AP19678106 Identification of novel germinal and somatic mutations in intracranial arteriovenous malformations

Relevance

Brain arteriovenous malformations (AVMs) are a serious cause of hemorrhagic stroke in children and adults, which very often leads to major disability mainly at working age or to death. Before the emergence of high-resolution diagnostic techniques, such as magnetic resonance imaging and computed tomography, as well as the development of multiomics technology, it was believed that cerebral AVMs could only be congenital. Currently, AVMs of the brain are divided into hereditary and sporadic forms, of which 95% are sporadic and about 3% are hereditary. To date, the mechanism of brain AVM formation has not been fully understood. This project is aimed at obtaining additional information about mutations and genes that have not been described before, which will allow to expand the understanding of the pathogenetic mechanisms of the disease.

The goal of the project is to identify new and rare germinal and somatic mutations responsible for the development of brain AVMs; to identify key genes and biological pathways involved in the progression of the disease based on RNA sequencing.

Multiome analysis – next generation sequencing (exomes) of blood and tissue lesions from the same patient, as well as RNA sequencing, allows us to identify potential germline and somatic brain AVM mutations and identify gene changes at the whole transcriptome level to further explore the mechanism of the disease.

The study of the mechanism of brain AVM formation will subsequently contribute to the development of new therapies, assessment of the prognosis of the disease, and the identification of risk groups.

The proposed project is fundamental. The project will contribute to scientific and technical development of the Republic of Kazakhstan, due to the use of modern molecular genetics research methods. The skills of working with annotated genomic data improve the skills of scientific staff of Kazakhstan in the field of molecular genetics and penalized medicine, especially of young scientists who will be involved in the project.

Objective

The aim of the project is to identify new and rare germinal and somatic mutations responsible for the development of AVM of the brain; identify key genes and biological pathways involved in the progression of AVM of the brain based on RNA sequencing.

Expected results

As a result of the project realization in accordance with the requirements of this documentation, it is expected to publish:

– at least three (3) articles and (or) reviews in peer-reviewed scientific journals indexed in the Science Citation Index Expanded of Web of Science database and (or) having a CiteScore percentile in the Scopus database of at least 35 (thirty-five). Presumably, one paper will be a case-report describing the comparison of mutation spectrum from blood and tissue from the same patient with AVM of the brain. Possible journals for publication are BMC Genomics data (Q2) (https://bmcgenomdata.biomedcentral.com/).  A publication devoted to the analysis of somatic KRAS/BRAF mutations in patients with AVM (presumably in Frontiers in Molecular Neuroscience, Scopus’ CiteScore, -8.4, Q1. The results are also expected to be presented in the Journal of Molecular Neuroscience, Scopus’ CiteScore, -3.1, Q3.

-as well as at least 1 (one) article or review in a peer-reviewed foreign or domestic periodical recommended by CQASES;

Other measurable results in accordance with the requirements of the competition documentation and the specifications of the project:

The proposed project is fundamental, but results obtained in the project will expand the understanding of the pathogenesis of this disease. It is relevant for the development of targeted therapies and personalized medicine for brain AVMs.

Results will be disseminated in the form of conference reports and publications in the periodicals of neighboring countries and further abroad.

Target consumers are scientific communities, university departments studying neuroscience, as well as specialists in biology.

This project represents adjacent fields of research – molecular genetics and molecular neuroscience.

The project will improve skills of scientists in the field of molecular biology and genetics; it will enhance their competence in working with annotated genomic data, given that this project involves more than 50% of young scientists.

Principal investigator

Project leader – Zholdybaeva Elena Vitalyevna, is a Candidate of Biological Sciences Hirsch index (h-index) – 9.

Members of the research group

Kulmambetova Gulmira, PhD – (h-индекс – 3) ResearcherID N-5975-2017, ORCID 0000-0001-8723-3752, AuthorID (56387533800);

Nurimanov Chingiz, PhD – doctoral student – (h-индекс -1);

Menlibaeva Karashash Kairatkyzy – (h-index -2);

Bekbaeva Ayazhan – (h-index -1);

Kurentai Botakoz Amandoskyzy;

Gusmaulemova Alua Dauren kyzy.

Results achieved

The formation of an experimental sample of brain AVMs (tissue and blood) has begun. Four patients meeting the inclusion criteria were selected. Venous blood samples and tissue from patients with AVMs are collected at the National Center for Neurosurgery directly by neurosurgeons who are participants in this project. The brain tissue was collected during surgery (AVM excision). From the collected blood and tissue samples from patients with AVM, DNA and RNA were isolated for further NGS sequencing.

From the collected venous blood samples of patients with AVM and brain tissue, DNA and RNA were isolated using a commercial kit according to the manufacturer’s instructions for further NGS sequencing. Quantitative DNA analysis was performed using a Nanodrop 1000 spectrophotometer (Thermo Fisher). To more accurately determine the DNA/RNA concentration, we used a Qubit 2.0 fluorometer, since nucleic acids will be used in the future for whole exome sequencing and transcriptional analysis.

For six samples (blood and tissue), full exome sequencing was performed on the basis of Macrogene (Korea). Bioinformatics analysis is currently underway. RNA samples were prepared to be sent to Macrogene for transcriptional analysis.