Funding: European Regional Development Fund (ERDF) “On Implementation of Activity 18.104.22.168 “Post-doctoral Research Aid” of the Specific Aid Objective 1.1.1 “To increase the research and innovative capacity of scientific institutions of Latvia and the ability to attract external financing, investing in human resources and infrastructure” of the Operational Programme “Growth and Employment”
Project Title: Evaluation the potential of virus-like particles carrying carbonic anhydrase IX as anti-cancer vaccine.
Project No.: 22.214.171.124/VIAA/4/20/728
Period: 1 January 2021 – 30 June 2023
Project costs: 111 505.00 EUR
Project implementer: Dr.biol. Zane Kalniņa
The aim of the project is to elaborate a VLP-based anti-carbonic anhydrase IX (CAIX) cancer vaccine candidate and evaluate its efficacy in a mouse model of breast cancer. CAIX is a membrane protein rarely found on the surface of normal cells, but frequently overexpressed in hypoxic tumours, which makes it an attractive tumour-specific target. Cell surface expression of CAIX allows to target it with CAIX-specific antibodies, leading to the antibody-dependent cellular cytotoxicity. Although several monoclonal anti-CAIX antibodies have been developed, they have proven to have limited efficacy in late clinical trials. Nevertheless, it is possible to induce anti-CAIX antibody production in the autologous host itself by overcoming the B cell tolerance against this self-antigen. To achieve this, we plan to use ssRNA bacteriophage virus-like particles (VLPs) with coupled mouse CAIX protein on their surfaces and use them as vaccines, thus overcoming the threshold of antigen-specific B cell activation.
Information published 04.01.2021.
1 January 2021 – 31 March 2021
At this stage of the project implementation, the latest scientific literature was studied, and a detailed experimental plan of the project was developed, which was presented to the project’s scientific consultant.
The project has started according to the planned schedule – work has been carried out on the implementation of WP1 and WP4 of the project. Within WP1, an experimental work has been started on the development and characterization of genetically modified mouse 4T1 breast cancer cell lines with the aim of establishing two cancer cell lines – one that consistently expresses carbonic anhydrase 9 (CAIX) and one in which this gene is inactivated.
Within WP4, a CPD training on the topic “CRISPR: Revolutionizing genome editing advanced certificate program” took place and a certificate of successful completion of the training course was obtained.
Information published 31.03.2021.
1 April 2021 – 30 June 2021
At this stage of the project implementation, the latest scientific literature was studied with the aim of proper planning of the expected in vivo experiments. The work on the project proposal preparation for the submission for local animal ethics committee has been initiated in order to receive a permit to perform the planned in vivo experiments.
The project has been carried out according to the planned schedule – mainly on the implementation of WP1 of the project. Within WP1, an experimental work has been continued on the characterization of the genetically modified mouse 4T1 breast cancer cell lines at protein level.
Information published 30.06.2021.
1 July 2021 – 30 September 2021
At this stage of the project, experimental work was carried out on the implementation of the first activity of the project – genetically modified mouse 4T1 breast cancer cell lines were characterized at the protein level. The preparation of a project application for the permission of the Animal Ethics Committee was continued for the planned work with experimental animals (activities 2 and 3), as well as the planning of experimental work, and student training for the work with cell and molecular biology methods. An experience exchange visit to the Experimental Animal Laboratory of the University of Tartu took place.
Information published 30.09.2021.
1 October 2021 – 31 December 2021
At the given period, work was carried out on the implementation of the first and second activities of the project. During this period, the work on the characterization of genetically modified 4T1 breast cancer cell lines at the protein level was completed. A project application for the permission of the Animal Protection Ethics Committee for the planned work with experimental animals was submitted, and an ethical permit for the planned experiments was received. Detailed protocols have been elaborated for the development of a triple-negative breast cancer model system in mice and for the vaccination of animals with VLP-mCAIX vaccine. Work was continued on the training of two students on cell and molecular biology methods.
Information published 30.12.2021.
1 |January 2022 – 31 March 2022
At the given period, work was carried out on the implementation of the first and second activities of the project. During this period, experimental work with laboratory animals was initiated and successfully completed – two experimental procedures were performed. First, the developed VLP-mCAIX vaccine in mice was shown to be successful in eliciting high-titer antibody responses against mouse CAIX protein. Second, an in vivo model of triple-negative breast cancer was successfully established – the 4T1 breast cancer cell lines developed during the previous project periods were implanted in the mammary fat pad of mice, after which the tumor growth rate was determined and compared between study groups. Work was continued on the training of two students on cell and molecular biology methods as well as on the work with experimental animals.
Information published 31.03.2022.
1 April 2022 – 30 June 2022
During this period, the nature of the antibody response elicited by the VLP-mCAIX vaccine was fully evaluated, and the developed VLP-mCAIX vaccine in mice was shown to elicit antibody responses with anti-cancer activity. Work on the treatment of tissue samples from the experimental animals has been completed and comprehensive histological analysis has begun. Training of two students to work with the methods of histology and molecular biology was continued; one of the students successfully defended the course work thesis on the evaluation of the antibody response elicited by the VLP-mCAIX vaccine in the mouse model. The results of this research project were presented in an oral report at the international scientific conference FEBS3+, which took place in Tallinn, Estonia on June 15-17. The preparation of a manuscript of a scientific publication on the development of a model system for CAIX research has been initiated.
Information published 30.06.2022.
1 July 2022 – 30 September 2022
During July and August no experimental work was pursued due to the official vacation. In September, during mobility visit, I attended Porto University where I participated in the training seminar and networking event “Train the trainers – Workshop on Teaching Experimental Design for Animal Experiments” organized by the FELASA Experimental Design Working Group. The preparation of a scientific publication manuscript on the development of a model system for CAIX research was continued. In parallel, the work on the planning of the experimental work, as well as the histological analysis of the lung tissues of experimental animals obtained during the previous project period was performed, which also included student training.
Information published 30.09.2022.
Progress of the project
1 October 2022 – 31 December 2022
During this time period, work was carried out on the implementation of project activities 1, 3 and 4. Several mobility and networking events took place in October and November – as part of the mobility, the Life Sciences Center of Vilnius University was visited, where experiments were conducted with experimental animals. Besides, I also participated in the management committee meeting of the COST action IMPROVE in Brussels, Belgium. Work continued on the preparation of a manuscript for a scientific publication on the creation of a model system for CAIX research. In parallel, the experimental work with laboratory animals was going on – an in vivo experiment was initiated and successfully completed, in which the developed VLP-mCAIX vaccine was tested in the elaborated triple-negative breast cancer model in mice. Starting from December 8, no experimental work was pursued due to the official vacation.
Information published 30.12.2022.