Professional Documents
Culture Documents
com
ScienceDirect
Angiotensin type 1 (AT1) receptors are key effector elements of two distinct pharmacological classes: angiotensin type 1
the renin–angiotensin system, mediating virtually all of the (AT1) and type 2 (AT2) receptors based on their differ-
classical physiological actions of angiotensin II. ential affinities for various nonpeptide antagonists. The
Pharmacological blockade of the AT1 receptor effectively AT1 receptor mediates virtually all of the classically
lowers blood pressure in a substantial proportion of patients recognized physiological actions of Ang II eliciting pow-
with hypertension, indicating the pivotal role of these receptors erful regulation of homeostatic cardiovascular functions
in human hypertension. AT1 receptors are expressed in many while contributing to the pathogenesis of cardiovascular
different organ systems where they have myriad cellular and kidney diseases.
actions. However, several lines of evidence have suggested
that direct actions of AT1 receptors in kidney have a major role AT1 receptors are expressed in all of the key organ
in regulation of blood pressure and in the pathogenesis of systems involved in regulating blood pressure including
hypertension. Here we review recent studies suggesting that the kidney, vasculature, immune system, adrenal gland,
renal epithelium and vasculature may be key cellular targets, heart and both central and peripheral nervous system.
where AT1 receptor activation has powerful physiological Furthermore, the AT1 receptor-associated proteins,
impact. We will also examine novel regulatory mechanisms by ATRAP and ARAP1, have been shown to be important
peptides associated with the C-terminus of the AT1 receptor. in determining the density of AT1 receptors on the cell
Addresses surface by interacting with the carboxyl-terminus of the
1
Division of Nephrology, Department of Medicine, Duke University and AT1 receptor and regulating receptor trafficking and
Durham VA Medical Centers, Durham, NC 27710, USA degradation. This review will focus on the AT1 receptor
2
Cardiovascular and Metabolic Disorders Research Program, Duke-
and its associated proteins in the vasculature and kidney
NUS Graduate Medical School, Singapore 169857, Singapore
epithelia, highlighting key findings that enhance under-
Corresponding author: Coffman, Thomas M (tcoffman@duke.edu) standing RAS biology while pointing toward new strate-
gies for preventing and treating hypertension and
Current Opinion in Pharmacology 2015, 21:122–126
cardiovascular diseases.
This review comes from a themed issue on Cardiovascular and renal
Edited by Pernille BL Hansen and Boye L Jensen
AT1 receptors in kidney epithelia in blood
pressure control
For a complete overview see the Issue and the Editorial
The importance of AT1 receptors in the kidney in regu-
Available online 14th February 2015 lating renal sodium handling and blood pressure has been
http://dx.doi.org/10.1016/j.coph.2015.01.006 long recognized. Activation of AT1 receptors in the kid-
1471-4892/# 2015 Elsevier Ltd. All rights reserved. ney leads to renal vasoconstriction and increased sodium
reabsorption [2–4]. Kidney cross-transplantation studies
from our laboratory using wild-type and AT1A receptor-
deficient mice demonstrated a key contribution of AT1
receptors in the kidney to maintaining baseline blood
pressure [5]. In addition, we found that AT1 receptors in
Introduction the kidney are required for the full phenotype of Ang II-
The renin–angiotensin system (RAS) has a central role in dependent hypertension, promoting hypertension by
blood pressure regulation and fluid balance. While the driving renal sodium retention [6]. However, AT1 recep-
RAS is critical for maintaining normal circulatory homeo- tors are expressed by many cell lineages in the kidney
stasis, especially during episodes of sodium deprivation, including epithelial cells across the nephron and renal
abnormal activation can contribute to the development of vasculature. Our cross-transplantation experiments could
hypertension and end-organ damage. Sequential cleavage not distinguish the relative contributions of these indi-
of the substrate angiotensinogen by renin and angioten- vidual cell lineages to blood pressure control.
sin-converting enzyme (ACE) generates the octapeptide
hormone angiotensin II (Ang II), the major physiological- There is an abundant literature demonstrating effects of
ly active peptide of the RAS. Ang II triggers its biological Ang II acting via AT1 receptors to modulate sodium and
effects by stimulating receptors belonging to the large fluid reabsorption along the nephron, particularly in the
family of 7-transmembrane, G protein-coupled receptors proximal tubule [4,7]. As technology has advanced, more
(GPCRs) [1]. These receptors can be further divided into sophisticated tools are available to allow delineation of
AT1 receptor functions in specific cell lineages. Cell- hypertension. In this regard, mice lacking ACE in the
specific gene targeting using the Cre–loxP technique, kidney exhibited enhanced natriuresis and resistance to
which allows precise localization and timing of gene L-NAME-induced hypertension [14]. These mice also
deletions or insertions, has proven very useful for such had preserved glomerular filtration rate (GFR) during L-
physiological studies. Accordingly, we and others used NAME, along with reduced abundance of several key
Cre–loxP techniques to generate mice with cell-specific transporter moieties including NHE3, NaPi2, gENaC,
deletion AT1A receptors from epithelia in the proximal and the phosphorylated forms of NKCC2 and NCC.
tubule of the kidney [8,9]. Our studies showed that AT1 Altogether, these studies highlight the importance of
receptors in the proximal tubule play critical roles both in Ang II generation within the kidney, which then acts
basal blood pressure control and in the pathogenesis of on AT1 receptors on renal epithelium and other cell
Ang II-induced hypertension. This is achieved by modu- lineages, to promote the development of hypertension.
lating fluid and solute reabsorption by the proximal
tubule through controlling the abundance of key sodium Vascular AT1 receptors and end-organ
transporters including the Na+/H+ exchanger 3 (NHE3) damage in hypertension
and the Na+-Pi cotransporter (NaPi2) [8–10]. AT1 receptors are broadly expressed on the vasculature,
particularly in vascular smooth muscle cells (VSMCs) but
In addition to these effects in the proximal tubule, recent also in other cell types involved in vascular pathology
studies have also suggested that direct actions of Ang II including endothelium, adventitia, and macrophages.
on AT1 receptors in the collecting duct can affect blood Along with their actions to induce vasoconstriction and
pressure by directly stimulating the activity of the epi- increase peripheral vascular resistance, vascular AT1A
thelial sodium channel (ENaC) [11]. This study indicat- receptors have been implicated in aneurysm formation.
ed that aldosterone and Ang II act coordinately to increase For example, a role for AT1 receptors has been suggested
the number of functionally active channels during low in humans with Marfan’s syndrome and in non-syndromic
sodium feeding. Furthermore, during Ang II-dependent thoracic aortic aneurysms and dissections [15]. Likewise,
hypertension, ENaC becomes overactive and is not ef- Ang II appears to promote development of aortic aneur-
fectively suppressed by inhibition of the aldosterone– ysms in mouse models with hyperlipidemia and hyper-
mineralocorticoid receptor axis, indicating a major role for tension [15–18]. Specifically, Ang II infusion in mice
direct stimulation by Ang II [11]. leads to pronounced aortic dilation and dissections that
are highly restricted to the ascending aorta, and these
The intra-renal RAS: autonomous regulation effects are modified by genetic background and exagger-
of Ang II generation in the kidney ated in the presence of concomitant atherosclerosis
Previous work by the Navar group and others has docu- [18,19]. Aneurysm formation is prevented by the admin-
mented the existence of an intra-renal RAS with inde- istration of the AT1 receptor antagonist, losartan, indicat-
pendent control of Ang II generation compared to the ing a critical role for AT1 receptors in their pathogenesis
circulating system [12]. These studies suggest a feed- [15]. To define the cell lineage mediating these actions,
forward stimulation of Ang II generation during Ang II- Rateri and colleagues performed bone marrow transplants
dependent hypertension, at a time when there is sup- and cell-specific elimination of AT1A receptors from
pression of the circulating RAS. Recent studies by Gon- endothelium and vascular smooth muscle cells [20].
zalez-Villalobos et al. have clearly demonstrated the Using these approaches, they found that only depletion
importance of the intra-renal RAS in this process, utilizing of AT1A receptors from endothelium attenuated the
mice genetically modified such that they are unable to development of aneurysms, indicating that Ang II infu-
produce Ang II in the kidney [13] These studies high- sion promotes aortic aneurysms via stimulation of AT1A
light a fundamental requirement for de novo generation of receptors expressed by endothelium. However, the
Ang II by ACE in the kidney in the development of Ang mechanism of this surprising finding is not clear.
II-dependent hypertension. Specifically, mice without
renal ACE have a significantly blunted hypertensive To define the contribution of AT1A receptors in vascular
response to chronic Ang II infusion, despite marked smooth muscle cells to aortic remodeling during hyperten-
increases in circulating Ang II. This resistance to hyper- sion, Cre/loxP technology was used to generate mice from
tension was accompanied by alterations in key ion trans- VSMC using a Cre transgene expressed in larger conduit
porters such as Na+/K+/2Cl co-transporter (NKCC2), the vessels but not in resistance vessels such as glomerular
thiazide-sensitive sodium chloride co-transporter (NCC), arterioles [21]. Acute and chronic hypertensive responses
ENaC and pendrin, as well as the transporter activating to Ang II were unaffected in these mice, consistent with
kinases SPAK and OSR1 [13], indicating that intra-renal lack of excision of AT1 receptors from resistance vessels.
generation of Ang II controls the activation of key sodium However, vascular oxidative stress was significantly atten-
transporters and this is a key pathway for hypertension uated in mice lacking AT1 receptors in VSMCs, consistent
pathogenesis. This pathway for intra-renal generation of with a role for these receptors to promote generation reac-
Ang II was also shown to be important in another form of tive oxygen species in Ang II-dependent hypertension.
Nonetheless, despite the apparent reduction in oxidative cells [31]. The inhibitory effect of ATRAP on Ang II
stress, the extent of aortic medial expansion induced by Ang action has been demonstrated in vivo [31,32]. For exam-
II was not affected in the mice lacking AT1A receptors from ple, in transgenic mice overexpressing ATRAP in all
smooth muscle. Taken together, these studies suggest that tissues, both inflammatory femoral artery remodeling
actions of AT1A receptors in VSMCs are not required for after injury and cardiac hypertrophy after aortic banding
aortic remodeling in Ang II-dependent hypertension were significantly reduced [32]. These models of end-
[20,21]. organ injury are also attenuated by angiotensin receptor
blockers (ARBs) reflecting their dependence on AT1
Along with regulation by circulating vasoconstrictors such receptors. In another transgenic mouse line with broad
as Ang II, stretch-activated alterations in diameter of over-expression of ATRAP, baseline blood pressures
small peripheral arteries and arterioles is another impor- were unaffected. On the other hand, sodium retention
tant mechanism for controlling vascular tone. These so- and the severity of hypertension during chronic Ang II
called ‘myogenic responses’ allow dynamic autoregula- infusion were attenuated. This was associated with sup-
tion of blood flow through immediate adjustments in pressed renal NCC and aENaC activation [33]. Thus,
vessel diameters, providing pressure-sensitive control generalized over-expression of ATRAP appears to atten-
for maintaining blood flow in the presence of fluctuations uate AT1 receptor actions in the kidney in vivo, consistent
in perfusion pressure. A recent study by Schleifenbaum with its capacity to suppress AT1 receptor signaling.
and colleagues identified a major role for AT1 receptors in
myogenic responses in the systemic and renal vasculature. ATRAP loss-of-function models have also been generat-
Furthermore, these actions were ligand-independent, as ed. Oppermann and colleagues found high blood pres-
they could be detected in the absence of angiotensinogen sure, plasma volume expansion, and suppressed plasma
and, thus, did not require the presence of Ang II [22]. renin in ATRAP knockouts, a phenotype consistent with
Furthermore, this study suggested that the transient re- enhanced AT1 receptor activation in the kidney [31].
ceptor potential channel 6 (TRPC6) and several voltage- Ohsawa and colleagues also developed a line of mice with
gated K+ gene family (KCNQ) channel gene families, targeted ablation of the ATRAP gene. While baseline
previously implicated in myogenic responses, did not make blood pressures were unaffected in their ATRAP-defi-
a major contribution to the response. Instead, these effects cient mouse line, these animals also had exaggerations of
seemed to require an XE991-sensitive K+ channel in hypertension and plasma volume expansion with chronic
VSMC [22]. The more proximal signaling pathways Ang II infusion [34]. Expression and activity of the major
mediating this response are not clear, but previous work sodium transporter in the distal tubules, ENaC, was also
characterizing ligand-independent, stretch-activation of significantly enhanced by chronic Ang II infusion in
AT1A receptors in cardiac myocytes linked this pathway ATRAP knockout mice [34]. On the basis of these
to b-arrestin signaling [23]. studies, ATRAP seems to be a negative modulator of
AT1 receptors in the renal tubules and loss of ATRAP
Specific regulation of AT1 receptor signaling results in enhanced renal absorptive function, leading to
by the intra-cellular proteins ATRAP and volume expansion and hypertension.
ARAP1
The physiological actions of AT1 receptors are controlled The other AT1 receptor associated protein, ARAP1 also
by regulatory mechanisms typical of other GPCRs includ- binds to the C terminal domain of the receptor. However,
ing transcription, desensitization and receptor recycling ARAP1 promotes recycling of the AT1 receptor to the cell
[24]. However, recent studies indicate additional control membrane [26] thereby increasing AT1 receptor expres-
by two intra-cellular proteins: AT1 receptor-associated sion, and potentially enhancing AT1 receptor signaling,
protein, ATRAP [25] and ARAP1 [26], which interact actions that are in direct opposition to ATRAP. In this
with the carboxyl-terminal domain of the AT1 receptor. regard, transgenic mice overexpressing of ARAP1 in the
As with other GPCRs, the carboxyl terminus of the AT1 proximal tubule under the control of the KAP promoter
receptor is phosphorylated and plays an important role in developed salt-sensitivity and Ang II dependent hyper-
regulating receptor desensitization and internalization tension, which was accompanied by marked kidney hy-
[27]. pertrophy [35]. Expression of ARAP1 has been detected
in the renal vasculature of mice and humans [36], and its
ATRAP is a relatively small protein, encoding an open expression is suppressed by Ang II infusion. Thus, these
reading frame of 161 amino acids with a predicted mo- two proteins interacting directly with the carboxyl-termi-
lecular mass of 18 kDa. ATRAP promotes the internali- nus of the AT1 receptor have the capacity to provide
zation of the AT1 receptors in cultured VSMC [28,29] and opposing regulation of AT1 receptor activation and its
inhibits Ang II-mediated intracellular signaling [30]. physiological consequences. With more detailed under-
ATRAP is expressed at high levels in the proximal tubule standing of their functions, it is conceivable that these
of the kidney, particularly the brush border, with negligi- pathways might be exploited therapeutically in cardio-
ble expression in the renal vasculature or juxtaglomerular vascular diseases.
ligand-independent. Furthermore, this response depends on an unu- 31. Oppermann M, Gess B, Schweda F, Castrop H: ATRAP
sual voltage-gated K+ channel. deficiency increases arterial blood pressure and plasma
volume. J Am Soc Nephrol 2010, 21:468-477.
23. Rakesh K, Yoo B, Kim IM, Salazar N, Kim KS, Rockman HA: beta- The highest level of ATRAP expression is localized to the proximal tubule,
Arrestin-biased agonism of the angiotensin receptor induced suggesting that the kidney may be a key target of this peptide. ATRAP-
by mechanical stress. Sci Signal 2010, 3:ra46. deficient mice have high BP, increased plasma volume and suppressed
plasma renins. Thus, ATRAP seems to target AT1 receptors in the kidney
24. Ferguson SS: Evolving concepts in G protein-coupled receptor
to control blood pressure by modulating volume status.
endocytosis: the role in receptor desensitization and
signaling. Pharmacol Rev 2001, 53:1-24. 32. Oshita A, Iwai M, Chen R, Ide A, Okumura M, Fukunaga S, Yoshii T,
25. Daviet L, Lehtonen JY, Tamura K, Griese DP, Horiuchi M, Dzau VJ: Mogi M, Higaki J, Horiuchi M: Attenuation of inflammatory
Cloning and characterization of ATRAP, a novel protein that vascular remodeling by angiotensin II type 1 receptor-
interacts with the angiotensin II type 1 receptor. J Biol Chem associated protein. Hypertension 2006, 48:671-676.
1999, 274:17058-17062. 33. Wakui H, Tamura K, Masuda S, Tsurumi-Ikeya Y, Fujita M,
26. Guo DF, Chenier I, Tardif V, Orlov SN, Inagami T: Type Maeda A, Ohsawa M, Azushima K, Uneda K, Matsuda M et al.:
1 angiotensin II receptor-associated protein ARAP1 binds and Enhanced angiotensin receptor-associated protein in renal
recycles the receptor to the plasma membrane. Biochem tubule suppresses angiotensin-dependent hypertension.
Biophys Res Commun 2003, 310:1254-1265. Hypertension 2013, 61:1203-1210.
Transgenic mice over-expressing ATRAP exhibit resistance to Ang II-
27. Guo DF, Sun YL, Hamet P, Inagami T: The angiotensin II type induced hypertension with a concomitant increase in natriuresis and
1 receptor and receptor-associated proteins. Cell Res 2001, suppression of ENaC. These findings are also consistent with a role of
11:165-180. ATRAP to inhibit AT1 receptor actions in the kidney.
28. Cui T, Nakagami H, Iwai M, Takeda Y, Shiuchi T, Tamura K, 34. Ohsawa M, Tamura K, Wakui H, Maeda A, Dejima T, Kanaoka T,
Daviet L, Horiuchi M: ATRAP, novel AT1 receptor associated Azushima K, Uneda K, Tsurumi-Ikeya Y, Kobayashi R et al.:
protein, enhances internalization of AT1 receptor and inhibits Deletion of the angiotensin II type 1 receptor-associated protein
vascular smooth muscle cell growth. Biochem Biophys Res enhances renal sodium reabsorption and exacerbates
Commun 2000, 279:938-941. angiotensin II-mediated hypertension. Kidney Int 2014, 86:570-581.
ATRAP deficiency exacerbates Ang II mediated hypertension by promot-
29. Azuma K, Tamura K, Shigenaga A, Wakui H, Masuda S, Tsurumi- ing sodium retention via pathological activation of ENaC.
Ikeya Y, Tanaka Y, Sakai M, Matsuda M, Hashimoto T et al.: Novel
regulatory effect of angiotensin II type 1 receptor-interacting 35. Guo DF, Chenier I, Lavoie JL, Chan JS, Hamet P, Tremblay J,
molecule on vascular smooth muscle cells. Hypertension 2007, Chen XM, Wang DH, Inagami T: Development of hypertension
50:926-932. and kidney hypertrophy in transgenic mice overexpressing
ARAP1 gene in the kidney. Hypertension 2006, 48:453-459.
30. Tanaka Y, Tamura K, Koide Y, Sakai M, Tsurumi Y, Noda Y,
Umemura M, Ishigami T, Uchino K, Kimura K et al.: The novel 36. Doblinger E, Hocherl K, Mederle K, Kattler V, Walter S, Hansen PB,
angiotensin II type 1 receptor (AT1R)-associated protein Jensen B, Castrop H: Angiotensin AT1 receptor-associated
ATRAP downregulates AT1R and ameliorates cardiomyocyte protein Arap1 in the kidney vasculature is suppressed by
hypertrophy. FEBS Lett 2005, 579:1579-1586. angiotensin II. Am J Physiol Renal Physiol 2012, 302:F1313-F1324.