title: Hsa-miR-34a mediated repression of corticotrophin releasing hormone receptor 1 regulates pro-opiomelanocortin expression in patients with complex regional pain syndrome.
abstract: BACKGROUND
  Ketamine provides relief for a subset of patients with complex regional pain syndrome (CRPS). The poor responders had a lower body mass index (BMI) relative to responders. Regulation of proopiomelanocortin (POMC) expression is crucial in normal body weight homeostasis. The main objectives of this study were to investigate the mechanisms underlying lower BMI characterizing CRPS patients responding poorly to intravenous ketamine therapy and identify potential biomarkers for predicting response.
  
  
  METHODS
  We investigated POMC transcript levels in blood from CRPS patients grouped as responders and poor responders to ketamine therapy. Plasma levels of β-endorphin, ACTH and α-MSH were measured by ELISA. We previously identified differential expression of small noncoding microRNA hsa-miR-34a in blood between responders and poor responders. We investigated whether a 11-fold downregulation of hsa-miR-34a in poor responders relative to responders is contributing to the differences in POMC levels by targeting POMC regulator CRHR1. Binding of miR-34a to CRHR1 was assessed using reporter assay; changes in mRNA and protein levels of CRHR1 were used to determine the regulation of CRHR1 by miR-34a. RNA from blood of CRPS and control subjects were used for quantitative PCR for CRHR1.
  
  
  RESULTS
  Though ketamine treatment did not alter POMC expression, poor responders had higher levels of POMC mRNA than responders, both before and after treatment. Corticotropin-releasing hormone (CRH) is a key regulator of POMC expression and the biological effects are mediated through its receptor corticotrophin releasing hormone receptor 1 (CRHR1). We show that hsa-miR-34a is a negative regulator of CRHR1; overexpression of hsa-miR-34a in Jurkat cells resulted in reduction of CRH-mediated POMC expression. Poor responders had higher expression of CRHR1 transcripts than responders, indicating a regulatory role for miR-34a. In addition, we found positive correlations between the pretreatment levels of miR-34a to BMI and response to ketamine therapy.
  
  
  CONCLUSIONS
  Our findings indicate a mechanism by which hsa-miR-34a can regulate the CRH/CRHR1/POMC axis and may influence BMI. Studies in larger patient cohorts are required to confirm the biomarker utility of circulating hsa-miR-34a levels in predicting treatment response to ketamine therapy.
authors.affiliation: Pharmacology and Physiology, Drexel University College of Medicine, 245 North 15th Street, Mail Stop 488, Philadelphia, PA, 19102, USA. bbs39@drexel.edu.
authors.forename: Botros B
authors.initials: BB
authors.lastname: Shenoda
authors.affiliation: Neurology, Drexel University College of Medicine, Philadelphia, USA. Guillermo.Alexander@DrexelMed.edu.
authors.forename: Guillermo M
authors.initials: GM
authors.lastname: Alexander
authors.affiliation: Pharmacology and Physiology, Drexel University College of Medicine, 245 North 15th Street, Mail Stop 488, Philadelphia, PA, 19102, USA. seena.ajit@drexelmed.edu.
authors.forename: Seena K
authors.initials: SK
authors.lastname: Ajit
mesh_terms: D000324:Adrenocorticotropic Hormone / Q000097:blood; D015992:Body Mass Index; D020918:Complex Regional Pain Syndromes / Q000097:blood / Q000188:drug therapy / Q000235:genetics*; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D019169:Jurkat Cells; D007649:Ketamine / Q000627:therapeutic use; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D011333:Pro-Opiomelanocortin / Q000097:blood / Q000378:metabolism*; D018019:Receptors, Corticotropin-Releasing Hormone / Q000235:genetics / Q000378:metabolism*; D015203:Reproducibility of Results; D000521:alpha-MSH / Q000097:blood; D001615:beta-Endorphin / Q000097:blood; D000099052:CRF Receptor, Type 1
chemical_list: C522425:MIRN34 microRNA, human; D035683:MicroRNAs; D018019:Receptors, Corticotropin-Releasing Hormone; D000521:alpha-MSH; D000099052:CRF Receptor, Type 1; D001615:beta-Endorphin; D011333:Pro-Opiomelanocortin; D007649:Ketamine; D000324:Adrenocorticotropic Hormone