Periodic Reporting for period 2 - 4DHeart (4D analysis of heart development and regeneration using advanced light microscopy) Reporting period: 2019-01-01 to 2021-02-28 Summary of the context and overall objectives of the project Cardiovascular disease is a main cause of death in Europe (WHO factsheet Nº310, updated May 2017). An estimated 17.9 million people died from CVDs in 2016, representing 31% of all global deaths. During adulthood, coronary artery diseases cause myocardial infarction ultimately causing heart failure and being the leading cause of death worldwide. Congenital heart defects (CHD) are found in over 20% of perinatal deaths and severe CHD is found in approximately 3.0 per 1,000 live born children. Thus, there is need for research for better early disease diagnosis and the development of therapeutic strategies. In vivo imaging and the possibility to evaluate cellular behaviour in a developing, regenerating, living and beating heart is an invaluable tool to investigate the biological principles governing the cardiovascular system and those that impact cardiovascular health. This EID training network strengthens on-going European academia-industry partnerships and provided innovative training to Early Stage Researchers (ESRs) at the intersection of the fields of biology, microscopy, optics and bio-computing. Through this consortium, we have been able to improve our knowledge on how a heart is formed and is able to regenerate upon injury.The central focus of 4DHeart was to train experts on cardiovascular research through the use of live imaging. Light sheet Microscopy, high-throughput fluorescent light microscopy platforms, as well as magnetic resonance imaging (MRI) were used in combination with newly developed image processing and image quantification pipelines to decipher how a vertebrate heart is formed at the cellular level and monitor alterations of cardiac function with high accuracy. The supervisors of the training program were academic (CNIC, UBERN, IGBMC, CNRS) and industrial (Leica, BITPLANE; ACQUIFER/Ditabis) partners, experts on cardiovascular development, physics, bio-computing, optics, image analysis, screening platforms and big data handling. Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far 6 Early Stage Researchers (ESRs) from Portugal, Spain, Italy and Rumania were employed within this multidisciplinary and intersectoral network. They spent 50% of their PhD contract working in an academic environment and 50% at the premises of the industrial partners in Spain, Germany, France and/or Switzerland. They participated in various common training programs but also received tailored mentoring and training opportunities for their individual projects, that were highly complementary to each other. Through the exposure to this diverse work environments, the participation in a coherent network of academic institutions and enterprises, the diverse and purposely sought after soft-skill guided training transformed the trainees into capable, creative, entrepreneurial, and innovative researchers. They received a training that could allow then pursue an academic career and become future leaders in the field of cardiovascular imaging.The overarching goal of 4DHeart was to use live imaging to gain insight into cardiac development in preclinical animal models (zebrafish, mouse and rabbit). Each ESR project was dealing with a specific project that had two main challenges: on the one hand we aimed to answer a meaningful biological question and on the other, to pioneer in novel imaging tools. The implementation has been done by achieving the following objectives:Objective 1. Molecular control of the role of fluid flow forces on valve development. Objective 2. Quantitative description of the role of fluid forces on epicardium development. Objective 3. Comprehensive description of zebrafish valve development at cellular resolution. Objective 4. Comprehensive description of early mouse heart development at cellular resolution. Objective 5. Identification of compounds leading to epicardium activation. Objective 6. Generation of novel MRI-based algorithms to track dynamic tissue changes in vivo. Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far) The skills and experience ESRs accumulated with 4DHeart will also very attractive to the job market in other fields of research and industry, in particular the field of imaging processing and microscope manufacturers. Objective 6. Generation of novel MRI-based algorithms to track dynamic tissue changes in vivo. Objective 3. Comprehensive description of zebrafish valve development at cellular resolution. Objective 4. Comprehensive description of early mouse heart development at cellular resolution. Objective 1. Molecular control of the role of fluid flow forces on valve development. Objective 5. Identification of compounds leading to epicardium activation. Objective 2. Quantitative description of the role of fluid forces on epicardium development.
