The circulating cell-free microRNA profile in systemic sclerosis is distinct from both healthy controls and systemic lupus erythematosus.
OBJECTIVE
To evaluate the expression profile of cell-free circulating microRNA (miRNA) in systemic sclerosis (SSc), healthy controls (HC), and systemic lupus erythematosus (SLE).


METHODS
Total RNA was purified from plasma and 45 different, mature miRNA were measured using quantitative PCR assays after reverse transcription. Samples (n = 189) were from patients with SSc (n = 120), SLE (n = 29), and from HC (n = 40). Expression data were clustered by principal components analysis, and diagnostically specific miRNA profiles were developed by leave-one-out cross-validation. Diagnostic probability scores were derived from stepwise logistic regression.


RESULTS
Thirty-seven miRNA specificities were consistently detected and 26 of these were unaffected by SSc sample age and present in more than two-thirds of SSc samples. SSc cases showed a distinct expression profile with 14/26 miRNA significantly decreased (false discovery rate < 0.05) and 5/26 increased compared with HC. A 21-miRNA classifier gave optimum accuracy (80%) for discriminating SSc from both HC and SLE. The discrimination between HC and SSc (95% accuracy) was strongly driven by miRNA of the 17 ∼ 92 cluster and by miR-16, -223, and -638, while SLE and SSc differed mainly in the expression of miR-142-3p, -150, -223, and -638. Except for a weak correlation between anti-Scl-70 and miR-638 (p = 0.048), there were no correlations with other patient variables.


CONCLUSIONS
Circulating miRNA profiles are characteristic for SSc compared with both HC and SLE cases. Some of the predicted targets of the differentially regulated miRNA are of relevance for transforming growth factor-β signaling and fibrosis, but need to be validated in independent studies.
From the Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut; Department of Dermatology, Bispebjerg Hospital, and the Department of Rheumatology, Rigshospitalet, University of Copenhagen, Copenhagen; Department of Clinical Genetics, and the Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, Odense; Research Unit of Human Genetics, and Clinical Biochemistry, Institute of Clinical Research, University of Southern Denmark, Odense, Denmark.S.O. Steen, MSc; A.L. Carlsen, PhD, Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut; L.V. Iversen, MD, PhD, Department of Dermatology, Bispebjerg Hospital, University of Copenhagen; M. Burton, PhD, Department of Clinical Genetics, Odense University Hospital, and the Institute of Clinical Research, Research Unit of Human Genetics, University of Southern Denmark; C.T. Nielsen, MD, PhD; S. Jacobsen, MD, DMedSc, Department of Rheumatology, Rigshospitalet, University of Copenhagen; N.H.H. Heegaard, MD, DMedSc, DNatSc, Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut, and the Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, and the Institute of Clinical Research, Clinical Biochemistry, University of Southern Denmark.
Samantha O
SO
Steen
From the Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut; Department of Dermatology, Bispebjerg Hospital, and the Department of Rheumatology, Rigshospitalet, University of Copenhagen, Copenhagen; Department of Clinical Genetics, and the Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, Odense; Research Unit of Human Genetics, and Clinical Biochemistry, Institute of Clinical Research, University of Southern Denmark, Odense, Denmark.S.O. Steen, MSc; A.L. Carlsen, PhD, Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut; L.V. Iversen, MD, PhD, Department of Dermatology, Bispebjerg Hospital, University of Copenhagen; M. Burton, PhD, Department of Clinical Genetics, Odense University Hospital, and the Institute of Clinical Research, Research Unit of Human Genetics, University of Southern Denmark; C.T. Nielsen, MD, PhD; S. Jacobsen, MD, DMedSc, Department of Rheumatology, Rigshospitalet, University of Copenhagen; N.H.H. Heegaard, MD, DMedSc, DNatSc, Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut, and the Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, and the Institute of Clinical Research, Clinical Biochemistry, University of Southern Denmark.
Line V
LV
Iversen
From the Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut; Department of Dermatology, Bispebjerg Hospital, and the Department of Rheumatology, Rigshospitalet, University of Copenhagen, Copenhagen; Department of Clinical Genetics, and the Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, Odense; Research Unit of Human Genetics, and Clinical Biochemistry, Institute of Clinical Research, University of Southern Denmark, Odense, Denmark.S.O. Steen, MSc; A.L. Carlsen, PhD, Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut; L.V. Iversen, MD, PhD, Department of Dermatology, Bispebjerg Hospital, University of Copenhagen; M. Burton, PhD, Department of Clinical Genetics, Odense University Hospital, and the Institute of Clinical Research, Research Unit of Human Genetics, University of Southern Denmark; C.T. Nielsen, MD, PhD; S. Jacobsen, MD, DMedSc, Department of Rheumatology, Rigshospitalet, University of Copenhagen; N.H.H. Heegaard, MD, DMedSc, DNatSc, Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut, and the Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, and the Institute of Clinical Research, Clinical Biochemistry, University of Southern Denmark.
Anting Liu
AL
Carlsen
From the Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut; Department of Dermatology, Bispebjerg Hospital, and the Department of Rheumatology, Rigshospitalet, University of Copenhagen, Copenhagen; Department of Clinical Genetics, and the Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, Odense; Research Unit of Human Genetics, and Clinical Biochemistry, Institute of Clinical Research, University of Southern Denmark, Odense, Denmark.S.O. Steen, MSc; A.L. Carlsen, PhD, Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut; L.V. Iversen, MD, PhD, Department of Dermatology, Bispebjerg Hospital, University of Copenhagen; M. Burton, PhD, Department of Clinical Genetics, Odense University Hospital, and the Institute of Clinical Research, Research Unit of Human Genetics, University of Southern Denmark; C.T. Nielsen, MD, PhD; S. Jacobsen, MD, DMedSc, Department of Rheumatology, Rigshospitalet, University of Copenhagen; N.H.H. Heegaard, MD, DMedSc, DNatSc, Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut, and the Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, and the Institute of Clinical Research, Clinical Biochemistry, University of Southern Denmark.
Mark
M
Burton
From the Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut; Department of Dermatology, Bispebjerg Hospital, and the Department of Rheumatology, Rigshospitalet, University of Copenhagen, Copenhagen; Department of Clinical Genetics, and the Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, Odense; Research Unit of Human Genetics, and Clinical Biochemistry, Institute of Clinical Research, University of Southern Denmark, Odense, Denmark.S.O. Steen, MSc; A.L. Carlsen, PhD, Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut; L.V. Iversen, MD, PhD, Department of Dermatology, Bispebjerg Hospital, University of Copenhagen; M. Burton, PhD, Department of Clinical Genetics, Odense University Hospital, and the Institute of Clinical Research, Research Unit of Human Genetics, University of Southern Denmark; C.T. Nielsen, MD, PhD; S. Jacobsen, MD, DMedSc, Department of Rheumatology, Rigshospitalet, University of Copenhagen; N.H.H. Heegaard, MD, DMedSc, DNatSc, Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut, and the Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, and the Institute of Clinical Research, Clinical Biochemistry, University of Southern Denmark.
Christoffer T
CT
Nielsen
From the Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut; Department of Dermatology, Bispebjerg Hospital, and the Department of Rheumatology, Rigshospitalet, University of Copenhagen, Copenhagen; Department of Clinical Genetics, and the Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, Odense; Research Unit of Human Genetics, and Clinical Biochemistry, Institute of Clinical Research, University of Southern Denmark, Odense, Denmark.S.O. Steen, MSc; A.L. Carlsen, PhD, Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut; L.V. Iversen, MD, PhD, Department of Dermatology, Bispebjerg Hospital, University of Copenhagen; M. Burton, PhD, Department of Clinical Genetics, Odense University Hospital, and the Institute of Clinical Research, Research Unit of Human Genetics, University of Southern Denmark; C.T. Nielsen, MD, PhD; S. Jacobsen, MD, DMedSc, Department of Rheumatology, Rigshospitalet, University of Copenhagen; N.H.H. Heegaard, MD, DMedSc, DNatSc, Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut, and the Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, and the Institute of Clinical Research, Clinical Biochemistry, University of Southern Denmark.
Søren
S
Jacobsen
From the Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut; Department of Dermatology, Bispebjerg Hospital, and the Department of Rheumatology, Rigshospitalet, University of Copenhagen, Copenhagen; Department of Clinical Genetics, and the Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, Odense; Research Unit of Human Genetics, and Clinical Biochemistry, Institute of Clinical Research, University of Southern Denmark, Odense, Denmark.S.O. Steen, MSc; A.L. Carlsen, PhD, Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut; L.V. Iversen, MD, PhD, Department of Dermatology, Bispebjerg Hospital, University of Copenhagen; M. Burton, PhD, Department of Clinical Genetics, Odense University Hospital, and the Institute of Clinical Research, Research Unit of Human Genetics, University of Southern Denmark; C.T. Nielsen, MD, PhD; S. Jacobsen, MD, DMedSc, Department of Rheumatology, Rigshospitalet, University of Copenhagen; N.H.H. Heegaard, MD, DMedSc, DNatSc, Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut, and the Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, and the Institute of Clinical Research, Clinical Biochemistry, University of Southern Denmark. nhe@ssi.dk.
Niels H H
NH
Heegaard
D000328:Adult; D000368:Aged; D015415:Biomarkers / Q000097:blood; D003430:Cross-Sectional Studies; D003937:Diagnosis, Differential; D005260:Female; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000097:blood / Q000175:diagnosis* / Q000235:genetics; D008297:Male; D035683:MicroRNAs / Q000097:blood*; D008875:Middle Aged; D012595:Scleroderma, Systemic / Q000097:blood / Q000175:diagnosis* / Q000235:genetics; D055815:Young Adult
D015415:Biomarkers; D035683:MicroRNAs
CLASSIFICATION; DIAGNOSIS; SYSTEMIC LUPUS ERYTHEMATOSUS; SYSTEMIC SCLEROSIS; microRNA