Periodic Reporting for period 1 - HIV VCC Interference (Characterisation of single molecule dynamics within HIV-1 reservoirs as a target for interference with virus persistence and immune evasion) Reporting period: 2019-01-01 to 2020-12-31 Summary of the context and overall objectives of the project Human Immunodeficiency Virus type 1 (HIV-1) still remains a significant cause of illness and death around the world. One of the reasons HIV-1 infection is difficult to treat is the virus’s ability to create hidden reservoirs in cells. These virus reservoirs, termed Virus-Containing Compartments (VCCs) are inaccessible to the immune system and help to spread the infection to different parts of the body. Moreover, VCCs are unaffected by current HIV-1 therapies, and thus remain a major obstacle in the development of successful HIV-1 cure strategies. This project has utilized advanced fluorescence microscopy techniques to study the dynamic behaviour of lipid and protein molecules inside VCC membranes of living cells of the immune system infected with HIV-1. This behaviour was then compared to the behaviour of the same molecules on the cell surface. Results of this project have shown that VCCs exhibit a distinct behaviour of lipids inside VCC at the sites of interaction between individual viruses and cell membranes as compared to the rest of the cell membranes. The interference with this unique behaviour may thus be utilised to design a new type of a drug therapy that reduces the ability of immune cells to create virus reservoirs and results have shown that drugs that alter the behaviour of lipid membranes in cells appear to reduce the ability of infected cells to form VCCs. Overall, this project has utilised advanced microscopy techniques to improve the understanding of VCC characteristics and thus provide a possible future avenue for new therapies that target the issue of HIV-1 immune system avoidance and persistence in infected person’s body. Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far The initial work for this project has focussed on establishing the advanced microscopy based technique required for the observation of single molecule dynamics inside VCCs of living cells of the immune system as well as optimising the cell culture and labelling conditions to ensure the acquisition of high quality data. The project has successfully achieved this goal and the platform for these observations was established utilising the equipment available at the collaborator’s laboratory. The project then focussed on the acquisition of the microscopy data for investigation of the behaviour of lipid and protein molecules on VCCs and to compare it with the behaviour of the same molecules at cell surface. The project has successfully acquired data indicating that behaviour of these lipid molecules is very distinct from the rest of the cell. Finally, the project assessed the possibility to exploit the existence to this distinct behaviour to interfere with VCC formation by the use of lipid behaviour modulating drugs. The results have shown that drugs that alter behaviour of lipid membranes in cells appear to reduce the ability of infected cells to form VCCs. While the progress of the project has been negatively impacted by the ongoing SARS-CoV-2 pandemic and associated lockdowns the project has managed to achieve most of its research objectives. The results of this project will be disseminated via scientific publications and conferences and its results may provide a possible future avenue for the design of new therapies that target the issue of HIV-1 persistence in infected person’s bodyAside from scientific goals this project has also focussed on the creation of public engagement opportunities to provide accessible and engaging information on how scientists utilise advanced microscopes to study viruses. To this end the project has created Virtual Reality based interactive educational activity (https://www.youtube.com/watch?v=Q6sdHp1T388) that will be disseminated as a stand-alone product as well as used at future science festivals and open days to provide a novel interactive and education experience. Moreover, the project has conducted a series of webinars and live on line demonstrations on the use of advanced microscopy for virus research as part of the University of Vic Science Week (https://youtu.be/vXdh6SBEUtQ) as well as Catalan Researchers’ Night (part of EU Researchers’ Night) (https://youtu.be/743dvynmhZM). 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 has advanced the state of the art in the field of the field of HIV-1 therapy as its results the possibility of a new lipid behaviour targeting therapeutic approaches which may lead to the reduction of HIV-1 latent reservoirs and thus HIV-1 persistence. Moreover project’s results also represent an important contribution to the state of the art in the field of microscopy as they demonstrate, for the first time, the possibility of the advanced microscopy techniques to study of the dynamic behaviour of single molecules inside compartments of the living cells. The results obtained by this project increase the understanding of the molecular details on formation and maintenance of HIV-1 containing VCCs and may provide a future avenue for new HIV-1 therapies that specifically target the issue of HIV-1 immune system avoidance and persistence in infected person’s body. Overview of the main result of the project
