Periodic Reporting for period 1 - rDNAstress (Novel insights into DNA damage and stress responses in the nucleolus: Mechanisms and relevance for genomic (in)stability and cancer)
Periodo di rendicontazione: 2019-01-01 al 2020-12-31
The Action "rDNAstress" contributes to target one of the priority challenges defined by the European Union: Health, demographic changes and well-being. The main objective of this challenge is health-improving during life and increasing the welfare of society. Importantly, the proposed project is generating in-depth knowledge about genome instability in human cells to understand cancer disease. Hopefully, this project's results will help extend our knowledge about a minimal region of the human genome, named ribosomal DNA, which is essential to genome maintenance and could be a potential tool for targeting cancer cells. Furthermore, the functional role of the ribosomal DNA to synthesize ribosomes, main machinery for protein synthesis, which is overstimulated in cancer cells, makes this proposal attractive to improve and develop new approaches and strategies for cancer treatments. Indeed, the proposal has developed and improved new methods and innovative technologies to uncover new factors involved in genome stability allow us to improve in the diagnosis and prognosis, opening the possibility of more personalized medicine.
The overall objective of this action has been to assess rDNA damage and the ensuing genomic instability in human cells exposed to insults such as radiation, chemotherapy drugs, replication and transcription stress, ribosome biogenesis stress, and oncogenes, identify and functionally characterize signalling and repair factors that guard nucleolar integrity and function, and investigate how tumour cells may evade such control of nucleolar (replication) stress. To achieve these goals, we have employed innovative cellular models, molecular genetic, cell biology, and imaging technologies and functional assays. The expected results are providing novel insights into genome integrity maintenance and cellular stress responses and may inspire the design of future strategies to treat or even prevent cancer.
Unfortunately, during this active period, the current COVID-10 pandemia has impacted our society, our lives and also in our project execution due to the continuous outbreak in Denmark and strict restrictions in the research centre environment. To solve this situation, we have depth in some previous results obtained before COVID-19 outbreak, and we get promising results that they will publish soon, hopefully.
The overall objective of this action has been to assess rDNA damage and the ensuing genomic instability in human cells exposed to insults such as radiation, chemotherapy drugs, replication and transcription stress, ribosome biogenesis stress, and oncogenes, identify and functionally characterize signalling and repair factors that guard nucleolar integrity and function, and investigate how tumour cells may evade such control of nucleolar (replication) stress. To achieve these goals, we have employed innovative cellular models, molecular genetic, cell biology, and imaging technologies and functional assays. The expected results are providing novel insights into genome integrity maintenance and cellular stress responses and may inspire the design of future strategies to treat or even prevent cancer.
Unfortunately, during this active period, the current COVID-10 pandemia has impacted our society, our lives and also in our project execution due to the continuous outbreak in Denmark and strict restrictions in the research centre environment. To solve this situation, we have depth in some previous results obtained before COVID-19 outbreak, and we get promising results that they will publish soon, hopefully.
Work was conducted via 3 main work packages (WPs). WP1 focused to the study of spatiotemporal nucleolar dynamics of DDR and ribosome biogenesis factors in response to stress. We took advantage of live cell imaging and DNA damage approaches to demonstrate that DDR proteins upon ribosomal DNA damage require active transcription to move DNA breaks to the perifery of the nucleoli to be repaired properly. This WP1 comprise a 1 manuscript currently in preparation but we cannot be shown in conferences due to COVID-19 pandemia.
WP2 addressed Oncogene-induced rDNA damage and genomic instability. We obtained promising results form overexpression of c-myc oncogene in a novel factor called RPAIN which we got data about its possible role in the DNA damage response upon oncogene stress in nucleoli. The investigation on these preliminary results was stopped due to the pandemic situation in order to focus on other results with more advance data to be published. On the other hand, in this WP2 we discovered that RNA transcription is essential to accurate repair of DSBs in whole-genome via homologous recombination. Currently, these results are under review in Nature Cell Biology journal, and they were shown in 2 international conferences (Germany and Spain) in 2019 and 2020 (previous COVID-19). The WP3 was focused on the detailed functional analysis of the role of selected factors in response to insult-induced and oncogene-induced nucleolar stress. We studied the functional synergy among RNA polymerase II inhibitors and olaparib in response to radiation as a source of DNA damage. Results form this approach include new strategies using a promising drug called THZ1, in combination with some novel drugs currently used in some type of breast cancer. This result has been shown in the conferences previously cited.
WP2 addressed Oncogene-induced rDNA damage and genomic instability. We obtained promising results form overexpression of c-myc oncogene in a novel factor called RPAIN which we got data about its possible role in the DNA damage response upon oncogene stress in nucleoli. The investigation on these preliminary results was stopped due to the pandemic situation in order to focus on other results with more advance data to be published. On the other hand, in this WP2 we discovered that RNA transcription is essential to accurate repair of DSBs in whole-genome via homologous recombination. Currently, these results are under review in Nature Cell Biology journal, and they were shown in 2 international conferences (Germany and Spain) in 2019 and 2020 (previous COVID-19). The WP3 was focused on the detailed functional analysis of the role of selected factors in response to insult-induced and oncogene-induced nucleolar stress. We studied the functional synergy among RNA polymerase II inhibitors and olaparib in response to radiation as a source of DNA damage. Results form this approach include new strategies using a promising drug called THZ1, in combination with some novel drugs currently used in some type of breast cancer. This result has been shown in the conferences previously cited.
This MSCA allowed the Fellow to develop agility with many different research methodologies and we set up cutting-edge techniques to visualize molecular machinery related to DNA stress metabolism. We have generated important new knowledge about ribosomal genome instability upon DNA damage suing CRISPR technology and also radiation. Besides, we have added to the field new techniques to study homologous recombination as the main repair machinery to avoid genome instability and we have characterized a novel RNA polymerase II inhibitor, called THZ1, which our objective will be to establish several research lines to carry on preclinical studies and clinical trials for cancer treatments.
For this reason, our MSCA is increased and improved tools to cancer fight and also added new drugs to be investigated with potential use in cancer treatments and new approaches on currently radiotherapy therapies.
For this reason, our MSCA is increased and improved tools to cancer fight and also added new drugs to be investigated with potential use in cancer treatments and new approaches on currently radiotherapy therapies.