Background: This study aimed to develop a superior aptamer-based therapeutic for targeted glioblastoma intervention by conducting a comparative analysis of two DNA aptamers: the original U2 sequence, selected against glioblastoma cells exhibiting high EGFRvIII expression, and its modified, shortened, and stabilized variant, Gol1. Methods: The effects of the investigated aptamers on primary human glioblastoma cells with graded receptor expression levels and on a rat 101/8 glioblastoma tissue model were rigorously studied. Results: The results demonstrated the significant superiority of the stabilized Gol1 aptamer, which exhibited exceptional binding affinity for the EGFRvIII receptor. Pronounced antiproliferative and antimigratory effects against EGFRvIII-positive human tumor cells, ultimately inducing complete cell death. Transcriptomic analysis revealed a sophisticated dual mechanism of action for Gol1: the specific activation of neuronal differentiation genes concurrent with the suppression of key alternative splicing factors. Crucially, in vivo confirmation showed highly selective accumulation of the FAM-labeled Gol1 aptamer exclusively within tumor tissue, with a maximum concentration gradient observed in the invasive border zone and a complete absence of accumulation in intact brain parenchyma. Conclusions: These comprehensive findings confirm that the Gol1 aptamer constitutes a highly promising and versatile platform for developing novel targeted theranostic strategies against glioblastoma, offering a precise approach for both diagnostic imaging and therapeutic intervention.

Glial cell line-derived neurotrophic factor (GDNF) is a highly conserved neurotrophic factor that is critical for organismal development and survival. GDNF is known for promoting the survival and differentiation of dopaminergic neurons, preventing apoptosis in mature neurons. These properties make GDNF a candidate for therapeutic applications, particularly in neurodegenerative diseases such as Parkinson's disease (PD). Despite promising preclinical results in rodent and non-human primate models, clinical trials have failed to replicate these findings, necessitating further investigation into the underlying causes. In prior studies, our team developed a chimeric mGDNF/GFP protein with notable neuroinductive properties. However, further characterization of mGDNF, excluding GFP, led to the discovery of alternative transcript variants, resulting in the identification of several small peptides. Among them, the peptide djGDNF47 (47shGDNF) demonstrated neuroinductive activity in vitro on cell models of the PC12 line and dorsal embryonic ganglia. This activity was observed both when synthesized in transgenic HEK293 cells expressing the cloned djGDNF47 and in the form of a synthesized analog. This finding suggests the existence of alternative splicing mechanisms that generate short neurotrophic peptides. Our current research focuses on djGDNF47 (47shGDNF), a bioactive GDNF peptide with promising therapeutic potential for treating neurodegenerative diseases such as PD.

Background: High-grade gliomas are treatment-resistant and prone to aggressive recurrence. Although radiation therapy is a fundamental treatment, it often fails to eradicate tumors and can enhance the migratory potential of surviving cells, promoting relapse. Anti-proliferative aptamers are novel agents that show promise, but their combination with radiation therapy and their effects on invasive phenotypes require further investigation. Objectives: This study evaluated the effects of ionizing radiation on the viability and migration of human glioma cells, both alone and in combination with the anti-proliferative aptamer bi-(AID-1-T). The study aimed to determine whether the aptamer could enhance the efficacy of radiotherapy and counteract ionizing radiation-induced pro-migratory effects. Methods: The study was conducted on cell cultures of primary and relapsed human glioma. The effects of combined radiation (single dose of 20 Gy) and the bi-(AID-1-T) aptamer (10 μM) were assessed using the MTS assay, Transwell analysis, immunocytochemistry and transcriptome analysis. Results: Ionizing radiation alone reduced proliferation in primary gliomas, but increased proliferation in recurrent cultures. Ionizing radiation also increased migration in both types of gliomas. Combining ionizing radiation with the bi-(AID-1-T) aptamer produced a synergistic effect: it significantly reduced cell proliferation and migration, and suppressed the ionizing radiation-induced migratory enhancement, more effectively than either treatment alone. Transcriptome analysis revealed that combination treatment decreased the expression of pro-proliferative and migratory genes (e.g., PDPN, CDH3), while increasing the expression of anti-migratory (RND3) and pro-apoptotic genes (e.g., XAF1, SEMA3A). Thus, combination treatment significantly reduces tumor cell proliferation and migration; however, further studies on surviving cells are needed.

Glioblastoma remains an incurable malignancy, driving the continuous search for novel therapeutic strategies. A significant challenge in this pursuit is the transition from in vitro models to in vivo efficacy. Building on our previously patented GQIcombi strategy (Patent RU2820200C2), this study investigates its efficacy in vivo using the rat 101/8 glioblastoma model. We evaluated various administration regimens of the GQIcombi cocktail, which was previously effective against patient-derived human glioma cell cultures. Although we attempted to reduce the dosing frequency to optimize the protocol, our results confirm that sequential exposure to the GQIcombi components is necessary to effectively inhibit tumor growth in vivo, mirroring its requirement in cell cultures.

The development of new drugs in nuclear medicine for diagnosis or treatment (chemotherapy) of brain tumors, in particular gliomas, is inextricably linked with the use of tumor models in animals (usually rats). OBJECTIVE:To compare the widely used glioma cell model C6 and the new experimental tissue model of glioblastoma 101.8. MATERIAL AND METHODS:A comparison was made of the diagnostic and morphological characteristics of the presented glioma models in two groups of animals with intracranially implanted tissue strain of experimental glioblastoma 101.8 (n=4) and the C6 glioma cell line (n=4) throughout the tumor development cycle within the rat brain. To monitor the progress of tumor growth and development, each animal underwent repeated diagnostic studies using PET-CT with 18F-FDG and 18F-FET to assess the metabolic activity and volume of the tumor. Also MRI images were collected. After the end of data acquisition, a histological examination of the tumor was carried out. RESULTS:The tissue model of glioblastoma 101.8 demonstrated rapid growth and pronounced accumulation of the tracers in all animals during the tumor observation cycle. Formation of intratumoral necrosis and signs of disruption of the blood-brain barrier (BBB) were detected. In PET-CT studies of animals with a transplanted C6 tumor, no visible necrosis in the tumor structure was observed. Tumor growth was less rapid than in the case of model 101.8. The obtained morphological characteristics of 101.8 tumors transplanted into the rat brain demonstrated similar properties observed in real clinical conditions in patients with glioblastoma of the brain (necrosis, neovascularization, multiple pseudopalisade structures). CONCLUSIONS:Tumor model 101.8 can be recommended for scientific research as it most closely reproduces the diagnostic and morphological features of a human glioblastoma.

Current therapy protocols fail to cure high-grade gliomas and prevent recurrence. Therefore, novel approaches need to be developed. A re-programing of glioma cell fate is an alternative attractive way to stop tumor growth. The two-step protocol applies the antiproliferative GQ bi-(AID-1-T) and small molecule inducers with BDNF to trigger neural differentiation into terminally differentiated cells, and it is very effective on GB cell cultures. This original approach is a successful example of the “differentiation therapy”. To demonstrate a versatility of this approach, in this publication we have extended a palette of cell cultures to gliomas of II, III and IV Grades, and proved an applicability of that version of differential therapy for a variety of tumor cells. We have justified a sequential mode of adding of GQIcombi components to the glioma cells. We have shown a significant retardation of tumor growth after a direct injection of GQIcombi into the tumor in rat brain, model 101/8. Thus, the proposed strategy of influencing on cancer cell growth is applicable to be further translated for therapy use.

Restoring visual function after damage or complete destruction of the optic nerve in adult patients has many natural barriers to neuroregeneration. Research to restore vision has focused on maintaining retinal ganglion cells (RGCs), stimulating axonal growth toward the brain, and restoring their proper synaptic connections. Unfortunately, mammalian RGC axons under normal conditions do not regenerate after injury and ultimately die. In this review, we summarize the currently known mechanisms of RGC survival and axonal regeneration in mammals, including specific intrinsic signaling pathways, key transcription factors, reprogramming genes, inflammation-related regeneration factors, and stem cell therapy. We also review the current understanding of the phenomena impeding optic nerve regeneration and possible ways to overcome these obstacles. The most important research results obtained in recent decades may be informative for the development of methods for treating the damaged visual system.

High-grade gliomas are considered an incurable disease. Despite all the various therapy options available, patient survival remains low, and the tumor usually returns. Tumor resistance to conventional therapy and stimulation of the migratory activity of surviving cells are the main factors that lead to recurrent tumors. When developing new treatment approaches, the effect is most often evaluated on standard and phenotypically depleted cancer cell lines. Moreover, there is much focus on the anti-proliferative effect of such therapies without considering the possible stimulation of migratory activity. In this paper, we studied how glioma cell migration changes after exposure to bi-(AID-1-T), an anti-proliferative aptamer. We investigated the effect of this aptamer on eight human glioma cell cultures (Grades III and IV) that were derived from patients' tumor tissue; the difference between primary and recurrent tumors was taken into account. Despite its strong anti-proliferative activity, bi-(AID-1-T) was shown to induce migration of recurrent tumor cells. This result shows the importance of studying the effect of therapeutic molecules on the invasive properties of glioma tumor cells in order to reduce the likelihood of inducing tumor recurrence.

Glioblastoma (GBM) is a malignant tumor with an average survival of 15–16 months with standard treatment; however, cases of successful treatment provide hope that a better understanding of the pathology will improve prognoses. Glial tumors contain clonogenic cells (cells capable of forming colonies in a culture medium) with high proliferative potential and descendants with a wide range of differentiation possibilities; these clonogenic cells are currently regarded as glioma stem cells (GSC). In normal and pathological conditions, the adult brain includes zones containing proliferating neural stem cells (NSC) and their descendants, i.e., progenitor cells which have started to differentiate. One such zone, lying on the lateral wall of the lateral ventricle and termed the subventricular zone (SVZ) of the lateral ventricle, has received extensive attention because of its importance in gliomagenesis. Numerous studies have demonstrated that the intense exchange of signaling molecules and cells between the GBM and the SVZ leads to accelerated tumor growth and an increased risk of relapse. Research results point to the possibility of developing new and more effective strategies to combat this dangerous disease, taking account of knowledge of the role of the SVZ in the development of this pathology.

Background: High-grade gliomas remain a virtually incurable form of brain cancer. Current therapies are unable to completely eradicate the tumor, and the tumor cells that survive chemotherapy or radiation therapy often become more aggressive and resistant to further treatment, leading to inevitable relapses. While the antiproliferative effects of new therapeutic molecules are typically the primary focus of research, less attention is given to their influence on tumor cell migratory activity, which can play a significant role in recurrence. A potential solution may lie in the synergistic effects of multiple drugs on the tumor. Objectives: In this study, we investigated the effect of combined exposure to bi-(AID-1-T), an anti-proliferative aptamer, and its analog bi-(AID-1-C), on the migratory activity of human GBM cells. Results: We examined the effects of various sequences of adding bi-(AID-1-T) and bi-(AID-1-C) on five human GBM cell cultures. Our findings indicate that certain sequences significantly reduced the ability of tumor cells to migrate and proliferate. Additionally, the expression of Nestin, PARP1, L1CAM, Caveolin-1, and c-Myc was downregulated in human GBM cells that survived exposure, suggesting that the treatment had a persistent antitumor effect on these cells.

Glioblastoma (GBM) is a malignant tumor with an average survival rate of 15–16 months with standard treatment; however, cases of successful treatment provide hope that a better understanding of the pathology will improve prognosis. Glial tumors contain clonogenic cells (cells capable of forming colonies in a culture medium) with a high proliferative potential, and their descendants have a wide range of possible differentiation; these clonogenic cells are currently considered as glioma stem cells (GSCs). In normal and pathological conditions, there are zones in the adult brain that contain proliferating neural stem cells (NSCs) and their descendants – progenitor cells that have begun to differentiate. One such zone lying on the lateral wall of the lateral ventricle, called the subventricular zone of the lateral ventricle (SVZ), has attracted much attention due to its importance for gliomagenesis. Numerous studies have shown that the intense exchange of signaling molecules and cells between the GBM and the SVZ leads to accelerated tumor growth and an increased risk of relapse. Research results indicate the possibility of developing new, more effective strategies to combat this dangerous disease, taking into account knowledge about the role of SVZ in the development of this pathology.

Many human diseases including cancer, degenerative and autoimmune disorders, diabetes and others are multifactorial. Pharmaceutical agents acting on a single target do not provide their efficient curation. Multitargeted drugs exhibiting pleiotropic pharmacological effects have certain advantages due to the normalization of the complex pathological processes of different etiology. Extracts of medicinal plants (EMP) containing multiple phytocomponents are widely used in traditional medicines for multifactorial disorders' treatment. Experimental studies of pharmacological potential for multicomponent compositions are quite expensive and time-consuming. In silico evaluation of EMP the pharmacological potential may provide the basis for selecting the most promising directions of testing and for identifying potential additive/synergistic effects. Multiphytoadaptogen (MPhA) containing 70 major phytocomponents of different chemical classes from 40 medicinal plant extracts has been studied in vitro, in vivo and in clinical researches. Antiproliferative and anti-tumor activities have been shown against some tumors as well as evidence-based therapeutic effects against age-related pathologies. In addition, the neuroprotective, antioxidant, antimutagenic, radioprotective, and immunomodulatory effects of MPhA were confirmed. Analysis of the PASS profiles of the biological activity of MPhA phytocomponents showed that most of the predicted anti-tumor and anti-metastatic effects were consistent with the results of laboratory and clinical studies. Antimutagenic, immunomodulatory, radioprotective, neuroprotective and anti-Parkinsonian effects were also predicted for most of the phytocomponents. Effects associated with positive effects on the male and female reproductive systems have been identified too. Thus, PASS and PharmaExpert can be used to evaluate the pharmacological potential of complex pharmaceutical compositions containing natural products

Radiation therapy induces double-stranded DNA breaks in tumor cells, which leads to their death. A fraction of glioblastoma cells repair such breaks and reinitiate tumor growth. It was necessary to identify the relationship between high radiation doses and the proliferative activity of glioblastoma cells, and to evaluate the contribution of DNA repair pathways, homologous recombination (HR), and nonhomologous end joining (NHEJ) to tumor-cell recovery. We demonstrated that the GO1 culture derived from glioblastoma cells from Patient G, who had previously been irradiated, proved to be less sensitive to radiation than the Sus\fP2 glioblastoma culture was from Patient S, who had not been exposed to radiation before. GO1 cell proliferation decreased with radiation dose, and MTT decreased to 35% after a single exposure to 125 Gγ. The proliferative potential of glioblastoma culture Sus\fP2 decreased to 35% after exposure to 5 Gγ. At low radiation doses, cell proliferation and the expression of RAD51 were decreased; at high doses, cell proliferation was correlated with Ku70 protein expression. Therefore, HR and NHEJ are involved in DNA break repair after exposure to different radiation doses. Low doses induce HR, while higher doses induce the faster but less accurate NHEJ pathway of double-stranded DNA break repair.

The review analyzes some parameters of CBA mice-males as model of spontaneous carcinogenesis characterizing adhesive and adaptive disorders. A weakening of the hepatocytes mutual adhesiveness force was noted already in early ontogenesis (5–10 days of postnatal development). This violation persisted and enhanced during hepatocarcinogenesis. A decrease of the β2 leukocyte integrins LFA-1 and Mac-1 expression on peripheral blood cells as well as an increase of the interleukins 6 and 10 in blood serum were determined during ontogenesis. It is significant for weakening the liver cells contact interactions (mutual adhesiveness) as well as immunity effectors and tumor cells interactions. A disbalance of the adaptive reactions and life quality important components was revealed in the CBA mice-males ontogenesis. Number of dopaminergic neurons and the neurogenesislevel in CBA micemales were decreasing. This does not contradict the dynamics of chronic stress and the aging process: an increase in the catabolic stress hormone corticosterone, a decrease in the anabolic hormone testosterone in the blood serum, a decrease in motor activity, signs of cachexia and alopecia, as well as a violation of immunological parameters.CBA mice-males with an assessment of parameters characterizing adhesive and adaptive disorders during spontaneous carcinogenesis (the hepatocytes mutual adhesiveness forсe, the expression of β2 leukocyte integrins LFA-1 and Mac-1 on peripheral blood cells, the content of interleukins 6, 10, corticosterone and testosterone in blood serum, the number of dopaminergic neurons in the midbrain during ontogenesis) as well as the frequency and size of tumours, lifespan and somatic status of animals can be used as a scientifically- and evidence-based test system to study cytostatic drugs as well as non-toxic geroprotective medications for prevention and treatment of cancer in individuals with an increased risk of malignant neoplasms developing especially hepatocellular carcinoma.

Cancer cell reprogramming based on treatment with G-quadruplex, having antiproliferative power, along with small molecules able to develop iPSCs into neurons, could create a novel approach to diminish the chance of glioblastoma recurrence and circumvent tumor resistance to conventional therapy. In this research, we have tested several combinations of factors to affect both total cell cultures, derived from tumor tissue of patients after surgical resection and two subfractions of this cell culture after dividing them into CD133-enriched and CD133-depleted populations (assuming CD133 to be a marker of glioblastoma stem-like cells). CD133(+) and CD133(-) cells exhibit different responses to the same combinations of factors; CD133(+) cells have stem-like properties and are more resistant. Therefore, the ability to affect CD133(+) cells provides a possibility to circumvent resistance to conventional therapy and to build a promising strategy for translation to improve the treatment of patients with glioblastoma.

Glial cell line-derived neurotrophic factor (GDNF) has a wide range of actions and positively affects viability, proliferative activity and migratory ability of cells in nervous system. That is why GDNF is being considered as a therapeutic molecule in the treatment of neurodegenerative diseases, in particular Parkinson's disease. However, GDNF has the same effect on high-grade glioma cells promoting their growth, resistance to therapy and dissemination. Expression of this factor in tissues and cultures of gliomas is up to five times higher than in intact brain matter. It was revealed that epigenetic modifications in GDNF gene promoter contribute to overexpression. Target suppression of GDNF gene transcription slows down growth of glioma and decreases cell migration. This review is devoted to the effect of GDNF on glioma cells, causes and consequences of its overexpression. Further analysis of expression and function of various GDNF isoforms in glial tumors may be valuable to develop new treatment methods for these dangerous diseases.

The review presents data which provides evidence for the internal relationship between the stages of rodent audiogenic seizures and post-ictal catalepsy with the general pattern of animal reaction to the dangerous stimuli and/or situation. The wild run stage of audiogenic seizure fit could be regarded as an intense panic reaction, and this view found support in numerous experimental data. The phenomenon of audiogenic epilepsy probably attracted the attention of physiologists as rodents are extremely sensitive to dangerous sound stimuli. The seizure proneness in this group shares common physiological characteristics and depends on animal genotype. This concept could be the new platform for the study of epileptogenesis mechanisms.

According to one of the theories, gliomas can occur as a result of dysregulation of stem cell division in the subventricular region of the brain. The CD133 membrane marker is a characteristic of both normal and tumor neural stem cells therefore it can be used to isolate a stem cell population from tumor tissue. Tumor cells actively proliferate which suggests that their possible differentiation may be achieved by inhibiting of their division as these two processes are mutually exclusive. For this purpose, G-quadruplex oligonucleotides together with neural-inducers such as a brain-derived neurotrophic factor (BDNF) may be used. Five cell cultures obtained from human glioblastoma tissues were analyzed for expression of CD133 using RT-qPCR. From cell culture with the highest level of CD133 using immunomagnetic separation CD133+ and CD133- cultures were received. CD133fr/peGFP-c1 recombinant DNA consisted of a CD133 second extracellular loop fragment and a peGFP-c1 vector was constructed to determine the localization of prominin-1, that is known as CD133 when found on cell membrane, using confocal microscopy. On chosen cell cultures an oligonucleotide bi-(AID-1-T) and its combination with BDNF were tested. The mechanism of GQ’s action is cytostatic and its non-toxicity properties were proved by flow cytometry. For evaluating the proliferative activity of cells MTT assay was performed on 10th and 20th days after exposure to the factors. Cell culture G01 was chosen for further research as it had the highest level of the CD133. Colocalization of CD133 and GFP demonstrated a membrane localization of CD133 in cells with high expression level of this marker. MTT assay on 10th day after exposure to bi-(AID-1-T) as well as its combination with BDNF on cell culture G01 CD133- showed total inhibition of cell proliferation. The same combinations tested on G01 CD133+ cell culture demonstrated no difference in proliferative activity. After 20 days after exposure to bi-(AID-1-T) and combination of bi-(AID-1-T) with BDNF the significant decrease of G01 CD133+ cells’ proliferation was observed. When tested on whole glioblastoma cell culture G01 these combinations also showed significant inhibition of cell proliferation. We showed that glioblastoma cells upon transfection with recombinant DNA, that contains a fragment of CD133, mainly have a membrane localization of this marker. It was observed that CD133+ cells are more stable to external influence that can be a proof of the fact that CD133 is charactered for glioblastoma stem cells. We tested the effect of an GQ bi-(AID-1-T) and its combination with BDNF and showed that BDNF is necessary for blocking proliferation of glioblastoma cells. Altogether, the results may be used for further research as it reveals a potential treatment for patients with glioblastoma. Grant №075-15-2020-809 (13.1902.21.0030).