Method for validating the inhibition of glioblastoma invasiveness by MitoQ based on single-cell data analysis

Description

2. Inhibition of glioblastoma invasiveness by MitoQ. Content of the invention The present invention aims to provide a method for validating the inhibition of glioblastoma invasiveness by MitoQ based on the analysis of single-cell data. This method allows, by means of animal models, imaging detection, single-cell classification and functional validations, to highlight the inhibitory effect of MitoQ on the increase in glioblastoma invasiveness induced by radiotherapy, and to provide a standardized validation system for its clinical translation. 10. I.Establishment of the orthotopic model of glioblastoma in naked mice and allocation into groups An orthotopic model of glioblastoma is established in naked mice, which are randomly allocated into four groups: a) Control group: Group Sham, treated with physiological saline, without administration15 of drug or irradiation; b) Group treated with MitoQ: Group D, receiving only treatment with MitoQ; c) Group irradiated with X-rays: Group R, receiving only X-ray irradiation; d) Combined treatment group MitoQ + X-ray irradiation: Group D+R, receiving20 an administration of MitoQ combined with X-ray irradiation. II. Tumor Volume Detection The tumor volume of naked mice in each group was measured by magnetic resonance imaging. The results show that group D+R has the lowest BE2026 / 7290 3 tumor volume, indicating that MitoQ can significantly enhance the cytotoxic effect of X-rays on the lioblastoma, with a radiosensitizing effect. III.Identification of subpopulations of tumor cells by single-cell analysis The tumor tissues are subjected to single-cell sequencing and a classification analysis, making it possible to divide the glioblastoma cells into two categories: 5 a) Sub-populations of tumor cells in the proliferative phase: Including clusters F2 and F5; b) Subpopulations of tumor cells with invasive and migratory phenotype: Comprising clusters F1, F3, F8 and F9. IV. Inducing effect of radiotherapy on tumor invasiveness10 Comparing the R group to the Sham group, a significant increase in the proportion of sub-cell populations with an invasive and migratory phenotype is observed in the R group, which confirms that X-ray radiotherapy can induce an increase in the invasiveness of glioblastoma and constitutes a major cause of tumor recurrence. V.Validation of the inhibition of invasiveness and the radiosensitizing effect of MitoQ15 By comparing group D+R to group R: a) The proportion below of cell populations with invasive and migratory phenotype is significantly reduced in group D+R; b) The invasion and migration capacities of cells with invasive and migratory phenotype are significantly decreased in group D+R;20 The above results demonstrate that MitoQ exerts a radiosensitizing effect by inhibiting the increase in tumor invasiveness induced by X-rays. Compared to the prior art, the present invention has the following beneficial effects: BE2026 / 7290 4 The present invention, by optimizing existing methods for validating the inhibition of glioblastoma invasiveness by MitoQ based on the analysis of single-cell data, allows for the first time the identification, at the single-cell level, of the key cell subpopulations involved in glioblastoma invasion and inhibited by MitoQ; the validation method exhibits high precision and good reproducibility.Furthermore, the present invention demonstrates that the radiosensitization mechanism of MitoQ consists of inhibiting the increase in tumor invasiveness induced by radiotherapy, thus providing a new strategy for improving the radiosensitivity of glioblastoma. Moreover, the present invention establishes a standardized validation system integrating an animal model, imaging techniques, single-cell classification, and functional validations, which can be extended to the categorization and validation of other radiosensitizing agents.Description of the figures Figure 1 is a schematic diagram illustrating the orthotopic model of glioblastoma in naked mice in the method for validating the inhibition of glioblastoma invasiveness by MitoQ based on single-cell data analysis according to the present invention; Figure 2 represents the magnetic resonance images of the tumor volume of naked mice in each group in the method for validating the inhibition of glioblastoma invasiveness by MitoQ based on single-cell data analysis according to the present invention; Figure 3 is a flowchart illustrating the method for validating the inhibition of glioblastoma invasiveness by MitoQ based on single-cell data analysis according to the present invention. BE2026 / 7290 5 Specific embodiments The present invention will be described below clearly and completely in combination with the.