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Circulating MiR-21 expression is upregulated after 30 days of head-down tilt bed rest

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Abstract

Summary

Relative expression of miR-21-5p in serum was upregulated in response to 30 days of bed rest, and miRNA fold changes were positively associated with serum calcium changes.

Introduction

Circulating miRNAs (c-miRNAs) have potential as biomarkers of cellular activity, and they may play a role in cell-to-cell communication. The purpose of this study was to examine c-miRNA and bone marker responses to a 30-day six-degree head-down bed rest protocol at an ambient 0.5% CO2.

Methods

Eleven participants (6 males/5 females, 25–50 years) had fasting blood draws taken 3 days before and immediately after completing the 30-day bed rest protocol at the Institute for Aerospace Medicine in Germany. Serum relative expression of miRNAs associated with bone function (miR-21-5p, -100-5p, -125b-5p, -126-3p) were analyzed using qPCR, and serum bone markers were quantitated using ELISA.

Results

Serum bone markers, sclerostin, and calcium significantly increased (p ≤ 0.036), and total hip aBMD significantly decreased (p = 0.003) post bed rest. Serum miR-21-5p relative expression was significantly upregulated (p = 0.018) post bed rest. Fold changes in miR-126-3p (r = 0.82, p = 0.002) and miR-21-5p (r = 0.62, p = 0.042) were positively correlated with absolute change in serum calcium. There were no sex differences in miRNA responses; women had greater percent increases in TRAP5b (37.3% vs. 16.9% p = 0.021) and greater percent decreases in total hip aBMD (− 2.15% vs. − 0.69%, p = 0.034) than men.

Conclusion

c-miR-21-5p has potential as a biomarker of bone resorption and bone loss in an unloading condition. The upregulation of miR-21-5p may reflect an increase in osteoclast activity after bed rest, which is corroborated by the increase in TRAP5b.

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Data availability

The data used to support the findings of this study are restricted by the NASA IRB in order to protect participant privacy. Data may be available in aggregate form from the corresponding author upon request.

Abbreviations

aBMD:

areal bone mineral density

BDC-14:

baseline data collection 14 days prior to bed rest

BDC-3:

baseline data collection 3 days prior to bed rest

BFLBM:

bone free lean body mass

BMC:

bone mineral content

BMD:

bone mineral density

BMP:

Bone Morphogenetic Protein

BMPR/BMPR2 BMP:

Receptor/BMP Receptor 2

Bone ALP:

Bone-specific Alkaline Phosphatase

cDNA:

Complementary Deoxyribonucleic Acid

Cel-miR-39-3p:

Caenorhabditis elegans microRNA-39-3p

c-miRNA:

circulating microRNA

CO2 :

carbon dioxide

Cq:

Quantification Cycle

ELISA:

Enzyme-linked Immunosorbent Assay

FM:

fat mass

MAPK:

Mitogen-Activated Protein Kinase

miRNA:

microRNA

MMP13:

Matrix Metalloproteinase 13

OSX:

Osterix

PDCD4:

Programmed Cell Death Protein 4

P1NP:

N-terminal propeptide of type 1 procollagen

PTH:

Parathyroid Hormone

qPCR:

Quantitative Real-Time Polymerase Chain Reaction

R+0:

recovery last day of bed rest

R+11:

recovery 11 days post bed rest

RNA:

Ribonucleic Acid

RUNX2:

Runt-Related Transcription Factor 2

SMAD1:

Small Mothers Against Decapentaplegic 1

SMAD7:

Small Mothers Against Decapentaplegic 7

TGF-β:

Transforming Growth Factor-Beta

TRAP5b:

Tartrate-resistant acid phosphatase 5b

UniSp4:

Uni-Spike-In 4

25-OH:

25-hydroxy Vitamin D

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Funding

This study was funded by a National Aeronautics and Space Administration (NASA) research grant (#NNX16AR26G) awarded to C.J Ade and D.A. Bemben.

Author information

Authors and Affiliations

  1. Department of Health and Exercise Science, University of Oklahoma, 1401 Asp Avenue, Norman, OK, 73019, USA

    D.A. Bemben, B.S. Baker & S.R. Buchanan

  2. Department of Orthopedic Surgery, University of Missouri, Columbia, MO, 65212, USA

    B.S. Baker

  3. Department of Health and Human Performance, University of Texas Rio Grande Valley, Edinburg, TX, 78539, USA

    S.R. Buchanan

  4. Department of Kinesiology, Kansas State University, Manhattan, KS, 66506, USA

    C.J. Ade

Authors
  1. D.A. Bemben
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  2. B.S. Baker
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Contributions

DAB wrote the first draft of the manuscript, contributed to the conception and design of the study, supervised the bone marker assays, and performed the data analyses for the bone and miRNA variables. CJA contributed to the conception and design of the study and revised the manuscript. SRB conducted assays, contributed to the data analyses, and revised the manuscript. BSB conducted assays and revised the manuscript.

Corresponding author

Correspondence to D.A. Bemben.

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This study was approved by the NASA Institutional Review Board.

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Supplementary information

Supplementary Figure 1

Individual Percent Change Responses to Bed Rest for Total Hip aBMD (Panel A), Serum Calcium (Panel B), Serum TRAP5b (Panel C), and Serum Sclerostin (Panel D). (n=10 for Total Hip aBMD; n=11 for bone markers) (PNG 83 kb)

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Bemben, D., Baker, B., Buchanan, S. et al. Circulating MiR-21 expression is upregulated after 30 days of head-down tilt bed rest. Osteoporos Int 32, 1369–1378 (2021). https://doi.org/10.1007/s00198-020-05805-2

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