Cryo-CLEM in situ imaging of adenovirus stepwise disassembly

Project Information

CryoCLEMAV

Grant agreement ID: 101107060

DOI

10.3030/101107060

Project closed

EC signature date 29 March 2023

Start date 16 January 2024

End date 15 January 2026

Funded under

• Marie Skłodowska-Curie Actions (MSCA)

Total cost

No data

EU contribution

€ 165 312,96

Coordinated by

AGENCIA ESTATAL CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS
Spain

Periodic Reporting for period 1 - CryoCLEMAV (Cryo-CLEM in situ imaging of adenovirus stepwise disassembly)

Reporting period: 2024-01-16 to 2026-01-15

Summary of the context and overall objectives of the project

Adenoviruses (AdVs) are very contagious viruses widespread in vertebrated. Human AdVs infections mainly affect children and immune-supressed people. On the other hand, AdV-derived vectors, nanocarriers and vaccines have turn out to be efficient therapeutic tools. Understanding AdVs disassembly mechanisms is vital to advance research in antiviral drugs, gene therapies, nanocarriers and vaccines. AdV particles’ metastability is key for surviving environmental conditions and cell defences, but at the same time, these particles should disassemble at the appropriate time and place during virus entry. The particle size and the speed of the process challenges the visualization of these events inside the cell. CryoCLEMAV aims at providing unprecedented structural insight on the process of virus entry and disassembly by combining virology with the cutting-edge microscopy techniques: correlative light and electron microscopy (cryo-CLEM), cryo-electron tomography (cryo-ET), FIB-milling and subtomogram averaging. This approach will allow to recapitulate the process with a previously unreachable resolution and specificity in a near native context of the host cell. CryoCLEMAV is an ambitious project whose success will be firmly supported by merging my expertise in virology and super resolution microscopy with the host lab knowledge and resources in cryo-EM techniques and adenovirus biology. Outcomes of this project will contribute to the development of several cryo-electron microscopy techniques and to the field of adenovirus biology, also impacting in the expanding field of adenovirus-derived therapeutic tools. All together will advance the research work of my host group, benefit the host organization (CNB-CSIC), and boost my professional career.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

During the course of the fellowship, the project focused on investigating virus–host interactions using advanced imaging approaches. The research combined fluorescence microscopy, super-resolution microscopy, and cryo-electron microscopy to study the mechanisms of viral infection at different spatial scales.
A major part of the work was devoted to characterizing the entry pathway of adenoviruses into host cells. Using advanced fluorescence microscopy and complementary structural approaches, the project investigated the cellular mechanisms involved in viral entry and the intracellular trafficking of viral particles. These studies provided new insights into the molecular steps that govern adenovirus infection and its interaction with host cellular components.
In addition to these biological discoveries, the project contributed to the application of advanced imaging workflows integrating multiple microscopy techniques. These methodological approaches enabled the visualization of virus–host interactions with high spatial resolution and can be applied to study other viral systems and cellular processes.
Overall, the project generated new knowledge on virus–host interactions, particularly regarding adenovirus entry mechanisms. These findings represent an important contribution to the fields of virology, structural biology, and advanced microscopy.

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 project advances the current understanding of virus–host interactions by combining advanced fluorescence microscopy, super-resolution microscopy, and cryo-electron microscopy to investigate viral infection mechanisms at multiple spatial scales.
One of the main contributions of the project is the characterization of the adenovirus entry pathway using complementary imaging approaches. While previous studies have described aspects of adenovirus infection, the integration of advanced microscopy techniques provides a more detailed view of the cellular processes involved in viral entry and intracellular trafficking.
Beyond the biological discoveries, the project also demonstrates the value of integrating multiple imaging techniques to study complex virus–host interactions. The methodological framework applied in the project provides new opportunities for investigating viral infection processes with high spatial resolution.
Further research may extend these approaches to other viral systems and explore the functional consequences of virus-induced genome reorganization. These advances may ultimately contribute to the development of new antiviral strategies and improved viral vector technologies.