Project acronym DYNACOM
Project From Genome Integrity to Genome Plasticity:
Dynamic Complexes Controlling Once per Cell Cycle Replication
Researcher (PI) Zoi Lygerou
Host Institution (HI) PANEPISTIMIO PATRON
Call Details Starting Grant (StG), LS3, ERC-2011-StG_20101109
Summary Accurate genome duplication is controlled by multi-subunit protein complexes which associate with chromatin and dictate when and where replication should take place. Dynamic changes in these complexes lie at the heart of their ability to ensure the maintenance of genomic integrity. Defects in origin bound complexes lead to re-replication of the genome across evolution, have been linked to DNA-replication stress and may predispose for gene amplification events. Such genomic aberrations are central to malignant transformation.
We wish to understand how once per cell cycle replication is normally controlled within the context of the living cell and how defects in this control may result in loss of genome integrity and provide genome plasticity. To this end, live cell imaging in human cells in culture will be combined with genetic studies in fission yeast and modelling and in silico analysis.
The proposed research aims to:
1. Decipher the regulatory mechanisms which act in time and space to ensure once per cell cycle replication within living cells and how they may be affected by system aberrations, using functional live cell imaging.
2. Test whether aberrations in the licensing system may provide a selective advantage, through amplification of multiple genomic loci. To this end, a natural selection experiment will be set up in fission yeast .
3. Investigate how rereplication takes place along the genome in single cells. Is there heterogeneity amongst a population, leading to a plethora of different genotypes? In silico analysis of full genome DNA rereplication will be combined to single cell analysis in fission yeast.
4. Assess the relevance of our findings for gene amplification events in cancer. Does ectopic expression of human Cdt1/Cdc6 in cancer cells enhance drug resistance through gene amplification?
Our findings are expected to offer novel insight into mechanisms underlying cancer development and progression.
Summary
Accurate genome duplication is controlled by multi-subunit protein complexes which associate with chromatin and dictate when and where replication should take place. Dynamic changes in these complexes lie at the heart of their ability to ensure the maintenance of genomic integrity. Defects in origin bound complexes lead to re-replication of the genome across evolution, have been linked to DNA-replication stress and may predispose for gene amplification events. Such genomic aberrations are central to malignant transformation.
We wish to understand how once per cell cycle replication is normally controlled within the context of the living cell and how defects in this control may result in loss of genome integrity and provide genome plasticity. To this end, live cell imaging in human cells in culture will be combined with genetic studies in fission yeast and modelling and in silico analysis.
The proposed research aims to:
1. Decipher the regulatory mechanisms which act in time and space to ensure once per cell cycle replication within living cells and how they may be affected by system aberrations, using functional live cell imaging.
2. Test whether aberrations in the licensing system may provide a selective advantage, through amplification of multiple genomic loci. To this end, a natural selection experiment will be set up in fission yeast .
3. Investigate how rereplication takes place along the genome in single cells. Is there heterogeneity amongst a population, leading to a plethora of different genotypes? In silico analysis of full genome DNA rereplication will be combined to single cell analysis in fission yeast.
4. Assess the relevance of our findings for gene amplification events in cancer. Does ectopic expression of human Cdt1/Cdc6 in cancer cells enhance drug resistance through gene amplification?
Our findings are expected to offer novel insight into mechanisms underlying cancer development and progression.
Max ERC Funding
1 531 000 €
Duration
Start date: 2012-02-01, End date: 2017-01-31
Project acronym HIVBIOCHIP
Project A POINT-OF-CARE BIOCHIP FOR HIV MONITORING IN THE DEVELOPING WORLD
Researcher (PI) Nikolaos Chronis
Host Institution (HI) "NATIONAL CENTER FOR SCIENTIFIC RESEARCH ""DEMOKRITOS"""
Call Details Starting Grant (StG), LS7, ERC-2011-StG_20101109
Summary HIV/AIDS is one of the most destructive pandemics in human history, responsible for more than 25 million deaths. More than 30 million people live with limited or no access to therapeutic treatments, mainly due to the high cost of highly active antiretroviral therapies (HAART) and current diagnostic tests as well as due to the lack of basic infrastructure (e.g. lack of electricity, no trained personnel) that can support these tests. The need for innovative, inexpensive diagnostic instrumentation technology that can be used in resource-limited settings is immediate.
While programs that offer free HAART are being implemented in resource-limited settings, no diagnostic tests are available for evaluating the efficacy of HAART provided for the reasons mentioned above. Efficient management of HAART requires monitoring the course of HIV infection over time. The World Health Organization (WHO) recommends the CD4 T-cell count test for monitoring the clinical status of HIV individuals in resource-limited settings.
We propose to develop a portable, inexpensive, MEMS (MicroElectroMechanical Systems)-based, imaging system for counting the absolute number of CD4 cells from 1 l of whole blood. We use the term ‘imaging system’ to denote the different approach we follow for counting CD4 cells: rather the reading one by one singles cells (as it is done with flow cytometry), our system can image simultaneously thousands of individual cells, pre-assembled on the surface of a biochip. Although the proposed imaging system can replace current expensive cell counting instrumentation, our goal is to develop a system that can reach the end-user wherever limited infrastructure is present and no access to a hospital or clinic is possible. Such technology will not only enable to monitor the efficacy of an individual’s HAART in the developing world, but it will make more medicines available by identifying patients who need a treatment from patients who do not need it.
Summary
HIV/AIDS is one of the most destructive pandemics in human history, responsible for more than 25 million deaths. More than 30 million people live with limited or no access to therapeutic treatments, mainly due to the high cost of highly active antiretroviral therapies (HAART) and current diagnostic tests as well as due to the lack of basic infrastructure (e.g. lack of electricity, no trained personnel) that can support these tests. The need for innovative, inexpensive diagnostic instrumentation technology that can be used in resource-limited settings is immediate.
While programs that offer free HAART are being implemented in resource-limited settings, no diagnostic tests are available for evaluating the efficacy of HAART provided for the reasons mentioned above. Efficient management of HAART requires monitoring the course of HIV infection over time. The World Health Organization (WHO) recommends the CD4 T-cell count test for monitoring the clinical status of HIV individuals in resource-limited settings.
We propose to develop a portable, inexpensive, MEMS (MicroElectroMechanical Systems)-based, imaging system for counting the absolute number of CD4 cells from 1 l of whole blood. We use the term ‘imaging system’ to denote the different approach we follow for counting CD4 cells: rather the reading one by one singles cells (as it is done with flow cytometry), our system can image simultaneously thousands of individual cells, pre-assembled on the surface of a biochip. Although the proposed imaging system can replace current expensive cell counting instrumentation, our goal is to develop a system that can reach the end-user wherever limited infrastructure is present and no access to a hospital or clinic is possible. Such technology will not only enable to monitor the efficacy of an individual’s HAART in the developing world, but it will make more medicines available by identifying patients who need a treatment from patients who do not need it.
Max ERC Funding
1 986 000 €
Duration
Start date: 2012-06-01, End date: 2017-05-31
Project acronym SET-NET
Project Enzymatic and genomic targets of histone modifying enzymes and their role in liver metabolism and hepatocarcinogenesis
Researcher (PI) Ioannis Talianidis
Host Institution (HI) BIOMEDICAL SCIENCES RESEARCH CENTER ALEXANDER FLEMING
Call Details Advanced Grant (AdG), LS4, ERC-2011-ADG_20110310
Summary "In this proposal we will study the role of histone methytransferases Set9, PR-SET7, Smyd2, Smyd3 and the demethylase LSD1 in hepatocarcinogenesis and in the regulation of hepatic metabolic pathways.
The hypothesis raised here suggests that the function of histone modifying enzymes is realized through 4 overlapping regulatory layers: i. via modifications of chromatin, ii. via modifications of recently identified transcription factor substrates, iii. via influencing the hepatic transcription factor crossregulatory circuitry, and iv. via modification of each other.
To dissect the role of these regulatory layers and determine their contribution in the control of hepatic metabolic pathways, and in the development of hepatocellular carcinoma, our activities will involve: i. Generation of relevant KO and transgenic mouse models, ii. Functional characterization of novel non-histone and histone substrates, iii. Analysis of novel cross-regulatory protein modifications affecting the activity of the enzymes and iv. the identification of their genomic targets and associated chromatin modifications using global approaches.
The work is expected to provide important insights into a previously unanticipated network of protein methylation-directed regulatory modules, potentially operating in multiple biological pathways such as liver development, metabolism, apoptosis and carcinogenesis. The functioning of such network would be of high biological importance, with far-reaching implications in drug development, rivaling those of phosphorylation or acetylation regulated processes."
Summary
"In this proposal we will study the role of histone methytransferases Set9, PR-SET7, Smyd2, Smyd3 and the demethylase LSD1 in hepatocarcinogenesis and in the regulation of hepatic metabolic pathways.
The hypothesis raised here suggests that the function of histone modifying enzymes is realized through 4 overlapping regulatory layers: i. via modifications of chromatin, ii. via modifications of recently identified transcription factor substrates, iii. via influencing the hepatic transcription factor crossregulatory circuitry, and iv. via modification of each other.
To dissect the role of these regulatory layers and determine their contribution in the control of hepatic metabolic pathways, and in the development of hepatocellular carcinoma, our activities will involve: i. Generation of relevant KO and transgenic mouse models, ii. Functional characterization of novel non-histone and histone substrates, iii. Analysis of novel cross-regulatory protein modifications affecting the activity of the enzymes and iv. the identification of their genomic targets and associated chromatin modifications using global approaches.
The work is expected to provide important insights into a previously unanticipated network of protein methylation-directed regulatory modules, potentially operating in multiple biological pathways such as liver development, metabolism, apoptosis and carcinogenesis. The functioning of such network would be of high biological importance, with far-reaching implications in drug development, rivaling those of phosphorylation or acetylation regulated processes."
Max ERC Funding
2 499 600 €
Duration
Start date: 2012-01-01, End date: 2016-12-31
