Genome-Wide Sequencing Reveals MicroRNAs Downregulated in Cerebral Cavernous Malformations

Abstract

Cerebral cavernous malformations (CCM) are vascular lesions associated with loss-of-function mutations in one of the three genes encoding KRIT1 (CCM1), CCM2, and PDCD10. Recent understanding of the molecular mechanisms that lead to CCM development is limited. The role of microRNAs (miRNAs) has been demonstrated in vascular pathologies resulting in loss of tight junction proteins, increased vascular permeability and endothelial cell dysfunction. Since the relevance of miRNAs in CCM pathophysiology has not been elucidated, the primary aim of the study was to identify the miRNA-mRNA expression network associated with CCM. Using small RNA sequencing, we identified a total of 764 matured miRNAs expressed in CCM patients compared to the healthy brains. The expression of the selected miRNAs was validated by qRT-PCR, and the results were found to be consistent with the sequencing data. Upon application of additional statistical stringency, five miRNAs (let-7b-5p, miR-361-5p, miR-370-3p, miR-181a-2-3p, and miR-95-3p) were prioritized to be top CCM-relevant miRNAs. Further in silico analyses revealed that the prioritized miRNAs have a direct functional relation with mRNAs, such as MIB1, HIF1A, PDCD10, TJP1, OCLN, HES1, MAPK1, VEGFA, EGFL7, NF1, and ENG, which are previously characterized as key regulators of CCM pathology. To date, this is the first study to investigate the role of miRNAs in CCM pathology. By employing cutting edge molecular and in silico analyses on clinical samples, the current study reports global miRNA expression changes in CCM patients and provides a rich source of data set to understand detailed molecular machinery involved in CCM pathophysiology.

Transcriptome clarifies mechanisms of lesion genesis versus progression in models of Ccm3 cerebral cavernous malformations

“…Circulating miRNome have been a major interest to the research community during recent decades and rigorous scientific effort have led major medical breakthroughs in the oligonucleotide research field [31]. To our knowledge, only one study has sequenced the intra-lesional miRNome of resected human CCM lesions [25]. Our group has also previously identified circulating miRNAs in human patients [36].…”

Recent advances in cerebral cavernous malformation research

“…Several proteins already implicated in CCM lesions were found to be targets of these miRNAs including MLLT4, VEGFA, MAPK1, RAC1, RHOA, FOXO1, ENG, SMURF1, and HEYL [17,83,99,106,126,139] . It was concluded that three miRNAs (let-7b-5p, miR-361-5p, and miR-370-3p) can potentially be involved in the pathogenesis of CCM [172] .…”

<p>Protective Role of Astrocyte-Derived Exosomal <em>microRNA-361</em> in Cerebral Ischemic-Reperfusion Injury by Regulating the <em>AMPK/mTOR</em> Signaling Pathway and Targeting <em>CTSB</em></p>

“… 15 miR-361-5p has been identified as one of the top five cerebral cavernous malformations-relevant miRNAs. 16 miRNAs are well known to induce translational repression by binding to their complementary target mRNAs. 17 The present study identified cathepsin B ( CTSB ) as a target of miR-361 .…”