title: Erythropoietin-mediated expression of placenta growth factor is regulated via activation of hypoxia-inducible factor-1α and post-transcriptionally by miR-214 in sickle cell disease.
abstract: Placental growth factor (PlGF) plays an important role in various pathological conditions and diseases such as inflammation, cancer, atherosclerosis and sickle cell disease (SCD). Abnormally high PlGF levels in SCD patients are associated with increased inflammation and pulmonary hypertension (PHT) and reactive airway disease; however, the transcriptional and post-transcriptional mechanisms regulating PlGF expression are not well defined. Herein, we show that treatment of human erythroid cells and colony forming units with erythropoietin (EPO) increased PlGF expression. Our studies showed EPO-mediated activation of HIF-1α led to subsequent binding of HIF-1α to hypoxia response elements (HREs) within the PlGF promoter, as demonstrated by luciferase transcription reporter assays and ChIP analysis of the endogenous gene. Additionally, we showed miR-214 post-transcriptionally regulated the expression of PlGF as demonstrated by luciferase reporter assays using wild-type (wt) and mutant PlGF-3'-UTR constructs. Furthermore, synthesis of miR-214, located in an intron of DNM3 (dynamin 3), was transcriptionally regulated by transcription factors, peroxisome proliferator-activated receptor-α (PPARα) and hypoxia-inducible factor-1α (HIF-1α). These results were corroborated in vivo wherein plasma from SCD patients and lung tissues from sickle mice showed an inverse correlation between PlGF and miR-214 levels. Finally, we observed that miR-214 expression could be induced by fenofibrate, a Food and Drug Administration (FDA) approved PPARα agonist, thus revealing a potential therapeutic approach for reduction in PlGF levels by increasing miR-214 transcription. This strategy has potential clinical implications for several pathological conditions including SCD.
authors: Departments of Biochemistry and Molecular Biology, Keck School of Medicine of University of Southern California, Los Angeles, CA 90033, U.S.A.
authors: Caryn S
authors: CS
authors: Gonsalves
authors: Departments of Biochemistry and Molecular Biology, Keck School of Medicine of University of Southern California, Los Angeles, CA 90033, U.S.A.
authors: Chen
authors: C
authors: Li
authors: Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, U.S.A.
authors: Marthe-Sandrine Eiymo Mwa
authors: MS
authors: Mpollo
authors: Division of Hematology, City of Hope, Duarte, CA 91010, U.S.A.
authors: Vinod
authors: V
authors: Pullarkat
authors: Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, U.S.A.
authors: Punam
authors: P
authors: Malik
authors: Molecular Microbiology and Immunology, Keck School of Medicine of University of Southern California, Los Angeles, CA 90033, U.S.A.
authors: Stanley M
authors: SM
authors: Tahara
authors: Departments of Biochemistry and Molecular Biology, Keck School of Medicine of University of Southern California, Los Angeles, CA 90033, U.S.A. vkalra@usc.edu.
authors: Vijay K
authors: VK
authors: Kalra
mesh_terms: D020413:3' Untranslated Regions / Q000187:drug effects; D000755:Anemia, Sickle Cell / Q000097:blood / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D003433:Crosses, Genetic; D041905:Erythroid Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D015672:Erythroid Precursor Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D004921:Erythropoietin / Q000494:pharmacology* / Q000627:therapeutic use; D017930:Genes, Reporter / Q000187:drug effects; D006397:Hematinics / Q000494:pharmacology* / Q000627:therapeutic use; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D008168:Lung / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D035683:MicroRNAs / Q000097:blood / Q000378:metabolism*; D009154:Mutation; D000072483:Placenta Growth Factor; D011257:Pregnancy Proteins / Q000819:agonists* / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000187:drug effects; D034622:RNA Interference; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D020413:3' Untranslated Regions; C497442:HIF1A protein, human; D006397:Hematinics; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; C547397:MIRN214 microRNA, human; D035683:MicroRNAs; C000608036:PGF protein, human; C000608037:Pgf protein, mouse; D011257:Pregnancy Proteins; D011994:Recombinant Proteins; D004921:Erythropoietin; D000072483:Placenta Growth Factor
keywords: dynamin 3 opposite strand (DNM3os); fenofibrate; hypoxia-inducible factor-1α (HIF-1α); microRNA (miRNA); peroxisome proliferator-activated receptor-α (PPARα); placenta growth factor