title: Immunophenotypic Profiling of Erythroid Progenitor-Derived Extracellular Vesicles in Diamond-Blackfan Anaemia: A New Diagnostic Strategy.
abstract: Diamond-Blackfan Anaemia (DBA) is a rare inherited anaemia caused by heterozygous mutations in one of 13 ribosomal protein genes. Erythroid progenitors (BFU-E and CFU-E) in bone marrow (BM) show a proapoptotic phenotype. Suspicion of DBA is reached after exclusion of other forms of BM failure syndromes. To improve DBA diagnosis, which is confirmed by mutation analysis, we tested a new approach based on the study of extracellular vesicles (EVs) isolated from plasma by differential centrifugations and analysed by flow cytometry. We chose CD34, CD71 and CD235a markers to study erythroid EVs. We characterised the EVs immunophentoypic profiles of 13 DBA patients, 22 healthy controls and 16 patients with other haematological diseases. Among the three EVs clusters we found, only the CD34+/CD71low population showed statistically significant differences between DBA patients and controls (p< 0.05). The absence of this cluster is in agreement with the low levels of BFU-E found in DBA patients. The assessment of ROC curves demonstrated the potential diagnostic value of this population. We suggest that this assay may be useful to improve DBA diagnosis as a quicker and less invasive alternative to BM BFU-E culture analysis.
authors.affiliation: Department of Health Sciences, University of Eastern Piedmont, Novara, Italy.
authors.forename: Serena
authors.initials: S
authors.lastname: Macrì
authors.affiliation: Department of Health Sciences, University of Eastern Piedmont, Novara, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Pavesi
authors.affiliation: Department of Health Sciences, University of Eastern Piedmont, Novara, Italy.
authors.forename: Rossella
authors.initials: R
authors.lastname: Crescitelli
authors.affiliation: Department of Health Sciences, University of Eastern Piedmont, Novara, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Aspesi
authors.affiliation: Chemical Clinical Analysis laboratory, SCDU, Azienda Universitaria Ospedaliera Maggiore della Carità, Novara, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Vizziello
authors.affiliation: Department of Public Health and Pediatric Sciences, University of Turin, Turin, Italy.
authors.forename: Carlotta
authors.initials: C
authors.lastname: Botto
authors.affiliation: Department of Pediatric Hematology, San Gerardo's Hospital, Monza, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Corti
authors.affiliation: Pediatric Onco-Hematology, Regina Margherita Children's Hospital, Turin, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Quarello
authors.affiliation: Chemical Clinical Analysis laboratory, SCDU, Azienda Universitaria Ospedaliera Maggiore della Carità, Novara, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Notari
authors.affiliation: Department of Public Health and Pediatric Sciences, University of Turin, Turin, Italy.
authors.forename: Ugo
authors.initials: U
authors.lastname: Ramenghi
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Louisville, Louisville, Kentucky, United States of America.
authors.forename: Steven Robert
authors.initials: SR
authors.lastname: Ellis
authors.affiliation: Department of Health Sciences, University of Eastern Piedmont, Novara, Italy.
authors.forename: Irma
authors.initials: I
authors.lastname: Dianzani
mesh_terms: D000293:Adolescent; D000328:Adult; D029503:Anemia, Diamond-Blackfan / Q000097:blood / Q000175:diagnosis* / Q000235:genetics; D015703:Antigens, CD / Q000097:blood; D018952:Antigens, CD34 / Q000097:blood; D015415:Biomarkers / Q000097:blood; D002648:Child; D002675:Child, Preschool; D003114:Colony-Forming Units Assay; D015672:Erythroid Precursor Cells / Q000378:metabolism*; D000067128:Extracellular Vesicles / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry; D006021:Glycophorins / Q000032:analysis; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D007223:Infant; D008297:Male; D008875:Middle Aged; D012372:ROC Curve; D011990:Receptors, Transferrin / Q000097:blood; D055815:Young Adult
chemical_list: D015703:Antigens, CD; D018952:Antigens, CD34; D015415:Biomarkers; C068126:CD71 antigen; D006021:Glycophorins; D011990:Receptors, Transferrin