Periodic Reporting for period 2 - V.A. Cure (A multidisciplinary approach towards sustainable improvement in rare diseases care uniting Europe's top class vascular research to find new treatment strategies for vascular anomalies)
Okres sprawozdawczy: 2021-03-01 do 2023-02-28
Vascular anomalies (VAs) are a group of rare diseases defined by blood- or lymph vessel dysfunction causing chronic pain, disabilities, and even sudden death. Effective therapeutic treatments are lacking and many patients require life-long clinical management by multidisciplinary medical teams. Management is often partial, leaving patients with chronic pain, dysfunction, and lowered quality of life. The inherent challenge in the field is the scarcity of clinical knowledge and expertise due to the limited number of patients.
The V.A. Cure network (7 academic institutions, 2 companies, and supported by 9 partner organisations across Europe) aimed to uncover core mechanisms of disease initiation and maintenance and leverage this information for establishing novel therapeutic strategies for VAs.
14 ESRs were working on the project’s objectives:
(1) identification of novel genes involved in VAs in patients,
(2) dissection of molecular mechanisms behind the diseases by in vitro modelling,
(3) in-depth analysis of tissular mechanisms in pathophysiological conditions through in vivo models,
(4) pre-clinical testing of identified treatment strategies.
The consortium has made major advances in the fundamental understanding of vascular diseases, which provide leads for the development of new therapies for multiple VAs. In addition, new techniques and models were developed that will be of great value for future vascular research, into vascular anomalies and other vascular diseases.
The 14 ESRs have successfully performed their individual projects. With the acquired biological knowledge, technical and soft skills and the network built up during V.A.Cure they will be able to continue their career in the path of their choice.
Furthermore, new collaborations in the project have proven fruitful and many of them will be continued after the project. Collaborations outside the network have also been initiated. The project has thus strengthened the vascular research field in Europe, both in terms of scientific quality and international collaboration.
The V.A. Cure network (7 academic institutions, 2 companies, and supported by 9 partner organisations across Europe) aimed to uncover core mechanisms of disease initiation and maintenance and leverage this information for establishing novel therapeutic strategies for VAs.
14 ESRs were working on the project’s objectives:
(1) identification of novel genes involved in VAs in patients,
(2) dissection of molecular mechanisms behind the diseases by in vitro modelling,
(3) in-depth analysis of tissular mechanisms in pathophysiological conditions through in vivo models,
(4) pre-clinical testing of identified treatment strategies.
The consortium has made major advances in the fundamental understanding of vascular diseases, which provide leads for the development of new therapies for multiple VAs. In addition, new techniques and models were developed that will be of great value for future vascular research, into vascular anomalies and other vascular diseases.
The 14 ESRs have successfully performed their individual projects. With the acquired biological knowledge, technical and soft skills and the network built up during V.A.Cure they will be able to continue their career in the path of their choice.
Furthermore, new collaborations in the project have proven fruitful and many of them will be continued after the project. Collaborations outside the network have also been initiated. The project has thus strengthened the vascular research field in Europe, both in terms of scientific quality and international collaboration.
We identified novel disease-causing genes in patients affected by CMDV (Capillary Malformation with Dilated Veins) and lymphoedema. The generated single cell transcriptomic data led to the discovery of a new mechanism in Lymphatic Malformations, involving paracrine cross-talk between endothelial and immune cells.
Multiple cellular models of various vascular anomalies were generated and validated. Ex vivo and in vitro 2D and 3D models were made with newly identified and known VA causative mutations. Cellular models were also made starting from iPS cells (induced pluripotent stem cells), for which a protocol was developed for differentiation to endothelial cells. The models have been used to investigate cellular and molecular mechanisms in the vascular lesion formation.
A tailor-made VA-on-a-chip microfluidic culture system was designed and fabricated, allowing for real time control of critical parameters (eg flow and shear stress). The VA-on-a-chip has been further developed to be used with multiple cellular models in collaboration with other partners.
In vivo models (mouse and zebrafish) were generated and validated with newly identified and known VA causative mutations. These were used for functional and molecular studies and they also enable pre-clinical testing of drugs. In addition, new techniques, protocols and analysis tools were developed to investigate the models.
The newly developed models were used to identify and test potential therapies for VAs. The newly identified mechanism in LM was shown to be a new therapeutic opportunity. Drugs already in use for some VAs, were found to be effective on cellular models of other VAs. Screening and testing of other approved drugs on iPS-derived ECs also gave promising results. The discovery of the role of a polycomb complex in CCM (cerebral cavernous malformation) provides a novel promising target for pharmacological intervention. Also in CCM, the misregulation of a signalling pathway was discovered and an inhibitor of this pathway was shown to have a curative effect in an in vivo model.
The screening and testing of cell penetrating peptides (CPPs) to enhance tissue specific targeting of therapeutic modalities proved challenging, but good progress has been made and the work is continued.
The results of the project have been broadly disseminated towards the relevant international fields. 9 scientific papers have so far been published,33 other publications are in preparation. ESRs also held oral and poster presentations at scientific conferences.
The network organised a big 4-day international conference (VAC2023), attended by 220 people from 25 countries. All ESRs of the project presented their work at the conference. The work and results of the project were also communicated to the general public through a website and social media. A project video was made, as well a an “elevator pitch” (short video) by each ESR. Most results of the project are or will be published in journals with open access, to the benefit of the scientific community. Several datasets are put in open repositories.
Finnadvance is currently commercialising the vasculature-on-chip platform as a tool for vascular model development. The ESR at Finnadvance has filed 24 patents on novel technologies related to micro fluidic chip development (one already granted).
Multiple cellular models of various vascular anomalies were generated and validated. Ex vivo and in vitro 2D and 3D models were made with newly identified and known VA causative mutations. Cellular models were also made starting from iPS cells (induced pluripotent stem cells), for which a protocol was developed for differentiation to endothelial cells. The models have been used to investigate cellular and molecular mechanisms in the vascular lesion formation.
A tailor-made VA-on-a-chip microfluidic culture system was designed and fabricated, allowing for real time control of critical parameters (eg flow and shear stress). The VA-on-a-chip has been further developed to be used with multiple cellular models in collaboration with other partners.
In vivo models (mouse and zebrafish) were generated and validated with newly identified and known VA causative mutations. These were used for functional and molecular studies and they also enable pre-clinical testing of drugs. In addition, new techniques, protocols and analysis tools were developed to investigate the models.
The newly developed models were used to identify and test potential therapies for VAs. The newly identified mechanism in LM was shown to be a new therapeutic opportunity. Drugs already in use for some VAs, were found to be effective on cellular models of other VAs. Screening and testing of other approved drugs on iPS-derived ECs also gave promising results. The discovery of the role of a polycomb complex in CCM (cerebral cavernous malformation) provides a novel promising target for pharmacological intervention. Also in CCM, the misregulation of a signalling pathway was discovered and an inhibitor of this pathway was shown to have a curative effect in an in vivo model.
The screening and testing of cell penetrating peptides (CPPs) to enhance tissue specific targeting of therapeutic modalities proved challenging, but good progress has been made and the work is continued.
The results of the project have been broadly disseminated towards the relevant international fields. 9 scientific papers have so far been published,33 other publications are in preparation. ESRs also held oral and poster presentations at scientific conferences.
The network organised a big 4-day international conference (VAC2023), attended by 220 people from 25 countries. All ESRs of the project presented their work at the conference. The work and results of the project were also communicated to the general public through a website and social media. A project video was made, as well a an “elevator pitch” (short video) by each ESR. Most results of the project are or will be published in journals with open access, to the benefit of the scientific community. Several datasets are put in open repositories.
Finnadvance is currently commercialising the vasculature-on-chip platform as a tool for vascular model development. The ESR at Finnadvance has filed 24 patents on novel technologies related to micro fluidic chip development (one already granted).
Several new targets for treatments of VAs were identified. These targets will be further investigated to validate the effectiveness. Also, some drugs already in use for other VAs or other diseases, were found to be effective in cellular VA models. These will be validated in vivo. If successful, the route to clinical trials can be short, because the drugs are already approved for use in humans.
The consortium has used cutting-edge technologies to generate relevant new models of multiple VAs: Ex vivo cell lines, in vitro 2D and 3D cellular models, iPS-EC lines and in vivo models (mouse and zebrafish). These models have been and will be used for mechanistic studies and to test novel therapeutic drugs. The iPS-EC lines offer a new and alternative cell source for in vitro assay development and characterization of genes involved in VAs.
Innovative technical progress has been made in the field of imaging. New methods and protocols were developed for intravital multi-photon imaging, visualizing protein interactions in 3D tissue, high-resolution endothelial cell morphometrics and imaging of eyes by light-sheet fluorescence microscopy. These tools are valuable for the vascular research field.
The identification and testing of novel endothelium specific targeting peptides has made great progress and the work will be continued. These CPPs have great potential to increase the efficiency of drugs while reducing the side effects.
The VA-on-a-chip is a unique tool for vascular model development. It can be used for mechanistic studies and for testing of potential drugs and offers the possibility to reduce animal experiments. Finnadvance is commercialising the platform and extending the methodologies to other disease areas.
14 ESRs have worked on the project. One obtained a PhD, the others are on track to do so in the coming year. Besides scientific knowledge and skills, they acquired many soft skills (collaboration, presentation, organising and chairing meetings, writing,..), and they built an international network.
The consortium has used cutting-edge technologies to generate relevant new models of multiple VAs: Ex vivo cell lines, in vitro 2D and 3D cellular models, iPS-EC lines and in vivo models (mouse and zebrafish). These models have been and will be used for mechanistic studies and to test novel therapeutic drugs. The iPS-EC lines offer a new and alternative cell source for in vitro assay development and characterization of genes involved in VAs.
Innovative technical progress has been made in the field of imaging. New methods and protocols were developed for intravital multi-photon imaging, visualizing protein interactions in 3D tissue, high-resolution endothelial cell morphometrics and imaging of eyes by light-sheet fluorescence microscopy. These tools are valuable for the vascular research field.
The identification and testing of novel endothelium specific targeting peptides has made great progress and the work will be continued. These CPPs have great potential to increase the efficiency of drugs while reducing the side effects.
The VA-on-a-chip is a unique tool for vascular model development. It can be used for mechanistic studies and for testing of potential drugs and offers the possibility to reduce animal experiments. Finnadvance is commercialising the platform and extending the methodologies to other disease areas.
14 ESRs have worked on the project. One obtained a PhD, the others are on track to do so in the coming year. Besides scientific knowledge and skills, they acquired many soft skills (collaboration, presentation, organising and chairing meetings, writing,..), and they built an international network.