Endothelial Colony-Forming Cells in Patients With VWD, AVWS and Healthy Subjects
Trial statusRecruiting
Trial phaseNot listed
Trial typeObservational
Biological sexAll
Age16+
SponsorFondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico
About this trial
The goal of this observational study is to learn how endothelial colony-forming cells (ECFCs) behave in people with von Willebrand disease (VWD), acquired von Willebrand syndrome (AVWS), and in healthy individuals.
Eligibility criteria
This trial accepts healthy volunteersQualifiers
No prior diagnosis of VWD, bleeding disorders, or thrombotic disorders.
Willingness to donate blood for study procedures.
Ability and willingness to provide written informed consent.
Age ≥ 18 years.
Disqualifiers
Pregnancy.
Anemia, as determined at screening or based on medical history.
Trial population
The study will include the enrollment of subjects with a previous diagnosis of VWD or AVWS and a group of healthy volunteers (controls). Patients will be selected among those referred at Angelo Bianchi Bonomi Haemophilia and Thrombosis Center, with a previous diagnosis of VWD or AVWS. Healthy volunteers, defined as individuals without a previous diagnosis of bleeding or thrombotic disorders will be enrolled in equal number to cases with the aim of creating a ECFCs reference group. At enrollment, controls will undergo an interview to confirm their health status, they will be asked to sign the informed consent and then they will undergo a blood withdrawal necessary to confirm plasma VWF levels and to isolate ECFCs as already described for patients.
Trial design
Design model
Cohort
Time perspective
Cross-sectional
Treatments tested in this trial
Blood sample collection for VWF measurements in plasma
Diagnostic testPlasma samples will be collected for the measurement of VWF levels.
Blood sample collection for ECFC isolation and characterization
Other interventionBlood samples will be collected to isolate ECFCs and perform their subsequent characterization.
Genetic testing of VWF
Other interventionAn additional blood sample will be collected for VWF genetic testing in patients with VWD without prior molecular characterization.
Treatment groups
48 Participants
are divided into 2 treatment groupsGroup A: von willebrand disease/ Acquired von Willebrand disease patients3 interventions
Group B: Healthy donors2 interventions
Trial outcomes
Primary outcomes
1
ECFCs isolation and characterization
Assessment of differences between ECFCs derived from patients with VWD or AVWS and healthy controls. Basal and stimulated VWF production and secretion by ECFCs, including basal VWF levels in culture media and cell lysates and stimulated VWF release following exposure to biological or chemical compounds, measured by ELISA (IU/dL) and compared between patients with VWD/AVWS and healthy controls. VWF mRNA expression levels in ECFCs, assessed by quantitative real-time PCR and expressed as ΔΔCt, compared between patients with VWD/AVWS and healthy controls.
Time frame
42 weeks from enrollment startSecondary outcomes
1
Correlation of ECFC VWF Antigen Protein and mRNA Levels with Plasma and Platelet VWF Antigen/Activity Levels and Molecular Defect.
Correlation analyses between ECFC-derived VWF parameters and circulating VWF levels in corresponding patients with VWD, AVWS, and healthy controls. ECFC-derived VWF antigen levels in culture media and cell lysates (IU/dL), measured by ELISA, and VWF mRNA expression levels in ECFCs (ΔΔCt), assessed by quantitative real-time PCR, will be correlated with plasma and platelet VWF antigen and activity levels (IU/dL). All ECFC-derived and circulating VWF parameters will additionally be analyzed in relation to the specific underlying molecular defect.
Time frame
42 weeks from enrollment start2
Comparison of angiogenic properties of ECFCs from patients with VWD/AVWS and healthy controls
Angiogenic properties of ECFCs will be assessed using an in vitro Matrigel tube formation assay and compared between patients with type 2 or type 3 VWD or AVWS and healthy controls. Quantitative parameters will include total tube length (μm), number of tubes, number of branches, and number of meshes.
Time frame
42 weeks from enrollment start3
Evaluation of Allele-Specific siRNA to Restore Normal VWF Levels in Type 2A VWD
Allele-specific small interfering RNA (siRNA) will be evaluated in endothelial ECFCs derived from a patient with type 2A VWD to assess whether selective inhibition of the mutant allele restores normal VWF expression and secretion. All analyses will be performed before and after siRNA transfection. The following parameters will be assessed: VWF mRNA expression levels, assessed by quantitative real-time PCR and expressed as ΔΔCt (relative expression); VWF protein levels in ECFC culture media, measured by ELISA (IU/mL).
Time frame
42 weeks from enrollment startSponsors and contacts
Click on the lead sponsor to view all of their trials.
Source ClinicalTrials.gov