Periodic Reporting for period 1 - ZIDOMS (Using Zebrafish as a novel tool to Improve the Diagnosis and Outcome of Marfan Syndrome)
Reporting period: 2018-09-01 to 2020-08-31
Marfan syndrome (MFS) is a rare genetic disease, affecting an estimated 1:5,000 to 1:10,000 people. It is caused by defects in the FBN1 gene, which codes for the fibrillin-1 protein. This protein plays an important role in the structure and support of many different tissue types and organs throughout the body. Therefore, MFS can cause a range of symptoms affecting the skeleton, the eyes, and particularly the heart and blood vessels. MFS can lead to a widening of the aorta, the biggest artery in the body, as well as tearing and eventually catastrophic rupture of this blood vessel.
To date, no cure exists for MFS, and patients can only be treated symptomatically by surgery or blood pressure-lowering medication. On the other hand, the consequences of a new sequence variant in the FBN1 gene which is identified during genetic testing cannot be predicted for each patient based on the currently available information. Changes in the FBN1 code might have no effects, could lead to mild or moderate symptoms, or might even cause a quick progression to life-threatening conditions. The lack of effective targeted therapy for MFS, as well as the uncertainty regarding the course of the disease places a heavy burden on the quality of life of MFS patients, and increases the costs for clinical management.
In order to address these issues, there is a strong need for flexible models of MFS which can mirror the complexity of this disease. Mouse models already exist but they are limited by the number of conditions that can be tested efficiently. Zebrafish have been used more and more in preclinical research, since the effects of many different drugs or genetic manipulations can be tested rapidly in this model. The overall goal of this project is to generate a new zebrafish model for MFS, in order to boost the development of a more personalized approach to the management of the disease. More specifically, the main objectives of the project are to validate the new zebrafish MFS model, to develop an innovative system to test the effects of specific human FBN1 gene defects in the zebrafish model, and to use a novel approach to test a large number of pharmaceutical molecules in this model in order to discover new treatment options for MFS patients.
Another important objective of this project is to foster the career development of the MSCA researcher. A major advantage of the interaction with the mentor, a clinician, is the strong link to the clinical aspects of MFS. This helps to clearly identify areas of research which have a strong relevance to patient care, and therefore have a great potential to realize maximal impact beyond the scope of the fundamental research project.
At the end of the MSCA project we have been able to validate a new zebrafish model of MFS, and we have uncovered the specific roles of the different zebrafish fibrillin genes in the cardiovascular system.
To date, no cure exists for MFS, and patients can only be treated symptomatically by surgery or blood pressure-lowering medication. On the other hand, the consequences of a new sequence variant in the FBN1 gene which is identified during genetic testing cannot be predicted for each patient based on the currently available information. Changes in the FBN1 code might have no effects, could lead to mild or moderate symptoms, or might even cause a quick progression to life-threatening conditions. The lack of effective targeted therapy for MFS, as well as the uncertainty regarding the course of the disease places a heavy burden on the quality of life of MFS patients, and increases the costs for clinical management.
In order to address these issues, there is a strong need for flexible models of MFS which can mirror the complexity of this disease. Mouse models already exist but they are limited by the number of conditions that can be tested efficiently. Zebrafish have been used more and more in preclinical research, since the effects of many different drugs or genetic manipulations can be tested rapidly in this model. The overall goal of this project is to generate a new zebrafish model for MFS, in order to boost the development of a more personalized approach to the management of the disease. More specifically, the main objectives of the project are to validate the new zebrafish MFS model, to develop an innovative system to test the effects of specific human FBN1 gene defects in the zebrafish model, and to use a novel approach to test a large number of pharmaceutical molecules in this model in order to discover new treatment options for MFS patients.
Another important objective of this project is to foster the career development of the MSCA researcher. A major advantage of the interaction with the mentor, a clinician, is the strong link to the clinical aspects of MFS. This helps to clearly identify areas of research which have a strong relevance to patient care, and therefore have a great potential to realize maximal impact beyond the scope of the fundamental research project.
At the end of the MSCA project we have been able to validate a new zebrafish model of MFS, and we have uncovered the specific roles of the different zebrafish fibrillin genes in the cardiovascular system.
Three main research objectives were defined for the project, and work was performed within work packages (WP) 1-6 toward achieving significant progress for each.
As a first objective, we wanted to validate a zebrafish model for MFS which can be used for functional genetic testing as well as for drug discovery. Despite initial setbacks, which showed that the zebrafish fibrillin genes do not behave exactly like the three human fibrillin genes, we have been able to generate a novel zebrafish model which recapitulates several relevant characteristics of the human disease. During the generation of this new model, we learned more about the role of the different zebrafish fibrillins in the cardiovascular system both during development as well as in adult stages. A manuscript describing the systematic study of the physiological role of the zebrafish fibrillin genes is currently under preparation and will be submitted for publication soon.
The second major objective was to use the new zebrafish MFS model to test the functional consequences of specific defects identified in human FBN1 genes during genetic testing of patients. The implementation of this aspect was hampered on one hand due to the delays experienced with the validation of the zebrafish MFS model. On the other hand, specific experiments planned to realign the proposed strategy with the new validated zebrafish MFS model unfortunately suffered severe disruptions due to the Covid-19 emergency situation which arose in the final quarter of the MSCA project. Nevertheless, efforts have been launched in order to achieve this objective to test the effects of human fibrillin-1 gene in our zebrafish models.
The third objective was to generate a new screenable zebrafish MFS model. This can be used on one hand to predict the effects of a number of MFS drugs on patients with different FBN1 gene defects, and on the other hand to discover new potential treatments by testing a large number of pharmaceutical molecules. During the project period we were able to successfully confirm that our proposed approach using a transgenic zebrafish line in combination with our new MFS model is feasible for evaluating the effects of chemical molecules which might have an effect on MFS symptoms. We have already tested a number of chemicals using this approach, and have some encouraging results which will be followed up on in future projects.
Besides the research aspect, within the framework of WP7 I have completed multiple courses on transferable skills which have improved my career prospects. The work performed in this project has also been disseminated (WP8) by posters and oral presentations on national and international conferences. During the course of the project I have co-authored 6 peer-reviewed publications in scientific journals, describing the technological and research progress made in our research group. Several publications relate to parallel research on preclinical models of MFS performed in our research group, which I supervised within the framework of increasing my scientific leadership skills and transition to academic independence. The action activities were also communicated outside of the common scientific channels (WP9), by the organization of a presentation and demonstration for the general public. I have also participated in events geared toward coaching and guiding new prospective MSCA candidates.
As a first objective, we wanted to validate a zebrafish model for MFS which can be used for functional genetic testing as well as for drug discovery. Despite initial setbacks, which showed that the zebrafish fibrillin genes do not behave exactly like the three human fibrillin genes, we have been able to generate a novel zebrafish model which recapitulates several relevant characteristics of the human disease. During the generation of this new model, we learned more about the role of the different zebrafish fibrillins in the cardiovascular system both during development as well as in adult stages. A manuscript describing the systematic study of the physiological role of the zebrafish fibrillin genes is currently under preparation and will be submitted for publication soon.
The second major objective was to use the new zebrafish MFS model to test the functional consequences of specific defects identified in human FBN1 genes during genetic testing of patients. The implementation of this aspect was hampered on one hand due to the delays experienced with the validation of the zebrafish MFS model. On the other hand, specific experiments planned to realign the proposed strategy with the new validated zebrafish MFS model unfortunately suffered severe disruptions due to the Covid-19 emergency situation which arose in the final quarter of the MSCA project. Nevertheless, efforts have been launched in order to achieve this objective to test the effects of human fibrillin-1 gene in our zebrafish models.
The third objective was to generate a new screenable zebrafish MFS model. This can be used on one hand to predict the effects of a number of MFS drugs on patients with different FBN1 gene defects, and on the other hand to discover new potential treatments by testing a large number of pharmaceutical molecules. During the project period we were able to successfully confirm that our proposed approach using a transgenic zebrafish line in combination with our new MFS model is feasible for evaluating the effects of chemical molecules which might have an effect on MFS symptoms. We have already tested a number of chemicals using this approach, and have some encouraging results which will be followed up on in future projects.
Besides the research aspect, within the framework of WP7 I have completed multiple courses on transferable skills which have improved my career prospects. The work performed in this project has also been disseminated (WP8) by posters and oral presentations on national and international conferences. During the course of the project I have co-authored 6 peer-reviewed publications in scientific journals, describing the technological and research progress made in our research group. Several publications relate to parallel research on preclinical models of MFS performed in our research group, which I supervised within the framework of increasing my scientific leadership skills and transition to academic independence. The action activities were also communicated outside of the common scientific channels (WP9), by the organization of a presentation and demonstration for the general public. I have also participated in events geared toward coaching and guiding new prospective MSCA candidates.
The research activities performed in this project have led to the development of the very first validated zebrafish model of MFS. This represents a major advance in the field since this will allow us as well as other scientists to study the mechanisms driving the disease processes in MFS from a completely new point of view. This model also enables the testing of genetic and pharmacologic modifiers on an unprecedented scale. In the long term it can be expected that the results obtained using this model might have an impact in the broader field of cardiovascular and/or heritable connective tissue disease, since disease mechanisms and modulators of MFS might also be relevant for other, more common disorders.
Another important output of the research efforts in this project lies in the technological advances that were made, enabling improved resolution and throughput of zebrafish cardiovascular phenotyping. The newly developed techniques will be applicable for other zebrafish models of disease and will therefore have an impact beyond the field of MFS research.
Another important output of the research efforts in this project lies in the technological advances that were made, enabling improved resolution and throughput of zebrafish cardiovascular phenotyping. The newly developed techniques will be applicable for other zebrafish models of disease and will therefore have an impact beyond the field of MFS research.