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CONTRIBUTION OF NEXT GENERATION FLOW (NGF) CYTOMETRY IN PRIMARY IMMUNE THROMBOCYTOPENIA (ITP): UTILITY FOR THE DIFFERENTIAL DIAGNOSIS WITH MYELODYSPLASTIC SYNDROMES
Author(s): ,
Tomas Jose Gonzalez-Lopez
Affiliations:
Department of Hematology,Hospital Universitario de Burgos,Burgos,Espagne;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spanien;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spagna;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spain;Department of Hematology,Hospital Universitario de Burgos,Burgos,España;Department of Hematology,Hospital Unive
,
Sergio Matarraz
Affiliations:
Cancer Research Center (IBMCC, USAL-CSIC), Cytometry Service (NUCLEUS) and Department of Medicine,University of Salamanca,Salamanca,Espagne;Cancer Research Center (IBMCC, USAL-CSIC), Cytometry Service (NUCLEUS) and Department of Medicine,University of Salamanca,Salamanca,Spanien;Cancer Research Center (IBMCC, USAL-CSIC), Cytometry Service (NUCLEUS) and Department of Medicine,University of Salamanc
,
Isabel Caparros
Affiliations:
Department of Hematology,Hospital Virgen de la Victoria,Málaga,Espagne;Department of Hematology,Hospital Virgen de la Victoria,Málaga,Spanien;Department of Hematology,Hospital Virgen de la Victoria,Málaga,Spagna;Department of Hematology,Hospital Virgen de la Victoria,Málaga,Spain;Department of Hematology,Hospital Virgen de la Victoria,Málaga,España;Department of Hematology,Hospital Virgen de la Vi
,
Maria Fernanda Lopez-Fernandez
Affiliations:
Department of Hematology,Complexo Hospitalario Universitario A Coruña,A Coruña,Espagne;Department of Hematology,Complexo Hospitalario Universitario A Coruña,A Coruña,Spanien;Department of Hematology,Complexo Hospitalario Universitario A Coruña,A Coruña,Spagna;Department of Hematology,Complexo Hospitalario Universitario A Coruña,A Coruña,Spain;Department of Hematology,Complexo Hospitalario Universi
,
Maria Eva Mingot-Castellano
Affiliations:
Department of Hematology,Complejo Hospitalario Regional de Málaga,Málaga,Espagne;Department of Hematology,Complejo Hospitalario Regional de Málaga,Málaga,Spanien;Department of Hematology,Complejo Hospitalario Regional de Málaga,Málaga,Spagna;Department of Hematology,Complejo Hospitalario Regional de Málaga,Málaga,Spain;Department of Hematology,Complejo Hospitalario Regional de Málaga,Málaga,España
,
Ana Jimenez Martin
Affiliations:
Department of Hematology,Hospital Universitario Ramón y Cajal,Madrid,Espagne;Department of Hematology,Hospital Universitario Ramón y Cajal,Madrid,Spanien;Department of Hematology,Hospital Universitario Ramón y Cajal,Madrid,Spagna;Department of Hematology,Hospital Universitario Ramón y Cajal,Madrid,Spain;Department of Hematology,Hospital Universitario Ramón y Cajal,Madrid,España;Department of Hemat
,
Fernando Fernandez-Fuertes
Affiliations:
Department of Hematology,Hospital Universitario Insular de Gran Canaria,Las Palmas de Gran Canaria,Espagne;Department of Hematology,Hospital Universitario Insular de Gran Canaria,Las Palmas de Gran Canaria,Spanien;Department of Hematology,Hospital Universitario Insular de Gran Canaria,Las Palmas de Gran Canaria,Spagna;Department of Hematology,Hospital Universitario Insular de Gran Canaria,Las Palm
,
José Ignacio Sánchez Gallego
Affiliations:
Cancer Research Center (IBMCC, USAL-CSIC), Cytometry Service (NUCLEUS) and Department of Medicine,University of Salamanca,Salamanca,Espagne;Cancer Research Center (IBMCC, USAL-CSIC), Cytometry Service (NUCLEUS) and Department of Medicine,University of Salamanca,Salamanca,Spanien;Cancer Research Center (IBMCC, USAL-CSIC), Cytometry Service (NUCLEUS) and Department of Medicine,University of Salamanc
,
Pilar Leoz
Affiliations:
Cancer Research Center (IBMCC, USAL-CSIC), Cytometry Service (NUCLEUS) and Department of Medicine,University of Salamanca,Salamanca,Espagne;Cancer Research Center (IBMCC, USAL-CSIC), Cytometry Service (NUCLEUS) and Department of Medicine,University of Salamanca,Salamanca,Spanien;Cancer Research Center (IBMCC, USAL-CSIC), Cytometry Service (NUCLEUS) and Department of Medicine,University of Salamanc
,
Beatriz Cuevas Ruiz
Affiliations:
Department of Hematology,Hospital Universitario de Burgos,Burgos,Espagne;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spanien;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spagna;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spain;Department of Hematology,Hospital Universitario de Burgos,Burgos,España;Department of Hematology,Hospital Unive
,
Maria Victoria Cuevas Ruiz
Affiliations:
Department of Hematology,Hospital Universitario de Burgos,Burgos,Espagne;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spanien;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spagna;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spain;Department of Hematology,Hospital Universitario de Burgos,Burgos,España;Department of Hematology,Hospital Unive
,
Cristina Martin Marín
Affiliations:
Department of Public Health,Consejeria de Sanidad, Junta de Castilla y León,Valladolid,Espagne;Department of Public Health,Consejeria de Sanidad, Junta de Castilla y León,Valladolid,Spanien;Department of Public Health,Consejeria de Sanidad, Junta de Castilla y León,Valladolid,Spagna;Department of Public Health,Consejeria de Sanidad, Junta de Castilla y León,Valladolid,Spain;Department of Public He
,
Juan Olazabal Herrero
Affiliations:
Department of Hematology,Hospital Universitario de Burgos,Burgos,Espagne;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spanien;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spagna;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spain;Department of Hematology,Hospital Universitario de Burgos,Burgos,España;Department of Hematology,Hospital Unive
,
Pilar de Vicente Cámara
Affiliations:
Department of Hematology,Hospital Universitario de Burgos,Burgos,Espagne;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spanien;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spagna;Department of Hematology,Hospital Universitario de Burgos,Burgos,Spain;Department of Hematology,Hospital Universitario de Burgos,Burgos,España;Department of Hematology,Hospital Unive
Alberto Orfao
Affiliations:
Cancer Research Center (IBMCC, USAL-CSIC), Cytometry Service (NUCLEUS) and Department of Medicine,University of Salamanca,Salamanca,Espagne;Cancer Research Center (IBMCC, USAL-CSIC), Cytometry Service (NUCLEUS) and Department of Medicine,University of Salamanca,Salamanca,Spanien;Cancer Research Center (IBMCC, USAL-CSIC), Cytometry Service (NUCLEUS) and Department of Medicine,University of Salamanc
(Abstract release date: 05/12/22) EHA Library. Jose Gonzalez-Lopez T. 06/10/22; 358504; P1647
Tomas Jose Gonzalez-Lopez
Tomas Jose Gonzalez-Lopez
Contributions
Abstract
Presentation during EHA2022: All (e)Poster presentations will be made available as of Friday, June 10, 2022 (09:00 CEST) and will be accessible for on-demand viewing until Monday, August 15, 2022 on the Congress platform.

Abstract: P1647

Type: Poster presentation

Session title: Platelet disorders

Background
Primary immune thrombocytopenia (ITP) is an immune-mediated acquired disorder characterized by impaired production and increased destruction of platelets with an elevated risk of bleeding. At present, diagnosis of primary ITP still remains one of exclusion with a need to discard other causes of isolated thrombocytopenia, in the absence of robust and accurate clinical and laboratory diagnostic criteria. For the diagnosis of ITP, a bone marrow (BM) study may be useful to differentiate between ITP and other diseases, such as myelodysplastic syndromes (MDS). Next generation flow (NGF) has emerged as a potential useful tool in these settings.

Aims
In FCR-PTI-2017-01 study we prospectively evaluated the potential utility of NGF analysis of BM for more accurate diagnosis of ITP vs MDS.

Methods
Multicenter, prospective, Spanish study to investigate morphological and/or immunophenotypic profiles among BM cell compartments from patients ≥ 18 years with newly diagnosed ITP or MDS. A multiparameter characterization of multiple compartments of BM cells was performed using the 8-color panel designed and validated by the EuroFlow Consortium for the diagnosis and characterization of MDS and acute myeloid leukaemia. NGF BM blinded results were then compared with the BM cytomorphological diagnosis.

Results
62 patients, median age 71 years, 59% male, were included. Main comorbidities were neoplasms (19%), diabetes (17%), chronic respiratory disease (12%) and cardiac insufficiency (10%). Median platelet count at diagnosis was 45 x 109/µL (range: 4-98 x 109/µL) and 20.6% of patients had bleeding at diagnosis. By cytomorphology, expert hematologists were able to conclude an ITP or MDS diagnosis in only 31 cases of the 53 samples that could be analyzed (58.5%) [ITP (n=18), MDS (n=10) or inconclusive, i.e. unclassifiable, (n= 25)]. 62 BM samples evaluated by NGF were allocated in 4 immunophenotypic groups attending to: maturation blockades, abnormal antigen expression and cross lineage markers. We observed normal phenotype cases (n=10), isolated (unilineage) alterations (n=24), mild multilineage (>1) alterations (n=20) and severe multilineage (MDS-like) phenotypes (n=8). Cytomorphology diagnosed our cases as ITP, MDS or unclassifiable with a median number of alterations observed by BM NGF of 4 (IQR, 3-6), 2 (IQR, 1-6) and 4 (IQR, 3-5) respectively. For ITP cytomorphology group, NGF demonstrated numerous BM alterations being monocytic alterations (n=17, 94%) the most frequent finding observed. MDS presumed cases were also associated with monocytic alterations (n=5, 50%) with a frequent decrease in neutrophil precursors (n=4, 40%). On the contrary, when cytomorphology was not capable to establish a diagnosis, NGF showed a mixture of alterations with no clear predominance of none of them (table 1). 

Conclusion
Limitations of BM cytomorphology when facing an isolated thrombocytopenia were demonstrated here. However, normal or unilineage BM NGF alterations may lead us to an ITP diagnosis while mild or severe multilineage BM phenotypes may correlate good with MDS.

Keyword(s): Cytometry, Diagnosis, Immune thrombocytopenia (ITP), Myelodysplastic syndrome

Presentation during EHA2022: All (e)Poster presentations will be made available as of Friday, June 10, 2022 (09:00 CEST) and will be accessible for on-demand viewing until Monday, August 15, 2022 on the Congress platform.

Abstract: P1647

Type: Poster presentation

Session title: Platelet disorders

Background
Primary immune thrombocytopenia (ITP) is an immune-mediated acquired disorder characterized by impaired production and increased destruction of platelets with an elevated risk of bleeding. At present, diagnosis of primary ITP still remains one of exclusion with a need to discard other causes of isolated thrombocytopenia, in the absence of robust and accurate clinical and laboratory diagnostic criteria. For the diagnosis of ITP, a bone marrow (BM) study may be useful to differentiate between ITP and other diseases, such as myelodysplastic syndromes (MDS). Next generation flow (NGF) has emerged as a potential useful tool in these settings.

Aims
In FCR-PTI-2017-01 study we prospectively evaluated the potential utility of NGF analysis of BM for more accurate diagnosis of ITP vs MDS.

Methods
Multicenter, prospective, Spanish study to investigate morphological and/or immunophenotypic profiles among BM cell compartments from patients ≥ 18 years with newly diagnosed ITP or MDS. A multiparameter characterization of multiple compartments of BM cells was performed using the 8-color panel designed and validated by the EuroFlow Consortium for the diagnosis and characterization of MDS and acute myeloid leukaemia. NGF BM blinded results were then compared with the BM cytomorphological diagnosis.

Results
62 patients, median age 71 years, 59% male, were included. Main comorbidities were neoplasms (19%), diabetes (17%), chronic respiratory disease (12%) and cardiac insufficiency (10%). Median platelet count at diagnosis was 45 x 109/µL (range: 4-98 x 109/µL) and 20.6% of patients had bleeding at diagnosis. By cytomorphology, expert hematologists were able to conclude an ITP or MDS diagnosis in only 31 cases of the 53 samples that could be analyzed (58.5%) [ITP (n=18), MDS (n=10) or inconclusive, i.e. unclassifiable, (n= 25)]. 62 BM samples evaluated by NGF were allocated in 4 immunophenotypic groups attending to: maturation blockades, abnormal antigen expression and cross lineage markers. We observed normal phenotype cases (n=10), isolated (unilineage) alterations (n=24), mild multilineage (>1) alterations (n=20) and severe multilineage (MDS-like) phenotypes (n=8). Cytomorphology diagnosed our cases as ITP, MDS or unclassifiable with a median number of alterations observed by BM NGF of 4 (IQR, 3-6), 2 (IQR, 1-6) and 4 (IQR, 3-5) respectively. For ITP cytomorphology group, NGF demonstrated numerous BM alterations being monocytic alterations (n=17, 94%) the most frequent finding observed. MDS presumed cases were also associated with monocytic alterations (n=5, 50%) with a frequent decrease in neutrophil precursors (n=4, 40%). On the contrary, when cytomorphology was not capable to establish a diagnosis, NGF showed a mixture of alterations with no clear predominance of none of them (table 1). 

Conclusion
Limitations of BM cytomorphology when facing an isolated thrombocytopenia were demonstrated here. However, normal or unilineage BM NGF alterations may lead us to an ITP diagnosis while mild or severe multilineage BM phenotypes may correlate good with MDS.

Keyword(s): Cytometry, Diagnosis, Immune thrombocytopenia (ITP), Myelodysplastic syndrome

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