Small RNA Sequencing Reveals MicroRNAs That Modulate Angiotensin II Effects in Vascular Smooth Muscle Cells

Wen, Jia; Reddy, Marpadga A.; Chen, Zhuo; Putta, Sumanth; Lanting, Linda; Kato, Mitsuo; Park, Sun-Hee; Chandra, Manasa; Wang, Charles; Tangirala, Rajendra K.; Natarajan, Rama

doi:10.1074/jbc.m111.322669

Cited by 75 publications

(77 citation statements)

References 58 publications

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“…Aortic VSMC were isolated from 10–12 weeks old type 2 diabetic db/db mice and control db/+ mice (Jackson laboratories, Bar Harbor, ME) and cultured as described 12, 33 . RNA extraction and gene expression analysis were performed by quantitative Realtime PCR (RT-qPCR) 28, 33 . Small RNA-seq (smRNA-seq) was performed once, with one sample per group using Illumina protocols.…”

Section: Methodsmentioning

confidence: 99%

“…SI). Transient transfection of VSMC, immunoblotting, luciferase assays, cell proliferation and migration assays were performed as described earlier with some modifications 28, 33 . All the experiments were repeated at least twice with replicates.…”

Section: Methodsmentioning

confidence: 99%

“…Vascular injury was found to downregulate miR-143/145, but upregulate miR-21, miR-146a, and miR-222 and promote synthetic phenotype 18, 23–30 . Angiotensin II induced miR-132 in VSMC which was associated with increased inflammatory gene expression 28 . Some miRNAs have been implicated in inflammatory and fibrotic gene regulation related to diabetic vascular complications 31, 32 .…”

Section: Introductionmentioning

confidence: 99%

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Regulation of Vascular Smooth Muscle Cell Dysfunction Under Diabetic Conditions by miR-504

Reddy

Das

Chen

et al. 2016

ATVB

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Objectives Diabetes accelerates pro-atherogenic and pro-inflammatory phenotype of VSMC associated with vascular complications. Evidence shows that microRNAs (miRNAs) play key roles in VSMC functions, but their role under diabetic conditions is unclear. We profiled miRNAs in VSMC from diabetic mice and examined their role in VSMC dysfunction. Approach and Results High throughput small RNA-sequencing identified 135 differentially expressed miRNAs in VSMC from type-2 diabetic db/db mice (db/dbVSMC) versus non-diabetic db/+ mice. Several of these miRNAs were known to regulate VSMC functions. We further focused on miR-504, because it was highly upregulated in db/dbVSMC, and its function in VSMC is unknown. miR-504 and its host gene Fgf13 were significantly increased in db/dbVSMC and in aortas from db/db mice. Bioinformatics analysis predicted that miR-504 targets including signaling adaptor Grb10 and transcription factor Egr2 could regulate growth factor signaling. We experimentally validated Grb10 and Egr2 as novel targets of miR-504. Overexpression of miR-504 in VSMC inhibited contractile genes, and enhanced ERK1/2 activation, proliferation and migration. These effects were blocked by miR-504 inhibitors. Grb10 knockdown mimicked miR-504 functions and increased inflammatory genes. Egr2 knockdown inhibited anti-inflammatory Socs1 and increased pro-inflammatory genes. Furthermore, high-glucose and palmitic acid upregulated miR-504 and inflammatory genes, but downregulated Grb10. Conclusions Diabetes mis-regulates several miRNAs including miR-504 that can promote VSMC dysfunction. Since changes in many of these miRNAs are sustained in diabetic VSMC even after in vitro culture, they may be involved in metabolic memory of vascular complications. Targeting such mechanisms could offer novel therapeutic strategies for diabetic complications.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Regulation of Vascular Smooth Muscle Cell Dysfunction Under Diabetic Conditions by miR-504

Reddy

Das

Chen

et al. 2016

ATVB

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“…It is well known that the biology of the glomerular compartment is heavily influenced by the renin-angiotensin system (RAS) that regulates local intrarenal hemodynamics, which apparently worsens the pathological outcome of a given glomerular disease. Prime examples where the activation of the RAS or ANG II contributes to or amplifies the injury are diabetic nephropathy and cardiovascular smooth cell dysfunctions, as reported in several studies, reviews, and perspectives (13,24,35,49,62,70). In any event, it is conceivable that the activation of the RAS besides causing perturbation in the glomerular compartment may also modulate the tubulointerstitial injury since some of the RAS components are believed to be expressed within this compartment (33,37).…”

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Modulation of angiotensin II-induced inflammatory cytokines by the Epac1-Rap1A-NHE3 pathway: implications in renal tubular pathobiology

Xie

Joladarashi

Dudeja

et al. 2014

American Journal of Physiology-Renal Physiology

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Besides the glomerulus, the tubulointerstitium is often concomitantly affected in certain diseases, e.g., diabetic nephropathy, and activation of the renin-angiotensin system, to a certain extent, worsens its outcome because of perturbations in hemodynamics and possibly tubuloglomerular feedback. Certain studies suggest that pathobiology of the tubulointerstitium is influenced by small GTPases, e.g., Rap1. We investigated the effect of ANG II on inflammatory cytokines, while at the same time focusing on upstream effector of Rap1, i.e., Epac1, and some of the downstream tubular transport molecules, i.e., Na/H exchanger 3 (NHE3). ANG II treatment of LLC-PK1 cells decreased Rap1a GTPase activity in a time- and dose-dependent manner. ANG II treatment led to an increased membrane translocation of NHE3, which was reduced with Epac1 and PKA activators. ANG II-induced NHE3 translocation was notably reduced with the transfection of Rap1a dominant positive mutants, i.e., Rap1a-G12V or Rap1a-T35A. Transfection of cells with dominant negative Rap1a mutants, i.e., Rap1a-S17A, or Epac1 mutant, i.e., EPAC-ΔcAMP, normalized ANG II-induced translocation of NHE3. In addition, ANG II treatment led to an increased expression of inflammatory cytokines, i.e., IL-1β, IL-6, IL-8, and TNF-α, which was reduced with Rap1a-G12V or Rap1a-T35A transfection, while it reverted to previous comparable levels following transfection of Rap1a-S17A or EPAC-ΔcAMP. ANG II-induced expression of cytokines was reduced with the treatment with NHE3 inhibitor S3226 or with Epac1 and PKA activators. These data suggest that this novel Epac1-Rap1a-NHE3 pathway conceivably modulates ANG II-induced expression of inflammatory cytokines, and this information may yield the impetus for developing strategies to reduce tubulointertstitial inflammation in various renal diseases.

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“…They found that miR-195 (alone of the miR-15 family) was increased in aneurysmal aortic tissue from Angiotensin II(AngII)-treated apoE−/− mice. AngII has previously been shown to induce or inhibit miR-15a, -15b, -16-1 and -16-2 in either rat or human smooth muscle cells (SMC), but not miR-195 10, 11 . Further, while significant downregulation of miR-15a, miR-195 and miR-497 has been observed in tissue from dissected human thoracic aorta when compared with normal aorta, Pahl et al’s expression profiling of human AAA tissue did not identify any differentially regulated miR-15 family members 12, 13 .…”

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Small RNA Sequencing Reveals MicroRNAs That Modulate Angiotensin II Effects in Vascular Smooth Muscle Cells

Cited by 75 publications

References 58 publications

Regulation of Vascular Smooth Muscle Cell Dysfunction Under Diabetic Conditions by miR-504

Regulation of Vascular Smooth Muscle Cell Dysfunction Under Diabetic Conditions by miR-504

Modulation of angiotensin II-induced inflammatory cytokines by the Epac1-Rap1A-NHE3 pathway: implications in renal tubular pathobiology

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