Differential regulation of miRNA and mRNA expression in the myocardium of Nrf2 knockout mice
Abstract Background Nuclear Factor Erythroid-derived 2-like 2 (Nrf2) senses oxidative environments and/or stress and initiates a cytoprotective response through transcriptional activation of antioxidant and detoxification genes. Several preclinical studies suggest that Nrf2 combats oxidative stress...
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doaj-1b04d53c3c9848f7b020ef00d4ae7a982020-11-24T22:25:14ZengBMCBMC Genomics1471-21642017-07-0118111610.1186/s12864-017-3875-3Differential regulation of miRNA and mRNA expression in the myocardium of Nrf2 knockout miceJustin M. Quiles0Madhusudhanan Narasimhan1Gobinath Shanmugam2Brett Milash3John R. Hoidal4Namakkal S. Rajasekaran5Cardiac Aging & Redox Signaling Laboratory, Division of Molecular & Cellular Pathology, Department of Pathology, University of Alabama at BirminghamDepartment of Pharmacology and Neuroscience, Texas Tech University Health Sciences CenterCardiac Aging & Redox Signaling Laboratory, Division of Molecular & Cellular Pathology, Department of Pathology, University of Alabama at BirminghamHuntsman Cancer InstituteDivision of PulmonaryCardiac Aging & Redox Signaling Laboratory, Division of Molecular & Cellular Pathology, Department of Pathology, University of Alabama at BirminghamAbstract Background Nuclear Factor Erythroid-derived 2-like 2 (Nrf2) senses oxidative environments and/or stress and initiates a cytoprotective response through transcriptional activation of antioxidant and detoxification genes. Several preclinical studies suggest that Nrf2 combats oxidative stress underlying a variety of pathologies. Despite Nrf2 deficits linked to functional abnormalities in many organ systems, the transcriptional network resulting from Nrf2 deficiency in the heart has remained elusive. Moreover, cross-talk between microRNAs (miRNAs) and cardiac Nrf2 signaling is unknown. Here, we utilized next generation RNA sequencing (RNAseq) to unbiasedly profile basal mRNA and miRNA expression in Nrf2 knockout (Nrf2−/−) hearts. Results RNAseq of mRNA revealed 152 differentially expressed genes (DEGs) in the Nrf2−/−myocardium, of which 129 were downregulated. Grouping of DEGs based on biological function and real-time qPCR validation indicated that DEGs were enriched for; mitochondrial genome and bioenergetics, oxidoreductase capacity, cardiac development, and chaperone activity. Interestingly, RNAseq analysis uncovered 27 significantly altered miRNAs, of which 11 were upregulated and 16 were downregulated in Nrf2−/− hearts. Expression changes were validated for 12 miRNAs using specific primer assays in real-time and revealed a significant decrease in miR-10b-5p, miR-674-3p, miR-3535, and miR-378c while miR-30b-5p, miR-208a-5p, miR-350-3p, and miR-582-5p, and miR-1249-3p levels were increased. High throughput data were integrated using prediction algorithms, and these in silico analyses discovered potential recognition elements within 39 repressed mRNAs which matched the seed sequence for 4 upregulated miRNAs; miR-30b-5p, miR-208a-5p, miR-350-3p, and miR-582-5p. Conclusion These high-throughput data reveal transcriptome-wide effects of myocardial Nrf2 deficiency. Further, our results suggest that Nrf2 may directly or indirectly regulate a sub-set of cardiac miRNAs in the basal setting. This comprehensive analysis is the first evidence to demonstrate a plausible regulatory cross-talk among cardiac miRNAs and the Nrf2 transcriptional network, and provides valuable candidates to examine in future mechanistic and preclinical studies.http://link.springer.com/article/10.1186/s12864-017-3875-3Nrf2microRNAmRNAHeartRNA sequencingDifferential expression |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Justin M. Quiles Madhusudhanan Narasimhan Gobinath Shanmugam Brett Milash John R. Hoidal Namakkal S. Rajasekaran |
spellingShingle |
Justin M. Quiles Madhusudhanan Narasimhan Gobinath Shanmugam Brett Milash John R. Hoidal Namakkal S. Rajasekaran Differential regulation of miRNA and mRNA expression in the myocardium of Nrf2 knockout mice BMC Genomics Nrf2 microRNA mRNA Heart RNA sequencing Differential expression |
author_facet |
Justin M. Quiles Madhusudhanan Narasimhan Gobinath Shanmugam Brett Milash John R. Hoidal Namakkal S. Rajasekaran |
author_sort |
Justin M. Quiles |
title |
Differential regulation of miRNA and mRNA expression in the myocardium of Nrf2 knockout mice |
title_short |
Differential regulation of miRNA and mRNA expression in the myocardium of Nrf2 knockout mice |
title_full |
Differential regulation of miRNA and mRNA expression in the myocardium of Nrf2 knockout mice |
title_fullStr |
Differential regulation of miRNA and mRNA expression in the myocardium of Nrf2 knockout mice |
title_full_unstemmed |
Differential regulation of miRNA and mRNA expression in the myocardium of Nrf2 knockout mice |
title_sort |
differential regulation of mirna and mrna expression in the myocardium of nrf2 knockout mice |
publisher |
BMC |
series |
BMC Genomics |
issn |
1471-2164 |
publishDate |
2017-07-01 |
description |
Abstract Background Nuclear Factor Erythroid-derived 2-like 2 (Nrf2) senses oxidative environments and/or stress and initiates a cytoprotective response through transcriptional activation of antioxidant and detoxification genes. Several preclinical studies suggest that Nrf2 combats oxidative stress underlying a variety of pathologies. Despite Nrf2 deficits linked to functional abnormalities in many organ systems, the transcriptional network resulting from Nrf2 deficiency in the heart has remained elusive. Moreover, cross-talk between microRNAs (miRNAs) and cardiac Nrf2 signaling is unknown. Here, we utilized next generation RNA sequencing (RNAseq) to unbiasedly profile basal mRNA and miRNA expression in Nrf2 knockout (Nrf2−/−) hearts. Results RNAseq of mRNA revealed 152 differentially expressed genes (DEGs) in the Nrf2−/−myocardium, of which 129 were downregulated. Grouping of DEGs based on biological function and real-time qPCR validation indicated that DEGs were enriched for; mitochondrial genome and bioenergetics, oxidoreductase capacity, cardiac development, and chaperone activity. Interestingly, RNAseq analysis uncovered 27 significantly altered miRNAs, of which 11 were upregulated and 16 were downregulated in Nrf2−/− hearts. Expression changes were validated for 12 miRNAs using specific primer assays in real-time and revealed a significant decrease in miR-10b-5p, miR-674-3p, miR-3535, and miR-378c while miR-30b-5p, miR-208a-5p, miR-350-3p, and miR-582-5p, and miR-1249-3p levels were increased. High throughput data were integrated using prediction algorithms, and these in silico analyses discovered potential recognition elements within 39 repressed mRNAs which matched the seed sequence for 4 upregulated miRNAs; miR-30b-5p, miR-208a-5p, miR-350-3p, and miR-582-5p. Conclusion These high-throughput data reveal transcriptome-wide effects of myocardial Nrf2 deficiency. Further, our results suggest that Nrf2 may directly or indirectly regulate a sub-set of cardiac miRNAs in the basal setting. This comprehensive analysis is the first evidence to demonstrate a plausible regulatory cross-talk among cardiac miRNAs and the Nrf2 transcriptional network, and provides valuable candidates to examine in future mechanistic and preclinical studies. |
topic |
Nrf2 microRNA mRNA Heart RNA sequencing Differential expression |
url |
http://link.springer.com/article/10.1186/s12864-017-3875-3 |
work_keys_str_mv |
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