Development and Prospective Validation of a Standardized Flow Cytometric Assay of Peripheral Blood Neutrophil Myeloperoxidase Expression for Ruling Out Myelodysplastic Syndromes.

NCT04399018 | Active Not Recruiting

• 2 other identifiers: 38RC19.4252019-A03341-56
• Study Type: observational
• Target: 103
• Locations: 1 country
• Sites: 1

Brief Summary

Myelodysplastic syndromes (MDS) comprise a heterogeneous group of clonal bone marrow neoplasms that predominate in the elderly, with a median age at diagnosis of 70 years. The diagnosis of MDS relies on peripheral blood cytopenia and morphologic dysplasia for one or more hematopoietic cell lineage. Cytopenia is evidenced with hemogram while dysplasia requires bone marrow aspirate, which is an invasive procedure . Considering the low prevalence of disease among subjects referred for suspected MDS, many patients are exposed to unnecessary bone marrow aspiration-related discomfort and harms. Therefore, an objective assay based on a peripheral blood sample that accurately discriminates MDS from other cytopenia etiologies is highly desirable. We have previously developed and refined a flow cytometric analysis protocol for quantifying neutrophil MPO expression in peripheral blood at three university-affiliated hospitals (i.e., Clermont-Ferrand, Saint-Etienne, and Grenoble) (Raskovalova et al, Hematologica 2019). We found that the robust coefficient of variation (RCV, computed as the robust standard deviation divided by the median) within an individual subject was the best parameter in discriminating patients with versus without MDS. Although promising, flow cytometric analysis of neutrophil MPO expression in peripheral blood is technically complex, time consuming, and not standardized. Hence, its performance requires specific expertise and the results show substantial variability. A single ready-to-use tube with lyophilized antibodies would have the potential to standardize the measurement of neutrophil MPO expression in peripheral blood across laboratories, with results available within 30-60 min in routine practice. In this study, the investigators hypothesize that a standardized and semi-automatic flow cytometric assay of neutrophil MPO expression in peripheral blood could accurately rule out MDS and obviate the need for bone marrow aspiration and biopsy, with sensitivity and negative predictive value estimates approaching 100%. In this observational diagnostic accuracy study, burden will be null for recruited patients. No specific intervention is assigned to participants. All diagnostic testing, procedures, and medication ordering are performed at the discretion of attending physicians. A test result will have no impact on patient management. .Compliance with current guidelines disseminated by the French Haute Autorité de Santé (HAS) will be advocated for the diagnostic work-up of patients with suspected MDS. No follow-up visits are planned in this cross-sectional study.

Conditions

Myelodysplastic Syndromes

Keywords

Diagnostic accuracy study; Flow cytometry analysis; Myeloperoxidase; Peripheral blood sample

Outcome Measures

Primary Outcomes (1)

• Reference diagnosis of MDS or CMML established by bone marrow examination

  The primary outcome is the reference diagnosis of MDS or CMML established by bone marrow examination (with Perls staining) by two independent experienced hematopathologists blinded to the index test results. Disagreements will be solved by a third hematopathologist. Bone marrow aspirate will be repeated within 4 to 6 months for patients with idiopathic cytopenia of uncertain significance (ICUS) or non-conclusive bone marrow examination.

  Baseline

Secondary Outcomes (4)

• Intra-laboratory coefficient of variation for intra-individual RCV, computed as the standard deviation multiplied by 100 and divided by the mean (intra- and inter-assay precision)

  Baseline

• Relative change from baseline, expressed in percentages for intra-individual RCV

  Baseline

• Inter-laboratory coefficient of variation for intra-individual RCV

  Baseline

• Negative predictive value point estimates (along with 95% confidence interval) of neutrophil myeloperoxidase expression in peripheral blood for the diagnosis of MDS or CMML

  Baseline

Interventions

Diagnostic Test: Flow cytometry analysis of neutrophil myeloperoxidase expression OTHER

Flow cytometry analysis of neutrophil myeloperoxidase expression in peripheral blood samples will be performed within 24 h of MDS diagnostic evaluation and blinded to the reference standard. Peripheral blood samples will be collected in 5 ml (EDTA) anticoagulant plastic tubes and processed within 24 h maximum of collection. Blood sample will be stained according to the manufacturers' recommendations with a lyophilized cocktail ("LyotubeTM ready-to-use", BD Bioscience). At least 10,000 neutrophils will be acquired on a 3-laser, 8-color BD FACSCanto-II TM flow cytometer (BD Biosciences, San José, CA). Each marker will be expressed as median, geometric and arithmetic mean, regular and robust coefficient of variation.

Eligibility Criteria

• Age: 18 Years - 100 Years
• Sex: all
• Healthy Volunteers: No
• Age Groups: Adult (18-64), Older Adult (65+)
• Sampling Method: Probability Sample

Study Population

Eligible participants are unselected consecutive adults referred for suspected MDS. Suspicion of MDS is based on medical history and peripheral blood cytopenia. To be eligible, patients will be required to meet all five inclusion criteria and none of the exclusion criteria.

You may qualify if:

• Age at enrollment ≥18 years
• Clinical suspicion of MDS
• Indication for bone marrow examination
• ≥1 peripheral blood cytopenia defined by hemoglobin concentration \<12 g/dL for female and \<13g/dL for male patients, platelet count \<150 x109/L, absolute neutrophil count \<1.8 x109/L
• Inpatient and outpatient care patients

You may not qualify if:

• Refusal to participate
• History of or active documented MDS
• Enrollment in intensive or critical care unit
• Incarcerated or individuals protected by French regulation (Article L1121.5 and following, Code de la Santé Publique)
• Not affiliated with social security system
• Previous enrollment in the study.

Sponsors & Collaborators

• University Hospital, Grenoble: lead

Study Sites (1)

Chu Grenoble Alpes

Grenoble, 38043, France

Location

Related Publications (2)

• Planta C, Scheffen L, Jacob MC, Szymanski G, Chevalier S, Tondeur S, Bulabois B, Meunier M, Lefebvre C, Gonnet N, Garban F, Molina L, Paradis C, Seigneurin A, Chiriac R, Merle R, Labarere J, Park S, Raskovalova T. Flow cytometric lyophilised reagent tube assay for peripheral blood neutrophil myeloperoxidase expression to rule out myelodysplastic neoplasms at a university hospital: a diagnostic accuracy study. BMJ Open. 2025 Aug 22;15(8):e095640. doi: 10.1136/bmjopen-2024-095640.

  PMID: 40846335 DERIVED

• Raskovalova T, Scheffen L, Jacob MC, Chevalier S, Tondeur S, Bulabois B, Meunier M, Szymanski G, Lefebvre C, Planta C, Dumestre-Perard C, Gonnet N, Garban F, Merle R, Park S, Labarere J. Flow cytometry lyophilised-reagent tube for quantifying peripheral blood neutrophil myeloperoxidase expression in myelodysplastic syndromes (MPO-MDS-Develop): protocol for a diagnostic accuracy study. BMJ Open. 2022 Oct 7;12(10):e065850. doi: 10.1136/bmjopen-2022-065850.

  PMID: 36207039 DERIVED

MeSH Terms

Conditions

Myelodysplastic Syndromes

Condition Hierarchy (Ancestors)

Bone Marrow Diseases; Hematologic Diseases; Hemic and Lymphatic Diseases

Study Officials

• Tatiana Raskovalova, MD

  Centre Hospitalier Universitaire Grenoble Alpes

  STUDY CHAIR

• Richard Veyrat-Masson, MD

  Centre Hospitalier Universitaire Clermont Ferrand

  PRINCIPAL INVESTIGATOR

• Carmen Aanei, MD

  Centre Hospitalier Universitaire Saint-Étienne

  PRINCIPAL INVESTIGATOR

Study Design

• Study Type: observational
• Observational Model: COHORT
• Time Perspective: CROSS SECTIONAL
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Study Record Dates

• First Submitted: May 18, 2020
• First Posted: May 22, 2020
• Study Start: July 27, 2020
• Primary Completion: August 1, 2024
• Study Completion: December 1, 2024
• Last Updated: March 19, 2024

Record last verified: 2024-03

Data Sharing

• IPD Sharing: Will share
• Shared Documents: STUDY PROTOCOL, SAP
• Time Frame: No later than 3 years after final acceptance of the primary study paper.
• Access Criteria: De-identified data will be available for individual participant data meta-analysis purpose. Researchers should submit a methodologically sound proposal that complies with the Preferred Reporting Items for Systematic Review and Meta-Analysis Protocols (PRISMA-P) 2015 statement. Proposals should be directed to TRaskovalova@chu-grenoble.fr. Data requestors will need to sign a data access agreement.

Locations

FR (1)