will represent a considerable leap in the understanding of CRISPR adaptation with respect to the process, potential applications, and the intriguing evolutionary significance.
Max ERC Funding
2 000 000 €
Duration
Start date: 2019-12-01, End date: 2024-11-30
Project acronym EcoBox
Project Ecosystem in a box: Dissecting the dynamics of a defined microbial community in vitro
Researcher (PI) Karoline FAUST
Host Institution (HI) KATHOLIEKE UNIVERSITEIT LEUVEN
Call Details Starting Grant (StG), LS2, ERC-2018-STG
Summary The dynamics of microbial communities may be driven by the interactions between community members, controlled by the environment, shaped by immigration or random events, influenced by evolutionary processes or result from an interplay of all these factors. This project aims to improve our understanding of how community structure and the environment impact community dynamics. Towards this aim, a defined in vitro community of human gut bacteria will be assembled, since their genomes are available and their metabolism is comparatively well resolved.
In the first step, we will quantify the intrinsic variability of community dynamics and look for alternative stable states. Next, we will systematically vary community structure as well as nutrient supply and monitor their effects on the dynamics. Finally, we will measure model parameters, evaluate to what extent different community models predict observed community dynamics and validate the models by identifying and experimentally validating keystone species.
Studies of microbial community dynamics are hampered by the cost of obtaining densely sampled time series in replicates and by the difficulty of community manipulation. We will address these challenges by setting up an in vitro system for parallel and automated cultivation in well-controlled conditions and by working with defined communities, where every community member is known.
The proposed project will discern how external factors and community structure drive community dynamics and encode this knowledge in mathematical models. Moreover, the project has the potential to transform our view on alternative microbial communities and their interpretation. In addition, the project will extend our knowledge of human gut microorganisms and their interactions. These insights will ease the design of defined gut communities optimized for therapeutic purposes.
Summary
The dynamics of microbial communities may be driven by the interactions between community members, controlled by the environment, shaped by immigration or random events, influenced by evolutionary processes or result from an interplay of all these factors. This project aims to improve our understanding of how community structure and the environment impact community dynamics. Towards this aim, a defined in vitro community of human gut bacteria will be assembled, since their genomes are available and their metabolism is comparatively well resolved.
In the first step, we will quantify the intrinsic variability of community dynamics and look for alternative stable states. Next, we will systematically vary community structure as well as nutrient supply and monitor their effects on the dynamics. Finally, we will measure model parameters, evaluate to what extent different community models predict observed community dynamics and validate the models by identifying and experimentally validating keystone species.
Studies of microbial community dynamics are hampered by the cost of obtaining densely sampled time series in replicates and by the difficulty of community manipulation. We will address these challenges by setting up an in vitro system for parallel and automated cultivation in well-controlled conditions and by working with defined communities, where every community member is known.
The proposed project will discern how external factors and community structure drive community dynamics and encode this knowledge in mathematical models. Moreover, the project has the potential to transform our view on alternative microbial communities and their interpretation. In addition, the project will extend our knowledge of human gut microorganisms and their interactions. These insights will ease the design of defined gut communities optimized for therapeutic purposes.
Max ERC Funding
1 493 899 €
Duration
Start date: 2019-01-01, End date: 2023-12-31
Project acronym EPICROP
Project Dissecting epistasis for enhanced crop productivity
Researcher (PI) Sebastian Soyk
Host Institution (HI) UNIVERSITE DE LAUSANNE
Call Details Starting Grant (StG), LS2, ERC-2018-STG
Summary A major goal in plant biology is to understand how naturally occurring genetic variation leads to quantitative differences in economically important traits. Efforts to navigate the genotype-to-phenotype map are often focused on linear genetic interactions. As a result, crop breeding is mainly driven by loci with predictable additive effects. However, it has become clear that quantitative trait variation often results from perturbations of complex genetic networks. Thus, understanding epistasis, or interactions between genes, is key for our ability to predictably improve crops. To meet this challenge, this project will reveal and dissect epistatic interactions in gene regulatory networks that guide stem cell differentiation in the model crop tomato. In the first aim, I will utilize exhaustive allelic series for epistatic MADS-box genes that quantitatively regulate flower and fruit production as an experimental model system to study fundamental principles of epistasis that can be applied to other genetic networks. Genome-wide transcript profiling will be used to reveal molecular signatures of epistasis and potential targets for predictable crop improvement by advanced CRISPR/Cas9 gene editing technology. Further, my preliminary data suggests that epistasis is widespread and important across major productivity traits in tomato. Thus, in a second aim, I will access this untapped resource of cryptic genetic variation by sensitizing a tomato diversity panel for weak epistatic effects from unknown natural modifier loci of stem cell differentiation using trans-acting CRISPR/Cas9 editing cassettes. This screen represents a new approach to mutagenesis in plants with potential to reveal cryptic variation in other system. The outcomes of this project will advance our knowledge in a fundamental area of plant genome biology, help uncover and understand the functional architecture of epistasis, and have potential to bring significant improvements to agriculture.
Summary
A major goal in plant biology is to understand how naturally occurring genetic variation leads to quantitative differences in economically important traits. Efforts to navigate the genotype-to-phenotype map are often focused on linear genetic interactions. As a result, crop breeding is mainly driven by loci with predictable additive effects. However, it has become clear that quantitative trait variation often results from perturbations of complex genetic networks. Thus, understanding epistasis, or interactions between genes, is key for our ability to predictably improve crops. To meet this challenge, this project will reveal and dissect epistatic interactions in gene regulatory networks that guide stem cell differentiation in the model crop tomato. In the first aim, I will utilize exhaustive allelic series for epistatic MADS-box genes that quantitatively regulate flower and fruit production as an experimental model system to study fundamental principles of epistasis that can be applied to other genetic networks. Genome-wide transcript profiling will be used to reveal molecular signatures of epistasis and potential targets for predictable crop improvement by advanced CRISPR/Cas9 gene editing technology. Further, my preliminary data suggests that epistasis is widespread and important across major productivity traits in tomato. Thus, in a second aim, I will access this untapped resource of cryptic genetic variation by sensitizing a tomato diversity panel for weak epistatic effects from unknown natural modifier loci of stem cell differentiation using trans-acting CRISPR/Cas9 editing cassettes. This screen represents a new approach to mutagenesis in plants with potential to reveal cryptic variation in other system. The outcomes of this project will advance our knowledge in a fundamental area of plant genome biology, help uncover and understand the functional architecture of epistasis, and have potential to bring significant improvements to agriculture.
Max ERC Funding
1 499 903 €
Duration
Start date: 2019-08-01, End date: 2024-07-31
Project acronym EpiRIME
Project Epigenetic Reprogramming, Inheritance and Memory: Dissect epigenetic transitions at fertilisation and early embryogenesis
Researcher (PI) Nicola Iovino
Host Institution (HI) MAX-PLANCK-GESELLSCHAFT ZUR FORDERUNG DER WISSENSCHAFTEN EV
Call Details Consolidator Grant (CoG), LS2, ERC-2018-COG
Summary During gametogenesis, germ cells undergo profound chromatin reorganisation, condensation and transcriptional shutdown. Upon fertilization, gamete chromatin is epigenetically reprogrammed, generating a totipotent zygote that can give rise to all cell types of the adult organism. The maternal factors that reprogram gametes to totipotency are unknown. The current dogma suggests that the parental epigenetic information must be erased in order to establish totipotency.
In contrast, we have recently discovered that maternal gametes transmit the epigenetic H3K27me3 histone modification to the next generation (Zenk et al., Science, 2017) adding to increasing evidence suggesting that gametes convey more than just DNA to the offspring. Nevertheless, the underlying mechanisms and the impact of epigenetic inheritance through the gametes are not yet fully resolved. Critically, the mechanisms and impact of (i) paternal gamete reprogramming, (ii) paternal epigenetic inheritance and (iii) de novo establishment of the zygotic epigenome remain essentially unknown.
The objective of this proposal is to unravel the fundamental principles underlying these three major epigenetic transitions in vivo in Drosophila.
We will achieve our objective via three aims: (i) We will investigate the mechanisms underlying the reprogramming of sperm chromatin at fertilization. Specifically, we will determine the nature and extent of the contributions of two proteins essential for sperm chromatin reprogramming (ii) We will examine the mechanism of histone H3K27me3 inheritance through the paternal germline (iii) We will genetically dissect the de novo establishment of constitutive heterochromatin in the newly formed zygote.
Our investigations of these epigenetic transitions are expected to reveal novel insights into the first steps in the formation of life, and to ultimately advance reproductive and regenerative medicine.
Summary
During gametogenesis, germ cells undergo profound chromatin reorganisation, condensation and transcriptional shutdown. Upon fertilization, gamete chromatin is epigenetically reprogrammed, generating a totipotent zygote that can give rise to all cell types of the adult organism. The maternal factors that reprogram gametes to totipotency are unknown. The current dogma suggests that the parental epigenetic information must be erased in order to establish totipotency.
In contrast, we have recently discovered that maternal gametes transmit the epigenetic H3K27me3 histone modification to the next generation (Zenk et al., Science, 2017) adding to increasing evidence suggesting that gametes convey more than just DNA to the offspring. Nevertheless, the underlying mechanisms and the impact of epigenetic inheritance through the gametes are not yet fully resolved. Critically, the mechanisms and impact of (i) paternal gamete reprogramming, (ii) paternal epigenetic inheritance and (iii) de novo establishment of the zygotic epigenome remain essentially unknown.
The objective of this proposal is to unravel the fundamental principles underlying these three major epigenetic transitions in vivo in Drosophila.
We will achieve our objective via three aims: (i) We will investigate the mechanisms underlying the reprogramming of sperm chromatin at fertilization. Specifically, we will determine the nature and extent of the contributions of two proteins essential for sperm chromatin reprogramming (ii) We will examine the mechanism of histone H3K27me3 inheritance through the paternal germline (iii) We will genetically dissect the de novo establishment of constitutive heterochromatin in the newly formed zygote.
Our investigations of these epigenetic transitions are expected to reveal novel insights into the first steps in the formation of life, and to ultimately advance reproductive and regenerative medicine.
Max ERC Funding
1 997 500 €
Duration
Start date: 2019-07-01, End date: 2024-06-30
Project acronym Host-TB
Project Using CRISPR genome screens and dual transcriptome analyses to dissect host-pathogen interactions in tuberculosis
Researcher (PI) Sergey NEJENTSEV
Host Institution (HI) STICHTING VUMC
Call Details Advanced Grant (AdG), LS2, ERC-2018-ADG
Summary With more than 10 million cases annually, tuberculosis (TB) remains a global health problem. TB epidemic is exacerbated by the spread of multidrug-resistant TB. Host-directed therapies (HDTs) can improve immune mechanisms by augmenting the ability of host cells to kill M. tuberculosis (Mtb) or by modulating the immune response to prevent excessive inflammation, cell death and tissue damage. Progress with HDT development has been slowed down by the limited understanding of host-pathogen interactions during Mtb infection. Screens of the whole human genome can identify novel genes involved in the immune responses to Mtb infection and susceptibility to TB. Previously, we successfully used genome-wide association studies to identify human genes associated with susceptibility to TB. Here, we will for the first time use the groundbreaking CRISPR technology to screen the human genome in macrophages infected with Mtb and discover genes that are critically involved in host-pathogen interactions. Then, we will comprehensively characterise pathways that mediate impacts of these genes on both the human macrophage and the intracellular Mtb bacilli using dual transcriptome analyses and high-throughput microscopy assays. This novel approach will dissect crucial mechanisms of host-pathogen interaction during Mtb infection and will point to new targets for HDTs of TB.
Summary
With more than 10 million cases annually, tuberculosis (TB) remains a global health problem. TB epidemic is exacerbated by the spread of multidrug-resistant TB. Host-directed therapies (HDTs) can improve immune mechanisms by augmenting the ability of host cells to kill M. tuberculosis (Mtb) or by modulating the immune response to prevent excessive inflammation, cell death and tissue damage. Progress with HDT development has been slowed down by the limited understanding of host-pathogen interactions during Mtb infection. Screens of the whole human genome can identify novel genes involved in the immune responses to Mtb infection and susceptibility to TB. Previously, we successfully used genome-wide association studies to identify human genes associated with susceptibility to TB. Here, we will for the first time use the groundbreaking CRISPR technology to screen the human genome in macrophages infected with Mtb and discover genes that are critically involved in host-pathogen interactions. Then, we will comprehensively characterise pathways that mediate impacts of these genes on both the human macrophage and the intracellular Mtb bacilli using dual transcriptome analyses and high-throughput microscopy assays. This novel approach will dissect crucial mechanisms of host-pathogen interaction during Mtb infection and will point to new targets for HDTs of TB.
Max ERC Funding
2 160 926 €
Duration
Start date: 2019-06-01, End date: 2024-05-31
Project acronym ImmuNiche
Project Identifying spatial determinants of immune cell fate commitment
Researcher (PI) Dominic GRÜN
Host Institution (HI) MAX-PLANCK-GESELLSCHAFT ZUR FORDERUNG DER WISSENSCHAFTEN EV
Call Details Consolidator Grant (CoG), LS2, ERC-2018-COG
Summary The complex architecture of the mammalian hematopoietic system has been studied for decades, yet the molecular mechanisms underlying cell fate commitment remain poorly understood. Although cytokine signals are major determinants of hematopoietic cell fate, only few niches of stem and progenitor cells have been characterized. Here, we propose to resolve the cell type composition of mouse bone marrow by integrating single-cell RNA-seq with high-resolution spatial analysis of gene expression in tissue sections, visualizing ~250 cell type specific markers by multiplexed single-molecule RNA fluorescent in situ hybridization. This approach will reveal preferential co-localization of hematopoietic cells with other bone marrow-resident cell types, and globally predict niches of hematopoietic progenitor sub-types to pinpoint microenvrionmental determinants of hematopoietic cell fate. Based on this reference we will investigate the role of the microenvironment in the pathogenesis of myelodysplastic syndromes (MDS), representing one of the most frequent blood cell malignancies, commonly giving rise to leukaemia with poor prognosis. To investigate conservation of microenvironmental determinants of normal and malignant hematopoietic differentiation in human, we will apply the same strategy to healthy donors and human MDS patients and compare predicted cell types, differentiation trajectories, and niche interactions to those derived from healthy mice and murine MDS models. This approach will reveal conserved niche regulators of cell fate commitment involved in disease pathogenesis, which we will functionally analyse in murine models to identify novel therapeutic targets for prevention and treatment of MDS. Our approach represents a blueprint for investigating human malignancies of under-characterized tissues by applying cutting-edge high-resolution techniques in combination with advanced computational methods to jointly analyse the murine model and human patient microenvironment.
Summary
The complex architecture of the mammalian hematopoietic system has been studied for decades, yet the molecular mechanisms underlying cell fate commitment remain poorly understood. Although cytokine signals are major determinants of hematopoietic cell fate, only few niches of stem and progenitor cells have been characterized. Here, we propose to resolve the cell type composition of mouse bone marrow by integrating single-cell RNA-seq with high-resolution spatial analysis of gene expression in tissue sections, visualizing ~250 cell type specific markers by multiplexed single-molecule RNA fluorescent in situ hybridization. This approach will reveal preferential co-localization of hematopoietic cells with other bone marrow-resident cell types, and globally predict niches of hematopoietic progenitor sub-types to pinpoint microenvrionmental determinants of hematopoietic cell fate. Based on this reference we will investigate the role of the microenvironment in the pathogenesis of myelodysplastic syndromes (MDS), representing one of the most frequent blood cell malignancies, commonly giving rise to leukaemia with poor prognosis. To investigate conservation of microenvironmental determinants of normal and malignant hematopoietic differentiation in human, we will apply the same strategy to healthy donors and human MDS patients and compare predicted cell types, differentiation trajectories, and niche interactions to those derived from healthy mice and murine MDS models. This approach will reveal conserved niche regulators of cell fate commitment involved in disease pathogenesis, which we will functionally analyse in murine models to identify novel therapeutic targets for prevention and treatment of MDS. Our approach represents a blueprint for investigating human malignancies of under-characterized tissues by applying cutting-edge high-resolution techniques in combination with advanced computational methods to jointly analyse the murine model and human patient microenvironment.
Max ERC Funding
1 997 500 €
Duration
Start date: 2019-07-01, End date: 2024-06-30
