The project is dedicated to an urgent problem in medicine and public health of colorectal cancer, namely the study of the causes of its occurrence and pathogenesis. Colorectal cancer is a multifactorial, oncological disease, the etiology of which is not fully understood. There are many risk factors that contribute to carcinogenesis, such as age, polyps in the intestine, increased consumption of red meat, obesity, smoking, alcohol consumption. Recently, this number of factors has been supplemented by infection with the bacterium Fusobacterium nucleatum, which according to research results, is actively involved in the process of carcinogenesis. However, the response to risk factors varies between geographical regions and ethnic groups, which can affect the prevalence and prognosis of the disease.
The idea of the project is to study Fusobacterium nucleatum and associated colorectal cancer based on modern molecular genetic methods. The results of early studies revealed the association of Fusobacterium nucleatum in the development of colorectal cancer, the bacterium promotes tissue malignancy, inducing inflammation and the host immune response in the microenvironment of the intestinal tract. Mechanisms of pathomorphogenesis development and involvement of genes involved in this process require additional experimental work. Also, the study of the complete genome of the bacterium may reveal new factors affecting the pathogenicity of the microorganism.
To study the genome-wide profile of Fusobacterium nucleatum associated with colorectal cancer and to study the expression of the inflammatory response in Kazakhstani patients.
As a result of the implementation of this project, genes contributing to the invasion of Fusobacterium nucleatum, genes of resistance to drug therapy against bacteria will be identified, a database of the full-genome sequence of Fusobacterium nucleatum isolated from Kazakhstani patients with colorectal cancer will be created, the correlation of the body’s immune response to invasion by bacteria will be studied. Thus, the data obtained will serve as a basis for studies of pathogenesis and carcinogenesis due to the presence of Fusobacterium nucleatum, all this will allow using the results as a tool for diagnosis, prognosis and correction of therapy. Quantitative and qualitative characteristics in the study will confirm the validity and significance of the results.
This project has a fundamental direction. The target consumers are biological subjects, clinicians, oncologists, and geneticists.
The results obtained during the project in the future may be relevant for personalized medicine in the field of oncology, selection and correction of therapy and preventive measures.
This study in its original form is fundamental in nature, requiring further research in the future in order to be able to use the results in practical medicine.
During the implementation of the project, new bacterial genes that were not described earlier may be identified, which will expand the understanding of the pathophysiological pathways of the disease. A full-genome sequence of bacteria isolated from Kazakhstani patients, not previously described, will replenish the databases. Associative relationships that have been validated in the Kazakh population can be used in medical and genetic counseling in the future.
Gulmira Nigmetzhanovna Kulmambetova, PhD, scientific advisor of project – Senior Researcher – National Scientific Laboratory of Biotechnology for Collective Use, National Center of Biotechnology, (Astana, Kazakhstan). The direction 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. Interned, performed research, was a postdoctoral fellow in Pharmacogenomics at Duke University, Center for Applied Genomics and Precision Medicine, Department of Medicine, Durham, North Carolina, (USA); Institute of Physico-Chemical Medicine, Moscow (Russia); Institute of Physico-Chemical Biology and Fundamental Medicine, Novosibirsk (Russia); Federal Polytechnic School, Lausanne (Switzerland). She was the scientific advisor of project within the grant funding of the KN MES RK: “Study of candidate genes involved in the development of subarachnoid hemorrhage (family forms) based on full-exome sequencing”, “Obtaining a strain of recombinant CagA protein producer and development of screening technology of genetic analysis to determine antibiotic resistance of H.pylori”, also within the framework of scientific and technical programs “DNA chips for the diagnosis of infectious diseases”, “Development of a comprehensive test system for predicting the development of human gastric cancer based on the factors of Helicobacter pylori virulence and polymorphism of the interleukin 1ß gene”, “The role of cytokine gene polymorphisms in the risk of gastric cancer”. He has 5 innovative patents in the field of microbiology and genetics. Participated in the research project “Creation of an individual genetic passport based on the genetic diversity of the peoples of Kazakhstan”. He is the author of more than 90 scientific papers, including 6 publications in foreign journals with an impact factor. According to Web of Science, Kulmambetova G.N. has a Hirsch index (h-index) equal to 3 (https://www.scopus.com/authid/detail.uri?authorId=56387533800 ). Participated in international conferences. She was the chairman of the dissertation Council of Kokshetau State University named after Sh.Ualikhanov, Kokshetau, is also involved as an expert by the National Center for State Scientific and Technical Information, as well as a reviewer of scientific publications of the international scientific journal “Turkish Journal of Gastroenterology”. The project will provide leadership, execution of the main tasks, external interaction, ethical issues, analysis, statistical analysis of data and interpretation of results, preparation of publications and reports.
The basis of the research group of the project, including the project manager, are young researchers of the National Scientific Laboratory of Biotechnology for Collective Use, the National Center of Biotechnology:
Researcher 1 – Mamlin Meiram Askarovich – oncologist surgeon in the field of coloproctology. Bolashak Scholarship holder under the Internship program at the Republican Cancer Center (2014), Kazan, Russian Federation. He completed trainings and internships in the following areas: Transanal minimally invasive surgery for early rectal cancers (2017), St. Yana, Bruges, Belgium; Introduction to the residency program (2018), UPMC, Pittsburgh, USA; Laparoscopic Coloproctology (2015, 4 weeks), the First Sechenov Moscow State Medical University. The course of P.V. Tsarkov, Moscow, Russian Federation; Laparoscopic oncoloproctology (2016), State Medical Institution “St. Petersburg Clinical Scientific and Practical Center for Specialized Types of Medical Care (Oncological)”; Course of I.L. Chernikov, St. Petersburg, Russian Federation; Laparoscopic oncoloproctology (2017), N.N. Petrov NMIC, course of A.M. Karachun, G. St. Petersburg, Russian Federation; Gastric Cancer Surgery (2018), N.N. Petrov NMIC, A.M. Karachun course, St. Petersburg, Russian Federation. It will carry out surgical intervention to patients, sampling of biological material, participation in the design of scientific publications and reports.
Researcher 2 – Shalekenov Sanzhar Bulatovich (Hirsch index – 3). Candidate of Medical Sciences, Phd – Doctor der Medizin (Germany). Currently, he is the Deputy Chairman of the Board for Scientific and Educational Activities. He completed trainings and internships in the following areas: 2007. “Management in urology, issues of oncourology and endourology”, University Clinic named after Friedrich–Schiller, Urological Clinic, Germany. Jena; 2009. “Questions of oncourology” – University Clinic named after Friedrich–Schiller, Urological Clinic, Germany. Jena; 2010 “Laparoscopic urology, robot–assisted operations in urology at Da Vinci, operative andrology”, University Clinic Tor Vergata – Clinic of Urology, Italy, Rome (certificate); 2011 “Organization of urological service, oncourology, laparoscopic methods of treatment in urology”, University Clinic, Germany, Regensburg; 2013 – 2014 “Endovideosurgery in Urology”, University Clinic of Urology, Germany, Regensburg (scholarship under the Bolashak program); He will be engaged in recruiting patients, forming cohorts of patients for research, participating in the design of scientific publications and reports.
Researcher 3 – Khamzina Saule Sansyzbaevna oncologist surgeon in the field of coloproctology. She has completed trainings and internships in the following areas: “Laparoscopic coloproctology” (2014), I.M. Sechenov First Moscow State Medical University, P.V. Tsarkov course, Moscow, Russian Federation; “General Surgery”, “Colorectal Surgery” (2020), Seoul National University Hospital, Seoul, Republic of Korea. It will carry out surgical intervention to patients, sampling of biological material, participation in the design of scientific publications and reports.
Researcher 4 – Kozhakhmetov Arman Maratovich surgeon-oncologist. It will carry out surgical intervention to patients, sampling of biological material, participation in the design of scientific publications and reports.
Researcher 5 – Kurentai 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. He is proficient in the methods of nucleic acid extraction, PCR, real-time PCR, Sanger sequencing.
Researcher 6 – Gusmaulemova Alua Daurenkyzy – Master of 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 the methods of nucleic acid extraction, PCR, real-time PCR, Sanger sequencing.
At this stage of the project implementation, a sample of biomaterial from Kazakhstani patients with colorectal cancer continues to be created. Diagnosis of biomaterial (biopsy) for colorectal cancer is carried out histologically. Fresh carcinoma tissues, adjacent normal tissues and distant normal tissues (at a distance of 10 cm) from each patient are included in the sample according to the inclusion and exclusion criteria. The sample preparation stage includes extraction of DNA and RNA molecules from a biopsy, followed by qualitative and quantitative characterization. DNA molecules are used to detect Fusobacterium nucleatum in biopsy by means of a species-specific gene by CT-PCR. The RNA molecules from the biopsy are used to analyze the expression of inflammatory response genes (interleukins) using digital PCR. The cultivation of Fusobacterium nucleatum is carried out on selective media under anaerobic conditions with subsequent genetic identification of strains by means of the 16S rRNA gene by the Sanger method. A collection of Fusobacterium nucleatum strains is being formed from clinical material. Genome-wide sequencing of Fusobacterium nucleatum strains is carried out on the high-performance MiSeq platform. Based on the results of genome-wide sequencing of a clinical strain of Fusobacterium nucleatum, an article was published in the Scopus database (Scopus) the CiteScore percentile index (Sitescore) is at least 35.