Periodic Reporting for period 1 - TVISTOFF (Tick-Virus Interactions Shape persistence and Transmission OF Flavivirus pathogen in tick vector)
Reporting period: 2021-09-01 to 2023-08-31
The project entitled “Tick-Virus Interactions Shape persistence and Transmission OF Flavivirus pathogen in tick vector” is concerned with the role of structural protein E of the tick-borne encephalitis virus and tick neurosecretion system in the virus persistence and transmission in tick vector. The research goals suggested the development of novel tools that would be instrumental in studying how the virus persists in ticks and how ticks harbor the virus. Such goals are important due to the lack of information on virus-tick interactions. Moreover, tick-borne encephalitis poses a health risk not only in Slovakia but also in several other European countries. Despite an available and effective vaccination, cases of tick-borne encephalitis have been on the rise and include severe cases with neurodegeneration and up to 2% fatality rate.
So far, most of the research on tick-borne encephalitis has involved a virus-mouse model, whereas virus-tick models have been understudied. Needless to say, it is true that the manipulation of a genetically modified live virus and experimentally infected ticks might be challenging as it is interdisciplinary and constitutes a biohazard, for which many institutions are not equipped. Society is often interested in fundamental questions regarding virus transmission in ticks, such as speed of transmission, and what tick species participate in it and how. Virus-tick interactions have ever been an essential part of virus circulation in nature about which we have had only fragmented information. Therefore, the fellow took advantage of available facilities and combined various interdisciplinary approaches based on his skills and knowledge to identify aspects of interactions between viruses and ticks in a comprehensive way for society.
Objectives of this MSCA have been to (i) construct and characterize a chimeric virus with mutated E protein and a virus carrying a fluorescent reporter gene; (ii) set up and optimize an artificial membrane system for tick feeding (without an animal) to study the speed of virus transfer; (iii) characterize tick biogenic amines, neuropeptide and their receptors in response to tick-borne encephalitis virus. A parallel aim of this MSCA is to set up an innovative research program and to advance the development of the fellow towards independent research leadership.
So far, most of the research on tick-borne encephalitis has involved a virus-mouse model, whereas virus-tick models have been understudied. Needless to say, it is true that the manipulation of a genetically modified live virus and experimentally infected ticks might be challenging as it is interdisciplinary and constitutes a biohazard, for which many institutions are not equipped. Society is often interested in fundamental questions regarding virus transmission in ticks, such as speed of transmission, and what tick species participate in it and how. Virus-tick interactions have ever been an essential part of virus circulation in nature about which we have had only fragmented information. Therefore, the fellow took advantage of available facilities and combined various interdisciplinary approaches based on his skills and knowledge to identify aspects of interactions between viruses and ticks in a comprehensive way for society.
Objectives of this MSCA have been to (i) construct and characterize a chimeric virus with mutated E protein and a virus carrying a fluorescent reporter gene; (ii) set up and optimize an artificial membrane system for tick feeding (without an animal) to study the speed of virus transfer; (iii) characterize tick biogenic amines, neuropeptide and their receptors in response to tick-borne encephalitis virus. A parallel aim of this MSCA is to set up an innovative research program and to advance the development of the fellow towards independent research leadership.
Work was carried out through 4 work packages (WPs). WP1 comprised research studies with efforts to generate reporter and mutated viruses and the fellow delivered 2 conference contributions and 1 journal paper with 2 additional journal manuscripts underway. WP2 aimed to set up and optimize in vitro tick feeding system, a tool to study reporter and mutated viruses without a direct influence of host immunity. The optimization of the system has been problematic and requires further development, which is underway. In WP3, the fellow conducted a bioinformatic search as well as expression studies for neuropeptide and biogenic amines’ receptors that yielded 1 journal paper.
Furthermore, the involvement of the studied molecules in virus transmission has been investigated along with the progressing development of the other versions of reporter viruses. This has laid the foundation for further publications. WP4 focused on the transfer of knowledge and communication of the research. The fellow delivered several public engagement activities to popularize science (Open Lab Day at the partner institution, and 2023 European Researchers’ Night) and gave several interviews about the topic of this MSCA (News media RTVS, Academia journal). The research was communicated at one national and one international conference. As for the transfer of knowledge, the research was discussed and managed regularly with the supervisors, and financial management coordinated with the members of the project division. During the duration of the project, the fellow supervised three undergraduate students, one of whom is currently a PhD student in his group at the host institution.
Furthermore, the fellow provided supervision and mentoring to early-career researchers. During the project, the fellow as a PI received a national grant (VEGA #2/0040/23) with the highest score (98 points out of 100) securing €75,000 to further develop his research. The results of this MSCA project are reported in a paper about how a reporter tick-borne encephalitis virus survives in several biological models including tick cells and live ticks. Another journal paper describes the spatial and temporal expression of the receptor for small neuropeptide F, likely involved in the modulation of viral load in ticks as it will be described in our forthcoming papers. Next journal articles in preparation will report on other reporter virus designs as well as on how specific mutations in viral envelope protein affect virus replication in ticks. In addition to the papers published during this project, the other data generated, yet incomplete, will enhance coming publications in the near future.
Furthermore, the involvement of the studied molecules in virus transmission has been investigated along with the progressing development of the other versions of reporter viruses. This has laid the foundation for further publications. WP4 focused on the transfer of knowledge and communication of the research. The fellow delivered several public engagement activities to popularize science (Open Lab Day at the partner institution, and 2023 European Researchers’ Night) and gave several interviews about the topic of this MSCA (News media RTVS, Academia journal). The research was communicated at one national and one international conference. As for the transfer of knowledge, the research was discussed and managed regularly with the supervisors, and financial management coordinated with the members of the project division. During the duration of the project, the fellow supervised three undergraduate students, one of whom is currently a PhD student in his group at the host institution.
Furthermore, the fellow provided supervision and mentoring to early-career researchers. During the project, the fellow as a PI received a national grant (VEGA #2/0040/23) with the highest score (98 points out of 100) securing €75,000 to further develop his research. The results of this MSCA project are reported in a paper about how a reporter tick-borne encephalitis virus survives in several biological models including tick cells and live ticks. Another journal paper describes the spatial and temporal expression of the receptor for small neuropeptide F, likely involved in the modulation of viral load in ticks as it will be described in our forthcoming papers. Next journal articles in preparation will report on other reporter virus designs as well as on how specific mutations in viral envelope protein affect virus replication in ticks. In addition to the papers published during this project, the other data generated, yet incomplete, will enhance coming publications in the near future.
Knowledge about interactions between tick-borne encephalitis virus and its tick vector has been very limited so far. Therefore, this MSCA pioneered the way reporter viruses could be used as a tool to study interactions between viruses and ticks, showing that the design of a reporter virus is crucial. Also, data on the tick neurosecretion system indicate that some compounds might be responsive to a viral infection in ticks. Although some mechanistic studies will have to be completed, the data have shed light on the roles of tick neurophysiology during the persistence and replication of the virus.
During this project, the fellow developed and mastered several advanced methodologies by which some essential datasets were generated. The fellow and his PhD student are generating novel data about how reporter virus designs affect the functionality of such viruses in live dormant as well as feeding ticks. Such information is instrumental in making this tool work. Besides, working together they are dissecting the mechanistic role of individual candidate neurosecretory compounds in response to the tick-borne encephalitis virus.
Impacts of this MSCA: focus on the production of tools and models to help decipher intricate relationships between viruses and their vectors; potentiate use of such tools for screening antiviral candidates not only in vitro but also in live ticks in vivo; increasing family of compounds to be tested as potential anti-tick vaccines; an overarching impact is an enhanced public perception of the complex relationship between ticks and viruses.
During this project, the fellow developed and mastered several advanced methodologies by which some essential datasets were generated. The fellow and his PhD student are generating novel data about how reporter virus designs affect the functionality of such viruses in live dormant as well as feeding ticks. Such information is instrumental in making this tool work. Besides, working together they are dissecting the mechanistic role of individual candidate neurosecretory compounds in response to the tick-borne encephalitis virus.
Impacts of this MSCA: focus on the production of tools and models to help decipher intricate relationships between viruses and their vectors; potentiate use of such tools for screening antiviral candidates not only in vitro but also in live ticks in vivo; increasing family of compounds to be tested as potential anti-tick vaccines; an overarching impact is an enhanced public perception of the complex relationship between ticks and viruses.