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Differential miRNA Expression in Cells and Matrix Vesicles in Vascular Smooth Muscle Cells from Rats with Kidney Disease.

Chaturvedi P, Chen NX, O'Neill K, McClintick JN, Moe SM, Janga SC - PLoS ONE (2015)

Bottom Line: The percentage of miRNA to total RNA was increased in MV compared to VSMC.We found several processes including vascular smooth muscle contraction, response to hypoxia and regulation of muscle cell differentiation to be enriched.In conclusion, our results demonstrate that miRs are concentrated in MV from calcifying VSMC, and that important functions and pathways are affected by the miRs dysregulation between calcifying VSMC and the MV they produce.

View Article: PubMed Central - PubMed

Affiliation: Department of Biohealth Informatics, School of Informatics and Computing, Indiana University Purdue University, Indianapolis, Indiana, United States of America.

ABSTRACT
Vascular calcification is a complex process and has been associated with aging, diabetes, chronic kidney disease (CKD). Although there have been several studies that examine the role of miRNAs (miRs) in bone osteogenesis, little is known about the role of miRs in vascular calcification and their role in the pathogenesis of vascular abnormalities. Matrix vesicles (MV) are known to play in important role in initiating vascular smooth muscle cell (VSMC) calcification. In the present study, we performed miRNA microarray analysis to identify the dysregulated miRs between MV and VSMC derived from CKD rats to understand the role of post-transcriptional regulatory networks governed by these miRNAs in vascular calcification and to uncover the differential miRNA content of MV. The percentage of miRNA to total RNA was increased in MV compared to VSMC. Comparison of expression profiles of miRNA by microarray demonstrated 33 miRs to be differentially expressed with the majority (~ 57%) of them down-regulated. Target genes controlled by differentially expressed miRNAs were identified utilizing two different complementary computational approaches Miranda and Targetscan to understand the functions and pathways that may be affected due to the production of MV from calcifying VSMC thereby contributing to the regulation of genes by miRs. We found several processes including vascular smooth muscle contraction, response to hypoxia and regulation of muscle cell differentiation to be enriched. Signaling pathways identified included MAP-kinase and wnt signaling that have previously been shown to be important in vascular calcification. In conclusion, our results demonstrate that miRs are concentrated in MV from calcifying VSMC, and that important functions and pathways are affected by the miRs dysregulation between calcifying VSMC and the MV they produce. This suggests that miRs may play a very important regulatory role in vascular calcification in CKD by controlling an extensive network of post-transcriptional targets.

No MeSH data available.


Related in: MedlinePlus

miRNA is concentrated in MV compared to VSMC from CKD rats.Total RNA from VSMC or MV was isolated from CKD rats and quantification performed using Agilent 2100 Bioanalyzer total and Small RNA kit. The results demonstrated that the total RNA level (A) is greater but miRNA levels are lower in VSMC compared to MV (B). n = 3, Data were expressed as mean±SEM, *p<0.05, MV vs. VSMC.
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pone.0131589.g002: miRNA is concentrated in MV compared to VSMC from CKD rats.Total RNA from VSMC or MV was isolated from CKD rats and quantification performed using Agilent 2100 Bioanalyzer total and Small RNA kit. The results demonstrated that the total RNA level (A) is greater but miRNA levels are lower in VSMC compared to MV (B). n = 3, Data were expressed as mean±SEM, *p<0.05, MV vs. VSMC.

Mentions: To determine the total RNA and miRNA concentration in MV compared to VSMC which they are originated, total RNA was isolated from MV and VSMC and the relative concentrations of total RNA and miRNA determined by Agilent 2100 Bioanalyzer total RNA and Small RNA kits, respectively. The results demonstrated that total RNA concentration is 4 times greater in VSMC than that in MV (Fig 2A). In contrast, percent of miRNA of total RNA in MV is 4 times greater than that in VSMC (Fig 2B), confirming that miRNA are concentrated in MV as they are in exosomes.


Differential miRNA Expression in Cells and Matrix Vesicles in Vascular Smooth Muscle Cells from Rats with Kidney Disease.

Chaturvedi P, Chen NX, O'Neill K, McClintick JN, Moe SM, Janga SC - PLoS ONE (2015)

miRNA is concentrated in MV compared to VSMC from CKD rats.Total RNA from VSMC or MV was isolated from CKD rats and quantification performed using Agilent 2100 Bioanalyzer total and Small RNA kit. The results demonstrated that the total RNA level (A) is greater but miRNA levels are lower in VSMC compared to MV (B). n = 3, Data were expressed as mean±SEM, *p<0.05, MV vs. VSMC.
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Related In: Results  -  Collection

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pone.0131589.g002: miRNA is concentrated in MV compared to VSMC from CKD rats.Total RNA from VSMC or MV was isolated from CKD rats and quantification performed using Agilent 2100 Bioanalyzer total and Small RNA kit. The results demonstrated that the total RNA level (A) is greater but miRNA levels are lower in VSMC compared to MV (B). n = 3, Data were expressed as mean±SEM, *p<0.05, MV vs. VSMC.
Mentions: To determine the total RNA and miRNA concentration in MV compared to VSMC which they are originated, total RNA was isolated from MV and VSMC and the relative concentrations of total RNA and miRNA determined by Agilent 2100 Bioanalyzer total RNA and Small RNA kits, respectively. The results demonstrated that total RNA concentration is 4 times greater in VSMC than that in MV (Fig 2A). In contrast, percent of miRNA of total RNA in MV is 4 times greater than that in VSMC (Fig 2B), confirming that miRNA are concentrated in MV as they are in exosomes.

Bottom Line: The percentage of miRNA to total RNA was increased in MV compared to VSMC.We found several processes including vascular smooth muscle contraction, response to hypoxia and regulation of muscle cell differentiation to be enriched.In conclusion, our results demonstrate that miRs are concentrated in MV from calcifying VSMC, and that important functions and pathways are affected by the miRs dysregulation between calcifying VSMC and the MV they produce.

View Article: PubMed Central - PubMed

Affiliation: Department of Biohealth Informatics, School of Informatics and Computing, Indiana University Purdue University, Indianapolis, Indiana, United States of America.

ABSTRACT
Vascular calcification is a complex process and has been associated with aging, diabetes, chronic kidney disease (CKD). Although there have been several studies that examine the role of miRNAs (miRs) in bone osteogenesis, little is known about the role of miRs in vascular calcification and their role in the pathogenesis of vascular abnormalities. Matrix vesicles (MV) are known to play in important role in initiating vascular smooth muscle cell (VSMC) calcification. In the present study, we performed miRNA microarray analysis to identify the dysregulated miRs between MV and VSMC derived from CKD rats to understand the role of post-transcriptional regulatory networks governed by these miRNAs in vascular calcification and to uncover the differential miRNA content of MV. The percentage of miRNA to total RNA was increased in MV compared to VSMC. Comparison of expression profiles of miRNA by microarray demonstrated 33 miRs to be differentially expressed with the majority (~ 57%) of them down-regulated. Target genes controlled by differentially expressed miRNAs were identified utilizing two different complementary computational approaches Miranda and Targetscan to understand the functions and pathways that may be affected due to the production of MV from calcifying VSMC thereby contributing to the regulation of genes by miRs. We found several processes including vascular smooth muscle contraction, response to hypoxia and regulation of muscle cell differentiation to be enriched. Signaling pathways identified included MAP-kinase and wnt signaling that have previously been shown to be important in vascular calcification. In conclusion, our results demonstrate that miRs are concentrated in MV from calcifying VSMC, and that important functions and pathways are affected by the miRs dysregulation between calcifying VSMC and the MV they produce. This suggests that miRs may play a very important regulatory role in vascular calcification in CKD by controlling an extensive network of post-transcriptional targets.

No MeSH data available.


Related in: MedlinePlus