(Craiova) Identification and clinical validation of biomarkers for long-term outcome after cerebral ischemia
Romania (FASMR) - Univesity of Medicine and Pharmacy Craiova, Craiova
Functional Sciences
Prof. Dr. Aurel Popa-Wagner
Prof. Dr. Aurel Popa-Wagner; Dr. Andrei Gresita
4 weeks
Cities/Months Jan Feb Mar Apr May Jun Jul Augt Sep Oct Nov Dec
No No No No No Yes Yes Yes Yes No No No
Type of Research Project
- Clinical Project with Laboratory work
What is the background of the project?
Ischemic stroke is an acute disease which often results in severe long-term physical disability, depression, and cognitive decline. Patients at risk for these late consequences of stroke are not duly diagnosed and treated due to the lack of reliable biomarkers. Extracellular vesicles derived from neurons (ND-EVs) enter the blood stream and have been shown to reflect neuronal health. Previous work by iBioStroke partners found that MicroRNA 21 (MIR21) and MicroRNA 223 (MIR223) genes are hypoxia inducible and secreted into extracellular vesicles (EVs) both in stroke models and stroke patients that were significantly associated with clinical outcome (measured by modified Rankin Scale, mRS) in post-stroke patients at 90 days. In addition, two Genome Wide Association studies have found genetic polymorphisms associated with stroke recovery.
What is the aim of the project?
This proposal aims at (1) isolating ND-EVs released into blood and cerebrospinal fluid (CSF) in aged mice and rats and patients following ischemic stroke, (2) using proteomics and transcriptomics to identify novel ND-EV-based biomarkers for the prediction of stroke outcome, (3) validate ND-EV-based biomarkers in two highly stratified stroke patient populations, and (4) cross-validate ND-EV-based biomarkers found in aged animal models in a cohort of stroke patients.
What techniques and methods are used?
Evaluation of ND-EV proteome and transcriptome during long-term recovery from transient focal cerebral ischemia in aged rodents (Partner 2 and 3) Transient focal cerebral ischemia in mice will be induced using the 60-minute filament middle cerebral artery occlusion model (MCAo) we recently developed that allows a unique long-term survival protocol [Lourbopoulos et al, JCBFM 2015]. This procedure will allow the mice to survive for one, three, or twelve months (n=25/group). Sham operated mice/rats will be used as controls for each time point (n=25/group). One group of unhandled mice (n=25) will be observed for twelve months as another control to exclude effects of surgery and anesthesia. Accordingly, a total of 175 mice (seven groups with 25 mice each) will be investigated in the current project. Brains will be fixed by perfusion fixation and processed for floating sections high-resolution 3D immunohistochemistry (Zeiss LSM 800 with Airyscan). Stroke-related histopathological changes on the cellular level (microglia and astrocyte activation assessed by artificial intelligence-based algorithms, neuronal survival, white matter injury, and blood-brain barrier integrity) will be correlated with morphological (magnetic resonance imaging, MRI) and behavioral outcomes (neurological function, learning, memory, depression-like behavior) and with the results obtained by liquid biopsy. All experiments will be fully randomized and blinded.
What is the role of the student?
- If the project includes “lab work”
- the student will take active part in the practical aspect of the project
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
Mice will undergo MCAo or sham surgery and will then be investigated monthly for brain morphology (infarct size, atrophy, edema) by MRI and for neurological function (motor-sensory function, learning, memory) by behavioral tests (mNSS, Barnes Maze). At defined, functionally-relevant time-points (7, 28, 56 and 90 days post-stroke), peripheral blood samples will be harvested from the femoral vein. At 90 days post-MCAO, CSF will also be collected.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Preliminary readings provided my the tutor.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will have the opportunity to present the results together with the supervisor at a conference
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts:
- Medical students
- Graduated students (less than 6 months)
- Pre-Medical students from the American-British system
- Students in biomedical fields
- Surugiu R; Olaru A; Hermann DM; Glavan D; Catalin B; Popa-Wagner A. Recent Advances in Mono- and Combined Stem Cell Therapies of Stroke in Animal Models and Humans. Int J Mol Sci. 2019 Nov 29;20(23). pii: E6029. doi: 10.3390/ijms20236029. Review. PubMed PMID: 31795466.