Platelet Glycoproteins in Inherited Thrombocytopathy: Association With Aggregation Studies and Bleeding Severity

• ClinicalTrials.gov Identifier: NCT03648190
• Recruitment Status: Recruiting
• First Posted: August 27, 2018
• Last Update Posted: August 28, 2018
• Sponsor: Assiut University
• Information provided by (Responsible Party): Mohammed Ashraf, Assiut University

Tracking Information

• First Submitted Date: August 21, 2018
• First Posted Date: August 27, 2018
• Last Update Posted Date: August 28, 2018
• Estimated Study Start Date: December 1, 2018
• Estimated Primary Completion Date: December 30, 2019 (Final data collection date for primary outcome measure)

Current Primary Outcome Measures (submitted: August 24, 2018)

Establishment of a protocol for Flowcytometry device to be used in routinely diagnosing cases with inherited platelets function defects. [ Time Frame: 6 Weeks ]

Determine the ability of flowcytometry to assay (determine the levels of expression of each surface marker in percent) platelets surface glycoproteins (CD 41, CD 61 and CD 42b) using BD FACSCalibur flowcytometry instrument in patients with inherited thrombocytopathies and its correlation with bleeding severity of these patients with establishment of a protocol for it to be used in routinely diagnosing these cases.

Original Primary Outcome Measures (submitted: August 23, 2018)

Establishment of a protocol for Flowcytometry device to be used in routinely diagnosing cases with inherited platelets function defects. [ Time Frame: 6 Weeks ]

Determine the ability of flowcytometry to assay platelets surface glycoproteins (CD 41, CD 61 and CD 42b) using BD FACSCalibur flowcytometry instrument in patients with inherited thrombocytopathies and its correlation with bleeding severity of these patients with establishment of a protocol for it to be used in routinely diagnosing these cases.

• Change History: Complete list of historical versions of study NCT03648190 on ClinicalTrials.gov Archive Site
• Current Secondary Outcome Measures: Not Provided
• Original Secondary Outcome Measures: Not Provided
• Current Other Pre-specified Outcome Measures: Not Provided
• Original Other Pre-specified Outcome Measures: Not Provided

Descriptive Information

• Brief Title: Platelet Glycoproteins in Inherited Thrombocytopathy: Association With Aggregation Studies and Bleeding Severity
• Official Title: Evaluation of Platelet Surface Glycoproteins in Patients With Inherited Thrombocytopathy: Association With Aggregation Studies and Bleeding Severity

Brief Summary

Disorders of platelet function are characterized by variable mucocutaneous bleeding manifestations and excessive hemorrhage following surgical procedures or trauma. Generally, most patients have mild to moderate bleeding manifestations with a prolonged bleeding time.

Platelet aggregation and secretion studies using platelet-rich plasma (PRP) provide evidence for platelet dysfunction but are neither predictive of severity of clinical manifestations nor the molecular mechanisms.

Glanzmann's thrombasthenia (GT) is a rare autosomal recessive genetic bleeding syndrome characterized by defects in platelet aggregometry. The clinical phenotype of patients with GT is variable. Some suffer from severe bleeding, while others have only mild bleeding. Some studies found bleeding severity in GT was influenced by the abundance and functioning of platelet receptors involved in aggregation and adhesion.

In addition to a complete medical history, a GT diagnosis requires a comprehensive laboratory workup, including platelet aggregation analysis, and a confirmation by flowcytometry or western blotting with monoclonal antibodies that recognize the GPIIb/IIIa complex.

Platelet flow cytometry is an emerging tool in diagnostic and therapeutic hematology. It is eminently suited to study the expression of platelet surface receptors both qualitatively as well as quantitatively.

Aim of the study:-

• Determine the role of flowcytometry as a quantitative measurement tool of platelets surface glycoproteins in patients with inherited thrombocytopathies and its correlation with bleeding severity of these patients.
• To compare the efficacy, advantages and disadvantages between platelets flowcytometry and aggregometer in diagnosing various inherited thrombocytopathies.

Detailed Description

This study is a case-control observational study to be done at Clinical pathology department, Assiut University hospital, Assiut University, Egypt.

Patients with inherited qualitative platelets defect with clinical manifestations in the form of mucocutaneous bleeding or hemorrhage will be included in the study. Individuals with similar age and sex distribution to the patient group will act as controls. Control group shouldn't take medications or anti-platelet drugs for the preceding two weeks. They should have normal platelet count and morphology. Bleeding patients with acquired bleeding, coagulation defects, and those on anti-platelet drugs will be excluded from the study.

All Patients with inherited platelets function disorders will be subjected to:-

I - Careful history and clinical examination data collecting including:

Clinical history of bleeding such as (Sites, Severity and frequency of bleeding, trauma related events, history of surgical procedures, history of menorrhagia in females, history of packed red cell/ platelets transfusion.

II - Family history such as consanguinity, bleeding complications in any parents/ siblings.

III - Grading of bleeding severity: according to World Health Organization (WHO) bleeding assessment scale from grade 1 to grade 4.

The following routine investigations will be done:-

1. Complete blood count (CBC) analysis.
2. Prothrombin time, concentration and international normalized ratio (INR) assay.
3. Activated partial thromboplastin time assay.
4. Fibrinogen level and thrombin time assay.

The following specific investigations:-

1. Platelets aggregation tests by aggregometer using four different agonists (ADP, Collagen, Arachidonic acid and ristocetin) on Bio/Data PAP-8E.
2. Flowcytometry analysis of platelets receptors (CD 41, CD 61 and CD 42b) on BD FACSCalibur flowcytometry instrument.

Data collection and analysis will be done by computer program SPSS version 21 Chicago USA. Data expressed as mean ±SD, mean±SE, number and percentage. Using Manwhitny test to determine the significance for numeric variable and chisquare to determine the significance for non-parametric variable. ROC curve was done to determine the area under curve (AUC), sensitivity and specificity for each marker.

• Study Type: Observational
• Study Design: Observational Model: Case-Control; Time Perspective: Cross-Sectional
• Target Follow-Up Duration: Not Provided
• Biospecimen: Not Provided
• Sampling Method: Probability Sample
• Study Population: Bleeding patients suffering from inherited qualitative platelets defect, with the exclusion of coagulation defects, acquired bleeding disorders and anti-platelet drugs intake. Both male and females are eligible with no age limits.

Condition

• Bleeding
• Platelet Dysfunction

Intervention

Diagnostic Test: Flowcytometry analysis of platelets surface receptors

BD FACSCalibur flowcytometry instrument.

Platelet flow cytometry is an emerging tool in diagnostic and therapeutic hematology. It is eminently suited to study the expression of platelet surface receptors both qualitatively as well as quantitatively.

Flow cytometry rapidly measures the specific characteristics of a large number of cells in suspension. Typically, the cells are labeled with fluorescently conjugated monoclonal antibodies (mAbs).

Other Name: Platelets aggregation tests by aggregometer

Study Groups/Cohorts

• Inherited qualitative platelets defect

  Clinical manifestations in the form of mucocutaneous bleeding or hemorrhage.

  Bleeding patients with acquired bleeding disorders, coagulation defects, and those on antiplatelet drugs will be excluded from the study.

  Intervention: Diagnostic Test: Flowcytometry analysis of platelets surface receptors
• Control
  Normal healthy participants, with no manifestations of bleeding disorders.
  Intervention: Diagnostic Test: Flowcytometry analysis of platelets surface receptors

Publications *

• Zhou L, Jiang M, Shen H, You T, Ding Z, Cui Q, Ma Z, Yang F, Xie Z, Shi H, Su J, Cao L, Lin J, Yin J, Dai L, Wang H, Wang Z, Yu Z, Ruan C, Xia L. Clinical and molecular insights into Glanzmann's thrombasthenia in China. Clin Genet. 2018 Aug;94(2):213-220. doi: 10.1111/cge.13366. Epub 2018 May 22.
• Harrison P. Assessment of platelet function in the laboratory. Hamostaseologie. 2009 Jan;29(1):25-31.
• Saboor M, Moinuddin M, Ilyas S. New horizons in platelets flow cytometry. Malays J Med Sci. 2013 Mar;20(2):62-6.

Recruitment Information

• Recruitment Status: Recruiting
• Estimated Enrollment (submitted: August 23, 2018): 50
• Original Estimated Enrollment: Same as current
• Estimated Study Completion Date: August 30, 2020
• Estimated Primary Completion Date: December 30, 2019 (Final data collection date for primary outcome measure)

Eligibility Criteria

Inclusion Criteria:

• Bleeding patients.

Exclusion Criteria:

• Coagulation defects, and those on anti-platelet drugs.

Sex/Gender

• Sexes Eligible for Study: All

• Ages: Child, Adult, Older Adult
• Accepts Healthy Volunteers: Yes

Contacts

Contact: Mohammed Ashraf, M.Sc; +201002867611; M1111a1111@yahoo.com

• Listed Location Countries: Egypt

Administrative Information

• NCT Number: NCT03648190
• Other Study ID Numbers: 17200237
• Has Data Monitoring Committee: No

U.S. FDA-regulated Product

• Studies a U.S. FDA-regulated Drug Product: No
• Studies a U.S. FDA-regulated Device Product: No

IPD Sharing Statement

• Plan to Share IPD: Undecided

• Responsible Party: Mohammed Ashraf, Assiut University
• Study Sponsor: Assiut University
• Collaborators: Not Provided

Investigators

• Principal Investigator: Hanan Galal, MD; Assiut University
• Study Chair: Madleen Adel, MD; Assiut University
• Study Chair: Eman Nasr-Eldin, MD; Assiut University
• Study Director: Mohammed Ashraf, M.Sc; Assiut University

• PRS Account: Assiut University
• Verification Date: August 2018