title: A Sex-Specific MicroRNA-96/5-Hydroxytryptamine 1B Axis Influences Development of Pulmonary Hypertension.
abstract: BACKGROUND
  Females are predisposed to pulmonary arterial hypertension (PAH); evidence suggests that serotonin, mutations in the bone morphogenetic protein receptor (BMPR) II gene, and estrogens influence development of PAH. The 5-hydroxytryptamine 1B receptor (5-HT1BR) mediates human pulmonary artery smooth muscle cell (hPASMC) proliferation.
  
  
  OBJECTIVE
  We aimed to determine whether selected microRNAs (miRNAs) expressed in PASMCs are influenced by sex, BMPR-II mutations, and estrogens, and contribute to PASMC proliferation in PAH.
  
  
  METHODS
  Expression levels of miRNAs targeting genes related to PAH, estrogen, and serotonin were determined by quantitative RT-PCR in hPASMCs and mouse PASMCs harboring a heterozygous mutation in BMPR-II (BMPR-II(R899X+/-) PASMCs). miRNA-96 targets 5-HT1BR and was selected for further investigation. miRNA target validation was confirmed by luciferase reporter assay. Precursor miRNA-96 was transfected into hPASMCs to examine effects on proliferation and 5-HT1BR expression. The effect of a miRNA-96 mimic on the development of hypoxic pulmonary hypertension in mice was also assessed.
  
  
  RESULTS
  miRNA-96 expression was reduced in BMPR-II(R899X+/-) PASMCs from female mice and hPASMCs from female patients with PAH; this was associated with increased 5-HT1BR expression and serotonin-mediated proliferation. 5-HT1BR was validated as a target for miRNA-96. Transfection of precursor miRNA-96 into hPASMCs reduced 5-HT1BR expression and inhibited serotonin-induced proliferation. Restoration of miRNA-96 expression in pulmonary arteries in vivo via administration of an miRNA-96 mimic reduced the development of hypoxia-induced pulmonary hypertension in the mouse.
  
  
  CONCLUSIONS
  Increased 5-HT1BR expression may be a consequence of decreased miRNA-96 expression in female patient PASMCs, and this may contribute to the development of PAH.
authors: 1 Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom; and.
authors: Emma
authors: E
authors: Wallace
authors: 2 School of Clinical Medicine, University of Cambridge, Cambridge, United Kingdom.
authors: Nicholas W
authors: NW
authors: Morrell
authors: 2 School of Clinical Medicine, University of Cambridge, Cambridge, United Kingdom.
authors: Xudong D
authors: XD
authors: Yang
authors: 2 School of Clinical Medicine, University of Cambridge, Cambridge, United Kingdom.
authors: Lu
authors: L
authors: Long
authors: 1 Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom; and.
authors: Hannah
authors: H
authors: Stevens
authors: 1 Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom; and.
authors: Margaret
authors: M
authors: Nilsen
authors: 1 Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom; and.
authors: Lynn
authors: L
authors: Loughlin
authors: 1 Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom; and.
authors: Kirsty M
authors: KM
authors: Mair
authors: 1 Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom; and.
authors: Andrew H
authors: AH
authors: Baker
authors: 1 Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom; and.
authors: Margaret R
authors: MR
authors: MacLean
mesh_terms: D000818:Animals; D052006:Bone Morphogenetic Protein Receptors, Type II / Q000378:metabolism; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D004195:Disease Models, Animal; D005260:Female; D020022:Genetic Predisposition to Disease; D006801:Humans; D006976:Hypertension, Pulmonary / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000378:metabolism; D011651:Pulmonary Artery / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D044302:Receptor, Serotonin, 5-HT1B / Q000378:metabolism*; D012727:Sex Characteristics; D015398:Signal Transduction / Q000502:physiology
chemical_list: D035683:MicroRNAs; D044302:Receptor, Serotonin, 5-HT1B; D052006:Bone Morphogenetic Protein Receptors, Type II
keywords: animal models; estrogen; microRNA; pulmonary hypertension; serotonin