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Information × Registration Number 0224U032972, (0124U004210) , R & D reports Title A comprehensive study of 3D model systems to elucidate the mechanisms of CHI3L1 isoforms influence on glioblastoma progression popup.stage_title Створення низькотемпреатурного стоку культур нейронів та макроглії фетального мозку людини та збірка векторної конструкції CRISPR/Cas9 для нокауту транскриптів гена CHI3L1 Head Skrypkina Inessa Ya., Кандидат біологічних наук Registration Date 13-12-2024 Organization Institute of Molecular Biology and Genetics of NAS of Ukraine popup.description1 The aim of the proposed project is to determine the functional role of the chitinase-like protein CHI3L1 isoforms in the development and progression of glioblastomas, the impact on the main cellular components of the microenvironment (macrophages and neurons) using innovative 3D cell model systems. popup.description2  As a result of the implementation of the first stage of the project, methods were developed to obtain viable neuronal cell cultures from cryo-frozen human fetal brain tissues. It was confirmed that the cell cultures obtained from human fetal brain tissue samples correspond to the phenotype of neurons and macroglia and can be successfully used to implement further experimental tasks at the next stage of this project. A low-temperature bank of the obtained cultures was created in a quantity sufficient for further stages of work. A strategy to knock out transcriptional isoforms of the human CHI3L1 gene using CRISPR/Cas9 was developed and applied to the U-87 glioblastoma cell line, where this protein is highly expressed. Conversely, to investigate the effect of this promising target for anti-oncology therapy, derivatives were obtained with overexpression of the short or long CHI3L1 isoforms in U-251 glioblastoma cells, where it is absent. Derivatives of U-251 cells with selectively overexpressed CHI3L1-L and CHI3L1-S transcripts were obtained by lentiviral/retroviral transduction. The results of this stage will have a significant impact on further research and editing of the processes of glioblastoma origin and progression. They will become the basis for further improvement of brain tumor models, as well as for tasks related to genome editing. Product Description popup.authors Anopriienko Olha V. Areshkov Pavlo O. Bukreieva Tetiana V. Nesterenko Yuliia A. Rybachuk Oksana А. Shloma Anastasiia R. popup.nrat_date 2024-12-13 Close