title: MIR144 and MIR451 regulate human erythropoiesis via RAB14.
abstract: Expression levels of MIR144 and MIR451 increase during erythropoiesis, a pattern that is conserved from zebrafish to humans. As these two miRs are expressed from the same polycistronic transcript, we manipulated MIR144 and MIR451 in human erythroid cells individually and together to investigate their effects on human erythropoiesis. Inhibition of endogenous human MIR451 resulted in decreased numbers of erythroid (CD71(hi) CD235a(hi) CD34(-) ) cells, consistent with prior studies in zebrafish and mice. In addition, inhibition of MIR144 impaired human erythroid differentiation, unlike in zebrafish and mouse studies where the functional effect of MIR144 on erythropoiesis was minimal. In this study, we found RAB14 is a direct target of both MIR144 and MIR451. As MIR144 and MIR451 expression increased during human erythropoiesis, RAB14 protein expression decreased. Enforced RAB14 expression phenocopied the effect of MIR144 and/or MIR451 depletion, whereas shRNA-mediated RAB14 knockdown protected cells from MIR144 and/or MIR451 depletion-mediated erythropoietic inhibition. RAB14 knockdown increased the frequency and number of erythroid cells, increased β-haemoglobin expression, and decreased CBFA2T3 expression during human erythropoiesis. In summary, we utilized MIR144 and MIR451 to identify RAB14 as a novel physiological inhibitor of human erythropoiesis.
authors.affiliation: Departments of Physiology and Pediatrics, Center for Stem Cell Biology & Regenerative Medicine, Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, MD, USA.
authors.forename: MinJung
authors.initials: M
authors.lastname: Kim
authors.forename: Yee Sun
authors.initials: YS
authors.lastname: Tan
authors.forename: Wen-Chih
authors.initials: WC
authors.lastname: Cheng
authors.forename: Tami J
authors.initials: TJ
authors.lastname: Kingsbury
authors.forename: Shelly
authors.initials: S
authors.lastname: Heimfeld
authors.forename: Curt I
authors.initials: CI
authors.lastname: Civin
mesh_terms: D045744:Cell Line, Tumor; D015672:Erythroid Precursor Cells / Q000166:cytology / Q000187:drug effects; D004920:Erythropoiesis / Q000502:physiology*; D004921:Erythropoietin / Q000494:pharmacology; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D004915:Leukemia, Erythroblastic, Acute / Q000473:pathology; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics; D014161:Transduction, Genetic; D025521:Tumor Suppressor Proteins / Q000096:biosynthesis / Q000235:genetics; D020691:rab GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*
chemical_list: C115744:CBFA2T3 protein, human; C554293:MIRN144 microRNA, human; C531201:MIRN451 microRNA, human; D035683:MicroRNAs; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; D012097:Repressor Proteins; D025521:Tumor Suppressor Proteins; D004921:Erythropoietin; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; C553411:Rab14 protein, human; D020691:rab GTP-Binding Proteins
keywords: MIR144; MIR451; RAB14; erythropoietic inhibitor; human erythropoiesis