Projects / Programmes source: ARIS

Identification of novel glioblastoma biomarkers for non-invasive liquid biopsy

Research activity

Code Science Field Subfield
3.03.00  Medical sciences  Neurobiology   

Code Science Field
3.01  Medical and Health Sciences  Basic medicine 
Liquid biopsy, novel biomarkers, glioblastoma, microarray, ALYREF, DPYSL2, FREM2, TUFM, TRIM28, VIM, CRMP1, SPRY1, NAP1L1, miRNA, lncRNA
Evaluation (rules)
source: COBISS
Researchers (1)
no. Code Name and surname Research area Role Period No. of publicationsNo. of publications
1.  33735  PhD Neja Šamec  Biochemistry and molecular biology  Head  2020 - 2023  82 
Organisations (1)
no. Code Research organisation City Registration number No. of publicationsNo. of publications
1.  0381  University of Ljubljana, Faculty of Medicine  Ljubljana  1627066  45,315 
Glioblastoma is the most common, aggressive and lethal form of brain tumor. Due to appearance of non-specific symptoms, disease diagnosis in early developmental stages is highly unlikely. First line of diagnosis is tumor detection with magnetic resonance imaging (MRI) that is also used for monitoring disease progression or shrinkage after/during treatment. However, MRI does not differentiate between actual tumor progression and necrotic tissue after surgery, and has limitations in terms of resolution about 2-3 mm. Therefore, the tumor is present in the brain even before we can macroscopically detect it. Moreover, in cases where a single biopsy sample is obtained, the entire tumor heterogeneity cannot be captured. In this project we will determine expression changes of circulating blood biomarkers from 58 plasma samples of patients with different grades of glioma compared with 38 healthy individuals. In the study, we will examine a group of biomarkers, ALYREF, DPYSL2, FREM2, TUFM, TRIM28, VIM, CRMP1, SPRY1 and NAP1L1, that were already identified as differentially expressed in glioma tissue samples compared to lower grade gliomas and reference non-tumor brain samples in our previous research. The first step of the project will be optimization of protocols for isolation of extracellular vesicles and RNA from blood. I will proceed with the protocol that will result in higher yield and better quality of the isolated RNA. After the protocol will be optimized, I will prepare isolates of total RNA, miRNA and proteins from 58 glioma patients. Next, the determination and validation on proteomic and transcriptomic levels of selected previously over expressed glioma biomarkers ALYREF, DPYSL2, FREM2, TUFM, TRIM28, VIM, CRMP1, SPRY1 and NAP1L1 will follow. RNA expression levels will be determined with digital droplet PCR due to the high sensitivity of the method which is crucial for biological samples where the amounts of the RNA of interest are limited. At the proteomic level, we will study the expression of ALYREF, DPYSL2, FREM2, TUFM, TRIM28, VIM, CRMP1, SPRY1 and NAP1L1 proteins with western blot. For 8 glioblastoma patients who will be enrolled in chemotherapy treatment with temozolomide, we will follow the expression levels of the aforementioned gene and protein biomarkers to determine if they can be used as biomarkers for monitoring disease changes (progression or shrinkage) during treatment. In the second year or the running project, we will examine the effect of micro (mi)RNA and long non-coding (lnc)RNA on changes in expression of circulating blood biomarkers in 8 patients with glioblastoma compared to 8 healthy individuals. To show different expression levels of miRNA and lncRNA in plasma samples from glioblastoma patients compared to samples originating from healthy individuals, we will construct a mRNA-lncRNA-miRNA coexpression network. With this research, we will provide easily accessible non-invasive liquid biopsy biomarkers for early glioblastoma detection and possibly for future successful treatment monitoring.
Most important scientific results Interim report
Most important socioeconomically and culturally relevant results
Views history