M I N I R E V I E W

MicroRNAs in Cardiovascular Health: From Order to

Disorder
Denuja Karunakaran and Katey J. Rayner
University of Ottawa Heart Institute, Ottawa, Ontario K1Y4W7 Canada

In the last decade, microRNAs (miRNAs) have revolutionized how we understand metabolism and
disease. These small, 20- to 22-nucleotide RNA molecules fine-tune gene expression and can often
coordinate multiple genes in a single pathway. Given the multifactorial nature of cardiovascular dis-
ease, it is perhaps not surprising that miRNAs have been shown to orchestrate many aspects of disease
development, from modulating metabolic risk factors over a lifetime (eg, cholesterol and hormones)
to controlling the response to an acute cardiovascular event (eg, inflammation and hypoxia). In this
review, we discuss how miRNAs exert control over metabolic pathways that maintain vascular health
and, when these pathways go awry, how miRNAs can be targeted for therapeutic modulation.
(Endocrinology 154: 4000 4009, 2013)

n the three decades since the discovery and subsequent lar health and, when these pathways go awry, how miRNAs
I widespread use of statins as lipid-lowering agents, there
has been a paucity of new therapies to treat and manage
can be targeted for therapeutic modulation.

the risk factors for cardiovascular disease (1). Although

sudden death from cardiac events (ie, myocardial infarc- The miRNA Revolution
tion [MI]) has decreased dramatically over the past 20
years, a significant majority of the population in the west- The miRNAs were first discovered in 1993, when Victor
ern world continues to suffer from hyperlipidemia, over- Ambros and colleagues (3) undertook a genetic screen to
nutrition, and obesity, all risk factors for cardiometabolic identify essential genes for lineage commitment in developing
disease (2). In the past, drug development has largely fo- Caenorhabditis elegans. They unexpectedly found that a
cused on pharmacological inhibition of single targets, of- small antisense RNA (termed lin-4) was binding to and in-
ten using small molecules, as a means to modulate bio- hibiting the expression of a protein-coding transcript, LIN-
logical pathways that promote cardiovascular disease 14, which was required for commitment to the first postem-
and/or its associated complications. However, this ap- bryonic larval stage. Nearly a decade later, it was revealed
proach has been plagued with difficulties, owing largely to that antisense RNA was not just an anomaly in worms but
the lack of tissue specificity of small molecules as well as that it was highly conserved and even found in humans (4).
their often dangerous off-target effects. Hence, there Almost immediately, additional miRNAs were discovered in
remains an urgent need to refocus efforts in therapeutic both C. elegans and mammals, and so began the miRNA
development away from pharmacologic modulators revolution (57). To date, there have been over a thousand
and instead exploit newly uncovered molecules and mech- individual miRNAs annotated in the human genome, with
anisms. MicroRNAs (miRNAs) are now understood to be more added every year, and over 50% of the human genome
powerful modulators of biological pathways and have is estimated to be under miRNA control (8, 9).
generated considerable excitement as next-generation miRNAs begin as long, primary transcripts that are
therapy for a variety of diseases, including cardiometa- most commonly transcribed by RNA polymerase II from
bolic disease. In this review, we will discuss how miRNAs intergenic, intronic, or even exonic locations in the ge-
exert control over metabolic pathways that maintain vascu- nome (10). They are cleaved in the nucleus by the ribo-

ISSN Print 0013-7227 ISSN Online 1945-7170 Abbreviations: ABC, ATP-binding cassette transporters; EC, endothelial cell; eNOS, endo-
Printed in U.S.A. thelial NO synthase; ER, estrogen receptor-; HDL, high-density lipoprotein; KLF4, Krp-
Copyright 2013 by The Endocrine Society pel-like factor 4; LDL, low-density lipoprotein; MI, myocardial infarction; miRNA, mi-
Received March 31, 2013. Accepted August 26, 2013. croRNA; NFB, nuclear factor kappa-light-chain-enhancer of activated B cells; PI3K,
First Published Online September 5, 2013 phosphoinositide 3-kinase; pre-miRNA, precursor miRNA; SMC, smooth muscle cell;
VCAM-1, vascular cell adhesion molecule 1.

4000 endo.endojournals.org Endocrinology, November 2013, 154(11):4000 4009 doi: 10.1210/en.2013-1299

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doi: 10.1210/en.2013-1299 endo.endojournals.org 4001

nuclease III enzyme-complex Drosha/DGCR8 (DiGeorge LDL deposits in the subendothelial space, where it is sub-
Syndrome chromosomal region 8, also known as pasha) to jected to oxidative damage by the environment rich in reac-
form smaller precursor miRNA (pre-miRNA) products. This tive oxygen species and oxidizing enzymes like myeloperox-
microprocessor complex is exported from the nucleus where idase (16). Macrophages and other inflammatory cells
it is further cleaved by Dicer to form the mature, 20- to 22- present in atherosclerotic plaques are an abundant source of
nucleotide double-stranded miRNA molecules. One strand, both reactive oxygen species and myeloperoxidase, and their
termed the effector/functional strand, of the miRNA is persistence in lesions causes an accumulation of these oxi-
loaded into the RNA-induced silencing complex, which con- dized LDL products. The uptake of oxidized LDL through
sists of RNA binding proteins like the Argonautes (ie, Ago1 scavenger receptor CD36 is itself inflammatory, which
and Ago2) with cleavage activity. The RNA-induced silenc- causes additional recruitment and retention of macrophages
ing complex is guided to complementary regions in specific to the lesional area, propagating the inflammatory cascade
mRNA transcripts, most often (but not exclusively) located and ultimately lesion progression (17). Excess lipid accumu-
in 3-untranslated regions of target genes. Until recently, it lates in macrophage foam cells and, if not efficiently removed
was believed that the passenger strand (previously denoted through the cholesterol efflux pathway, largely mediated by
with an asterisk) was simply degraded and was thus disre- the cholesterol transporters ATP-binding cassette transport-
garded as nonfunctional; however, evidence is mounting that ers (ABC), ABCA1 and ABCG1, will lead to additional in-
these passenger strands can also be functional and bind flammatory activation and even cell death. On the other
mRNA transcripts under certain circumstances (11). Com- hand, HDL acts as an opposing force to LDL and can enter
plete complementary base pairing between the miRNA:m- the subendothelial space and act as an acceptor for the excess
RNA duplex results in destabilization or degradation of the cholesterol that is effluxed through ABCA1 and ABCG1.
mRNA, whereas partial base pairing results in deadenylation This cholesterol is then brought to the liver, taken up via
or translational inhibition (12, 13). Generally speaking, the scavenger receptor BI, and excreted through the bile and feces
consequence of miRNA:mRNA duplex formation is a down- in a process known as reverse cholesterol transport. HDL is
regulation of target protein expression, although exceptions made by the liver, intestine, and adipose tissue by ABCA1, yet
to this have been reported (14, 15). Frequently, the magni- it is the efficiency of reverse cholesterol transport that is be-
tude of protein repression enforced by miRNA is modest; lieved to underlie the benefits often ascribed to high levels of
that is, any one miRNA may cause only a 20% decrease in HDL and is now being touted as the most important metric
individual target gene protein expression under basal condi- of HDL function (18).
tions. However, miRNAs often repress multiple members of miRNAs have been shown to control lipoproteins and
the same pathway simultaneously, making their cumulative their pro-/antiatherogenic effects at many levels (Figure
impact on cellular function more potent and dramatic. Given 1A). The first miRNA demonstrated to specifically mod-
the multifactorial nature of cardiovascular disease, it is per- ulate cholesterol balance was miR-33. miR-33 is a lipid-
haps not surprising that miRNAs have been shown to or- responsive miRNA embedded in the introns of the sterol
chestrate many aspects of disease development, from mod- regulatory transcription factor genes SREBP-1 and
ulating metabolic risk factors over a lifetime (eg, cholesterol SREBP-2 and is cotranscribed along with its host genes
and hormones) to controlling the response to an acute car- under conditions that induce SREBP activation (19 21).
diovascular event (eg, inflammation and hypoxia). miR-33 represses the expression of ABCA1, ABCG1, and
NPC1, which ultimately prevents cholesterol export from
cells. Inhibition of miR-33 was shown to induce the ex-
Metabolic Risk Factor Regulation by pression of ABCA1 not only in the liver, where it increased
miRNAs circulating HDL levels but also in atherosclerotic plaque
macrophages where it promoted cholesterol removal from
Regulation of cholesterol and lipoproteins by miRNAs the plaque and decreased lesion size (22). Genetic deletion
Lipids are carried in the bloodstream by lipoproteins of miR-33 also resulted in sustained increases in HDL as
where they are shuttled to and from tissues as required. Low- well as a decrease in atherosclerosis progression in
density lipoprotein (LDL) delivers cholesterol and phospho- Apoe/ mice (23, 24). miR-33 can also control the fatty
lipids from the liver to tissues in need, whereas high-density acid oxidation and synthesis pathway, and modulation of
lipoprotein (HDL) ferries excess lipids away from peripheral miR-33 levels in nonhuman primates reduced circulating
tissues back to the liver for excretion. High levels of LDL very-low-density lipoprotein, another important risk fac-
cholesterol and low levels of HDL cholesterol are both in- tor for cardiometabolic disease (25). Another miRNA,
dependent risk factors for the development of atherosclerosis miR-144, was recently shown to modulate ABCA1 ex-
and its downstream complications. Consequently, excess pression and cholesterol efflux from hepatocytes, and in-

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4002 Karunakaran and Rayner miRNA and Cardiovascular Health Endocrinology, November 2013, 154(11):4000 4009

hibition of miR-144 in vivo increases hepatic ABCA1 and Estrogen and other hormones are under the
circulating HDL (26, 27). Unlike that of HDL, the influence of miRNAs
miRNA-mediated control of LDL cholesterol has been less Steroid hormones are an essential component of develop-
well studied. miR-122, the most abundant miRNA ex- ment and are required for homeostatic function of many
pressed in liver, controls both LDL and HDL cholesterol tissues, including the vessel wall. The importance of estrogen
levels, primarily by indirect modulation of cholesterol syn- and its cognate receptors, estrogen receptor- (ER) and
thesis pathways, although the exact mechanisms remain in- ER, on both the physiology and pathophysiology of the
completely understood (28 30). Recently, miR-27b was cardiovascular system have been well described (3234). In
identified as a potential regulatory hub of lipid metabolism in brief, estrogen promotes vasodilation through induction of
the liver, potentially controlling both HDL cholesterol and endothelial NO synthase (eNOS) by vascular endothelial
triglyceride levels (31). It is interesting to postulate that miR- cells (ECs), while it also prevents vascular smooth muscle cell
27b could be exerting control over an entire network of lipid (SMC) proliferation and contraction (35, 36). Moreover, es-
metabolism genes, although these observations have yet to be trogen has been shown to reduce cytokine expression in the
systematically validated in vivo. vessel wall (37) and has even been demonstrated to reduce

Figure 1. microRNAs in cardiovascular health. A, miRNAs controls lipid metabolism, lipoprotein clearance and their pro- or anti-atherogenic
effects at multiple sites. B, In the vessel wall, estrogen regulates miRNA function in vascular cells. C, miRNAs target multiple cell types and
functions within the vessel wall to modulate inflammatory responses and atherosclerosis progression. D, Several miRNAs have been shown to
intricately regulate cardiac fibrosis, hypertrophy and remodelling and repair post-ischemic injury or MI.

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doi: 10.1210/en.2013-1299
LDL and raise HDL through modulation of apolipoprotein
expression (32). These beneficial effects of estrogen underlie
the observation that premenopausal women, by and large,
have lower rates of cardiovascular disease than postmeno-
pausal women, and postmenopausal women are at greater
risk of dying from heart disease than age-matched men (38).
Although clinical trials have been confusing and often
fraught with controversy, most of the evidence supports the
notion that hormone replacement therapy immediately after
the onset of menopause protects against cardiovascular dis-
ease and its associated complications. Interestingly, estrogen
has been shown to control specific miRNA expression in vivo
and in vitro, and it is believed that ER is primarily respon-
sible for these effects (39). This is accomplished through di-
rect interaction of ER with the microprocessor complex
Drosha, which governs the cleavage of primary miRNA tran-
scripts to the pre-miRNA transcripts (39). Notably, ER-
mediated destabilization of Drosha decreased the expression
of miR-125a and miR-16, which then provided a positive-
feedback loop through derepression of ER-induced target
genes (39). This elegant study revealed the complexity of
estrogen and ER in the biogenesis of miRNA and began to
shed light on the multifaceted nature of estrogen action on
cellular function (Figure 1B). In the vessel wall, estrogen con-
trols the expression of miR-203, which then negatively reg-
ulates the cellular proliferation program through Abl1 and
p63 (40). Although these observations have yet to be con-
firmed in vivo, this was the first study demonstrating a role
for estrogen-mediated miRNA expression in controlling vas-
cular cell homeostasis, and full examination of the spectrum
of miRNA-mediated estrogen action in the vasculature is
anticipated.
Control of obesity, insulin signaling, and glucose
regulation by miRNAs
Type 2 diabetes is a chronic metabolic disorder caused by
a combination of altered insulin secretion and progressive
tissue insulin resistance. Diabetic complications can further
exacerbate cardiovascular risk factors such as hypertension,
dyslipidemia, obesity, and heart failure, all of which increase
the overall cardiovascular disease burden (2). In healthy in-
dividuals, high blood glucose levels stimulate pancreatic
-cells to secrete insulin, promoting the uptake of glucose by
peripheral target tissues such as skeletal muscle and adipose
tissue (41). Glucose homeostasis is stringently regulated by a
variety of interacting signaling pathways that synchronize
both gene and protein expression and function, and miRNAs
were recently identified as another level of posttranscrip-
tional regulation of these pathways (Figure 1A). The most
abundant miRNA in pancreatic endocrine cells, miR-375,
negatively regulates glucose-induced insulin secretion by di-
rectly repressing myotrophin (MTPN), an activator of NFB
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endo.endojournals.org 4003
(nuclear factor kappa-light-chain-enhancer of activated B
cells) and regulator of vesicular trafficking of insulin, and
3-phosphoinositide-dependent protein kinase-1 (Pdk1), an
integral constituent of the insulin-mediated PI3K-AKT/PKB
(phosphoinositide 3-kinase-AKT/protein kinase B) signaling
pathways (42, 43). miR-375 knockout mice have decreased
-cell mass and increased gluconeogenesis and
hepatic glucose output and are hyperglycemic, demonstrat-
ing the important role miR-375 has in regulating glucose
homeostasis (43). Interestingly, both diabetic patients (44)
and ob/ob diabetic mice (43) have elevated miR-375 expres-
sion and -cell mass but no significant changes in -cell mass,
suggesting miR-375 overexpression may contribute to the
observed decreased insulin and the resultant high blood glu-
cose levels, all contributing factors for type 2 diabetes. Ago2
has recently been implicated in regulating pancreatic -cell
exocytosis, because small interfering RNA knockdown of
Ago2 increased expression of miR-375 target genes gephyrin
and ywhaz, which enhanced insulin secretion in response to
glucose (45). Thus, miRNA-mediated control of pancreatic
gene expression is essential for proper insulin secretion and
homeostasis. Pancreatic -cell function is regulated in a dis-
tinct manner by miR-33. Cholesterol homeostasis in pancre-
atic -cells is maintained by ABCA1, and regulation of cel-
lular cholesterol content is required for proper insulin
secretion (46). Similar to macrophages and hepatocytes, as
mentioned above, overexpression of miR-33 also inversely
modulates ABCA1 expression in pancreatic -cells, and this
in turn corresponds to higher cholesterol levels and decreased
glucose-mediated insulin exocytosis from these cells (47). In-
hibition of miR-33 improved insulin secretion from islet cells
in vitro from hypercholesterolemic mice, suggesting that this
pathway is indeed impaired in these mice. miR-33 also mod-
ulates the tissue response to insulin by its regulation of insulin
receptor substrate 2 (IRS2), a critical component in mediat-
ing insulin-signaling pathways, and, when impaired, con-
tributes to insulin resistance (48). Although the effects of
miR33 inhibition therapy have yet to be tested in models of
insulin resistance or diabetes in vivo, it would stand to reason
that antimiR-33 would promote glucose-mediated insulin
exocytosis from -cells while simultaneously restoring insu-
lin-mediated signaling pathways in peripheral tissues and,
thus, improving the development of atherosclerosis and
diabetes.
In addition to its release from pancreatic -cells, miRNAs
modulate the downstream response to insulin by altering
components of the insulin signaling machinery in periph-
eral tissues. miR-103 and miR-107, which differ by only
one nucleotide, are necessary for insulin signaling through
the insulin receptor (49). Inactivation or silencing of miR-
103/107 upregulates caveolin-1, which in turn stabilizes
the insulin receptor and promotes insulin sensitivity in the
4004 Karunakaran and Rayner miRNA and Cardiovascular Health
liver and adipose tissue. Notably, the expression of miR-
103/107 is raised in the liver of both obese and type 2
diabetic mouse models (49, 50), and inactivation of these
miRNAs improves glucose homoeostasis in diabetic mice
(49). miR-143/145 expression is similarly upregulated in
the liver of obese and diabetic mice (51). miR-143 and
miR-145 target the AKT signaling pathway, and mice de-
ficient in the miR-143/145 cluster are protected from the
development of diet-induced insulin resistance. Taken to-
gether, these studies suggest that manipulating miRNA
clusters (rather than a single miRNA) could present po-
tential therapies to treat obesity and the development of
type 2 diabetes.
Control of Cardiovascular Inflammation
and Injury by miRNAs
Inflammation in the vessel wall is regulated by
multiple miRNAs
As described above, modified lipoproteins trapped in the
vessel wall instigate the inflammatory response that underlies
atherosclerosis and the dangerous development of the un-
stable, rupture-prone plaque. Apart from monocytes/mac-
rophages, other key cells involved in this inflammatory cas-
cade include ECs, vascular SMCs, leukocytes, and platelets.
ECs are considered the inflammatory gatekeepers of the ves-
sel wall, and resting ECs do not express inflammatory mark-
ers like VCAM-1 (vascular cell adhesion molecule-1). Upon
activation by cytokines, they increase their expression of
VCAM-1 and recruit inflammatory cells to the subendothe-
lial space, and it is the constant recruitment of these cells into
the intima that leads to the formation of large atherosclerotic
lesions. miR-126 in ECs directly targets the untranslated re-
gion of VCAM-1 to repress its expression, and inhibition of
miR-126 upregulates VCAM-1 expression and leukocyte
adherence to ECs (52). Interestingly, EC apoptotic bodies
shed microvesicles containing miR-126, which represses
RGS16 (regulator of G protein signaling 16) in arteries, de-
creases production of the inflammatory chemokine
CXCL12 (chemokine (C-X-C motif) ligand 12), decreases
recruitment of inflammatory cells, and overall reduces in-
flammation and stabilizes the atherosclerotic plaque (53).
miR-126 is now recognized as an important regulator of
vascular EC maintenance and is integral to the progression of
atherosclerosis (Figure 1C).
miR-155 was recently found to be upregulated in ath-
erosclerotic lesions in both mice and humans and is decreased
in the circulation of coronary artery disease patients (54, 55).
Despite these observations, the data supporting the exact role
of miR-155 in the progression of atherosclerosis is contra-
dictory to date. Transplantation of miR-155/ mice bone
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Endocrinology, November 2013, 154(11):4000 4009
marrow into high-fat diet-fed LDLR/ mice resulted in in-
creased atherosclerotic lesion development and increased le-
sional inflammation (55, 56). However, hyperlipidemic
apoE/ mice transplanted with miR-155/ bone marrow
displayed decreased macrophage content and reduced ath-
erosclerotic plaque size (55). In macrophage foam cells, miR-
155 targets Bcl6, an NFB antagonist, which promotes pro-
inflammatory signals such as CCL2 expression (55).
However, miR-155 is also required for early cell fate deci-
sions in inflammatory cells, and deficiency of miR-155 alters
the profile of monocytes and neutrophils in miR-155/ hy-
perlipidemic mice (56). The role of miR-155 in inflammatory
precursor cell fate vs advanced macrophage foam cells likely
underlies the discrepancy observed between the two mouse
models and highlights the dual role of miR-155 in promoting
atherosclerosis. Interestingly, miR-3425p was found to di-
rectly activate the expression of miR-155 and is upregulated
in early atherosclerotic lesions of apoE/ mice (57). This in
turn induces proinflammatory mediators such as Nos2 and
IL-6. Furthermore, miR-155 expression is upregulated in
ECs and appears to suppress angiotensin II type I receptor
expression in these cells, reducing the proinflammatory and
vascular remodeling roles of angiotensin II (59). The thera-
peutic potential of miR-155 will thus depend on the stage of
disease when therapy is initiated but will likely have benefi-
cial outcomes in the patient with advanced atherosclerotic
disease.
Encompassed in the same gene cluster but with little ho-
mology to each other, both miR-143 and miR-145 are highly
conserved and expressed in SMCs, and their expression is
markedly reduced in coronary artery disease patients (54, 60,
61). Apart from their role in the pancreas, miR-143/145 ex-
pression is also upregulated by transcription factors in vessel
wall SMCs, resulting in a combinatorial effect where miR-
143 inhibits SMC proliferation and miR-145 promotes SMC
differentiation through direct targeting of Krppel-like fac-
tor 4 (KLF4), myocardin, and Elk-1 (ELK1) as well as an-
giotensin converting enzyme (ACE) and its cognate receptor
(60, 61). Genetic knockdown of miR-143/145 results in the
development of spontaneous atherosclerotic lesions in old
mice (60). SMC-specific overexpression of miR-145 using
lentivirus in mice resulted in reduced plaque size, reduced
necrotic core, increased fibrous cap area, and increased
plaque collagen content, all of which may contribute to sta-
bilizing the plaque (62). Additionally, the vascular shear-
responsive transcription factor KLF2 upregulates the miR-
143/145 cluster in ECs, and these miRs are secreted into the
extracellular space, where they are thought to target SMCs
and thus reduce atherosclerosis lesion progression in
apoE/ mice (63). Thus, miR-143/145 regulates vascular
wall homeostasis from both intracellular and extracellular
doi: 10.1210/en.2013-1299
mechanisms, adding another layer of complexity in
miRNA-mediated gene regulation.
Exaggerated platelet responses or hyperactivity is thought
to contribute to the development of thrombotic and occlu-
sive events that lead to life-threatening MI. Because platelets
are anucleated and possess no genomic DNA and little
mRNA, there had been minimal focus on understanding
transcriptional regulation in platelet function until recently.
Landry et al (64) performed a comprehensive study of miR-
NAs in platelets, demonstrating the presence of 219 miRNAs
and Dicer and Ago2 complexes; however, understanding the
role of platelet miRNAs is in its infancy. miR-223 has been
implicated as a key regulator of P2Y12 (purinergic receptor
P2Y, G-protein coupled 12), which binds to soluble ADP
that is released from activated platelet granules to further
propagate the platelet aggregation response (64). Another
study further confirmed a functional role for platelet miR-
NAs, and here they implicated that miR-200b can repress
PRKAR2B (protein kinase cAMP-dependent type II-beta),
decreasing platelet responsiveness (65). It is possible that
platelet miRNAs not only serve as transcriptional regulators
within platelets but also secrete miRNAs into the circulation
to potentially target specific cells in response to vascular in-
jury (66). As a result, platelet-derived miRNAs may serve as
diagnostic biomarkers of platelet activation or post-anti-
platelet therapy (65, 67).
Cardiac injury and remodeling after MI: miRNAs
play a key role
The build-up and subsequent rupture of a vulnerable ath-
erosclerotic plaque leads to an MI, resulting in cardiac tissue
ischemia and necrosis. After restoration of blood flow, the
response to injury involves deposition of extracellular matrix
and the formation of a fibrotic scar in the damaged area. In
addition, inflammatory cells are recruited to the site of injury,
and angiogenesis is activated to restore blood flow to dam-
aged tissue. Several miRNAs have been shown to intricately
regulate cardiac fibrosis, hypertrophy, and remodeling and
repair post-ischemic injury or MI (Figure 1D). Perhaps the
most well-studied miRNA in the post-MI environment is
miR-21, whose expression is induced in a number of path-
ological conditions (68). The role of miR-21 in cardiac re-
modeling is controversial. Cholesterol-modified antisense
oligonucleotides against miR-21 inhibited ERK-MAPK ki-
nase pathways in cardiac fibroblasts through miR-21s spe-
cific target, Spry1, resulting in the attenuation of interstitial
fibrosis and cardiac hypertrophy and dysfunction in mice
(69). In contrast, another study used locked nucleic acid-
modified anti-miR oligonucleotides in vivo to inhibit miR-21
and demonstrated no difference in the pathological cardiac
remodeling in response to cardiac stress, which was further
confirmed in a miR-21 knockout model (70). The discrep-
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endo.endojournals.org 4005
ancies between these two studies could be attributable to the
anti-miR therapies used, which differ in their pharmacoki-
netic profiles, especially with respect to potency, toxicity,
tissue uptake, and off-target effects. The observation that
cardiac remodeling was unaffected with long-term repres-
sion of miR-21, such as in the knockout model, suggests that
there may be miRNA redundancy or compensatory mecha-
nisms at play. Additionally, inhibition of miR-21 in SMCs
upregulates phosphatase and tensin homolog (PTEN),
which in turn inhibits the PI3K/AKT pathways resulting in
decreased SMC proliferation and increased apoptosis, caus-
ing attenuation of neointima formation after angioplasty
(71). Caution must be taken in considering miR-21targeted
therapies for cardiovascular disease, due to the ubiquitous
expression of miR-21, which makes the development of tis-
sue- or cell-specific therapies essential.
Expressed solely in the heart, miR-208a has been shown
to regulate cardiac hypertrophy by repressing negative reg-
ulators (eg, thyroid hormone-associated protein 1 and myo-
statin) of muscle growth and hypertrophy while failing to
upregulate myosin heavy chain (MYH7), a key marker in
cardiac remodeling, (72, 73). Therapeutic inhibition of this
miRNA prevented the development of heart failure in rats via
the prevention of myosin switching in damaged hearts (74).
Interestingly, antimiR-208a also decreased circulating
miRNA levels in the serum, including miR-499, a biomarker
of heart injury. These data suggest that the beneficial effects
of miR-208a inhibition may be a synergistic effect of both
myosin upregulation and decreasing circulating miRNA. In
addition, miR-208a regulates systemic energy homeostasis
by negatively regulating MED13, and pharmacological in-
hibition of miR-208a confers protection to diet-induced obe-
sity by increasing MED13 expression and increasing energy
expenditure (75). The inhibition of miR-208a thus provides
a provocative strategy for both the repair of cardiac tissue
after MI while simultaneously improving metabolic energy
output to decrease overall cardiometabolic risk. Other miR-
NAs that are downregulated during cardiac hypertrophy in
both mice and human include miR-1 and miR-133 (76).
Knockout of both miR-133a isoforms (miR-133a1 and miR-
133a2) resulted in embryonic or postnatal lethality after se-
vere cardiac fibrosis, heart failure, and cardiac malforma-
tions (77). In contrast, transgenic overexpression of miR-
133a protected hearts from fibrosis after aortic constriction
(78). These studies support the notion that forced expression
of miR-133 may be a viable therapeutic option to reduce
cardiac hypertrophy. Interestingly, and excitingly, a recent
study demonstrated that antagonism of miR-133 induces the
phenotypic switching of skeletal muscle progenitor cells to
brown adipocytes, which increases whole-body thermogen-
esis and impedes the development of obesity (79). Thus, over-
expressing miR-133 and/or inhibiting miR-208a would
4006 Karunakaran and Rayner miRNA and Cardiovascular Health
likely have benefits in cardiac tissue as well as in other tissues,
decreasing other cardiometabolic risk factors such as obesity.
Damage to the myocardium by MI or ischemic injury
requires remodeling and healing processes, which includes
the generation of new blood vessels via angiogenesis. Both
ECs and SMCs are involved in these processes, and several
miRNAs have been implicated in regulating them. Inhibition
of miR-92a in mice, a component of the highly expressed
miR-1792 cluster, increased invading ECs and formation of
new blood vessels through their targets integrin-5 and
eNOS (80). Furthermore, systemic administration of the
miR-92a antagomir in an MI mouse model enhanced neo-
vascularization, functional recovery of damaged tissue, in-
creased left ventricular function, decreased cardiomyocyte
apoptosis, and reduced infarct size (80). More recently, miR-
92a knockdown has been shown to enhance KLF4 and KLF2
expression, which is thought to be protective against athero-
genesis (81). As indicated above, miR-126 not only increases
VCAM1 expression in ECs but also has been implicated in
maintaining vascular integrity and angiogenesis during de-
velopment through regulation of its targets, SPRED1 and
PIK3RS (82, 83). Many other miRNAs have been implicated
in the regulation of angiogenesis (see Table 1), but many of
them have not been studied in the context of cardiovascular
injury and repair.
Therapy for Cardiovascular Disease Based
on miRNAs
Only a decade has elapsed since the discovery of miRNAs
in humans, yet already a number of miRNA-based therapies
are gaining strides as potential therapeutics. This can be at-
tributed to the relatively rapid development of effective, po-
tent anti-miRNA targeting strategies that were quickly used
in vivo in preclinical models. Effective inhibition of miRNAs
can be achieved through single-stranded antisense oligonu-
cleotides, similar to those used for small interfering RNA
silencing of mRNA gene transcripts. Various chemical mod-
ifications have been used to stabilize these anti-miRNA mol-
ecules, improve their toxicity, and improve their tissue up-
take, reducing the effective dose for in vivo use (84). A
landmark study in 2005 by Krtzfeldt et al (30) used antago-
mirs (cholesterol-modified miRNA antisense molecules) in
mice to demonstrate that miR-122 could be effectively si-
lenced in vivo, revealing the importance of this miRNA-reg-
ulated pathway in lipid metabolism. Elmn et al (85) quickly
followed with miR-122 inhibition studies in African green
monkeys, which both confirmed the role of miR-122 in reg-
ulating lipid metabolism in vivo as well as highlighted the
therapeutic targetability of miRNAs in a model highly re-
lated to humans. To date, anti-miR122 remains the most
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Endocrinology, November 2013, 154(11):4000 4009
advanced miRNA-targeted therapeutic, primarily used in
patients suffering from hepatitis C infection. This is not re-
lated to the control of lipid metabolism genes by miR-122 but
instead is attributable to the hijacking of host hepatic miR-
122 for efficient viral replication and the relatively liver-spe-
cific expression of miR-122 (29). Overall, the safety profile
and efficacy of anti-miR122 (known as miraversen) has been
positive, with phase IIa results indicating that almost 50% of
patients treated with anti-miR122 had undetectable hepatitis
C viral loads after treatment (86). Although the future of
anti-miR122 for the treatment of hepatitis C infection looks
promising, its use for the treatment of hyperlipidemia awaits
further study. On the other hand, anti-miR33 is another
promising miRNA-targeting therapeutic for cardiometa-
bolic disease, which is quickly advancing through preclinical
testing for patients with atherosclerosis, and phase I trials are
projected to begin sometime in 2015 (84).
Summary and Future Directions
The observation that dysregulated miRNA expression
contributes on many levels to the pathogenesis of cardio-
vascular disease has led to considerable excitement for
their use as therapeutic targets. One could envision a pre-
ventative therapy that combines targeting of miR-33 and
miR-103/107 to improve lipid profiles and insulin resis-
tance while simultaneously targeting miR-126 and miR-
155 to reduce inflammation in the vessel wall. Or perhaps
a more acute delivery of anti-miR-208/21 and miR-133
mimics either systemically or locally after a myocardial
event could be used to improve the response to injury and
subsequent recovery. Although many of these miRNAs
likely will not enter clinical trials for years to come, the
realization that miRNAs represent such an untapped
source of therapeutic development has invigorated both
the academic and pharmaceutical communities alike, and
the therapeutic potential of these tiny RNAs is indeed
enormous.
Acknowledgments
Address all correspondence and requests for reprints to: Katey
Rayner, PhD, Assistant Professor, University of Ottawa Heart
InstituteBiochemistry, 40 Ruskin Street, H4211, Ottawa, On-
tario K1Y4W7, Canada. E-mail: krayner@ottawaheart.ca.
This study was funded by CIHR Operating Grant MOP
130365 (to K.J.R.) and UOHI Cardiovascular Genetics Research
Fellowship (to D.K.).
Disclosure Summary: The authors have nothing to disclose.
doi: 10.1210/en.2013-1299 endo.endojournals.org 4007

Table 1. miRNAs Regulating Cardiometabolic Health/Disease

Risk Factors and
miRNAs Target Cell/Tissue Effect Ref.
Cholesterol and
lipoproteins
miR-33 ABCA1, ABCG1, NPC1 Macrophages, liver HDL/cholesterol efflux 19 21
miR-144 ABCA1 Liver HDL/cholesterol efflux 26, 27
miR-122 ? (cholesterol synthesis pathways) Liver LDL, HDL 28 30
miR-27b Angptl3, Gpam Hepatocytes HDL, triglycerides 31
Hormones (eg, estrogen)
miR-16, miR-125a ER gene targets Breast cancer Estrogen-responsive gene changes 39
miR-203a Abl1, p63 Vascular SMCs SMC proliferation 40
Insulin resistance and
obesity
miR-375 NFB, PDK1 Pancreatic endocrine cells Insulin vesicular trafficking and 42, 43
exocytosis, - and -cell mass
Ago2 Gephyrin, ywhaz Pancreatic cell line, MIN6 Insulin exocytosis 45
miR-33 ABCA1, IRS2 Islet cells Cholesterol efflux, insulin 47, 48
exocytosis
miR-103, miR-107 Caveolin-1 Peripheral tissues (eg, liver, Stabilization of insulin receptor, 49, 50
adipose) insulin sensitivity
miR-143/145 AKT liver Insulin receptor signaling? 51
miR-208a MED13 Heart/systemic Energy expenditure 75
miR-133 Prdm16 Skeletal muscle progenitor cells Conversion to brown adipocytes, 78
whole-body thermogenesis
Vessel wall inflammation/
atherosclerosis
miR-126 VCAM-1, RGS16, SPRED1, ECs, arteries? Leukocyte adherence, 52, 53
PIK3R2/p85 atherosclerotic lesion
formation, CXCL12 production
miR-217 SIRTI, FOXO1 ECs Senescence 86
miR-155, miR-342-5p Bcl6, NFB Monocytes, macrophages, CCL2 expression, inflammatory 54 59
neutrophils, ECs, SMCs cell fate, angiotensin II receptor
expression
miR-143/145 KLF2, KLF4, Elk-1, ACE, ACE ECs, SMCs SMC proliferation and 54, 60 63
receptor differentiation
miR-223 P2Y12 Platelets Platelet activation and 64
aggregation
miR-200b PRKAR2B Platelets PKA activity, platelet 65
responsiveness
Cardiac injury/remodeling
after MI
miR-21 Spry1, PTEN Fibroblasts, SMCs Interstitial fibrosis, cardiac 69 71
hypertrophy and dysfunction,
cardiac remodeling, SMC
proliferation and apoptosis,
neointima formation
miR-208a Thyroid hormone-associated Cardiomyocytes Cardiac hypertrophy and 7274
protein 1, myostatin, MYH7, remodeling
miR-499
miR-133/133a1/2 RhoA, Cdc42 Cardiomyocytes Cardiac hypertrophy, cardiac 76
malformation, cardiac fibrosis,
heart failure
miR-92a Integrin-5, eNOS, KLF2, KLF4 ECs Angiogenesis, neovascularization, 80, 81
ventricular function,
cardiomyocyte apoptosis,
infarct size, atherogenesis
miR-126 SPRED1, PI3KS ECs Vascular integrity, angiogenesis 82, 83
miR-221/222 ECs Cell migration, angiogenesis 87
miR-24 GATA2, p21-activated kinase, ECs Apoptosis, vascularity, infarct size, 88
PAK4, BAD/Bcl-XL/Bcl-2 angiogenesis after MI
miR-320 Hsp20 Cardiomyocytes Apoptosis, infarct size post- 89
ischemic injury
miR-199b Dyrk1a ? Calcineurin/NFAT signaling, 90
cardiac hypertrophy, fibrosis,
heart failure
miR-15 Cell cycle checkpoint kinase 1 Cardiomyocytes Cardiomyocyte proliferation, post- 58, 91
ischemic injury or MI

Abbreviations: NPC1, Niemann-Pick disease type C1; Angptl3, angiopoietin-like 3; Gpam, glycerol-3-phosphate acyltransferase 1; MED13, mediator
complex subunit 13; Prdm16, PR domain containing 16; PKA, protein kinase A; SPRED1 Sprouty-related, EVH1 domain-containing protein 1; PIK3R2,
phosphoinositide-3-kinase, regulatory subunit 2; SIRTI, sirtuin 1; FOXO1, Forkhead box protein O1; Spry1, sprouty homolog 1; RhoA, Ras homolog gene
family, member A; Cdc42, cell division control protein 42 homolog; GATA2, GATA binding protein 2; BAD, Bcl-2-associated death promoter; Bcl, B-cell
leukemia/lymphoma; Hsp20, heat shock protein 20; Dyrk1a, dual specificity tyrosine-phosphorylation-regulated kinase 1A.

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4008 Karunakaran and Rayner miRNA and Cardiovascular Health
References
1. Tobert JA. Lovastatin and beyond: the history of the HMG-CoA
reductase inhibitors. Nat Rev Drug Discov. 2003;2(7):517526.
2. Bornfeldt KE, Tabas I. Insulin resistance, hyperglycemia, and ath-
erosclerosis. Cell Metab. 2011;14(5):575585.
3. Lee RC, Feinbaum RL, Ambros V. The C. elegans heterochronic
gene lin-4 encodes small RNAs with antisense complementarity to
lin-14. Cell. 1993;75(5):843 854.
4. Pasquinelli AE, Reinhart BJ, Slack F, et al. Conservation of the se-
quence and temporal expression of let-7 heterochronic regulatory
RNA. Nature. 2000;408(6808):86 89.
5. Lagos-Quintana M, Rauhut R, Lendeckel W, Tuschl T. Identifica-
tion of novel genes coding for small expressed RNAs. Science. 2001;
294(5543):853 858.
6. Lau NC, Lim LP, Weinstein EG, Bartel DP. An abundant class of tiny
RNAs with probable regulatory roles in Caenorhabditis elegans.
Science. 2001;294(5543):858 862.
7. Lee RC, Ambros V. An extensive class of small RNAs in Caeno-
rhabditis elegans. Science. 2001;294(5543):862 864.
8. Claverie JM. Fewer genes, more noncoding RNA. Science. 2005;
309(5740):1529 1530.
9. Pasquinelli AE. MicroRNAs and their targets: recognition, regula-
tion and an emerging reciprocal relationship. Nat Rev Genet. 2012;
13(4):271282.
10. Rodriguez A, Griffiths-Jones S, Ashurst JL, Bradley A. Identification
of mammalian microRNA host genes and transcription units. Ge-
nome Res. 2004;14(10A):19021910.
11. Mah SM, Buske C, Humphries RK, Kuchenbauer F. miRNA*: a
passenger stranded in RNA-induced silencing complex? Crit Rev
Eukaryot Gene Expr. 2010;20(2):141148.
12. Giraldez AJ, Mishima Y, Rihel J, et al. Zebrafish MiR-430 promotes
deadenylation and clearance of maternal mRNAs. Science. 2006;
312(5770):7579.
13. Wu L, Fan J, Belasco JG. MicroRNAs direct rapid deadenylation of
mRNA. Proc Natl Acad Sci U S A. 2006;103(11):4034 4039.
14. Lee S, Vasudevan S. Post-transcriptional stimulation of gene expres-
sion by microRNAs. Adv Exp Med Biol. 2013;768:97126.
15. Graff JW, Dickson AM, Clay G, McCaffrey AP, Wilson ME. Iden-
tifying functional microRNAs in macrophages with polarized phe-
notypes. J Biol Chem. 2012;287(26):21816 21825.
16. Heinecke JW. Oxidants and antioxidants in the pathogenesis of
atherosclerosis: implications for the oxidized low density lipopro-
tein hypothesis. Atherosclerosis. 1998;141(1):115.
17. Moore KJ, Tabas I. Macrophages in the pathogenesis of atheroscle-
rosis. Cell. 2011;145(3):341355.
18. Rader DJ, Tall AR. The not-so-simple HDL story: Is it time to revise
the HDL cholesterol hypothesis? Nat Med. 2012;18(9):1344 1346.
19. Rayner KJ, Surez Y, Dvalos A, et al. MiR-33 contributes to the
regulation of cholesterol homeostasis. Science. 2010;328(5985):
1570 1573.
20. Najafi-Shoushtari SH, Kristo F, Li Y, et al. MicroRNA-33 and the
SREBP host genes cooperate to control cholesterol homeostasis. Sci-
ence. 2010;328(5985):1566 1569.
21. Gerin I, Clerbaux LA, Haumont O, et al. Expression of miR-33 from
an SREBP2 intron inhibits cholesterol export and fatty acid oxida-
tion. J Biol Chem. 2010;285(44):3365233661.
22. Rayner KJ, Sheedy FJ, Esau CC, et al. Antagonism of miR-33 in mice
promotes reverse cholesterol transport and regression of atheroscle-
rosis. J Clin Invest. 2011;121(7):29212931.
23. Horie T, Baba O, Kuwabara Y, et al. MicroRNA-33 deficiency
reduces the progression of atherosclerotic plaque in ApoE/ mice.
J Am Heart Assoc. 2012;1(6):e003376.
24. Horie T, Ono K, Horiguchi M, et al. MicroRNA-33 encoded by an
intron of sterol regulatory element-binding protein 2 (Srebp2) regulates
HDL in vivo. Proc Natl Acad Sci U S A. 2010;107(40):1732117326.
25. Rayner KJ, Esau CC, Hussain FN, et al. Inhibition of miR-33a/b in
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 05 July 2016. at 23:43 For personal use only. No other uses without permission. . All rights reserved.
Endocrinology, November 2013, 154(11):4000 4009
non-human primates raises plasma HDL and lowers VLDL triglyc-
erides. Nature. 2011;478(7369):404 407.
26. de Aguiar Vallim TQ, Tarling EJ, Kim T, et al. MicroRNA-144
regulates hepatic ATP binding cassette transporter A1 and plasma
high-density lipoprotein after activation of the nuclear receptor
farnesoid X receptor. Circ Res. 2013;112(12):16021612.
27. Ramrez CM, Rotllan N, Vlassov AV, et al. Control of cholesterol
metabolism and plasma high-density lipoprotein levels by mi-
croRNA-144. Circ Res. 2013;112(12):15921601.
28. Esau C, Davis S, Murray SF, et al. miR-122 regulation of lipid me-
tabolism revealed by in vivo antisense targeting. Cell Metab. 2006;
3(2):8798.
29. Lanford RE, Hildebrandt-Eriksen ES, Petri A, et al. Therapeutic
silencing of microRNA-122 in primates with chronic hepatitis C
virus infection. Science. 2010;327(5962):198 201.
30. Krtzfeldt J, Rajewsky N, Braich R, et al. Silencing of microRNAs
in vivo with antagomirs. Nature. 2005;438(7068):685 689.
31. Vickers KC, Shoucri BM, Levin MG, et al. MicroRNA-27b is a
regulatory hub in lipid metabolism and is altered in dyslipidemia.
Hepatology. 2013;57(2):533542.
32. Mendelsohn ME, Karas RH. The protective effects of estrogen on the
cardiovascular system. N Engl J Med. 1999;340(23):18011811.
33. Knowlton AA, Lee AR. Estrogen and the cardiovascular system.
Pharmacol Ther. 2012;135(1):54 70.
34. Knowlton AA. Estrogen and cardiovascular disease: aging and estrogen
loss at the heart of the matter? Future Cardiol. 2012;8(1):9 12.
35. Darblade B, Pendaries C, Krust A, et al. Estradiol alters nitric oxide
production in the mouse aorta through the -, but not -, estrogen
receptor. Circ Res. 2002;90(4):413 419.
36. Nakamura Y, Suzuki T, Miki Y, et al. Estrogen receptors in ath-
erosclerotic human aorta: inhibition of human vascular smooth
muscle cell proliferation by estrogens. Mol Cell Endocrinol. 2004;
219(12):1726.
37. Chan GH, Fiscus RR. Exaggerated production of nitric oxide (NO)
and increases in inducible NO-synthase mRNA levels induced by the
pro-inflammatory cytokine interleukin-1 in vascular smooth mus-
cle cells of elderly rats. Exp Gerontol. 2004;39(3):387394.
38. Wilson PW, Garrison RJ, Castelli WP. Postmenopausal estrogen use,
cigarette smoking, and cardiovascular morbidity in women over 50.
The Framingham Study. N Engl J Med. 1985;313(17):1038 1043.
39. Yamagata K, Fujiyama S, Ito S, et al. Maturation of microRNA is
hormonally regulated by a nuclear receptor. Mol Cell. 2009;36(2):
340 347.
40. Zhao J, Imbrie GA, Baur WE, et al. Estrogen receptor-mediated
regulation of microRNA inhibits proliferation of vascular smooth
muscle cells. Arterioscler Thromb Vasc Biol. 2013;33(2):257265.
41. Ashcroft FM, Rorsman P. Diabetes mellitus and the -cell: the last
ten years. Cell. 2012;148(6):1160 1171.
42. El Ouaamari A, Baroukh N, Martens GA, Lebrun P, Pipeleers D, van
Obberghen E. miR-375 targets 3-phosphoinositide-dependent pro-
tein kinase-1 and regulates glucose-induced biological responses in
pancreatic -cells. Diabetes. 2008;57(10):2708 2717.
43. Poy MN, Hausser J, Trajkovski M, et al. miR-375 maintains normal
pancreatic - and -cell mass. Proc Natl Acad Sci U S A. 2009;
106(14):58135818.
44. ZhaoH,GuanJ,LeeHM,etal.Up-regulatedpancreatictissuemicroRNA-
375 associates with human type 2 diabetes through -cell deficit and islet
amyloid deposition. Pancreas. 2010;39(6):843846.
45. Tattikota SG, Sury MD, Rathjen T, et al. Argonaute2 regulates the
pancreatic -cell secretome. Mol Cell Proteomics. 2013;12(5):
1214 1225.
46. Brunham LR, Kruit JK, Pape TD, et al. -Cell ABCA1 influences
insulin secretion, glucose homeostasis and response to thiazolidin-
edione treatment. Nat Med. 2007;13(3):340 347.
47. Wijesekara N, Zhang LH, Kang MH, et al. miR-33a modulates
ABCA1 expression, cholesterol accumulation, and insulin secretion
in pancreatic islets. Diabetes. 2012;61(3):653 658.
doi: 10.1210/en.2013-1299
48. Dvalos A, Goedeke L, Smibert P,et al. miR-33a/b contribute to the
regulation of fatty acid metabolism and insulin signaling. Proc Natl
Acad Sci U S A. 2011;108(22):92329237.
49. Trajkovski M, Hausser J, Soutschek J, et al. MicroRNAs 103 and
107 regulate insulin sensitivity. Nature. 2011;474(7353):649 653.
50. Herrera BM, Lockstone HE, Taylor JM, et al. Global microRNA
expression profiles in insulin target tissues in a spontaneous rat
model of type 2 diabetes. Diabetologia. 2010;53(6):1099 1109.
51. Jordan SD, Krger M, Willmes DM, et al. Obesity-induced overex-
pression of miRNA-143 inhibits insulin-stimulated AKT activation
and impairs glucose metabolism. Nat Cell Biol. 2011;13(4):434 446.
52. Harris TA, Yamakuchi M, Ferlito M, Mendell JT, Lowenstein CJ.
MicroRNA-126 regulates endothelial expression of vascular cell ad-
hesion molecule 1. Proc Natl Acad Sci U S A. 2008;105(5):1516 1521.
53. Zernecke A, Bidzhekov K, Noels H, et al. Delivery of microRNA-
126 by apoptotic bodies induces CXCL12-dependent vascular pro-
tection. Sci Signal. 2009;2(100):ra81.
54. Fichtlscherer S, De Rosa S, Fox H, et al. Circulating microRNAs in
patients with coronary artery disease. Circ Res. 2010;107(5):677
684.
55. Nazari-Jahantigh M, Wei Y, Noels H, et al. MicroRNA-155 pro-
motes atherosclerosis by repressing Bcl6 in macrophages. J Clin
Invest. 2012;122(11):4190 4202.
56. Donners MM, Wolfs IM, Stoger LJ, et al. Hematopoietic miR155
deficiency enhances atherosclerosis and decreases plaque stability in
hyperlipidemic mice. PLoS One. 2012;7(4):e35877.
57. Wei Y, Nazari-Jahantigh M, Chan L, et al. The microRNA-3425p
fosters inflammatory macrophage activation through an Akt1- and
microRNA-155-dependent pathway during atherosclerosis. Circu-
lation. 2013;127(15):1609 1619.
58. Porrello ER, Johnson BA, Aurora AB, et al. MiR-15 family regulates
postnatal mitotic arrest of cardiomyocytes. Circ Res. 2011;109(6):
670 679.
59. Zhu N, Zhang D, Chen S, et al. Endothelial enriched microRNAs
regulate angiotensin II-induced endothelial inflammation and mi-
gration. Atherosclerosis. 2011;215(2):286 293.
60. Cordes KR, Sheehy NT, White MP, et al. miR-145 and miR-143
regulate smooth muscle cell fate and plasticity. Nature. 2009;
460(7256):705710.
61. Boettger T, Beetz N, Kostin S, et al. Acquisition of the contractile
phenotype by murine arterial smooth muscle cells depends on the
Mir143/145 gene cluster. J Clin Invest. 2009;119(9):2634 2647.
62. Lovren F, Pan Y, Quan A, et al. MicroRNA-145 targeted therapy
reduces atherosclerosis. Circulation. 2012;126(11 Suppl 1):S81
S90.
63. Hergenreider E, Heydt S, Trguer K, et al. Atheroprotective com-
munication between endothelial cells and smooth muscle cells
through miRNAs. Nat Cell Biol. 2012;14(3):249 256.
64. Landry P, Plante I, Ouellet DL, Perron MP, Rousseau G, Provost P.
Existence of a microRNA pathway in anucleate platelets. Nat Struct
Mol Biol. 2009;16(9):961966.
65. Nagalla S, Shaw C, Kong X, et al. Platelet microRNA-mRNA co-
expression profiles correlate with platelet reactivity. Blood. 2011;
117(19):5189 5197.
66. Diehl P, Fricke A, Sander L, et al. Microparticles: major transport
vehicles for distinct microRNAs in circulation. Cardiovasc Res.
2012;93(4):633 644.
67. Willeit P, Zampetaki A, Dudek K, et al. Circulating microRNAs as
novel biomarkers for platelet activation. Circ Res. 2013;112(4):
595 600.
68. Jazbutyte V, Thum T. MicroRNA-21: from cancer to cardiovascu-
lar disease. Curr Drug Targets. 2010;11(8):926 935.
69. Thum T, Gross C, Fiedler J, et al. MicroRNA-21 contributes to
myocardial disease by stimulating MAP kinase signalling in fibro-
blasts. Nature. 2008;456(7224):980 984.
70. Patrick DM, Montgomery RL, Qi X, et al. Stress-dependent cardiac
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 05 July 2016. at 23:43 For personal use only. No other uses without permission. . All rights reserved.
endo.endojournals.org 4009
remodeling occurs in the absence of microRNA-21 in mice. J Clin
Invest. 2010;120(11):39123916.
71. Ji R, Cheng Y, Yue J, et al. MicroRNA expression signature and
antisense-mediated depletion reveal an essential role of MicroRNA
in vascular neointimal lesion formation. Circ Res. 2007;100(11):
1579 1588.
72. Callis TE, Pandya K, Seok HY, et al. MicroRNA-208a is a regulator
of cardiac hypertrophy and conduction in mice. J Clin Invest. 2009;
119(9):27722786.
73. van Rooij E, Sutherland LB, Qi X, Richardson JA, Hill J, Olson EN.
Control of stress-dependent cardiac growth and gene expression by
a microRNA. Science. 2007;316(5824):575579.
74. Montgomery RL, Hullinger TG, Semus HM, et al. Therapeutic in-
hibition of miR-208a improves cardiac function and survival during
heart failure. Circulation. 2011;124(14):15371547.
75. Grueter CE, van Rooij E, Johnson BA, Det al. A cardiac microRNA
governs systemic energy homeostasis by regulation of MED13. Cell.
2012;149(3):671 683.
76. Car A, Catalucci D, Felicetti F, et al. MicroRNA-133 controls car-
diac hypertrophy. Nat Med. 2007;13(5):613 618.
77. Liu N, Bezprozvannaya S, Williams AH, et al. microRNA-133a regu-
lates cardiomyocyte proliferation and suppresses smooth muscle gene
expression in the heart. Genes Dev. 2008;22(23):32423254.
78. Matkovich SJ, Wang W, Tu Y, et al. MicroRNA-133a protects
against myocardial fibrosis and modulates electrical repolarization
without affecting hypertrophy in pressure-overloaded adult hearts.
Circ Res. 2010;106(1):166 175.
79. Yin H, Pasut A, Soleimani VD, et al. MicroRNA-133 controls
brown adipose determination in skeletal muscle satellite cells by
targeting Prdm16. Cell Metab. 2013;17(2):210 224.
80. Bonauer A, Carmona G, Iwasaki M, et al. MicroRNA-92a controls
angiogenesis and functional recovery of ischemic tissues in mice.
Science. 2009;324(5935):1710 1713.
81. Fang Y, Davies PF. Site-specific microRNA-92a regulation of Krup-
pel-like factors 4 and 2 in atherosusceptible endothelium. Arterio-
scler Thromb Vasc Biol. 2012;32(4):979 987.
82. Wang S, Aurora AB, Johnson BA, et al. The endothelial-specific
microRNA miR-126 governs vascular integrity and angiogenesis.
Dev Cell. 2008;15(2):261271.
83. Fish JE, Santoro MM, Morton SU, et al. miR-126 regulates angio-
genic signaling and vascular integrity. Dev Cell. 2008;15(2):272
284.
84. Broderick JA, Zamore PD. MicroRNA therapeutics. Gene Ther.
2011;18(12):1104 1110.
85. Elmn J, Lindow M, Schtz S, et al. LNA-mediated microRNA silenc-
ing in non-human primates. Nature. 2008;452(7189):896 899.
86. Franciscus A. Santaris Pharma A/S advances miravirsen, the first
microRNA-targeted drug to enter clinical trials, into Phase 2 to treat
patients infected with Hepatitis C virus. HCV Advocate website.
http://www.hcvadvocate.org/hepatitis/hepc/HCVDrugs.html. Ac-
cessed March 21, 2011.
87. Poliseno L, Tuccoli A, Mariani L, et al. MicroRNAs modulate the
angiogenic properties of HUVECs. Blood. 2006;108(9):3068
3071.
88. Fiedler J, Jazbutyte V, Kirchmaier BC, et al. MicroRNA-24 regulates
vascularity after myocardial infarction. Circulation. 2011;124(6):
720 730.
89. Ren XP, Wu J, Wang X, et al. MicroRNA-320 is involved in the
regulation of cardiac ischemia/reperfusion injury by targeting heat-
shock protein 20. Circulation. 2009;119(17):23572366.
90. da Costa Martins PA, Salic K, Gladka MM, et al. MicroRNA-199b
targets the nuclear kinase Dyrk1a in an auto-amplification loop
promoting calcineurin/NFAT signalling. Nat Cell Biol. 2010;
12(12):1220 1227.
91. Hullinger TG, Montgomery RL, Seto AG, et al. Inhibition of miR-15
protects against cardiac ischemic injury. Circ Res. 2012;110(1):
71 81.