Project acronym CELLDOCTOR
Project Quantitative understanding of a living system and its engineering as a cellular organelle
Researcher (PI) Luis Serrano
Host Institution (HI) FUNDACIO CENTRE DE REGULACIO GENOMICA
Call Details Advanced Grant (AdG), LS2, ERC-2008-AdG
Summary The idea of harnessing living organisms for treating human diseases is not new but, so far, the majority of the living vectors used in human therapy are viruses which have the disadvantage of the limited number of genes and networks that can contain. Bacteria allow the cloning of complex networks and the possibility of making a large plethora of compounds, naturally or through careful redesign. One of the main limitations for the use of bacteria to treat human diseases is their complexity, the existence of a cell wall that difficult the communication with the target cells, the lack of control over its growth and the immune response that will elicit on its target. Ideally one would like to have a very small bacterium (of a mitochondria size), with no cell wall, which could be grown in Vitro, be genetically manipulated, for which we will have enough data to allow a complete understanding of its behaviour and which could live as a human cell parasite. Such a microorganism could in principle be used as a living vector in which genes of interests, or networks producing organic molecules of medical relevance, could be introduced under in Vitro conditions and then inoculated on extracted human cells or in the organism, and then become a new organelle in the host. Then, it could produce and secrete into the host proteins which will be needed to correct a genetic disease, or drugs needed by the patient. To do that, we need to understand in excruciating detail the Biology of the target bacterium and how to interface with the host cell cycle (Systems biology aspect). Then we need to have engineering tools (network design, protein design, simulations) to modify the target bacterium to behave like an organelle once inside the cell (Synthetic biology aspect). M.pneumoniae could be such a bacterium. It is one of the smallest free-living bacterium known (680 genes), has no cell wall, can be cultivated in Vitro, can be genetically manipulated and can enter inside human cells.
Summary
The idea of harnessing living organisms for treating human diseases is not new but, so far, the majority of the living vectors used in human therapy are viruses which have the disadvantage of the limited number of genes and networks that can contain. Bacteria allow the cloning of complex networks and the possibility of making a large plethora of compounds, naturally or through careful redesign. One of the main limitations for the use of bacteria to treat human diseases is their complexity, the existence of a cell wall that difficult the communication with the target cells, the lack of control over its growth and the immune response that will elicit on its target. Ideally one would like to have a very small bacterium (of a mitochondria size), with no cell wall, which could be grown in Vitro, be genetically manipulated, for which we will have enough data to allow a complete understanding of its behaviour and which could live as a human cell parasite. Such a microorganism could in principle be used as a living vector in which genes of interests, or networks producing organic molecules of medical relevance, could be introduced under in Vitro conditions and then inoculated on extracted human cells or in the organism, and then become a new organelle in the host. Then, it could produce and secrete into the host proteins which will be needed to correct a genetic disease, or drugs needed by the patient. To do that, we need to understand in excruciating detail the Biology of the target bacterium and how to interface with the host cell cycle (Systems biology aspect). Then we need to have engineering tools (network design, protein design, simulations) to modify the target bacterium to behave like an organelle once inside the cell (Synthetic biology aspect). M.pneumoniae could be such a bacterium. It is one of the smallest free-living bacterium known (680 genes), has no cell wall, can be cultivated in Vitro, can be genetically manipulated and can enter inside human cells.
Max ERC Funding
2 400 000 €
Duration
Start date: 2009-03-01, End date: 2015-02-28
Project acronym EpigenomeProgramming
Project An experimental and bioinformatic toolbox for functional epigenomics and its application to epigenetically making and breaking a cancer cell
Researcher (PI) Christoph Bock
Host Institution (HI) CEMM - FORSCHUNGSZENTRUM FUER MOLEKULARE MEDIZIN GMBH
Call Details Starting Grant (StG), LS2, ERC-2015-STG
Summary Epigenetic alterations can be detected in all cancers and in essentially every patient. Despite their prevalence, the concrete functional roles of these alterations are not well understood, for two reasons: First, cancer samples tend to carry many correlated epigenetic alterations, making it difficult to statistically distinguish relevant driver events from those that co-occur for other reasons. Second, we lack tools for targeted epigenome editing that could be used to validate biological function in perturbation and rescue experiments.
The proposed project strives to overcome these limitations through experimental and bioinformatic methods development, with the ambition of making and breaking cancer cells in vitro by introducing defined sets of epigenetic alterations. We will focus on leukemia as our “model cancer” (given its low mutation rate, frequent defects in epigenetic regulators, and availability of excellent functional assays), but the concepts and methods are general. In Aim 1, we will generate epigenome profiles for a human knockout cell collection comprising 100 epigenetic regulators and use the data to functionally annotate thousands of epigenetic alterations observed in large cancer datasets. In Aim 2, we will develop an experimental toolbox for epigenome programming using epigenetic drugs, CRISPR-assisted recruitment of epigenetic modifiers for locus-specific editing, and cell-derived guide RNA libraries for epigenome copying. Finally, in Aim 3 we will explore epigenome programming (methods from Aim 2) of candidate driver events (predictions from Aim 1) with the ultimate goal of converting cancer cells into non-cancer cells and vice versa.
In summary, this project will establish a broadly applicable methodology and toolbox for dissecting the functional roles of epigenetic alterations in cancer. Moreover, successful creation of a cancer that is driven purely by epigenetic alterations could challenge our understanding of cancer as a genetic disease.
Summary
Epigenetic alterations can be detected in all cancers and in essentially every patient. Despite their prevalence, the concrete functional roles of these alterations are not well understood, for two reasons: First, cancer samples tend to carry many correlated epigenetic alterations, making it difficult to statistically distinguish relevant driver events from those that co-occur for other reasons. Second, we lack tools for targeted epigenome editing that could be used to validate biological function in perturbation and rescue experiments.
The proposed project strives to overcome these limitations through experimental and bioinformatic methods development, with the ambition of making and breaking cancer cells in vitro by introducing defined sets of epigenetic alterations. We will focus on leukemia as our “model cancer” (given its low mutation rate, frequent defects in epigenetic regulators, and availability of excellent functional assays), but the concepts and methods are general. In Aim 1, we will generate epigenome profiles for a human knockout cell collection comprising 100 epigenetic regulators and use the data to functionally annotate thousands of epigenetic alterations observed in large cancer datasets. In Aim 2, we will develop an experimental toolbox for epigenome programming using epigenetic drugs, CRISPR-assisted recruitment of epigenetic modifiers for locus-specific editing, and cell-derived guide RNA libraries for epigenome copying. Finally, in Aim 3 we will explore epigenome programming (methods from Aim 2) of candidate driver events (predictions from Aim 1) with the ultimate goal of converting cancer cells into non-cancer cells and vice versa.
In summary, this project will establish a broadly applicable methodology and toolbox for dissecting the functional roles of epigenetic alterations in cancer. Moreover, successful creation of a cancer that is driven purely by epigenetic alterations could challenge our understanding of cancer as a genetic disease.
Max ERC Funding
1 281 205 €
Duration
Start date: 2016-12-01, End date: 2021-11-30
Project acronym GameofGates
Project Solute carrier proteins as the gates managing chemical access to cells
Researcher (PI) Giulio SUPERTI-FURGA
Host Institution (HI) CEMM - FORSCHUNGSZENTRUM FUER MOLEKULARE MEDIZIN GMBH
Call Details Advanced Grant (AdG), LS2, ERC-2015-AdG
Summary Chemical exchange between cells and their environment occurs at cellular membranes, the interface where biology meets chemistry. Studying mechanisms of drug resistance, I realized that SoLute Carrier proteins (SLCs), not only represent the major class of small molecule transporters, but that they are encoded by one of the most neglected group of human genes. I identified a case where an SLC controls the activity of mTOR, suggesting that other SLCs may be involved in signalling. This formed the basis for the GameofGates project proposal. The name refers to SLCs as a metaphor for cellular gates coordinating access to resources following game rules that are largely unknown but worth learning, as the acquired knowledge could impact our understanding of cellular physiology and open avenues for innovative treatment of human diseases.
GameofGates (GoG) plans the investigation of SLC function by comprehensively and deeply charting the genetic and protein interaction landscape of SLCs in a human cell line, while monitoring fitness, drug sensitivity and metabolic state. GoG aims at functionally de-orphanize many SLCs by assessing hundreds of thousands of genetic interactions as well as thousands protein and drug interactions. I hypothesize that SLC action is linked to signalling pathways and plays an important role in integration of metabolism and cell regulation for successful homeostasis. I propose that whole circuits of SLCs may be linked to particular nutrient auxotrophy states and that knowledge of these dependencies could instruct assessment of vulnerabilities in cancer cells. In turn, these could be pharmacologically exploited using existing or future drugs. Overall, GoG should position enough pieces into functional and regulatory networks in the SLC puzzle game to facilitate future work and motivate the community to embrace investigation of SLCs as conveyers of metabolic and chemical integration of cell biology with physiology and, in a wider scope, ecology.
Summary
Chemical exchange between cells and their environment occurs at cellular membranes, the interface where biology meets chemistry. Studying mechanisms of drug resistance, I realized that SoLute Carrier proteins (SLCs), not only represent the major class of small molecule transporters, but that they are encoded by one of the most neglected group of human genes. I identified a case where an SLC controls the activity of mTOR, suggesting that other SLCs may be involved in signalling. This formed the basis for the GameofGates project proposal. The name refers to SLCs as a metaphor for cellular gates coordinating access to resources following game rules that are largely unknown but worth learning, as the acquired knowledge could impact our understanding of cellular physiology and open avenues for innovative treatment of human diseases.
GameofGates (GoG) plans the investigation of SLC function by comprehensively and deeply charting the genetic and protein interaction landscape of SLCs in a human cell line, while monitoring fitness, drug sensitivity and metabolic state. GoG aims at functionally de-orphanize many SLCs by assessing hundreds of thousands of genetic interactions as well as thousands protein and drug interactions. I hypothesize that SLC action is linked to signalling pathways and plays an important role in integration of metabolism and cell regulation for successful homeostasis. I propose that whole circuits of SLCs may be linked to particular nutrient auxotrophy states and that knowledge of these dependencies could instruct assessment of vulnerabilities in cancer cells. In turn, these could be pharmacologically exploited using existing or future drugs. Overall, GoG should position enough pieces into functional and regulatory networks in the SLC puzzle game to facilitate future work and motivate the community to embrace investigation of SLCs as conveyers of metabolic and chemical integration of cell biology with physiology and, in a wider scope, ecology.
Max ERC Funding
2 389 782 €
Duration
Start date: 2016-10-01, End date: 2021-09-30
Project acronym NONCODRIVERS
Project Finding noncoding cancer drivers
Researcher (PI) Nuria Lopez Bigas
Host Institution (HI) FUNDACIO INSTITUT DE RECERCA BIOMEDICA (IRB BARCELONA)
Call Details Consolidator Grant (CoG), LS2, ERC-2015-CoG
Summary Finding the mutations, genes and pathways directly involved in cancer is of paramount importance to understand the mechanisms of tumour development and devise therapeutic strategies to overcome the disease. Due to their role in cancer development and maintenance, the proteins encoded by cancer genes are candidate therapeutic targets. Indeed, in recent years we have witnessed the development of successful cancer-targeting therapies to counteract the effect of driver mutations. Although the coding part of the human genome has now largely been explored in the search for cancer driver mutations in most frequent cancer types, the extent of involvement of noncoding mutations in cancer development remains a mystery. The main challenges faced are: 1) the functional role of most noncoding regions is unknown, and 2) tumours often have thousands of somatic mutations, so that distinguishing cancer driver mutations from bystanders is like finding the proverbial needle in a haystack. To overcome these two challenges I propose to analyse the pattern of somatic mutations across thousands of tumours in noncoding regions to identify signals of positive selection. These signals are an indication that mutations in the region have been positively selected during tumour evolution and are thus directly involved in the tumour phenotype. The large scale analysis proposed here will allow us to create a catalogue of noncoding elements involved in different types of cancer upon mutations. We will study in detail a selected set of driver elements to uncover their specific function and role in the tumourigenic process. Furthermore, we will explore possibilities of counteracting their driver effect with targeted drugs. The results of this project may boost our understanding of the biological role of noncoding regions, help to unravel novel molecular causes of cancer and provide novel targeted therapeutic opportunities for cancer patients.
Summary
Finding the mutations, genes and pathways directly involved in cancer is of paramount importance to understand the mechanisms of tumour development and devise therapeutic strategies to overcome the disease. Due to their role in cancer development and maintenance, the proteins encoded by cancer genes are candidate therapeutic targets. Indeed, in recent years we have witnessed the development of successful cancer-targeting therapies to counteract the effect of driver mutations. Although the coding part of the human genome has now largely been explored in the search for cancer driver mutations in most frequent cancer types, the extent of involvement of noncoding mutations in cancer development remains a mystery. The main challenges faced are: 1) the functional role of most noncoding regions is unknown, and 2) tumours often have thousands of somatic mutations, so that distinguishing cancer driver mutations from bystanders is like finding the proverbial needle in a haystack. To overcome these two challenges I propose to analyse the pattern of somatic mutations across thousands of tumours in noncoding regions to identify signals of positive selection. These signals are an indication that mutations in the region have been positively selected during tumour evolution and are thus directly involved in the tumour phenotype. The large scale analysis proposed here will allow us to create a catalogue of noncoding elements involved in different types of cancer upon mutations. We will study in detail a selected set of driver elements to uncover their specific function and role in the tumourigenic process. Furthermore, we will explore possibilities of counteracting their driver effect with targeted drugs. The results of this project may boost our understanding of the biological role of noncoding regions, help to unravel novel molecular causes of cancer and provide novel targeted therapeutic opportunities for cancer patients.
Max ERC Funding
1 995 829 €
Duration
Start date: 2016-12-01, End date: 2021-11-30
Project acronym PIWI-Chrom
Project Understanding small RNA-mediated transposon control at the level of chromatin in the animal germline
Researcher (PI) Julius Brennecke
Host Institution (HI) INSTITUT FUER MOLEKULARE BIOTECHNOLOGIE GMBH
Call Details Consolidator Grant (CoG), LS2, ERC-2015-CoG
Summary Transposable elements—universal components of genomes—pose a major threat to genome integrity due to their mutagenic character. In all eukaryotic lineages small RNA pathways act as defense systems to protect the host genome against the activity of transposons. The central pathway in animals is the gonad-specific PIWI interacting RNA (piRNA) pathway, one of the most elaborate but also least understood small RNA silencing systems.
Here I propose to study the interplay between the piRNA pathway and chromatin biology in Drosophila with two aims: First, we will identify the factors and investigate the processes that underlie piRNA-guided silencing in the nucleus. Our objective is to understand how recruitment of an Argonaute protein to a nascent RNA mechanistically leads to the assembly of effector proteins that govern heterochromatin formation and transcriptional silencing. Second, we will study the biology of piRNA clusters, heterochromatic loci that encompass a library of transposon fragments and that act as the pathway’s memory system. Our goal is to uncover how a group of proteins—several of which are germline-specific variants of basic cellular factors—enable transcription within heterochromatin and control the downstream fate of the emerging non-coding RNAs.
Our work centers on the piRNA pathway in Drosophila ovaries, undeniably the model system at the forefront of the field. By combining the strength of fly genetics with the power of genome-wide approaches we will uncover how heterochromatin on the one hand governs silencing and how the piRNA pathway on the other hand exploits it to facilitate the transcription of piRNA precursors. This will reveal fundamental insights into the co-evolution of transposons and host genomes. At the same time, by studying the piRNA pathway’s intersection with chromatin biology and transcription, we expect to reveal new insights into basic principles of gene expression.
Summary
Transposable elements—universal components of genomes—pose a major threat to genome integrity due to their mutagenic character. In all eukaryotic lineages small RNA pathways act as defense systems to protect the host genome against the activity of transposons. The central pathway in animals is the gonad-specific PIWI interacting RNA (piRNA) pathway, one of the most elaborate but also least understood small RNA silencing systems.
Here I propose to study the interplay between the piRNA pathway and chromatin biology in Drosophila with two aims: First, we will identify the factors and investigate the processes that underlie piRNA-guided silencing in the nucleus. Our objective is to understand how recruitment of an Argonaute protein to a nascent RNA mechanistically leads to the assembly of effector proteins that govern heterochromatin formation and transcriptional silencing. Second, we will study the biology of piRNA clusters, heterochromatic loci that encompass a library of transposon fragments and that act as the pathway’s memory system. Our goal is to uncover how a group of proteins—several of which are germline-specific variants of basic cellular factors—enable transcription within heterochromatin and control the downstream fate of the emerging non-coding RNAs.
Our work centers on the piRNA pathway in Drosophila ovaries, undeniably the model system at the forefront of the field. By combining the strength of fly genetics with the power of genome-wide approaches we will uncover how heterochromatin on the one hand governs silencing and how the piRNA pathway on the other hand exploits it to facilitate the transcription of piRNA precursors. This will reveal fundamental insights into the co-evolution of transposons and host genomes. At the same time, by studying the piRNA pathway’s intersection with chromatin biology and transcription, we expect to reveal new insights into basic principles of gene expression.
Max ERC Funding
1 999 530 €
Duration
Start date: 2016-07-01, End date: 2021-06-30
Project acronym UNICODE
Project Evolution and Impact of Heterochromatin on a Young Drosophila Y chromosome
Researcher (PI) Qi Zhou
Host Institution (HI) UNIVERSITAT WIEN
Call Details Starting Grant (StG), LS2, ERC-2015-STG
Summary The transition from euchromatin to heterochromatin is a fundamental process that particularly reshaped the epigenomic landscape of Y chromosome. Its definitive genomic underpinning and broad functional impact are still unclear, as heterochromatin (e.g., that of human Y) is usually too repetitive to study. I have previously demonstrated that, the young Y (‘neo-Y’) chromosome of Drosophila miranda has just initiated such a transition, thus is a powerful model to unveil the evolution, regulation and functional interaction of heterochromatin. I showed that this neo-Y still harbours over 1800 genes, and only 20-50% of the sequences are transposable elements (TE). Over five years, I aim to: 1) precisely resolve the structure and insertion sites of TEs as a pre-requisite for studying heterochromatin, by combining state-of-art sequencing and bioinformatic techniques. 2) I will reveal the de novo heterochromatin formation triggered by TE insertions or the heterochromatin/euchromatin boundary shifts on the neo-Y, by comparing the binding profiles of histone modification hallmarks and insulator proteins of D. miranda to its sibling species D. pseudoobscura, which lacks the neo-Y. Such epigenomic changes have likely driven the exaptation or innovation of small RNA pathways that govern the TE mobility. 3) I will then identify the responsible small RNAs and their encoding loci, which are expected to have newly emerged or differentially expressed in D. miranda relative to D. pseudoobscura. 4) Finally, I will develop CRISPR/Cas9 in D. miranda to manipulate the expression of TEs encoding such small RNAs on the neo-Y, in order to scrutinize how TE/heterochromatin evolution on the Y would impact the chromatin landscape of the entire host genome. The combined aim of this multidisciplinary project is to generate a framework for understanding the basic mechanisms of how heterochromatin evolves; and open a new avenue toward the discovery of Y chromosome function beyond male determination.
Summary
The transition from euchromatin to heterochromatin is a fundamental process that particularly reshaped the epigenomic landscape of Y chromosome. Its definitive genomic underpinning and broad functional impact are still unclear, as heterochromatin (e.g., that of human Y) is usually too repetitive to study. I have previously demonstrated that, the young Y (‘neo-Y’) chromosome of Drosophila miranda has just initiated such a transition, thus is a powerful model to unveil the evolution, regulation and functional interaction of heterochromatin. I showed that this neo-Y still harbours over 1800 genes, and only 20-50% of the sequences are transposable elements (TE). Over five years, I aim to: 1) precisely resolve the structure and insertion sites of TEs as a pre-requisite for studying heterochromatin, by combining state-of-art sequencing and bioinformatic techniques. 2) I will reveal the de novo heterochromatin formation triggered by TE insertions or the heterochromatin/euchromatin boundary shifts on the neo-Y, by comparing the binding profiles of histone modification hallmarks and insulator proteins of D. miranda to its sibling species D. pseudoobscura, which lacks the neo-Y. Such epigenomic changes have likely driven the exaptation or innovation of small RNA pathways that govern the TE mobility. 3) I will then identify the responsible small RNAs and their encoding loci, which are expected to have newly emerged or differentially expressed in D. miranda relative to D. pseudoobscura. 4) Finally, I will develop CRISPR/Cas9 in D. miranda to manipulate the expression of TEs encoding such small RNAs on the neo-Y, in order to scrutinize how TE/heterochromatin evolution on the Y would impact the chromatin landscape of the entire host genome. The combined aim of this multidisciplinary project is to generate a framework for understanding the basic mechanisms of how heterochromatin evolves; and open a new avenue toward the discovery of Y chromosome function beyond male determination.
Max ERC Funding
1 971 846 €
Duration
Start date: 2016-08-01, End date: 2021-07-31
