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Imaging life from Molecules to cells - building knowledge on Cryo-electron microscopy methodologies

Periodic Reporting for period 1 - IMpaCT (Imaging life from Molecules to cells - building knowledge on Cryo-electron microscopy methodologies)

Reporting period: 2019-09-01 to 2020-11-30

Proteins are building blocks of life and they often form complexes with essential roles in the cell. Their structure and regulation allow for robust molecular homeostatic mechanisms. Therefore, access and expertise to modern methodologies in structural biology is essential for the study of life itself.
Dysfunction of proteins or their complexes are associated with a wide variety of pathologies, making them excellent drug target candidates. Many pharmaceutical drug discovery and development pipelines are based on the structural characterization of the components and their complex. Thus, knowledge of their 3D structures is essential for a rational drug design.
Extraordinary advances in instrumentation, sample preparation and image processing methods developed in recent years have brought forward the potential of direct imaging of individual molecules using Cryo-Electron Microscopy (CryoEM). With modern high-end instruments, structures of a wide range of macromolecular complexes can be determined at near-atomic resolution. In addition, an unprecedented level of detail in cell imaging can be obtained using cell tomography.
In order to bring the knowledge on CryoEM at UNL - ITQB NOVA, and consequently to Portugal, this project has established collaborations with three research Centres in Europe, with proven track record and expertise in the different aspects and applications of CryoEM: Spanish National Research Council - National Centre for Biotechnology (CSIC, Spain), University of Helsinki - Institute of Biotechnology (UH, Finland) and Weizmann Institute of Science (WIS, Israel). All with well-equipped CryoEM instruments and highly trained technical staff.
Through IMpaCT, several activities are planned, in which the collaborations will take the form of a network of close interactions between researchers from UNL, CSIC, UH and WIS. The ultimately goal will be the knowledge transfer in CryoEM methodologies from the partners to UNL researchers, allowing them to become independent and capable end-user of this methodology. Thus, IMpaCT will be a fundamental instrument to develop the applications of CryoEM to life sciences research at UNL and assist the establishment of an electron microscopy facility for life sciences in Portugal.
In the first 18 months of this project, several important activities have been developed to meet the proposed goals, despite the forced limitations caused by the pandemic outbreak. The Consortium has been actively seeking to put into action several mitigation actions to circumvent this unforeseen situation and some actions were prioritized to be organized as they will contribute to important achievements in the project. As such, important achievements and milestones were accomplished towards the goals of boosting the knowledge in the CryoEM methodologies within the UNL Early Researchers as well as in promoting network building in this field with relevant international experts.
An important goal is to provide a high-level training of our team members, in particular the UNL Early Career Researchers, to enable them to become skilled end-users of the CryoEM methodologies. In line with this, training program composed by 3 hands-on workshops was planned. From those, one was completed fully onsite, while the other two were partially completed in a virtual setting. Despite the success, the hands-on training for the two virtual workshops will be prioritized with organization of practical sessions once the travel restrictions are alleviated. Furthermore, 1 of the Early Career Exchange visits was successfully completed, with a 3-month exchange visit from a PhD student from the UNL to the UH partner. During this period, the PhD student had the chance to develop a close collaboration with the UH experts team members to further develop her skills and knowledge in the area of Single Particle Analysis by CryoEM.
Network building is another important goal within this project, as it will assist to place the UNL teams in the CryoEM roadmap, with clear benefits to the Structural and Cell Biology scientific communities in Portugal. From the several activities developed, we underline the organization of seminars with renowned international experts, dissemination activities targeting both the scientific community and lay audiences, and participation at international conferences. The adjustment to run several of our events in a virtual format has allowed to welcome the participation of a larger and more diverse scientific audience.
With these actions we aim to positively contribute to broadening the career options of early career researchers and other science-related staff, whereas the expected increase in new collaborative projects will create new job opportunities. Also, this will contribute to set the Portuguese Researchers on the CryoEM roadmap and assist the implementation of relevant facilities and capacities in Portugal.
There are important outcomes to emphasize from the network building promoted by IMpaCT, such as the IMpaCT Coordinators invitation to be part of the Iberian ESRF CryoEM “Beam” Allocation Group, in connection with the Cryo-EM@PT Network, and the invitation to one of the IMpaCT Coordinators to become a member of the Scientific Committee of the SPMICROS – Sept 2021.
The project started with a Portuguese Research community with a strong trajectory and background in X-rays crystallography, but with an almost non-existing one in CryoEM. Scientific projects, therefore, did not benefit from an integrated structural biology approach in which CryoEM is nowadays considered essential. This could be considered the “state of the start” at the onset of this special Twinning IMpaCT project.
It has been a very difficult period for all activities requiring personal interactions, however the project has been able to accommodate the new “remote operation” mode in all its training activities, extending its scientific network much beyond initial expectations, profiting from the enhanced availability of researchers to attend online events without the need to travel. In this way the Portuguese scientific community has benefitted from many more interactions than initially planned, a fact that will very positively impact the way scientific projects are conducted and designed, promoting international cooperation. By the end of the project we expect this influence will be cemented in new scientific collaborations, between partners and new institutions, though the ability to participate in experiments using additional cutting-edge instrumentation offered through Instruct-ERIC and ESRF.
More importantly, in the long term we expect this Twinning project will assist the purchase, set-up and development of the first CryoEM infrastructure in Portugal for biological research. We aim to promote the construction of fruitful and sustainable networks that will assist the establishment of national and international collaborations and the capacity to attract pertinent funding opportunities. Also, the project aims to contribute to a crucially and essential point which is the empowerment of the Early Career Researchers, in particular from the UNL, to become capable and autonomous end-users of this technology. It is essential to secure high level technical capacity to efficiently benefit from a future Portuguese CryoEM facility and to secure the position of the Portuguese Scientific community in the CryoEM roadmap.
IMpaCT team members participation at international scientific conferences
IMpaCT team members and the SAB members at the Project's Kick off meeting, Sept. 15-17, 2019
IMpaCT team members participation in several outreach events