title: Detection of hair-microRNAs as the novel potent biomarker: evaluation of the usefulness for the diagnosis of scleroderma.
abstract: BACKGROUND
  Serum microRNA levels are known as useful biomarkers for various diseases. Recent publication has indicated the existence of microRNAs in hair roots and hair shafts.
  
  
  OBJECTIVE
  In this study, we evaluated several methods for the extraction of hair microRNAs, and their usefulness for the diagnosis of scleroderma.
  
  
  METHODS
  A single hair root and 5 pieces of hair shafts were obtained from the occiput of each individual of 11 scleroderma patients and 13 normal subjects at the time of serum sampling. microRNA extraction from sera or hair roots was performed with commercially available kits. microRNAs were extracted from hair shafts using four different methods. microRNA expression was evaluated by PCR array and real-time PCR.
  
  
  RESULTS
  We demonstrated microRNAs in hair roots and hair shafts were detectable and quantitative using our method. We found the difference of microRNA levels in hair roots and hair shafts obtained from different places of head in each individual were within 2-fold, indicating the reproducibility of hair microRNA levels by our method. PCR array revealed microRNAs from sera, hair roots and hair shafts have different expression pattern, and can be independent biomarkers. Serum and hair root miR-196a levels were not significantly changed in scleroderma patients, while we found miR-196a levels in hair shafts were significantly decreased in scleroderma patients compared to those in normal subjects (p<0.05).
  
  
  CONCLUSIONS
  Hairs are more accessible than sera among human samples. microRNAs levels in hair roots or hair shafts may become effective and independent biomarkers.
authors.affiliation: Department of Dermatology and Plastic Surgery, Faculty of Life Sciences, Kumamoto University, Honjo 1-1-1, Kumamoto 860-8556, Japan.
authors.forename: Zhongzhi
authors.initials: Z
authors.lastname: Wang
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Jinnin
authors.forename: Hideo
authors.initials: H
authors.lastname: Kudo
authors.forename: Kuniko
authors.initials: K
authors.lastname: Inoue
authors.forename: Wakana
authors.initials: W
authors.lastname: Nakayama
authors.forename: Noritoshi
authors.initials: N
authors.lastname: Honda
authors.forename: Katsunari
authors.initials: K
authors.lastname: Makino
authors.forename: Ikko
authors.initials: I
authors.lastname: Kajihara
authors.forename: Satoshi
authors.initials: S
authors.lastname: Fukushima
authors.forename: Yuji
authors.initials: Y
authors.lastname: Inoue
authors.forename: Hironobu
authors.initials: H
authors.lastname: Ihn
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000378:metabolism*; D005260:Female; D020869:Gene Expression Profiling; D006197:Hair / Q000378:metabolism*; D018859:Hair Follicle / Q000378:metabolism; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000378:metabolism*; D008875:Middle Aged; D010865:Pilot Projects; D015203:Reproducibility of Results; D012595:Scleroderma, Systemic / Q000097:blood / Q000175:diagnosis* / Q000378:metabolism*
chemical_list: D015415:Biomarkers; C542543:MIRN196 microRNA, human; D035683:MicroRNAs
keywords: 3′ UTRs; 3′ untranslated regions; AUCs; CTGF; Hair; IL-17; ROC; SSc; Scleroderma; TGF-β; TNF; areas under curves; connective tissue growth factor; dcSSc; diffuse cutaneous SSc; interleukin-17; lcSSc; limited cutaneous SSc; miRNAs; microRNA; microRNAs; receiver operating characteristic; systemic sclerosis; transforming growth factor-β; tumor necrosis factor