Circulating miRNAs as Biomarkers of Acute Muscle Damage in Rats

Abstract

Skeletal muscle damage is an often-occurring event. Diagnosis using the classic blood marker creatine kinase sometimes yields unsatisfactory results due to great interindividual variability. Therefore, the identification of reliable biomarkers is important. Our aim was to detect and characterize circulating miRNAs in plasma in response to acute notexin-induced muscle damage in rats. Real-time quantitative RT-PCR profiling led to the identification of miRNAs that were highly increased in plasma in response to notexin injection into several muscles, namely miR-1-3p, -133a-3p, -133b-3p, -206-3p, -208b-3p, and -499-5p, as well as miR-378a-3p and miR-434-3p. Peak values of miRNAs appeared 12 hours after injury, and were contained both in the vesicular and nonvesicular fractions of plasma. Receiver operating characteristic curve analysis showed that circulating miRNAs could accurately discriminate between damaged and nondamaged tissues. Furthermore, we tested the robustness of expression profiles in slow- and fast-type fibers. Upon inducing damage in slow- or fast-type muscle, we found that the damaged-muscle phenotype had a very limited impact on the miRNA response. Similarly, the circulating miRNAs selected were not affected by hemolysis or platelets, two pre-analytical factors known to affect plasma miRNA profiles. Taken together, our results show that circulating muscle-specific miRNAs, miR-378a-3p and miR-434-3p, are robust and promising biomarkers of acute muscle damage in rats.

Circulating myomiRs: a new class of biomarkers to monitor skeletal muscle in physiology and medicine

“…To investigate the effect of an acute muscle injury in the absence of contraction on c‐miRNA levels, we studied plasma miRNA profiles in rats after an intramuscular injection of notexin, a myotoxin known to provoke massive myofibre necrosis. We found a substantial increase in circulating myomiRs that could efficiently discriminate between damaged and non‐damaged animals . Concomitant myofibre necrosis and a reduction in myomiRs within damaged muscle tissue suggest that a passive release by injured myofibres may account for high circulating levels.…”

Circulating levels of non-muscle-specific miRNAs in response to acute muscle damage in rat

“…Pick-&-Mix microRNA PCR panels (Exiqon) were used [1], 500 µL 2× PCR Master mix were combined with 6.25 µL cDNA and 493.75 µL sterile water. Then, 10 µL were distributed in each well.…”

“…Then, a set of miRNAs was selected based on detectability and alteration in response to injury, and was measured on individual samples. Muscle-specific miRNAs results have been described elsewhere [1]. Fig.…”

“…Blood collected 6 h, 12 h, 24 h or 48 h later for miRNA analysis Data source location Clamart, France Data accessibility Data related to previously published article (Siracusa et al, 2016) [1]…”

MicroRNAs, heart failure, and aging: potential interactions with skeletal muscle

“…Moreover, interference of the exosome biosynthesis process or disruption of exosomal membranes reduces extracellular miRNA content [59]. Most recently, it has been shown that serum miRNAs can be found in both vesicular and non-vesicular fractions in response to tissue damage in rats [95]. Although the manner in which miRNAs are transported in the circulation may be condition dependent, they appear to be abundantly expressed and likely exert effects in different tissue types.…”