Stroke, TIA, Stroke-mimics, Stroke BiomarkersElViS-ACSNCT06319742
Extracellular Vesicle Surface Markers In Acute Cerebrovascular Syndromes.
An observational study of Stroke, TIA, Stroke-mimics, Stroke Biomarkers, run by Ospedale Civico, Lugano at 1 site in Switzerland.
Trial summary
Clinical implication of eventual blood biomarkers for stroke diagnosis and prognosis would be limited, mainly because clinical evaluation and scales (providing stroke severity) or neuroimaging (providing accurate size of the lesion) are more reliable predictors for clinical outcome prediction. Read more about this study
- Who is it for?
- Ages 18 years and older
- Conditions the study is for
- Stroke, TIA, Stroke-mimics, Stroke Biomarkers
- What is being tested?
- Extracellular vescicles (EV) analysis
- How many people
- 200 participants
- Where
- 1 siteSwitzerland
- Study dates
- 18 Nov 2022to 31 Jan 2026
- Phase
- Observational
- Are there results?
- None posted in the registry
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 years and older
- All sexes
- No healthy volunteers
- Adult, Older adult
Study population: We will enroll all patients admitted to the stroke Units because of symptoms suggesting a cerebrovascular syndrome
Who may be able to join
- Affected by:
- Suspect ischemic minor stroke (acute loss of focal cerebral function of presumed vascular etiology and NIHSS ≤327, 28) within 48 hours of symptoms onset or II. TIA (defined as the acute onset of focal neurological symptoms lasting <24 h and presumed to be caused by brain ischemia at the time of referral)19 III. Symptoms mimicking I. or II. with but with high suspition of a non-ischemic condition being the cause of the event;
- 2) age ≥18 years; 3) if cerebral magnetic resonance imaging will be performed within 48-72 hours from admission; 4) if baseline CT/MRI is without hemorrhage; 5) if informed consent obtained.
Who cannot join
- ocular TIA (amaurosis fugax);
- contraindications for MRI;
- pregnancy;
- concomitant acute/chronic inflammatory disease (e.g., infections, autoimmune disease);
- concomitant hematological diseases;
- Central Nervous System infection within 30 days;
- serious head trauma within 30 days;
- major surgery within 90 days.
Trial locations and contacts
1 site in 1 country.
Switzerland1 site, 1 recruiting
- Recruiting
Neurocenter of Southern Switzerland, Ospedale Civico
Lugano, Canton Ticino, CH-6900
Carlo Cereda, MD, +41 91 811 6691, carlo.cereda@eoc.ch
Carlo Cereda
About this study
Clinical implication of eventual blood biomarkers for stroke diagnosis and prognosis would be limited, mainly because clinical evaluation and scales (providing stroke severity) or neuroimaging (providing accurate size of the lesion) are more reliable predictors for clinical outcome prediction. In clinical practice, it would be more useful to find a biomarker, which can help to orientate the physician in conditions in which the clinical picture and imaging provide a limited support.
Transient Ischemic Attacks (TIAs) represent a classical example for which a biomarker would be of interest to confirm and distinguish a brain ischemic process from a stroke mimic. Diagnostic biomarkers of TIA have been investigated, but none of the potential candidates reached enough accuracy for TIA diagnosis. Our group has found that Extracellular Vesicles (EVs) could be useful as biomarkers for detecting brain ischemia in patients with TIA because the EV-surface antigen profile appears to be different in patients with transient symptoms, adjudicated to be very likely caused by brain ischemia, compared to patients whose symptoms were less likely to due to brain ischemia. Our study has raised interest in the scientific community recognizing the promising role of of blood-derived EVs analysis in expanding the possibilities to correctly diagnose and classify TIA and stroke events, discriminate them from TIA or stroke mimics, with important future implications in management and therapy of the patients with acute ischemic cerebrovascular syndrome. the validity of our approach needs to be tested in a larger, prospective, multicenter study.
Read the detailed description
Characterization of extracellular vesicles Extracellular vesicles (EVs) will be characterized in pre-cleared serum samples (see transfer and managements of blood samples). EVs will be characterized by nanoparticle tracking analysis, western blot, bead-based flow cytometry (direct and inverse approach), and ELISA.
We will evaluate diameter and concentration of serum nanoparticles (NPs) by nanoparticle tracking analysis (NTA) using NanoSight LM10 (Malvern Instruments, United Kingdom) equipped with a 405 nm laser and Nanoparticle Tracking Analysis NTA 2.3 analytic software. A camera will record Brownian movement of EVs and size and number of EVs per mL will be calculated by Stokes-Einstein equation. Three videos of 60 s will be recorded for each analysis.
Western Blot analysis will be performed on protein lysate after EV immuno-capture (using beads covered with antibodies against EV specific tetraspanins, CD9, CD63 and CD81). Blots of representative samples will be incubated with primary antibodies against specific markers expressed on the surface or within EVs (TSG101, Syntenin-1, CD81, Alix) and potential contaminants (GRP94, Apolipoprotein A1, Apolipoprotein B48).
EV profiling will be performed using a standardized multiplex bead-based flow cytometric assay, using MACSPlex Human Exosome Kit (Miltenyi Biotec; Bergisch Gladbach, Germany), as previously described. 22 The detection of 37 different surface antigens commonly expressed on EV membrane (including markers from activated platelets, endothelium, and inflammatory cells) will be simultaneously performed. Background will be subtracted from the MFI value of each marker, and then normalized using by the mean fluorescence intensity levels (MFI) of CD9, CD63, and CD81. Levels of expression will be reported as normalized MFI (nMFI; %) for each EV-surface antigen (median value and interquartile range).
After an initial screening by multiplexed assay, single candidate EV biomarkers will be confirmed by either reverse flow cytometry and ELISA techniques.
The reverse flow cytometric assay will be performed by isolating EVs by capture beads coated with antibodies against CD9-CD63-CD81 (EpCam; JSR Micro) and then incubated with fluorochrome-conjugated antibodies against single EV surface antigens, associated to patient diagnosis during the initial screening by MACSPlex assay. Beads incubated with isotype-matched antibodies, and beads incubated in absence of vesicles will serve as controls. Candidate EV biomarkers will be confirmed also by ELISA technique, using specific antibodies as per manufacturer's instructions.
- Being tested
- Extracellular vescicles (EV) analysis
- Medical subject headings
- Stroke, Ischemic Attack, Transient
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
Extracellular vescicles (EV) analysisDiagnostic test
The patients undergo a blood sample for EV analyses: EVs will be characterized by nanoparticle tracking analysis, western blot, bead-based flow cytometry (direct and inverse approach), and ELISA.
We will evaluate diameter and concentration of serum nanoparticles (NPs) by nanoparticle tracking analysis (NTA) using NanoSight LM10. Western Blot analysis will be performed on protein lysate after EV immuno-capture (using beads covered with antibodies against EV specific tetraspanins, CD9, CD63 and CD81). EV profiling will be performed using a standardized multiplex bead-based flow cytometric assay, using MACSPlex Human Exosome Kit (Miltenyi Biotec; Bergisch Gladbach, Germany), as previously described. 22 The detection of 37 different surface antigens commonly expressed on EV membrane (including markers from activated platelets, endothelium, and inflammatory cells) will be simultaneously performed.
| Study group | Role | What they receive |
|---|---|---|
| Stroke | Not described Receives: Diagnostic Test: Extracellular vescicles (EV) analysis | |
| TIA | Not described Receives: Diagnostic Test: Extracellular vescicles (EV) analysis | |
| Stroke Mimic | Not described Receives: Diagnostic Test: Extracellular vescicles (EV) analysis |
What is being measured
The primary outcome is the main question the study is designed to answer.
TIA differentiation from TIA mimics
determine the performance of EV profiling (analysis of antigens expressed on vesicles surface), added to a structured clinical (PREDISC score) and imaging evaluation (brain MRI), to discriminate TIAs from TIA mimics (non ischemic events).
Measured: 72 hours from symptoms onset
Secondary outcomes (2)
TIA differentiation from Stroke
determine the performance of EV profiling to differentiate between TIAs (with negative MRI for acute lesion) and acute ischemic strokes (AIS, MRI confirmed).
Measured: acute phase, 3 and 12 months
Stroke etiology
determine the performance of EVs profiling to categorize pre-specified causes of ischemic events
Measured: acute phase, 3 and 12 months
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted20 Mar 2024
- Study start18 Nov 2022
- Primary completion31 Dec 2025 (estimated)
- Study completion31 Jan 2026 (estimated)
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Other trials for Stroke, TIA, Stroke-mimics, Stroke BiomarkersStudies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Cohort
- Time perspective
- Prospective
- Enrolment
- 200 participants (estimated)
Sponsor and investigators
- Lead sponsor
- Ospedale Civico, Lugano
- Sponsor type
- Other government
- Responsible party
- Principal investigator, Dr. med. Carlo Cereda, Ospedale Civico, Lugano
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT06319742
- Sponsor's protocol ID
- NSISTRCR_2201
- Acronym
- ElViS-ACS
- Status last verified
- Apr 2024
- First submitted
- 13 Mar 2024
- Last updated
- 18 Apr 2024
- Results posted
- No
- Data monitoring committee
- No
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- No
Study at a glance
From the registry record, last updated by the sponsor on 18 Apr 2024. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Extracellular Vesicle Surface Markers In Acute Cerebrovascular Syndromes.
- ClinicalTrials.gov ID
- NCT06319742
- Acronym / sponsor ID
- ElViS-ACS / NSISTRCR_2201
- Study type
- Observational
- Recruitment status
- Unknown status
- Enrolment
- 200 participants (estimated)
- Lead sponsor
- Ospedale Civico, LuganoOther government
- Study start
- 18 Nov 2022 (actual)
- Primary completion
- 31 Dec 2025 (estimated)
- Study completion
- 31 Jan 2026 (estimated)
- First posted
- 20 Mar 2024 (actual)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
Clinical implication of eventual blood biomarkers for stroke diagnosis and prognosis would be limited, mainly because clinical evaluation and scales (providing stroke severity) or neuroimaging (providing accurate size of the lesion) are more reliable predictors for clinical outcome prediction. In clinical practice, it would be more useful to find a biomarker, which can help to orientate the physician in conditions in which the clinical picture and imaging provide a limited support.
Transient Ischemic Attacks (TIAs) represent a classical example for which a biomarker would be of interest to confirm and distinguish a brain ischemic process from a stroke mimic. Diagnostic biomarkers of TIA have been investigated, but none of the potential candidates reached enough accuracy for TIA diagnosis. Our group has found that Extracellular Vesicles (EVs) could be useful as biomarkers for detecting brain ischemia in patients with TIA because the EV-surface antigen profile appears to be different in patients with transient symptoms, adjudicated to be very likely caused by brain ischemia, compared to patients whose symptoms were less likely to due to brain ischemia. Our study has raised interest in the scientific community recognizing the promising role of of blood-derived EVs analysis in expanding the possibilities to correctly diagnose and classify TIA and stroke events, discriminate them from TIA or stroke mimics, with important future implications in management and therapy of the patients with acute ischemic cerebrovascular syndrome. the validity of our approach needs to be tested in a larger, prospective, multicenter study.
Detailed description
Characterization of extracellular vesicles Extracellular vesicles (EVs) will be characterized in pre-cleared serum samples (see transfer and managements of blood samples). EVs will be characterized by nanoparticle tracking analysis, western blot, bead-based flow cytometry (direct and inverse approach), and ELISA.
We will evaluate diameter and concentration of serum nanoparticles (NPs) by nanoparticle tracking analysis (NTA) using NanoSight LM10 (Malvern Instruments, United Kingdom) equipped with a 405 nm laser and Nanoparticle Tracking Analysis NTA 2.3 analytic software. A camera will record Brownian movement of EVs and size and number of EVs per mL will be calculated by Stokes-Einstein equation. Three videos of 60 s will be recorded for each analysis.
Western Blot analysis will be performed on protein lysate after EV immuno-capture (using beads covered with antibodies against EV specific tetraspanins, CD9, CD63 and CD81). Blots of representative samples will be incubated with primary antibodies against specific markers expressed on the surface or within EVs (TSG101, Syntenin-1, CD81, Alix) and potential contaminants (GRP94, Apolipoprotein A1, Apolipoprotein B48).
EV profiling will be performed using a standardized multiplex bead-based flow cytometric assay, using MACSPlex Human Exosome Kit (Miltenyi Biotec; Bergisch Gladbach, Germany), as previously described. 22 The detection of 37 different surface antigens commonly expressed on EV membrane (including markers from activated platelets, endothelium, and inflammatory cells) will be simultaneously performed. Background will be subtracted from the MFI value of each marker, and then normalized using by the mean fluorescence intensity levels (MFI) of CD9, CD63, and CD81. Levels of expression will be reported as normalized MFI (nMFI; %) for each EV-surface antigen (median value and interquartile range).
After an initial screening by multiplexed assay, single candidate EV biomarkers will be confirmed by either reverse flow cytometry and ELISA techniques.
The reverse flow cytometric assay will be performed by isolating EVs by capture beads coated with antibodies against CD9-CD63-CD81 (EpCam; JSR Micro) and then incubated with fluorochrome-conjugated antibodies against single EV surface antigens, associated to patient diagnosis during the initial screening by MACSPlex assay. Beads incubated with isotype-matched antibodies, and beads incubated in absence of vesicles will serve as controls. Candidate EV biomarkers will be confirmed also by ELISA technique, using specific antibodies as per manufacturer's instructions.
Study design
- Observational model
- Cohort
- Time perspective
- Prospective
- Number of arms
- 3
| Arm | Type | Receives |
|---|---|---|
| Stroke | – | Diagnostic Test: Extracellular vescicles (EV) analysis |
| TIA | – | Diagnostic Test: Extracellular vescicles (EV) analysis |
| Stroke Mimic | – | Diagnostic Test: Extracellular vescicles (EV) analysis |
Who may qualify
- Ages
- 18 years and older
- Sex
- All
- Healthy volunteers
- Not accepted
- Age groups
- Adult, Older adult
- Study population
- We will enroll all patients admitted to the stroke Units because of symptoms suggesting a cerebrovascular syndrome
Key inclusion criteria (3 listed)
- Suspect ischemic minor stroke (acute loss of focal cerebral function of presumed vascular etiology and NIHSS ≤327, 28) within 48 hours of symptoms onset or II. TIA (defined as the acute onset of focal neurological symptoms lasting <24 h and presumed to be caused by brain ischemia at the time of referral)19 III. Symptoms mimicking I. or II. with but with high suspition of a non-ischemic condition being the cause of the event;
- 2) age ≥18 years; 3) if cerebral magnetic resonance imaging will be performed within 48-72 hours from admission; 4) if baseline CT/MRI is without hemorrhage; 5) if informed consent obtained.
Key exclusion criteria (8 listed)
- ocular TIA (amaurosis fugax);
- contraindications for MRI;
- pregnancy;
- concomitant acute/chronic inflammatory disease (e.g., infections, autoimmune disease);
- concomitant hematological diseases;
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Interventions
| Intervention | Type | Arms |
|---|---|---|
| Extracellular vescicles (EV) analysisThe patients undergo a blood sample for EV analyses: EVs will be characterized by nanoparticle tracking analysis, western blot, bead-based flow cytometry (direct and inverse approach), and ELISA. We will evaluate diameter and concentration of serum nanoparticles (NPs) by nanoparticle tracking analysis (NTA) using NanoSight LM10. Western Blot analysis will be performed on protein lysate after EV immuno-capture (using beads covered with antibodies against EV specific tetraspanins, CD9, CD63 and CD81). EV profiling will be performed using a standardized multiplex bead-based flow cytometric assay, using MACSPlex Human Exosome Kit (Miltenyi Biotec; Bergisch Gladbach, Germany), as previously described. 22 The detection of 37 different surface antigens commonly expressed on EV membrane (including markers from activated platelets, endothelium, and inflammatory cells) will be simultaneously performed. | Diagnostic test | Stroke, Stroke Mimic, TIA |
Primary outcome
TIA differentiation from TIA mimics
Time frame: 72 hours from symptoms onset
determine the performance of EV profiling (analysis of antigens expressed on vesicles surface), added to a structured clinical (PREDISC score) and imaging evaluation (brain MRI), to discriminate TIAs from TIA mimics (non ischemic events).
2 secondary outcomes
TIA differentiation from Stroke
Time frame: acute phase, 3 and 12 months
Stroke etiology
Time frame: acute phase, 3 and 12 months
Results and safety
Results are not available in the source record.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 18 Apr 2024.
- Individual participant dataSharing planned: No
Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part. This study is not recruiting at the moment.
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
An observational study of Stroke, TIA, Stroke-mimics, Stroke Biomarkers, run by Ospedale Civico, Lugano at 1 site in Switzerland.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 3 inclusion and 8 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: status not confirmed, as of the sponsor's last update on 18 Apr 2024. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in Switzerland. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
Results are not available in the source record.
How can a doctor refer a patient or contact the research team?
Contact Carlo Walter Cereda, PI (+41918116691, carlo.cereda@eoc.ch), or a site near the patient. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
TIA differentiation from TIA mimics (time frame: 72 hours from symptoms onset). It is what the study was designed to measure, not a finding.
Where can I read the original study information?
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 18 Apr 2024.