MicroRNA-140-5p and SMURF1 regulate pulmonary arterial hypertension
Alexander M.K. Rothman, … , David J. Rowlands, Allan Lawrie
Alexander M.K. Rothman, … , David J. Rowlands, Allan Lawrie
Published May 23, 2016
Citation Information: J Clin Invest. 2016;126(7):2495-2508. https://doi.org/10.1172/JCI83361.
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MicroRNA-140-5p and SMURF1 regulate pulmonary arterial hypertension

Abstract

Loss of the growth-suppressive effects of bone morphogenetic protein (BMP) signaling has been demonstrated to promote pulmonary arterial endothelial cell dysfunction and induce pulmonary arterial smooth muscle cell (PASMC) proliferation, leading to the development of pulmonary arterial hypertension (PAH). MicroRNAs (miRs) mediate higher order regulation of cellular function through coordinated modulation of mRNA targets; however, miR expression is altered by disease development and drug therapy. Here, we examined treatment-naive patients and experimental models of PAH and identified a reduction in the levels of miR-140-5p. Inhibition of miR-140-5p promoted PASMC proliferation and migration in vitro. In rat models of PAH, nebulized delivery of miR-140-5p mimic prevented the development of PAH and attenuated the progression of established PAH. Network and pathway analysis identified SMAD-specific E3 ubiquitin protein ligase 1 (SMURF1) as a key miR-140-5p target and regulator of BMP signaling. Evaluation of human tissue revealed that SMURF1 is increased in patients with PAH. miR-140-5p mimic or SMURF1 knockdown in PASMCs altered BMP signaling, further supporting these factors as regulators of BMP signaling. Finally, Smurf1 deletion protected mice from PAH, demonstrating a critical role in disease development. Together, these studies identify both miR-140-5p and SMURF1 as key regulators of disease pathology and as potential therapeutic targets for the treatment of PAH.

Authors

Alexander M.K. Rothman, Nadine D. Arnold, Josephine A. Pickworth, James Iremonger, Loredana Ciuclan, Robert M.H. Allen, Sabine Guth-Gundel, Mark Southwood, Nicholas W. Morrell, Matthew Thomas, Sheila E. Francis, David J. Rowlands, Allan Lawrie

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Figure 7

miR-140-5p and SMURF1 modulate BMP signaling in vitro, and SMURF1 is critical to the development of experimental PAH.

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miR-140-5p and SMURF1 modulate BMP signaling in vitro, and SMURF1 is cri...
(A–C) miR-140-5p mimic and SMURF1 siRNA augment BMP signaling in vitro. (A) PASMCs were transfected with SCR and miR-140-5p mimic or NTsiRNA and SMURF1 siRNA prior to stimulation with BMP4. At 72 hours, expression of native ID1 protein was increased in miR-140-5p mimic– and SMURF1 siRNA–treated cells (n = 4, mean ± SEM). (B) PASMCs were cotransfected with BRE construct and SCR or miR-140-5p prior to stimulation with BMP4. At 24 hours, BRE activity was increased in miR-140-5p mimic–treated cells. (C) PASMCs were cotransfected with BRE construct and NTsiRNA or SMURF1 siRNA prior to stimulation with BMP4. At 24 hours, BRE activity was increased in SMURF1 siRNA–treated cells. (B and C: n = 4, *P < 0.05, 2-tailed Mann-Whitney U test, mean ± SEM). (D–F) Genetic ablation of SMURF1 confers protection from PAH in the SuHx mouse model. (D) RVSP at week 3. (E) RVH at week 3. (F) Vessel muscularization at week 3. (D–F: n = 11–13 per group, *P < 0.05; **P < 0.01; ***P < 0.001, ****P < 0.0001, 1-way ANOVA with Tukey’s post-test correction, mean ± SEM).