Project acronym BrainBreaks
Project DNA Breaks Shape Neural Genome Heterogeneity
Researcher (PI) Pei-Chi Wei
Host Institution (HI) DEUTSCHES KREBSFORSCHUNGSZENTRUM HEIDELBERG
Country Germany
Call Details Starting Grant (StG), LS2, ERC-2020-STG
Summary Neural progenitor cells undergo tens of thousands of cell divisions to generate the 80 billion neurons in a human brain. In neural progenitor cells, replication stress can lead to recurrent DNA break clusters (RDCs). Joining of two RDC breaks may introduce somatic genomic diversity. On the other hand, unbalanced genomic mosaicism in neural progenitor cells may lead to brain cancer and neuropsychiatric disorders. This proposal will test whether cell-autonomous DNA lesions that accumulate during rapid progenitor division contribute to the genetic heterogeneity found across neuronal cell populations.
Aim 1 will elucidate how replication stress drives recurrent break clusters in the neural progenitor cell genome. We will evaluate whether chromatin loop extrusion mechanistically contributes to breakage repairs, and thus helps shape genomic structure variations.
Aim 2 will quantify the extent and impact of tissue-specific recurrent break clusters in the embryonic brain. I will create a mouse model to identify DNA breaks in temporal and cell-type-specific manner across the entire population of neuronal progenitor cells.
Aim 3 will evaluate whether replicative stress drives the recurrent genomic alteration in the RDC-containing gene during embryonic neurogenesis. We will investigate one of the RDC-containing gene Neurexin 1, where deletion or truncation results in neurological disorders.
By combining a powerful in vitro cell line-based tool, versatile in vivo mouse models, and cutting-edge multi-omics approaches, we will uncover the mechanisms that are critical to the fields of genomics and developmental neuroscience and may also provide valuable new insights into neuropsychiatric disorders and tumor biology.
Summary
Neural progenitor cells undergo tens of thousands of cell divisions to generate the 80 billion neurons in a human brain. In neural progenitor cells, replication stress can lead to recurrent DNA break clusters (RDCs). Joining of two RDC breaks may introduce somatic genomic diversity. On the other hand, unbalanced genomic mosaicism in neural progenitor cells may lead to brain cancer and neuropsychiatric disorders. This proposal will test whether cell-autonomous DNA lesions that accumulate during rapid progenitor division contribute to the genetic heterogeneity found across neuronal cell populations.
Aim 1 will elucidate how replication stress drives recurrent break clusters in the neural progenitor cell genome. We will evaluate whether chromatin loop extrusion mechanistically contributes to breakage repairs, and thus helps shape genomic structure variations.
Aim 2 will quantify the extent and impact of tissue-specific recurrent break clusters in the embryonic brain. I will create a mouse model to identify DNA breaks in temporal and cell-type-specific manner across the entire population of neuronal progenitor cells.
Aim 3 will evaluate whether replicative stress drives the recurrent genomic alteration in the RDC-containing gene during embryonic neurogenesis. We will investigate one of the RDC-containing gene Neurexin 1, where deletion or truncation results in neurological disorders.
By combining a powerful in vitro cell line-based tool, versatile in vivo mouse models, and cutting-edge multi-omics approaches, we will uncover the mechanisms that are critical to the fields of genomics and developmental neuroscience and may also provide valuable new insights into neuropsychiatric disorders and tumor biology.
Max ERC Funding
1 500 000 €
Duration
Start date: 2021-01-01, End date: 2025-12-31
Project acronym BugDrug
Project Bugs as Drugs: Understanding Microbial Interaction Networks to Prevent and Treat Infections
Researcher (PI) Christoph RATZKE
Host Institution (HI) EBERHARD KARLS UNIVERSITAET TUEBINGEN
Country Germany
Call Details Starting Grant (StG), LS2, ERC-2020-STG
Summary Our body is inhabited by an enormous number of microbes which are crucial for our health. The bacteria in our gut for example help us to digest food but can also make us sick in case of an infection. The overall composition and function of those communities are strongly shaped by the interactions of the microbes within them e.g. how the microbes influence each others growth. These interactions are especially important in the case of microbial infections. Since pathogens have to interact with the microbial communities that already inhabit the host, the native microbes can repel or support the pathogens and thus protect the host from diseases or even facilitate them. Although microbial assemblages are pivotal for our health we have currently no satisfying way to get a mechanistic understanding of them which would be crucial to specifically manipulate these communities e.g. for therapeutic interventions. The aim of this proposal is to develop a microscopy method that allows to obtain interaction networks within complex communities. Since bacteria can only share the same space if they tolerate each other but avoid each other in case of competition, interaction networks of complex communities can be derived from the spatial co-occurence of the bacteria that form them. With this technology I especially want to investigate how pathogens embed into the native gut microbial community of the model organism Caenorhabditis elegans. I want to understand how interactions between pathogens and native gut microbiota can protect a host from infections. Finally, I want to use this technology to identify bacteria in the native gut community that can outcompete pathogens and work as specific probiotic against microbial infections. This approach could revolutionize the usage of probiotics and offer completely new ways to prevent and treat infectious diseases, which are especially valuable in times where we see more and more pathogens become resistant against antibiotics.
Summary
Our body is inhabited by an enormous number of microbes which are crucial for our health. The bacteria in our gut for example help us to digest food but can also make us sick in case of an infection. The overall composition and function of those communities are strongly shaped by the interactions of the microbes within them e.g. how the microbes influence each others growth. These interactions are especially important in the case of microbial infections. Since pathogens have to interact with the microbial communities that already inhabit the host, the native microbes can repel or support the pathogens and thus protect the host from diseases or even facilitate them. Although microbial assemblages are pivotal for our health we have currently no satisfying way to get a mechanistic understanding of them which would be crucial to specifically manipulate these communities e.g. for therapeutic interventions. The aim of this proposal is to develop a microscopy method that allows to obtain interaction networks within complex communities. Since bacteria can only share the same space if they tolerate each other but avoid each other in case of competition, interaction networks of complex communities can be derived from the spatial co-occurence of the bacteria that form them. With this technology I especially want to investigate how pathogens embed into the native gut microbial community of the model organism Caenorhabditis elegans. I want to understand how interactions between pathogens and native gut microbiota can protect a host from infections. Finally, I want to use this technology to identify bacteria in the native gut community that can outcompete pathogens and work as specific probiotic against microbial infections. This approach could revolutionize the usage of probiotics and offer completely new ways to prevent and treat infectious diseases, which are especially valuable in times where we see more and more pathogens become resistant against antibiotics.
Max ERC Funding
1 494 437 €
Duration
Start date: 2021-01-01, End date: 2025-12-31
Project acronym CisTune
Project Regulatory Logic, Thresholds and Epigenetic Memory: How cis-regulatory landscapes tune gene activity during mammalian development
Researcher (PI) Edda SCHULZ
Host Institution (HI) MAX-PLANCK-GESELLSCHAFT ZUR FORDERUNG DER WISSENSCHAFTEN EV
Country Germany
Call Details Starting Grant (StG), LS2, ERC-2020-STG
Summary Development of multicellular organisms relies on differential gene activation in a single genome. In response to multiple quantitative signals, cell-type specific transcriptional programs are established that determine cell identify. Their perturbation can result in pathologies such as cancer. Large cis-regulatory landscapes integrate information on cell state, space and time to precisely tune the activity of developmental genes. How cis-regulatory landscapes decode multiple quantitative signals remains poorly understood. CisTune aims at gaining a functional and mechanistic understanding of how regulator levels are sensed, how the input from multiple regulators is integrated and how information is processed by cis-regulatory landscapes.
CisTune will use the Xist locus as a model, which controls X-chromosome inactivation, an essential developmental process in mammals. Xist's cis-regulatory landscape integrates multiple quantitative input signals that transmit information on sex and developmental time, to ensure up-regulation from one X chromosome in each female cell. In CisTune we will thus study an essential process in great depth to identify regulatory principles that control activity of the mammalian genome during development.
CisTune will use an interdisciplinary approach at the intersection of systems biology, epigenetics and gene regulation, where highly multiplexed perturbation experiments of endogenous genes are interpreted with the help of mathematical models. We will build on recent technological breakthroughs, including single-cell genomics and high-throughput CRISPR screens, which we will complement with a new approach to functionally link sequence elements to their input signals. CisTune has the potential to overcome challenges that have prevented mammalian quantitative biology of gene regulation to becoming more broadly applied and will set the stage for investigating gene regulation across multiple layers of complexity.
Summary
Development of multicellular organisms relies on differential gene activation in a single genome. In response to multiple quantitative signals, cell-type specific transcriptional programs are established that determine cell identify. Their perturbation can result in pathologies such as cancer. Large cis-regulatory landscapes integrate information on cell state, space and time to precisely tune the activity of developmental genes. How cis-regulatory landscapes decode multiple quantitative signals remains poorly understood. CisTune aims at gaining a functional and mechanistic understanding of how regulator levels are sensed, how the input from multiple regulators is integrated and how information is processed by cis-regulatory landscapes.
CisTune will use the Xist locus as a model, which controls X-chromosome inactivation, an essential developmental process in mammals. Xist's cis-regulatory landscape integrates multiple quantitative input signals that transmit information on sex and developmental time, to ensure up-regulation from one X chromosome in each female cell. In CisTune we will thus study an essential process in great depth to identify regulatory principles that control activity of the mammalian genome during development.
CisTune will use an interdisciplinary approach at the intersection of systems biology, epigenetics and gene regulation, where highly multiplexed perturbation experiments of endogenous genes are interpreted with the help of mathematical models. We will build on recent technological breakthroughs, including single-cell genomics and high-throughput CRISPR screens, which we will complement with a new approach to functionally link sequence elements to their input signals. CisTune has the potential to overcome challenges that have prevented mammalian quantitative biology of gene regulation to becoming more broadly applied and will set the stage for investigating gene regulation across multiple layers of complexity.
Max ERC Funding
1 494 375 €
Duration
Start date: 2021-08-01, End date: 2026-07-31
Project acronym DECIPHER
Project A computational framework to interpret the chemical language of the microbiome
Researcher (PI) Marnix MEDEMA
Host Institution (HI) WAGENINGEN UNIVERSITY
Country Netherlands
Call Details Starting Grant (StG), LS2, ERC-2020-STG
Summary "Humans, animals and plants are covered in microbes. Such microbiomes have a major impact on the health of their hosts and have been linked to traits like growth promotion, stress resilience, and diseases. However, the mechanisms underlying microbiome-host interactions remain poorly understood. Recent studies have shown that microbiome-associated phenotypes are often mediated by specific molecules, a ‘chemical language’ that enables microbes to interact with each other and with the host. The biosynthesis of these molecules is encoded in metabolic gene clusters (MGCs) that are subject to frequent horizontal transfer and are therefore highly strain-specific.
Current computational methods for analysing microbiomes largely focus on comparative taxonomic analyses and generic metabolism, and overlook this complex ""chemical dialog"". Indeed, no adequate methods are available to systematically study the roles of MGCs in microbiomes. At the same time, metabolomics data from microbiomes are full of ‘dark matter’: unknown molecules that cannot be traced to their producers. Here, I propose to develop the first comprehensive computational framework to study the chemical language of the microbiome.
In the past years, I have developed technologies that lay the foundation for this ERC project, including automated identification of MGCs, grouping them into families and annotating them using reference data. With DECIPHER, I will move my research to the next level, by developing new algorithms to link MGCs to their metabolic products and to predict their molecular and ecological functions in microbiomes. I will then apply this new framework in a systematic study of microbiome- associated phenotypes in plants and humans. Together, the innovations proposed here will fill a key gap in the analysis of microbiome function and pave the way toward precision-engineering of microbiomes with specific metabolic capabilities for designer soils and microbiome-based therapies."
Summary
"Humans, animals and plants are covered in microbes. Such microbiomes have a major impact on the health of their hosts and have been linked to traits like growth promotion, stress resilience, and diseases. However, the mechanisms underlying microbiome-host interactions remain poorly understood. Recent studies have shown that microbiome-associated phenotypes are often mediated by specific molecules, a ‘chemical language’ that enables microbes to interact with each other and with the host. The biosynthesis of these molecules is encoded in metabolic gene clusters (MGCs) that are subject to frequent horizontal transfer and are therefore highly strain-specific.
Current computational methods for analysing microbiomes largely focus on comparative taxonomic analyses and generic metabolism, and overlook this complex ""chemical dialog"". Indeed, no adequate methods are available to systematically study the roles of MGCs in microbiomes. At the same time, metabolomics data from microbiomes are full of ‘dark matter’: unknown molecules that cannot be traced to their producers. Here, I propose to develop the first comprehensive computational framework to study the chemical language of the microbiome.
In the past years, I have developed technologies that lay the foundation for this ERC project, including automated identification of MGCs, grouping them into families and annotating them using reference data. With DECIPHER, I will move my research to the next level, by developing new algorithms to link MGCs to their metabolic products and to predict their molecular and ecological functions in microbiomes. I will then apply this new framework in a systematic study of microbiome- associated phenotypes in plants and humans. Together, the innovations proposed here will fill a key gap in the analysis of microbiome function and pave the way toward precision-engineering of microbiomes with specific metabolic capabilities for designer soils and microbiome-based therapies."
Max ERC Funding
1 499 965 €
Duration
Start date: 2020-12-01, End date: 2025-11-30
Project acronym DNA_MICROSCOPY
Project In situ DNA sequencing-based microscopy for subcellular spatial transcriptomics
Researcher (PI) Ian Torao HOFFECKER
Host Institution (HI) KUNGLIGA TEKNISKA HOEGSKOLAN
Country Sweden
Call Details Starting Grant (StG), LS2, ERC-2020-STG
Summary New tools are needed in spatial transcriptomics, which uses imaging to resolve the positions of RNA in their native biological contexts to reveal molecular mechanisms underlying cell states and interactions. The developing embryo exhibits complex transcriptional regulation during the maternal-to-zygotic transition, when the reservoir of maternal RNA is phased out and the zygotic genes are turned on. Existing spatial sequencing approaches cannot simultaneously achieve subcellular resolution, whole transcriptome coverage, isotropic 3D resolution, and the parallel mapping of proteins and genetic regulatory elements that is desirable to form a mechanistic picture of transcriptional regulation in any system as complex is the embryo. The nascent field of DNA microscopy is perfectly suited for the high-throughput, multiplexed, molecular mapping needs of such problems. DNA microscopy uses carefully engineered in situ PCR, next-generation sequencing, and the mathematics of stochastic geometry instead of optics to convey microscale spatial information. In 2018, I developed a 2D DNA microscopy approach based on topological reconstruction of adjacent patches of barcoded DNA “polonies”. My work is among a few papers appearing just in the last year that together constitute a new field. I will adapt my 2D topological DNA microscopy method to one based on in situ whole-transcriptome sequencing in 3D, aiming to achieve subcellular resolution. I will also develop the mathematical basis of topological reconstruction and new computational tools to deal with the 3D data. Finally, I aim to incorporate the capability to localize other molecules in parallel with the transcriptome such as oligonucleotide-conjugated antibodies targeting specific transcription factors and other genetic regulatory elements. The technique, deployed on developing C. elegans embryos, will be used to study spatially dependent regulatory mechanisms such as the determination of cell polarity and fate during cleavage in greater breadth and depth than ever previously achieved. By being optics-free, this has the potential to overcome fundamental limitations imposed by traditional forms of microscopy, greatly expand the ease and throughput of spatial transcriptomics, and pave the way for routine use of hyper-multiplexed molecular imaging.
Summary
New tools are needed in spatial transcriptomics, which uses imaging to resolve the positions of RNA in their native biological contexts to reveal molecular mechanisms underlying cell states and interactions. The developing embryo exhibits complex transcriptional regulation during the maternal-to-zygotic transition, when the reservoir of maternal RNA is phased out and the zygotic genes are turned on. Existing spatial sequencing approaches cannot simultaneously achieve subcellular resolution, whole transcriptome coverage, isotropic 3D resolution, and the parallel mapping of proteins and genetic regulatory elements that is desirable to form a mechanistic picture of transcriptional regulation in any system as complex is the embryo. The nascent field of DNA microscopy is perfectly suited for the high-throughput, multiplexed, molecular mapping needs of such problems. DNA microscopy uses carefully engineered in situ PCR, next-generation sequencing, and the mathematics of stochastic geometry instead of optics to convey microscale spatial information. In 2018, I developed a 2D DNA microscopy approach based on topological reconstruction of adjacent patches of barcoded DNA “polonies”. My work is among a few papers appearing just in the last year that together constitute a new field. I will adapt my 2D topological DNA microscopy method to one based on in situ whole-transcriptome sequencing in 3D, aiming to achieve subcellular resolution. I will also develop the mathematical basis of topological reconstruction and new computational tools to deal with the 3D data. Finally, I aim to incorporate the capability to localize other molecules in parallel with the transcriptome such as oligonucleotide-conjugated antibodies targeting specific transcription factors and other genetic regulatory elements. The technique, deployed on developing C. elegans embryos, will be used to study spatially dependent regulatory mechanisms such as the determination of cell polarity and fate during cleavage in greater breadth and depth than ever previously achieved. By being optics-free, this has the potential to overcome fundamental limitations imposed by traditional forms of microscopy, greatly expand the ease and throughput of spatial transcriptomics, and pave the way for routine use of hyper-multiplexed molecular imaging.
Max ERC Funding
1 499 947 €
Duration
Start date: 2021-05-01, End date: 2026-04-30
Project acronym EpiRibo
Project Cross-evolutionary dissection of the functional plasticity of the ribosomal epitranscriptome
Researcher (PI) Schraga SCHWARTZ
Host Institution (HI) WEIZMANN INSTITUTE OF SCIENCE
Country Israel
Call Details Consolidator Grant (CoG), LS2, ERC-2020-COG
Summary A remarkable diversity of post-transcriptional modifications adorn the ribosomal RNA (rRNA) throughout all domains of life. While initially conceptualized as a constitutive component of the ribosome, recent evidence demonstrates that some modifications are only present on a fraction of cellular ribosomes, and our lab recently revealed unprecedented environmentally-mediated dynamics of an rRNA modification. This suggests that some rRNA modification may serve as a tunable layer for regulation of ribosome properties, facilitating adaptation to a new environment, stress or pathological state. Here we aim to systematically explore the extent to which rRNA modifications are tunable across evolution and highly diverse growth conditions, and to understand the consequences thereof. We seek to understand which rRNA modifications are subject to regulation, to uncover the contexts in which such regulation occurs, and to unravel the underlying mechanisms. Dissecting these questions requires an approach allowing systematic measurement of diverse rRNA modifications. We propose to establish a streamlined workflow, allowing multiplexed, precise, robust and cost-efficient systematic profiling of 19 distinct rRNA modifications in dozens of samples in a single experiment (Aim 1). In Aim 2, we will systematically measure rRNA modifications across the three domains of life, focusing on species that can thrive across two widely different physical/chemical gradients, where the potential for tunable rRNA modifications is particularly high. In Aim 3, we will combine gain- and loss-of-function approaches to systematically explore the functions RNA modifications can bestow on ribosomes. Collectively, EpiRibo addresses a fundamental open question regarding the extent of plasticity and regulatory potential present within the ribosome itself and encoded via its epitranscriptome, setting the stage for investigations in other RNA species, and in more complex and disease-related contexts.
Summary
A remarkable diversity of post-transcriptional modifications adorn the ribosomal RNA (rRNA) throughout all domains of life. While initially conceptualized as a constitutive component of the ribosome, recent evidence demonstrates that some modifications are only present on a fraction of cellular ribosomes, and our lab recently revealed unprecedented environmentally-mediated dynamics of an rRNA modification. This suggests that some rRNA modification may serve as a tunable layer for regulation of ribosome properties, facilitating adaptation to a new environment, stress or pathological state. Here we aim to systematically explore the extent to which rRNA modifications are tunable across evolution and highly diverse growth conditions, and to understand the consequences thereof. We seek to understand which rRNA modifications are subject to regulation, to uncover the contexts in which such regulation occurs, and to unravel the underlying mechanisms. Dissecting these questions requires an approach allowing systematic measurement of diverse rRNA modifications. We propose to establish a streamlined workflow, allowing multiplexed, precise, robust and cost-efficient systematic profiling of 19 distinct rRNA modifications in dozens of samples in a single experiment (Aim 1). In Aim 2, we will systematically measure rRNA modifications across the three domains of life, focusing on species that can thrive across two widely different physical/chemical gradients, where the potential for tunable rRNA modifications is particularly high. In Aim 3, we will combine gain- and loss-of-function approaches to systematically explore the functions RNA modifications can bestow on ribosomes. Collectively, EpiRibo addresses a fundamental open question regarding the extent of plasticity and regulatory potential present within the ribosome itself and encoded via its epitranscriptome, setting the stage for investigations in other RNA species, and in more complex and disease-related contexts.
Max ERC Funding
2 000 000 €
Duration
Start date: 2022-05-01, End date: 2027-04-30
Project acronym EPYC
Project Evolution of pro- and eukaryotic commensals within the human gut
Researcher (PI) Falk HILDEBRAND
Host Institution (HI) QUADRAM INSTITUTE BIOSCIENCE
Country United Kingdom
Call Details Starting Grant (StG), LS2, ERC-2020-STG
Summary The EPYC project will characterize the evolution of long-term human associated eukaryotes and prokaryotes, using colonization patterns in 3 human generations.
The gut microbiome is important for human health, supporting nutrition, pathogen defence and immune homeostasis, with more than 200 species inhabiting each human gut. In recent years metagenomics led to notable breakthroughs in describing this microbial diversity, yet 50-90% of species are typically present at too low abundance to be genome or strain resolved. Thus, most gut microbiome studies focused to date on dominant bacteria and very little is known of the highly diverse, yet low abundance, pro- and eukaryotes (elusive microbes). Importantly, elusive microbes are an inherent part of ecosystem successions persisting at different ages of the host. I propose that niche adaptation and persistence are key indicators of a taxa’s importance to the gut ecosystem and host health. I will determine which microbes persist for years within a human, or even a family for several generations. This should be reflected in microbial genetic adaption, also indicating which genes are likely important to successfully colonize the human gut.
I hypothesize that low abundance pro- and eukaryotes are adapted to persist for multiple generations in the human host, indicating their importance, despite being largely ignored so far.
To investigate this knowledge gap in EPYC, I will
(O1) Enable high-precision metagenomics of elusive microbes
(O2) Estimate pro- and eukaryotic strain persistence across three human generations
(O3) Describe the microbial genetics of gastrointestinal persistence
EPYC will develop the next-generation of high-resolution metagenomics of an extended taxonomic range, enabling me to research microbial evolution in the human gut.
Summary
The EPYC project will characterize the evolution of long-term human associated eukaryotes and prokaryotes, using colonization patterns in 3 human generations.
The gut microbiome is important for human health, supporting nutrition, pathogen defence and immune homeostasis, with more than 200 species inhabiting each human gut. In recent years metagenomics led to notable breakthroughs in describing this microbial diversity, yet 50-90% of species are typically present at too low abundance to be genome or strain resolved. Thus, most gut microbiome studies focused to date on dominant bacteria and very little is known of the highly diverse, yet low abundance, pro- and eukaryotes (elusive microbes). Importantly, elusive microbes are an inherent part of ecosystem successions persisting at different ages of the host. I propose that niche adaptation and persistence are key indicators of a taxa’s importance to the gut ecosystem and host health. I will determine which microbes persist for years within a human, or even a family for several generations. This should be reflected in microbial genetic adaption, also indicating which genes are likely important to successfully colonize the human gut.
I hypothesize that low abundance pro- and eukaryotes are adapted to persist for multiple generations in the human host, indicating their importance, despite being largely ignored so far.
To investigate this knowledge gap in EPYC, I will
(O1) Enable high-precision metagenomics of elusive microbes
(O2) Estimate pro- and eukaryotic strain persistence across three human generations
(O3) Describe the microbial genetics of gastrointestinal persistence
EPYC will develop the next-generation of high-resolution metagenomics of an extended taxonomic range, enabling me to research microbial evolution in the human gut.
Max ERC Funding
1 499 993 €
Duration
Start date: 2021-02-01, End date: 2026-01-31
Project acronym FateID
Project Learning from the past: linking ancestral epigenetic states to current cellular fates with single-cell multi-omic approaches
Researcher (PI) jop KIND
Host Institution (HI) KONINKLIJKE NEDERLANDSE AKADEMIE VAN WETENSCHAPPEN - KNAW
Country Netherlands
Call Details Consolidator Grant (CoG), LS2, ERC-2020-COG
Summary The establishment of cell type-specific transcriptional programs involves many interconnected regulatory mechanisms acting on different genomic scales. To dissect this multi-layered control of gene expression in detail, I will develop methods that a) measure multiple cellular outputs in single cells, and b) obtain that information in a time-resolved manner. This proposal outlines my approach to study early mouse development at numerous levels, including (but not limited to) transcription, chromatin context, and nuclear organization. In doing so, I expect to shed light on the mechanism behind cell fate specification and the epigenetic states that precede it.
I will develop a novel strategy to simultaneously profile many factors involved in gene regulation in the same cell. Its successful implementation will give insight into transcriptional control at unprecedented modality, revealing the causal relationships between histone modifications, spatial positioning within the nucleus, Polycomb group proteins, and others. Next, I will pursue several “molecular memory” strategies to obtain recordings of past regulatory states, followed by multi-omic readouts at later developmental times. Such retrospective analyses enable linking fate decisions to past molecular events in the same cell, thereby permitting careful reconstruction of the molecular trajectories underlying cell fate choice. Finally, these and previously developed single-cell methods will be applied to unravel the gene-regulatory mechanisms that govern lineage determination in early mouse development.
Summary
The establishment of cell type-specific transcriptional programs involves many interconnected regulatory mechanisms acting on different genomic scales. To dissect this multi-layered control of gene expression in detail, I will develop methods that a) measure multiple cellular outputs in single cells, and b) obtain that information in a time-resolved manner. This proposal outlines my approach to study early mouse development at numerous levels, including (but not limited to) transcription, chromatin context, and nuclear organization. In doing so, I expect to shed light on the mechanism behind cell fate specification and the epigenetic states that precede it.
I will develop a novel strategy to simultaneously profile many factors involved in gene regulation in the same cell. Its successful implementation will give insight into transcriptional control at unprecedented modality, revealing the causal relationships between histone modifications, spatial positioning within the nucleus, Polycomb group proteins, and others. Next, I will pursue several “molecular memory” strategies to obtain recordings of past regulatory states, followed by multi-omic readouts at later developmental times. Such retrospective analyses enable linking fate decisions to past molecular events in the same cell, thereby permitting careful reconstruction of the molecular trajectories underlying cell fate choice. Finally, these and previously developed single-cell methods will be applied to unravel the gene-regulatory mechanisms that govern lineage determination in early mouse development.
Max ERC Funding
2 000 000 €
Duration
Start date: 2021-10-01, End date: 2026-09-30
Project acronym FIND-seq
Project Functional Interrogation of Non-coding DNA Sequences in leukemia development and drug resistance
Researcher (PI) Davide Seruggia
Host Institution (HI) ST ANNA KINDERKREBSFORSCHUNG VEREIN
Country Austria
Call Details Starting Grant (StG), LS2, ERC-2020-STG
Summary Functional non-coding regions of the genome play a fundamental role in gene expression and are enriched for disease associated variants. Perturbation of non-coding regions harbouring disease-associated variants is now the rationale of ongoing clinical trials (e.g. NCT03432364), highlighting the translational potential of basic research in the non-coding space. However, our ability to systematically identify disease-associated functional elements in the non-coding genome, understand its grammar, and subsequently develop new therapies is limited. CRISPR-based pooled screens targeting non-coding elements in situ have been successful in uncovering complex gene regulatory architecture in a variety of biological systems. However, these approaches are limited to a few loci, lack of direct genotype-phenotype correlation, and do not target large chromatin structures that determine gene expression programs. To overcome these limitations, I propose a multi-scale approach platform that is generalizable to different cell types and phenotypes. Under this proposal, I will focus on the role of non-coding sequences in the context of blood malignancies. I will investigate non-coding sequences whose change in chromatin state (activation or repression) is associated with drug resistance in Chronic Myelogenous Leukemia (CML). I will study alterations in the chromatin structure (i.e. at chromatin loops or topologically associated domains) that are causal to imatinib resistance in CML. Finally, to learn enhancer grammar and mechanistically link non-coding variants to disease, I will focus on non-coding sequence variation in leukemia and dissect non-coding sequences at base pair resolution using dense mutagenesis coupled with long-reads sequencing. A deeper understanding of the non-coding regulatory architecture in diseases will provide the basis for development of innovative therapies targeting the non-coding genome.
Summary
Functional non-coding regions of the genome play a fundamental role in gene expression and are enriched for disease associated variants. Perturbation of non-coding regions harbouring disease-associated variants is now the rationale of ongoing clinical trials (e.g. NCT03432364), highlighting the translational potential of basic research in the non-coding space. However, our ability to systematically identify disease-associated functional elements in the non-coding genome, understand its grammar, and subsequently develop new therapies is limited. CRISPR-based pooled screens targeting non-coding elements in situ have been successful in uncovering complex gene regulatory architecture in a variety of biological systems. However, these approaches are limited to a few loci, lack of direct genotype-phenotype correlation, and do not target large chromatin structures that determine gene expression programs. To overcome these limitations, I propose a multi-scale approach platform that is generalizable to different cell types and phenotypes. Under this proposal, I will focus on the role of non-coding sequences in the context of blood malignancies. I will investigate non-coding sequences whose change in chromatin state (activation or repression) is associated with drug resistance in Chronic Myelogenous Leukemia (CML). I will study alterations in the chromatin structure (i.e. at chromatin loops or topologically associated domains) that are causal to imatinib resistance in CML. Finally, to learn enhancer grammar and mechanistically link non-coding variants to disease, I will focus on non-coding sequence variation in leukemia and dissect non-coding sequences at base pair resolution using dense mutagenesis coupled with long-reads sequencing. A deeper understanding of the non-coding regulatory architecture in diseases will provide the basis for development of innovative therapies targeting the non-coding genome.
Max ERC Funding
1 784 000 €
Duration
Start date: 2021-03-01, End date: 2026-02-28
Project acronym GENTE_Pop
Project GENETIC AND ENVIRONMENTAL BASIS OF NATURAL TRANSPOSITION AND ITS POTENTIAL TO CREATE ADAPTIVE VARIATION
Researcher (PI) Leandro Quadrana
Host Institution (HI) CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS
Country France
Call Details Starting Grant (StG), LS2, ERC-2020-STG
Summary Transposable elements (TEs) are powerful engines of genome evolution, as illustrated by their implication in the rewiring of regulatory networks and the creation of new cellular functions. Short-term consequences of TE mobilization can also be particularly dramatic given that TE insertions are a unique source of large effect mutations. However, there is a lack of knowledge about the contribution of ongoing transposition to within-species variation. This situation stems in large part from the repetitive nature of TEs, which complicates their analysis. Moreover, TE mobilization is typically rare and therefore new TE insertions tend to be missed in small-scale population studies. Hence, a major challenge in genomics is to determine the conditions leading to transposition in nature and the range of effects it generates. While most TE insertions are likely to be deleterious or neutral, it is widely proposed that because TE activity can be sensitive to the environment, transposition may in fact act as a major adaptive response of the genome to environmental changes.
Here, using large experimental and wild populations of the plant A. thaliana, I propose to leverage innovative genomic, molecular genetics and eco-evolutionary approaches to determine the Genetic x Environmental (GxE) map of heritable transposition and its contribution to the creation of adaptive variation.
Aim 1 is to identify the genetic and environmental factors that underpin TE mobilization by quantifying newly generated heritable insertions in thousands of genetically diverse individuals subjected to a range of environmental stressors. Aim 2 is to determine the fitness effects of these heritable TE insertions using multigenerational competition experiments and highly complex environments.
This project will greatly increase our understanding of the nature of the genetic variation TEs contribute to and our ability to predict the impact of ongoing transposition, notably in the context of climate change.
Summary
Transposable elements (TEs) are powerful engines of genome evolution, as illustrated by their implication in the rewiring of regulatory networks and the creation of new cellular functions. Short-term consequences of TE mobilization can also be particularly dramatic given that TE insertions are a unique source of large effect mutations. However, there is a lack of knowledge about the contribution of ongoing transposition to within-species variation. This situation stems in large part from the repetitive nature of TEs, which complicates their analysis. Moreover, TE mobilization is typically rare and therefore new TE insertions tend to be missed in small-scale population studies. Hence, a major challenge in genomics is to determine the conditions leading to transposition in nature and the range of effects it generates. While most TE insertions are likely to be deleterious or neutral, it is widely proposed that because TE activity can be sensitive to the environment, transposition may in fact act as a major adaptive response of the genome to environmental changes.
Here, using large experimental and wild populations of the plant A. thaliana, I propose to leverage innovative genomic, molecular genetics and eco-evolutionary approaches to determine the Genetic x Environmental (GxE) map of heritable transposition and its contribution to the creation of adaptive variation.
Aim 1 is to identify the genetic and environmental factors that underpin TE mobilization by quantifying newly generated heritable insertions in thousands of genetically diverse individuals subjected to a range of environmental stressors. Aim 2 is to determine the fitness effects of these heritable TE insertions using multigenerational competition experiments and highly complex environments.
This project will greatly increase our understanding of the nature of the genetic variation TEs contribute to and our ability to predict the impact of ongoing transposition, notably in the context of climate change.
Max ERC Funding
1 499 627 €
Duration
Start date: 2021-01-01, End date: 2025-12-31
