title: Comparison of the Proteome of Adult and Cord Erythroid Cells, and Changes in the Proteome Following Reticulocyte Maturation.
abstract: Cord blood stem cells are an attractive starting source for the production of red blood cells in vitro for therapy because of additional expansion potential compared with adult peripheral blood progenitors and cord blood banks usually being more representative of national populations than blood donors. Consequently, it is important to establish how similar cord RBCs are to adult cells. In this study, we used multiplex tandem mass tag labeling combined with nano-LC-MS/MS to compare the proteome of adult and cord RBCs and reticulocytes. 2838 unique proteins were identified, providing the most comprehensive compendium of RBC proteins to date. Using stringent criteria, 1674 proteins were quantified, and only a small number differed in amount between adult and cord RBC. We focused on proteins critical for RBC function. Of these, only the expected differences in globin subunits, along with higher levels of carbonic anhydrase 1 and 2 and aquaporin-1 in adult RBCs would be expected to have a phenotypic effect since they are associated with the differences in gaseous exchange between adults and neonates. Since the RBC and reticulocyte samples used were autologous, we catalogue the change in proteome following reticulocyte maturation. The majority of proteins (>60% of the 1671 quantified) reduced in abundance between 2- and 100-fold following maturation. However, ∼5% were at a higher level in RBCs, localized almost exclusively to cell membranes, in keeping with the known clearance of intracellular recycling pools during reticulocyte maturation. Overall, these data suggest that, with respect to the proteome, there is no barrier to the use of cord progenitors for the in vitro generation of RBCs for transfusion to adults other than the expression of fetal, not adult, hemoglobin.
authors.affiliation: From the ‡School of Biochemistry, University of Bristol, Bristol, UK;
authors.forename: Marieangela C
authors.initials: MC
authors.lastname: Wilson
authors.affiliation: From the ‡School of Biochemistry, University of Bristol, Bristol, UK; §Department of Biochemistry, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand;
authors.forename: Kongtana
authors.initials: K
authors.lastname: Trakarnsanga
authors.affiliation: From the ‡School of Biochemistry, University of Bristol, Bristol, UK;
authors.forename: Kate J
authors.initials: KJ
authors.lastname: Heesom
authors.affiliation: ¶Bristol Institute for Transfusion Sciences, National Health Service Blood and Transplant (NHSBT), Filton, Bristol, UK; ‖NIHR Blood and Transplant Research Unit, University of Bristol, Bristol, UK.
authors.forename: Nicola
authors.initials: N
authors.lastname: Cogan
authors.affiliation: ¶Bristol Institute for Transfusion Sciences, National Health Service Blood and Transplant (NHSBT), Filton, Bristol, UK; ‖NIHR Blood and Transplant Research Unit, University of Bristol, Bristol, UK.
authors.forename: Carole
authors.initials: C
authors.lastname: Green
authors.affiliation: From the ‡School of Biochemistry, University of Bristol, Bristol, UK; ¶Bristol Institute for Transfusion Sciences, National Health Service Blood and Transplant (NHSBT), Filton, Bristol, UK; ‖NIHR Blood and Transplant Research Unit, University of Bristol, Bristol, UK.
authors.forename: Ashley M
authors.initials: AM
authors.lastname: Toye
authors.affiliation: ¶Bristol Institute for Transfusion Sciences, National Health Service Blood and Transplant (NHSBT), Filton, Bristol, UK;
authors.forename: Steve F
authors.initials: SF
authors.lastname: Parsons
authors.affiliation: ¶Bristol Institute for Transfusion Sciences, National Health Service Blood and Transplant (NHSBT), Filton, Bristol, UK; ‖NIHR Blood and Transplant Research Unit, University of Bristol, Bristol, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Anstee
authors.affiliation: From the ‡School of Biochemistry, University of Bristol, Bristol, UK; ‖NIHR Blood and Transplant Research Unit, University of Bristol, Bristol, UK. jan.frayne@bristol.ac.uk.
authors.forename: Jan
authors.initials: J
authors.lastname: Frayne
mesh_terms: D000328:Adult; D024401:Carbonic Anhydrase I / Q000378:metabolism; D024402:Carbonic Anhydrase II / Q000378:metabolism; D002454:Cell Differentiation; D002853:Chromatography, Liquid; D041905:Erythroid Cells / Q000166:cytology* / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007231:Infant, Newborn; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D012156:Reticulocytes / Q000166:cytology* / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D020543:Proteome; D024401:Carbonic Anhydrase I; D024402:Carbonic Anhydrase II