title: Ineffective erythropoiesis in beta-thalassemia major is due to apoptosis at the polychromatophilic normoblast stage.
abstract: Beta-thalassemia major is characterized by ineffective erythropoiesis, although it is difficult to define the dynamics of this process from the static information revealed by analysis of bone marrow (BM) aspirates. We aimed to study the kinetics of sequential erythroid differentiation in beta-thalassemia major. We isolated the progenitor cells (CD34(+) and CD34(+)CD38(-) cells) from BM of thalassemia major patients and studied in vitro erythropoiesis. This is the first report of an in vitro study in human beta-thalassemia major from purified BM CD34(+) progenitor cells, using erythroid culture conditions, which allow unilineage differentiation to mature enucleated red blood cells. In contrast to normal donors, a high proportion of BM CD34(+) and CD34(+)CD38(-) progenitors from beta-thalassemia major coexpressed the late erythroid lineage-specific protein glycophorin A and generated a higher proportion of erythroid colonies. However, despite the marked increase in erythroid clonogenicity of the progenitor population, erythroid cultures initiated from beta-thalassemia major BM CD34(+) cells expanded 10- to 20-fold less than from normal BM. There were less viable cells during differentiation, specifically after the polychromatophilic normoblast stage. There was a progressive increase in the apoptotic erythroid progeny with differentiation, and apoptosis occurred predominantly at the polychromatophilic normoblast stage. In thalassemia major, BM progenitor cells show increased erythroid clonogenicity, increased expression of late erythroid lineage-specific proteins, and accelerated erythroid differentiation. However, despite the apparent increased erythroid commitment, ineffective erythropoiesis occurs due to apoptosis at the polychromatophil stage. Identification of the differentiation stage at which apoptosis occurs will permit further studies of the underlying mechanisms and target therapeutic strategies to improve red cell production.
authors.affiliation: Division of Hematology/Oncology, Childrens Hospital Los Angeles, Los Angeles, CA 90027, USA.
authors.forename: L A
authors.initials: LA
authors.lastname: Mathias
authors.forename: T C
authors.initials: TC
authors.lastname: Fisher
authors.forename: L
authors.initials: L
authors.lastname: Zeng
authors.forename: H J
authors.initials: HJ
authors.lastname: Meiselman
authors.forename: K I
authors.initials: KI
authors.lastname: Weinberg
authors.forename: A L
authors.initials: AL
authors.lastname: Hiti
authors.forename: P
authors.initials: P
authors.lastname: Malik
mesh_terms: D036541:ADP-ribosyl Cyclase; D051997:ADP-ribosyl Cyclase 1; D000328:Adult; D015703:Antigens, CD; D018952:Antigens, CD34 / Q000032:analysis; D000943:Antigens, Differentiation / Q000032:analysis; D017209:Apoptosis; D001854:Bone Marrow Cells / Q000473:pathology; D002454:Cell Differentiation; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D004906:Erythrocyte Count; D015672:Erythroid Precursor Cells / Q000737:chemistry / Q000378:metabolism / Q000473:pathology*; D004920:Erythropoiesis; D015870:Gene Expression; D006021:Glycophorins / Q000032:analysis / Q000235:genetics; D006801:Humans; D020287:In Situ Nick-End Labeling; D007223:Infant; D008562:Membrane Glycoproteins; D009244:NAD+ Nucleosidase / Q000032:analysis; D017086:beta-Thalassemia / Q000473:pathology*
chemical_list: D015703:Antigens, CD; D018952:Antigens, CD34; D000943:Antigens, Differentiation; D006021:Glycophorins; D008562:Membrane Glycoproteins; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; D009244:NAD+ Nucleosidase; D051997:ADP-ribosyl Cyclase 1