Project acronym CHROMATINPRINCIPLES
Project Principles of Chromatin Organization
Researcher (PI) Bas Van Steensel
Host Institution (HI) STICHTING HET NEDERLANDS KANKER INSTITUUT-ANTONI VAN LEEUWENHOEK ZIEKENHUIS
Call Details Advanced Grant (AdG), LS2, ERC-2011-ADG_20110310
Summary Chromatin is the ensemble of genomic DNA and hundreds of structural and regulatory proteins. Together these proteins govern the gene expression program of a cell. While biochemical and genetic approaches have tought us much about interactions between individual chromatin proteins, we still lack a “big picture” of chromatin: how is the entire interaction network of chromatin proteins organized?
My lab discovered that chromatin in Drosophila consists of a limited number of principal types that partition the genome into domains with distinct regulatory properties. Among these is BLACK chromatin, a novel repressive type of chromatin that covers nearly half of the fly genome. It is still largely unclear how these different chromatin types are formed, how they are targeted to specific genomic regions, and how they interact with each other.
Here, I propose a combination of systematic approaches aimed to gain insight into the basic mechanisms that drive the partioning of the genome into distinct chromatin types. New genomics techniques, developed in my laboratory, will be used to construct an integrated view of the interplay of more than one hundred representative chromatin proteins with each other and with sequence elements in the genome. Specifically, we will: (1) Study the genome-wide dynamic repositioning of chromatin domains during development in relation to gene regulation; (2) Use a novel and versatile parallel genome-wide reporter assay to dissect the interplay among DNA sequences and chromatin types; (3) Combine computational modeling with a high-throughput genome-wide assay to uncover the network of interactions responsible for the formation of the principal chromatin types; (4) Dissect the molecular architecture of BLACK chromatin and its role in gene repression.
The results will provide understanding of the basic principles that govern the structure and composition of chromatin, and reveal how the principal chromatin types together direct gene expression.
Summary
Chromatin is the ensemble of genomic DNA and hundreds of structural and regulatory proteins. Together these proteins govern the gene expression program of a cell. While biochemical and genetic approaches have tought us much about interactions between individual chromatin proteins, we still lack a “big picture” of chromatin: how is the entire interaction network of chromatin proteins organized?
My lab discovered that chromatin in Drosophila consists of a limited number of principal types that partition the genome into domains with distinct regulatory properties. Among these is BLACK chromatin, a novel repressive type of chromatin that covers nearly half of the fly genome. It is still largely unclear how these different chromatin types are formed, how they are targeted to specific genomic regions, and how they interact with each other.
Here, I propose a combination of systematic approaches aimed to gain insight into the basic mechanisms that drive the partioning of the genome into distinct chromatin types. New genomics techniques, developed in my laboratory, will be used to construct an integrated view of the interplay of more than one hundred representative chromatin proteins with each other and with sequence elements in the genome. Specifically, we will: (1) Study the genome-wide dynamic repositioning of chromatin domains during development in relation to gene regulation; (2) Use a novel and versatile parallel genome-wide reporter assay to dissect the interplay among DNA sequences and chromatin types; (3) Combine computational modeling with a high-throughput genome-wide assay to uncover the network of interactions responsible for the formation of the principal chromatin types; (4) Dissect the molecular architecture of BLACK chromatin and its role in gene repression.
The results will provide understanding of the basic principles that govern the structure and composition of chromatin, and reveal how the principal chromatin types together direct gene expression.
Max ERC Funding
2 495 080 €
Duration
Start date: 2012-03-01, End date: 2017-02-28
Project acronym CHROMATINREPAIRCODE
Project CHROMATIN-REPAIR-CODE: Hacking the chromatin code for DNA repair
Researcher (PI) Haico Van Attikum
Host Institution (HI) ACADEMISCH ZIEKENHUIS LEIDEN
Call Details Consolidator Grant (CoG), LS2, ERC-2013-CoG
Summary "Our cells receive tens of thousands of different DNA lesions per day. Failure to repair these lesions will lead to cell death, mutations and genome instability, which contribute to human diseases such as neurodegenerative disorders and cancer. Efficient recognition and repair of DNA damage, however, is complicated by the fact that genomic DNA is packaged, through histone and non-histone proteins, into a condensed structure called chromatin. The DNA repair machinery has to circumvent this barrier to gain access to the damaged DNA and repair the lesions. Our recent work suggests that chromatin-modifying enzymes (CME) help to overcome this barrier at sites of DNA damage. However, the identity of these CME, their mode of action and interconnections with DNA repair pathways remain largely enigmatic. The aim of this project is to systematically identify and characterize the CME that operate during DNA repair processes in both yeast and human cells. To reach this goal we will use a cross-disciplinary approach that combines novel and cutting-edge genomics approaches with bioinformatics, genetics, biochemistry and high-resolution microscopy. Epigenetics-IDentifier (Epi-ID) will be used as a tool to unveil novel CME, whereas RNAi-interference and genetic interaction mapping studies will pinpoint the CME that may potentially regulate repair of DNA damage. A series of functional assays will eventually characterize their role in distinct DNA repair pathways, focusing on those that counteract DNA strand breaks and replication stress. Together these studies will provide insight into how CME assist cells to repair DNA damage in chromatin and inform on the relevance of CME to maintain genome stability and counteract human diseases."
Summary
"Our cells receive tens of thousands of different DNA lesions per day. Failure to repair these lesions will lead to cell death, mutations and genome instability, which contribute to human diseases such as neurodegenerative disorders and cancer. Efficient recognition and repair of DNA damage, however, is complicated by the fact that genomic DNA is packaged, through histone and non-histone proteins, into a condensed structure called chromatin. The DNA repair machinery has to circumvent this barrier to gain access to the damaged DNA and repair the lesions. Our recent work suggests that chromatin-modifying enzymes (CME) help to overcome this barrier at sites of DNA damage. However, the identity of these CME, their mode of action and interconnections with DNA repair pathways remain largely enigmatic. The aim of this project is to systematically identify and characterize the CME that operate during DNA repair processes in both yeast and human cells. To reach this goal we will use a cross-disciplinary approach that combines novel and cutting-edge genomics approaches with bioinformatics, genetics, biochemistry and high-resolution microscopy. Epigenetics-IDentifier (Epi-ID) will be used as a tool to unveil novel CME, whereas RNAi-interference and genetic interaction mapping studies will pinpoint the CME that may potentially regulate repair of DNA damage. A series of functional assays will eventually characterize their role in distinct DNA repair pathways, focusing on those that counteract DNA strand breaks and replication stress. Together these studies will provide insight into how CME assist cells to repair DNA damage in chromatin and inform on the relevance of CME to maintain genome stability and counteract human diseases."
Max ERC Funding
1 999 575 €
Duration
Start date: 2014-03-01, End date: 2019-02-28
Project acronym CHROMATINRNA
Project The role of CpG island RNAs and Polycomb-RNA interactions in developmental gene regulation
Researcher (PI) Richard Gareth Jenner
Host Institution (HI) UNIVERSITY COLLEGE LONDON
Call Details Starting Grant (StG), LS2, ERC-2012-StG_20111109
Summary A great challenge in developmental biology research has been to understand how cell type specific expression programs are orchestrated through regulated access to chromatin. The interaction between non-coding RNAs and chromatin regulators is emerging as an exciting new research area with the potential to explain how chromatin modifications are targeted.
Polycomb repressive complex 2 (PRC2) modifies chromatin to maintain developmental regulator genes specific for other cell types in a repressed state and is essential for embryogenesis across Metazoa. We have recently determined that CpG islands targeted by PRC2 generate a class of short non-coding RNAs. The RNAs are produced independently from mRNA, indicative of hitherto uncharacterised transcriptional processes. Furthermore, we have found that the PRC2 subunit Suz12 is an RNA binding protein and directly interacts with these short RNAs and with other RNAs in cells. The role of ncRNA in targeting PRC2 to CpG islands and the importance of PRC2 RNA binding activity for development remains to be understood. Our aims are to:
1. Determine the functional properties of CpG-island RNAs by A. identifying their conserved features, B. determining their role in polycomb targeting of CpG islands and C. investigating whether such a role relates to the antagonism of polycomb targeting by DNA methylation.
2. Establish the biological role for Suz12 RNA binding activity by A. determining the structural determinants for Suz12 binding in vitro, B. verifying these features play a role in PRC2 RNA binding in cells and C. determining the role for PRC2-RNA interactions for polycomb function and development.
This work promises to characterise a potentially fundamental aspect of cell biology and will open a number of avenues for understanding the function of ncRNAs, the RNA binding activity of chromatin regulators, how transcription and chromatin structure are regulated, and how cell state is maintained and reshaped during development.
Summary
A great challenge in developmental biology research has been to understand how cell type specific expression programs are orchestrated through regulated access to chromatin. The interaction between non-coding RNAs and chromatin regulators is emerging as an exciting new research area with the potential to explain how chromatin modifications are targeted.
Polycomb repressive complex 2 (PRC2) modifies chromatin to maintain developmental regulator genes specific for other cell types in a repressed state and is essential for embryogenesis across Metazoa. We have recently determined that CpG islands targeted by PRC2 generate a class of short non-coding RNAs. The RNAs are produced independently from mRNA, indicative of hitherto uncharacterised transcriptional processes. Furthermore, we have found that the PRC2 subunit Suz12 is an RNA binding protein and directly interacts with these short RNAs and with other RNAs in cells. The role of ncRNA in targeting PRC2 to CpG islands and the importance of PRC2 RNA binding activity for development remains to be understood. Our aims are to:
1. Determine the functional properties of CpG-island RNAs by A. identifying their conserved features, B. determining their role in polycomb targeting of CpG islands and C. investigating whether such a role relates to the antagonism of polycomb targeting by DNA methylation.
2. Establish the biological role for Suz12 RNA binding activity by A. determining the structural determinants for Suz12 binding in vitro, B. verifying these features play a role in PRC2 RNA binding in cells and C. determining the role for PRC2-RNA interactions for polycomb function and development.
This work promises to characterise a potentially fundamental aspect of cell biology and will open a number of avenues for understanding the function of ncRNAs, the RNA binding activity of chromatin regulators, how transcription and chromatin structure are regulated, and how cell state is maintained and reshaped during development.
Max ERC Funding
1 499 094 €
Duration
Start date: 2013-09-01, End date: 2019-04-30
Project acronym CHROMATINSYS
Project Systematic Approach to Dissect the Interplay between Chromatin and Transcription
Researcher (PI) Nir Friedman
Host Institution (HI) THE HEBREW UNIVERSITY OF JERUSALEM
Call Details Advanced Grant (AdG), LS2, ERC-2013-ADG
Summary Epigenetic mechanisms play an important role in regulating and maintaining the functionality of cells and have been implicated in a wide range of human diseases. Histone proteins that form the protein core of nucleosomes are subject to a bewildering array of covalent and structural modifications, which can repress, permit, or promote transcription. These modifications can be added and removed by specialized complexes that are recruited by other covalent modifications, by transcription factors, or by the transcriptional machinery. Advances in genomics led to comprehensive mapping of the ``epigenome'' in a range of tissues and organisms. These maps established the tight connection between histone modifications and transcription programs. These static charts, however, are less successful at uncovering the underlying mechanisms, logic, and function of histone modifications in establishing and maintaining transcriptional programs. Our premise is that we can answer these basic questions by observing the effect of genetic perturbations on the dynamics of both chromatin state and transcriptional activity. We aim to dissect the chromatin-transcription system in a systematic manner by building on our extensive experience in modeling and analysis, and a unique high-throughput experimental system we established in my lab.
We plan to use the budding yeast model organism, which allows for
efficient genetic and experimental manipulations. We will combine two technologies: (1) high-throughput measurements of single-cell
transcriptional output using fluorescence reporters; and (2) high-throughput immunoprecipitation sequencing assays to map chromatin state. Measuring with these the dynamics of response to stimuli under different genetic backgrounds and using advanced stochastic network models, we will chart detailed mechanisms that are opaque to current approaches and elucidate the general principles that govern the interplay between chromatin and transcription.
Summary
Epigenetic mechanisms play an important role in regulating and maintaining the functionality of cells and have been implicated in a wide range of human diseases. Histone proteins that form the protein core of nucleosomes are subject to a bewildering array of covalent and structural modifications, which can repress, permit, or promote transcription. These modifications can be added and removed by specialized complexes that are recruited by other covalent modifications, by transcription factors, or by the transcriptional machinery. Advances in genomics led to comprehensive mapping of the ``epigenome'' in a range of tissues and organisms. These maps established the tight connection between histone modifications and transcription programs. These static charts, however, are less successful at uncovering the underlying mechanisms, logic, and function of histone modifications in establishing and maintaining transcriptional programs. Our premise is that we can answer these basic questions by observing the effect of genetic perturbations on the dynamics of both chromatin state and transcriptional activity. We aim to dissect the chromatin-transcription system in a systematic manner by building on our extensive experience in modeling and analysis, and a unique high-throughput experimental system we established in my lab.
We plan to use the budding yeast model organism, which allows for
efficient genetic and experimental manipulations. We will combine two technologies: (1) high-throughput measurements of single-cell
transcriptional output using fluorescence reporters; and (2) high-throughput immunoprecipitation sequencing assays to map chromatin state. Measuring with these the dynamics of response to stimuli under different genetic backgrounds and using advanced stochastic network models, we will chart detailed mechanisms that are opaque to current approaches and elucidate the general principles that govern the interplay between chromatin and transcription.
Max ERC Funding
2 396 450 €
Duration
Start date: 2014-01-01, End date: 2018-12-31
Project acronym CHROMOTHRIPSIS
Project Dissecting the Molecular Mechanism of Catastrophic DNA Rearrangement in Cancer
Researcher (PI) Jan Oliver Korbel
Host Institution (HI) EUROPEAN MOLECULAR BIOLOGY LABORATORY
Call Details Starting Grant (StG), LS2, ERC-2013-StG
Summary Recent cancer genome analyses have led to the discovery of a process involving massive genome structural rearrangement (SR) formation in a one-step, cataclysmic event, coined chromothripsis. The term chromothripsis (chromo from chromosome; thripsis for shattering into pieces) stands for a hypothetical process in which individual chromosomes are pulverised, resulting in a multitude of fragments, some of which are lost to the cell whereas others are erroneously rejoined. Compelling evidence was presented that chromothripsis plays a crucial role in the development, or progression of a notable subset of human cancers – thus, tumorigensis models involving gradual acquisitions of alterations may need to be revised in these cancers.
Presently, chromothripsis lacks a mechanistic basis. We recently showed that in childhood medulloblastoma brain tumours driven by Sonic Hedgehog (Shh) signalling, chromothripsis is linked with predisposing TP53 mutations. Thus, rather than occurring in isolation, chromothripsis appears to be prone to happen in conjunction with (or instigated by) gradually acquired alterations, or in the context of active signalling pathways, the inference of which may lead to further mechanistic insights. Using such rationale, I propose to dissect the mechanism behind chromothripsis using interdisciplinary approaches. First, we will develop a computational approach to accurately detect chromothripsis. Second, we will use this approach to link chromothripsis with novel factors and contexts. Third, we will develop highly controllable cell line-based systems to test concrete mechanistic hypotheses, thereby taking into account our data on linked factors and contexts. Fourth, we will generate transcriptome data to monitor pathways involved in inducing chromothripsis, and such involved in coping with the massive SRs occurring. We will also combine findings from all these approaches to build a comprehensive model of chromothripsis and its associated pathways.
Summary
Recent cancer genome analyses have led to the discovery of a process involving massive genome structural rearrangement (SR) formation in a one-step, cataclysmic event, coined chromothripsis. The term chromothripsis (chromo from chromosome; thripsis for shattering into pieces) stands for a hypothetical process in which individual chromosomes are pulverised, resulting in a multitude of fragments, some of which are lost to the cell whereas others are erroneously rejoined. Compelling evidence was presented that chromothripsis plays a crucial role in the development, or progression of a notable subset of human cancers – thus, tumorigensis models involving gradual acquisitions of alterations may need to be revised in these cancers.
Presently, chromothripsis lacks a mechanistic basis. We recently showed that in childhood medulloblastoma brain tumours driven by Sonic Hedgehog (Shh) signalling, chromothripsis is linked with predisposing TP53 mutations. Thus, rather than occurring in isolation, chromothripsis appears to be prone to happen in conjunction with (or instigated by) gradually acquired alterations, or in the context of active signalling pathways, the inference of which may lead to further mechanistic insights. Using such rationale, I propose to dissect the mechanism behind chromothripsis using interdisciplinary approaches. First, we will develop a computational approach to accurately detect chromothripsis. Second, we will use this approach to link chromothripsis with novel factors and contexts. Third, we will develop highly controllable cell line-based systems to test concrete mechanistic hypotheses, thereby taking into account our data on linked factors and contexts. Fourth, we will generate transcriptome data to monitor pathways involved in inducing chromothripsis, and such involved in coping with the massive SRs occurring. We will also combine findings from all these approaches to build a comprehensive model of chromothripsis and its associated pathways.
Max ERC Funding
1 471 964 €
Duration
Start date: 2014-04-01, End date: 2019-01-31
Project acronym CHROMREP
Project Dissecting the chromatin response to DNA damage in silenced heterochromatin regions
Researcher (PI) Aniek Janssen
Host Institution (HI) UNIVERSITAIR MEDISCH CENTRUM UTRECHT
Call Details Starting Grant (StG), LS2, ERC-2019-STG
Summary Cells are continuously exposed to insults that can break or chemically modify their DNA. To protect the DNA, cells have acquired an arsenal of repair mechanisms. Proper repair of DNA damage is essential for organismal viability and disease prevention. What is often overlooked is the fact that the eukaryotic nucleus contains many different chromatin domains that can each influence the dynamic response to DNA damage. Different chromatin environments are defined by specific molecular and biophysical properties, which could necessitate distinct chromatin responses to ensure safe DNA damage repair.
The aim of this proposal is to understand how diverse chromatin domains, and in particular the dense heterochromatin environment, shape the dynamic chromatin response to DNA damage.
I recently developed locus-specific DNA damage systems that allow for in-depth analysis of chromatin domain-specific repair responses in Drosophila tissue. I will employ these systems and develop new ones to directly observe heterochromatin-specific dynamics and repair responses. I will combine these systems and state-of-the art chromatin analysis with high-resolution live imaging to dissect the DNA damage-associated heterochromatin changes to determine their function in repair -kinetics, -dynamics and -pathway choice.
Deciphering the chromatin dynamics that regulate DNA damage repair in heterochromatin will have broad conceptual implications for understanding the role of these dynamics in other essential nuclear processes, such as replication and transcription. More importantly, understanding how chromatin proteins promote repair will be important in determining how cancer-associated mutations in these chromatin proteins impact genetic instability in tumours in the long run.
Summary
Cells are continuously exposed to insults that can break or chemically modify their DNA. To protect the DNA, cells have acquired an arsenal of repair mechanisms. Proper repair of DNA damage is essential for organismal viability and disease prevention. What is often overlooked is the fact that the eukaryotic nucleus contains many different chromatin domains that can each influence the dynamic response to DNA damage. Different chromatin environments are defined by specific molecular and biophysical properties, which could necessitate distinct chromatin responses to ensure safe DNA damage repair.
The aim of this proposal is to understand how diverse chromatin domains, and in particular the dense heterochromatin environment, shape the dynamic chromatin response to DNA damage.
I recently developed locus-specific DNA damage systems that allow for in-depth analysis of chromatin domain-specific repair responses in Drosophila tissue. I will employ these systems and develop new ones to directly observe heterochromatin-specific dynamics and repair responses. I will combine these systems and state-of-the art chromatin analysis with high-resolution live imaging to dissect the DNA damage-associated heterochromatin changes to determine their function in repair -kinetics, -dynamics and -pathway choice.
Deciphering the chromatin dynamics that regulate DNA damage repair in heterochromatin will have broad conceptual implications for understanding the role of these dynamics in other essential nuclear processes, such as replication and transcription. More importantly, understanding how chromatin proteins promote repair will be important in determining how cancer-associated mutations in these chromatin proteins impact genetic instability in tumours in the long run.
Max ERC Funding
1 499 404 €
Duration
Start date: 2019-12-01, End date: 2024-11-30
Project acronym CHROMTOPOLOGY
Project Understanding and manipulating the dynamics of chromosome topologies in transcriptional control
Researcher (PI) Thomas, Ivor Sexton
Host Institution (HI) CENTRE EUROPEEN DE RECHERCHE EN BIOLOGIE ET MEDECINE
Call Details Starting Grant (StG), LS2, ERC-2015-STG
Summary Transcriptional regulation of genes in eukaryotic cells requires a complex and highly regulated interplay of chromatin environment, epigenetic status of target sequences and several different transcription factors. Eukaryotic genomes are tightly packaged within nuclei, yet must be accessible for transcription, replication and repair. A striking correlation exists between chromatin topology and underlying gene activity. According to the textbook view, chromatin loops bring genes into direct contact with distal regulatory elements, such as enhancers. Moreover, we and others have shown that genomes are organized into discretely folded megabase-sized regions, denoted as topologically associated domains (TADs), which seem to correlate well with transcription activity and histone modifications. However, it is unknown whether chromosome folding is a cause or consequence of underlying gene function.
To better understand the role of genome organization in transcription regulation, I will address the following questions:
(i) How are chromatin configurations altered during transcriptional changes accompanying development?
(ii) What are the real-time kinetics and cell-to-cell variabilities of chromatin interactions and TAD architectures?
(iii) Can chromatin loops be engineered de novo, and do they influence gene expression?
(iv) What genetic elements and trans-acting factors are required to organize TADs?
To address these fundamental questions, I will use a combination of novel technologies and approaches, such as Hi-C, CRISPR knock-ins, ANCHOR tagging of DNA loci, high- and super-resolution single-cell imaging, genome-wide screens and optogenetics, in order to both study and engineer chromatin architectures.
These studies will give groundbreaking insight into if and how chromatin topology regulates transcription. Thus, I anticipate that the results of this project will have a major impact on the field and will lead to a new paradigm for metazoan transcription control.
Summary
Transcriptional regulation of genes in eukaryotic cells requires a complex and highly regulated interplay of chromatin environment, epigenetic status of target sequences and several different transcription factors. Eukaryotic genomes are tightly packaged within nuclei, yet must be accessible for transcription, replication and repair. A striking correlation exists between chromatin topology and underlying gene activity. According to the textbook view, chromatin loops bring genes into direct contact with distal regulatory elements, such as enhancers. Moreover, we and others have shown that genomes are organized into discretely folded megabase-sized regions, denoted as topologically associated domains (TADs), which seem to correlate well with transcription activity and histone modifications. However, it is unknown whether chromosome folding is a cause or consequence of underlying gene function.
To better understand the role of genome organization in transcription regulation, I will address the following questions:
(i) How are chromatin configurations altered during transcriptional changes accompanying development?
(ii) What are the real-time kinetics and cell-to-cell variabilities of chromatin interactions and TAD architectures?
(iii) Can chromatin loops be engineered de novo, and do they influence gene expression?
(iv) What genetic elements and trans-acting factors are required to organize TADs?
To address these fundamental questions, I will use a combination of novel technologies and approaches, such as Hi-C, CRISPR knock-ins, ANCHOR tagging of DNA loci, high- and super-resolution single-cell imaging, genome-wide screens and optogenetics, in order to both study and engineer chromatin architectures.
These studies will give groundbreaking insight into if and how chromatin topology regulates transcription. Thus, I anticipate that the results of this project will have a major impact on the field and will lead to a new paradigm for metazoan transcription control.
Max ERC Funding
1 500 000 €
Duration
Start date: 2016-06-01, End date: 2021-05-31
Project acronym cis-CONTROL
Project Decoding and controlling cell-state switching: A bottom-up approach based on enhancer logic
Researcher (PI) Stein Luc AERTS
Host Institution (HI) VIB
Call Details Consolidator Grant (CoG), LS2, ERC-2016-COG
Summary Cell-state switching in cancer allows cells to transition from a proliferative to an invasive and drug-resistant phenotype. This plasticity plays an important role in cancer progression and tumour heterogeneity. We have made a striking observation that cancer cells of different origin can switch to a common survival state. During this epigenomic reprogramming, cancer cells re-activate genomic enhancers from specific regulatory programs, such as wound repair and epithelial-to-mesenchymal transition.
The goal of my project is to decipher the enhancer logic underlying this canalization effect towards a common survival state. We will then employ this new understanding of enhancer logic to engineer synthetic enhancers that are able to monitor and manipulate cell-state switching in real time. Furthermore, we will use enhancer models to identify cis-regulatory mutations that have an impact on cell-state switching and drug resistance. Such applications are currently hampered because there is a significant gap in our understanding of how enhancers work.
To tackle this problem we will use a combination of in vivo massively parallel enhancer-reporter assays, single-cell genomics on microfluidic devices, computational modelling, and synthetic enhancer design. Using these approaches we will pursue the following aims: (1) to identify functional enhancers regulating cell-state switching by performing in vivo genetic screens in mice; (2) to elucidate the dynamic trajectories whereby cells of different cancer types switch to a common survival cell-state, at single-cell resolution; (3) to create synthetic enhancer circuits that specifically kill cancer cells undergoing cell-state switching.
Our findings will have an impact on genome research, characterizing how cellular decision making is implemented by the cis-regulatory code; and on cancer research, employing enhancer logic in the context of cancer therapy.
Summary
Cell-state switching in cancer allows cells to transition from a proliferative to an invasive and drug-resistant phenotype. This plasticity plays an important role in cancer progression and tumour heterogeneity. We have made a striking observation that cancer cells of different origin can switch to a common survival state. During this epigenomic reprogramming, cancer cells re-activate genomic enhancers from specific regulatory programs, such as wound repair and epithelial-to-mesenchymal transition.
The goal of my project is to decipher the enhancer logic underlying this canalization effect towards a common survival state. We will then employ this new understanding of enhancer logic to engineer synthetic enhancers that are able to monitor and manipulate cell-state switching in real time. Furthermore, we will use enhancer models to identify cis-regulatory mutations that have an impact on cell-state switching and drug resistance. Such applications are currently hampered because there is a significant gap in our understanding of how enhancers work.
To tackle this problem we will use a combination of in vivo massively parallel enhancer-reporter assays, single-cell genomics on microfluidic devices, computational modelling, and synthetic enhancer design. Using these approaches we will pursue the following aims: (1) to identify functional enhancers regulating cell-state switching by performing in vivo genetic screens in mice; (2) to elucidate the dynamic trajectories whereby cells of different cancer types switch to a common survival cell-state, at single-cell resolution; (3) to create synthetic enhancer circuits that specifically kill cancer cells undergoing cell-state switching.
Our findings will have an impact on genome research, characterizing how cellular decision making is implemented by the cis-regulatory code; and on cancer research, employing enhancer logic in the context of cancer therapy.
Max ERC Funding
1 999 660 €
Duration
Start date: 2017-06-01, End date: 2022-05-31
Project acronym CISREGVAR
Project Cis-regulatory variation: Using natural genetic variation to dissect cis-regulatory control of embryonic development
Researcher (PI) Eileen Eunice Furlong
Host Institution (HI) EUROPEAN MOLECULAR BIOLOGY LABORATORY
Call Details Advanced Grant (AdG), LS2, ERC-2012-ADG_20120314
Summary Embryonic development is very robust: In the midst of segregating mutations and fluctuating environments, a fertilized egg has the remarkable capacity to give rise to a precisely patterned embryo. The stereotypic progression of development is driven by tightly regulated programs of gene expression. However, this deterministic view from genetics is at odds with an emerging view of transcription from genomics as a “noisy” process, variable and changing both within and between individuals. How variable transcriptional programs can regulate robust embryonic development remains a long-standing question, which this proposal aims to address. By combining population genetics, genomics, and developmental genetics in Drosophila we will dissect the relationship between DNA sequence variation, transcription factor (TF) occupancy, and the regulatory control of developmental gene expression.
The backdrop for this work is extensive information generated by my lab on the location and function of over 12,000 developmental cis-regulatory elements, including accurate, predictive models of their spatio-temporal activity. To understand the impact of variation on transcription and development, we will make use of a powerful experimental resource – 192 sequenced Drosophila strains, collected from a highly genetically diverse wild population. The proposed research has three Specific Aims: 1) Perform the first high-resolution study associating SNPs and structural variants (eQTLs) with gene expression variation during embryonic development, 2) Quantify in vivo the relationship between cis-regulatory variation, TF occupancy, and gene expression, 3) Incorporate these data into an integrated, predictive model of transcription. These Aims, together with our cis-regulatory data, will offer unique, mechanistic insights into how cis-regulatory variation impacts developmental gene regulation, and into the molecular bases of robustness in developmental regulatory networks.
Summary
Embryonic development is very robust: In the midst of segregating mutations and fluctuating environments, a fertilized egg has the remarkable capacity to give rise to a precisely patterned embryo. The stereotypic progression of development is driven by tightly regulated programs of gene expression. However, this deterministic view from genetics is at odds with an emerging view of transcription from genomics as a “noisy” process, variable and changing both within and between individuals. How variable transcriptional programs can regulate robust embryonic development remains a long-standing question, which this proposal aims to address. By combining population genetics, genomics, and developmental genetics in Drosophila we will dissect the relationship between DNA sequence variation, transcription factor (TF) occupancy, and the regulatory control of developmental gene expression.
The backdrop for this work is extensive information generated by my lab on the location and function of over 12,000 developmental cis-regulatory elements, including accurate, predictive models of their spatio-temporal activity. To understand the impact of variation on transcription and development, we will make use of a powerful experimental resource – 192 sequenced Drosophila strains, collected from a highly genetically diverse wild population. The proposed research has three Specific Aims: 1) Perform the first high-resolution study associating SNPs and structural variants (eQTLs) with gene expression variation during embryonic development, 2) Quantify in vivo the relationship between cis-regulatory variation, TF occupancy, and gene expression, 3) Incorporate these data into an integrated, predictive model of transcription. These Aims, together with our cis-regulatory data, will offer unique, mechanistic insights into how cis-regulatory variation impacts developmental gene regulation, and into the molecular bases of robustness in developmental regulatory networks.
Max ERC Funding
2 260 116 €
Duration
Start date: 2013-09-01, End date: 2018-08-31
Project acronym CLEAR
Project Modulating cellular clearance to cure human disease
Researcher (PI) Andrea Ballabio
Host Institution (HI) FONDAZIONE TELETHON
Call Details Advanced Grant (AdG), LS2, ERC-2009-AdG
Summary Cellular clearance is a fundamental process required by all cells in all species. Important physiological processes, such as aging, and pathological mechanisms, such as neurodegeneration, are strictly dependent on cellular clearance. In eukaryotes, most of the cellular clearing processes occur in a specialized organelle, the lysosome. This project is based on a recent discovery, made in our laboratory, of a gene network, which we have named CLEAR, that controls lysosomal biogenesis and function and regulates cellular clearance. The specific goals of the project are: 1) the comprehensive characterization of the mechanisms underlying the CLEAR network, 2) the thorough understanding of CLEAR physiological function at the cellular and organism levels, 3) the development of strategies and tools to modulate cellular clearance, and 4) the implementation of proof-of-principle therapeutic studies based on the activation of the CLEAR network in murine models of human lysosomal storage disorders and of neurodegenerative diseases, such as Alzheimers s and Huntington s diseases. A combination of genomics, bioinformatics, systems biology, chemical genomics, cell biology, and mouse genetics approaches will be used to achieve these goals. Our goal is to develop tools to modulate cellular clearance and to use such tools to develop therapies to cure human disease. The potential medical relevance of this project is very high, particularly in the field of neurodegenerative disease. Therapies that prevent, ameliorate or delay neurodegeneration in these diseases would have a huge impact on human health.
Summary
Cellular clearance is a fundamental process required by all cells in all species. Important physiological processes, such as aging, and pathological mechanisms, such as neurodegeneration, are strictly dependent on cellular clearance. In eukaryotes, most of the cellular clearing processes occur in a specialized organelle, the lysosome. This project is based on a recent discovery, made in our laboratory, of a gene network, which we have named CLEAR, that controls lysosomal biogenesis and function and regulates cellular clearance. The specific goals of the project are: 1) the comprehensive characterization of the mechanisms underlying the CLEAR network, 2) the thorough understanding of CLEAR physiological function at the cellular and organism levels, 3) the development of strategies and tools to modulate cellular clearance, and 4) the implementation of proof-of-principle therapeutic studies based on the activation of the CLEAR network in murine models of human lysosomal storage disorders and of neurodegenerative diseases, such as Alzheimers s and Huntington s diseases. A combination of genomics, bioinformatics, systems biology, chemical genomics, cell biology, and mouse genetics approaches will be used to achieve these goals. Our goal is to develop tools to modulate cellular clearance and to use such tools to develop therapies to cure human disease. The potential medical relevance of this project is very high, particularly in the field of neurodegenerative disease. Therapies that prevent, ameliorate or delay neurodegeneration in these diseases would have a huge impact on human health.
Max ERC Funding
2 100 000 €
Duration
Start date: 2010-03-01, End date: 2015-02-28
