The project is dedicated to the study of colorectal cancer (CRC) factors, namely the study of the relationship of Fusobacterium nucleatum with tumor cells, changes in the protein composition of the affected colon tissue and changes in the tumor microenvironment. The disease colorectal cancer remains relevant worldwide due to its high mortality and prevalence. The etiology of colorectal cancer is multifaceted and is related to a person’s lifestyle, habits, and genetics. Among the risk factors contributing to carcinogenesis, infection with Fusobacterium nucleatum is of scientific interest because according to the research results, it is actively involved in the process of tumor growth and development. At the same time, a number of bacteria associated with cancer are also involved in the formation of a favorable microenvironment for tumor development. When malignancy of the colon tissue changes its protein composition, the study of this aspect may reveal any patterns and lead to interesting conclusions. The study of CRC factors is specific by geography and ethnic groups, which can affect the prevalence and prognosis of the disease.
RNA sequencing (RNAseq) is the most effective tool in studying the transcriptomic profile of pathogenic microorganisms involved in the development of colorectal cancer, as it reveals differential patterns of gene expression. The object of the study will be a clinical strain of Fusobacterium nucleatum isolated from a Kazakhstani patient with CRC, with the complete genome we studied earlier.
The study of the transcriptomic profile of Fusobacterium nucleatum modulated by colorectal cancer cells, the study of the proteomic profile of colon tissue and the abundance of bacteria associated with colorectal cancer.
As a result of this project, the transcriptomic profile of a clinical Fusobacterium nucleatum strain from a Kazakhstani patient modulated by colorectal cancer cells will be studied for the first time, as well as the proteomic profile of colon tissues characterizing precancerous conditions of the body and the abundance of bacteria associated with colorectal cancer. Thus, the data obtained will make it possible to understand the behavior of the bacterium when interacting with host cells; it will be established which genes are F. nucleatum are activated for invasion of host cells, which will provide insight into the fundamentals of the molecular process, and these results can be used to develop treatments for infection. The quantitative and qualitative characteristics in the study will confirm the validity and significance of the results.
Gulmira Nigmetzhanovna Kulmambetova, PhD, Project Leader – Leading Researcher – National Scientific Laboratory of Biotechnology for Collective Use, National Center of Biotechnology, (Astana, Kazakhstan). The field of scientific activity of G.N. Kulmambetova is biotechnology in medicine. The research work focuses on the application of methods of biotechnology, molecular biology, genetics, microbiology, and statistics for the analysis and interpretation of genetic and statistical data (logical, vectors, dataframe, variables, polymorphisms, mutations, sequences), as well as on the development of new methods and approaches for their analysis. According to the Web of Science, Kulmambetova G.N. has a Hirsch index (h-index) equal to 3 (https://www.scopus.com/authid/detail.uri?authorId=56387533800 ).
The research team of the project, including the project leader, is based on young researchers from the National Center for Biotechnology:
Researcher 1 – Kurentay Botakoz Amandoskyzy – Master of Biotechnology, laboratory assistant at the National Scientific Laboratory of Biotechnology for Collective Use, NCB. Graduated from L.N.Gumilyov Eurasian National University. She is proficient in nucleic acid extraction methods, PCR, real-time PCR, and Sanger sequencing.
Researcher 2 – Gusmaulemova Alua Daurenkyzy – Master’s degree in Virology, laboratory assistant at the National Scientific Laboratory of Biotechnology for Collective Use, NCB. Graduated from Al-Farabi Kazakh National University. He is proficient in nucleic acid extraction methods, PCR, real-time PCR, and Sanger sequencing.
Researcher 3 – Venera Tulegenovna Kumasheva, Master of Science at the Eurasian National University, Researcher at the National Research Center. In 2008, she received a bachelor’s degree in Biotechnology from the Eurasian National University. In 2010, she received a master’s degree in Biotechnology. From 2013 to 2014, she completed a scientific internship in flow cytometry at the Korea Research Institute of Bioscience and Biotechnology under the supervision of Professor Inpyo Choi (Daejeon, South Korea). He has 15 years of experience in cell and tissue engineering technologies. Employment in the project is 0.5.
2024
At this stage of the project, Fusobacterium nucleatum strains with the studied genome-wide sequence were cultivated. The cultivation of Fusobacterium nucleatum was carried out on a selective medium under anaerobic conditions. The conditions for in vitro cultivation of a colorectal cancer cell line with Fusobacterium nucleatum were optimized. A model of invasion of a cell line by a bacterium was tested. The prevalence of bacteria associated with colorectal cancer was also studied. A significant frequency distribution of F. nucleatum was found in the tumor and normal tissues of patients with CRC (P < 0.05). Statistical analysis using cycle threshold (Ct) values from qPCR data and clinical characteristics showed that tumor size, location, and consumption of processed meat significantly correlated with the prevalence of F. nucleatum (P < 0.05). The prevalence of B. fragilis and E. coli in tumor tissues was slightly higher than in normal tissues (P < 0.1), and consumption of processed/red meat influenced the prevalence of these bacteria (P < 0.05).