Systems-level regulation of microRNA networks by miR-130/301 promotes pulmonary hypertension
Thomas Bertero, … , Katherine A. Cottrill, Stephen Y. Chan
Thomas Bertero, … , Katherine A. Cottrill, Stephen Y. Chan
Published June 24, 2014
Citation Information: J Clin Invest. 2014;124(8):3514-3528. https://doi.org/10.1172/JCI74773.
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Systems-level regulation of microRNA networks by miR-130/301 promotes pulmonary hypertension

Abstract

Development of the vascular disease pulmonary hypertension (PH) involves disparate molecular pathways that span multiple cell types. MicroRNAs (miRNAs) may coordinately regulate PH progression, but the integrative functions of miRNAs in this process have been challenging to define with conventional approaches. Here, analysis of the molecular network architecture specific to PH predicted that the miR-130/301 family is a master regulator of cellular proliferation in PH via regulation of subordinate miRNA pathways with unexpected connections to one another. In validation of this model, diseased pulmonary vessels and plasma from mammalian models and human PH subjects exhibited upregulation of miR-130/301 expression. Evaluation of pulmonary arterial endothelial cells and smooth muscle cells revealed that miR-130/301 targeted PPARγ with distinct consequences. In endothelial cells, miR-130/301 modulated apelin-miR-424/503-FGF2 signaling, while in smooth muscle cells, miR-130/301 modulated STAT3-miR-204 signaling to promote PH-associated phenotypes. In murine models, induction of miR-130/301 promoted pathogenic PH-associated effects, while miR-130/301 inhibition prevented PH pathogenesis. Together, these results provide insight into the systems-level regulation of miRNA-disease gene networks in PH with broad implications for miRNA-based therapeutics in this disease. Furthermore, these findings provide critical validation for the evolving application of network theory to the discovery of the miRNA-based origins of PH and other diseases.

Authors

Thomas Bertero, Yu Lu, Sofia Annis, Andrew Hale, Balkrishen Bhat, Rajan Saggar, Rajeev Saggar, W. Dean Wallace, David J. Ross, Sara O. Vargas, Brian B. Graham, Rahul Kumar, Stephen M. Black, Sohrab Fratz, Jeffrey R. Fineman, James D. West, Kathleen J. Haley, Aaron B. Waxman, B. Nelson Chau, Katherine A. Cottrill, Stephen Y. Chan

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Figure 2

Upregulation of the miR-130/301 family by hypoxia is mediated by HIF-2α and POU5F1/OCT4.

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Upregulation of the miR-130/301 family by hypoxia is mediated by HIF-2α ...
(A) As demonstrated by RT-qPCR, hypoxia (0.2% O2 for 24 hours) upregulated the miR-130/301 family in both human PAECs and PASMCs as compared with normoxia (21% O2 for 24 hours). (B) In PAECs transfected with siRNA control (si-NC) or siRNA specific for HIF-1α (si-HIF-1α), hypoxia upregulated all miR-130/301 family members compared with normoxia. In contrast, in normoxia, miR-130/301 family members were downregulated during HIF-2α knockdown (si-HIF-2α) compared with control. Moreover, during HIF-2α knockdown, family members were not induced by hypoxia. (C) In normoxic PAECs, lentiviral transduction with a constitutively expressed HIF-1α transgene carrying a proline-to-alanine mutation (pHIF1) did not alter miRNA expression compared with transduction with empty vector (pEmpty) alone. In contrast, hypoxia upregulated all miR-130/301 family members compared with normoxia in the presence of pEmpty alone. (D) During HIF-2α (si-HIF-2α) or POU5F1/OCT4 (si-OCT4) knockdown in PAECs, miR-130/301 family members were not induced by hypoxia, in contrast to cells transfected with siRNA control (si-NC). In each panel, for each miRNA, mean expression in normoxic control groups (21% O2 in A; si-NC 21% O2 in B and D; pEmpty 21% O2 in C) was assigned a fold change of 1, to which relevant samples were compared. Data are presented as mean ± SD (*P < 0.05; **P < 0.01).