bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 094: Smooth Muscle PPARγ Mutation Causes Impaired Renal Blood Flow and Salt-Sensitive Hypertension
Authors: WU Jing , Agbor Larry N , Mukohda Masashi , Nair Anand R , Nakagawa Pablo , Morgan Donald A , Rahmouni Kamal , Gotlinger Katherine H , Schwartzman Michal L , Sigmund Curt D
Published: December 2018

Mutations in PPARγ cause hypertension (HT) while PPARγ activation lowers blood pressure (BP) in humans. To determine if vascular smooth muscle (VSM) PPARγ regulates salt sensitivity, we studied transgenic mice selectively expressing a HT-causing PPARγ mutant in VSM (S-P467L) and non-transgenic littermates (NT) fed a 4% high salt (HS) diet for 4 weeks. Salt equally suppressed plasma renin in both strains, but S-P467L mice exhibited increased systolic BP (S-P467L 136±3 mmHg vs NT 124±2 mmHg, p<0.01) and pulse wave velocity (3.1±0.1 vs 2.7±0.1 m/s, p<0.01) in response to HS. The salt-induced HT was not associated with changes in diastolic BP, sympathetic nerve activity, heart rate, or cardiac output. Thus, the pressor effect of HS was likely due to higher peripheral vascular resistance. HS-fed S-P467L mice developed impaired acetylcholine (ACh)- and sodium nitroprusside (SNP)-induced vasorelaxation in carotid (Max ACh relaxation: 31±4.9% vs 90±1.8%, p<0.01; Max SNP relaxation: 38±2.8% vs 89±2.6%, p<0.01) and basilar artery (Max ACh relaxation: -3.2±9.3% vs 57±5.9%, p<0.01). The impaired vasodilation rapidly developed after 3-day HS diet, preceding salt-induced BP elevation. Pre-incubation with a cyclooxygenase inhibitor indomethacin normalized ACh/SNP relaxation responses, and preliminary mass spectrometry indicated HS increased prostaglandin E2 in S-P467L aortas. HS-fed S-P467L mice had smaller renal artery luminal diameter (322±21 vs 389±22 μm, p<0.05) and blunted renal blood flow (36±3.6 vs. 50±6.4 μL/min/g, p<0.05). During the 4
th
week of HS diet, S-P467L mice produced 31% less nitrate/nitrite in 24 hour urine compared to NT controls (2.2±0.3 vs 3.2±0.4 μmol, p<0.05), suggesting blunted renal bioavailability of nitric oxide, a potent inhibitor of Na-K-2Cl cotransporter (NKCC2). This was associated with a declined capacity of HS-fed S-P467L mice to excrete an acute volume/Na
+
load, which was rescued by an NKCC2 inhibitor furosemide, but not by the Na-Cl-cotransporter inhibitor hydrochlorothiazide. Our data support the novel concept that smooth muscle PPARγ regulates systemic vascular resistance, renal perfusion and tubular sodium transport, and loss of these protective actions of PPARγ predisposes to salt sensitivity and hypertension.
Abstract 095: Renal Transcriptomic Analysis of Transgenic Mice Containing Human Cyp11b2 Gene
Authors: Mopidevi Brahmaraju , Sivankutty Indu , Perla Sravan , Jain Sudhir , Kumar Ashok
Published: December 2018

The hCYP11B2 gene encodes aldosterone synthase, the rate-limiting enzyme in the biosynthesis of aldosterone. Inappropriate excess levels of aldosterone induce positive sodium balance and predisposes to hypertension and other cardiovascular problems. Epidemiological studies have suggested that the variant -344T of the Cyp11B2 gene is associated with hypertension and myocardial hypertrophy. We have found that hCyp11B2 gene has three SNPs in 1 Kb of its promoter that are in complete linkage disequilibirium. These SNPs are T/C at -344, C/T at -470 and C/A at -663. Thus variant -344T always occurs with variants -470C, -663A (Hap-I), and variant -344C almost always occurs with variants, -470T, -663T (Hap-II). We have generated novel transgenic (TG) mice with the hCYP11B2 gene, targeted to the mHPRT locus, with either haplotype I or II variants. We observed increased baseline blood pressure in Hap I TG mice (Hap-I 117±2.5 vs. Hap-II 109±1.9 mm Hg, p<0.05), an effect accentuated by high-salt diet (Hap-I 135±2.6 vs. Hap-II 122±2.2 mm Hg, p<0.05). In order to identify the downstream genes that are involved in Cyp11B2 induced hypertension and kidney damage, we have performed transcriptome analysis in kidneys of TG animals using RNAseq. Our results indicate that the highly significant differentially expressed genes are involved in the neuro-inflammation signaling pathway (p=2.71E-10), complement system (p=2.83E-07) and IL-7 signaling pathway (p=1.43E-04) with Z scores 2.333, 0.816 and 2.714 respectively. Using Ingenuity pathway analysis, we have identified genes that are associated with renal fibrosis. We have also found that expression of CTSW, CCL5, CCR2, CX3CR1, IRF1, and IRF7 are increased by 12.27 fold (p=3.49E-05), 4.23 fold (p= 3.78E-08), 2.9 fold (p= 3.45E-11), 3.37 fold (p=2.60E-12), 2.1 fold (p= 3.20E-06) and 2.0 fold (p= 1.21E-04) respectively in kidneys of hypertensive animals. Our RNA seq results delineate the regulatory molecules involved in the aldosterone mediated nephrotoxicity. These findings not only provide the valuable information on the pathophysiology of kidney damage but also can be used to identify novel therapeutic targets to reduce high blood pressure and end organ damage.
Abstract 098: Aldosterone-Producing Cell Clusters in Essential Hypertension
Authors: Omata Kei , Kusaka Ryo , Yamazaki Yuto , Ito Sadayoshi , Satoh Fumitoshi , Rainey William , Tomlins Scott , Sasano Hironobu
Published: December 2018

Introduction:
Primary aldosteronism (PA) is one of the major causes of secondary hypertension and has two major subtypes. One is unilateral aldosterone-producing adenoma (APA), which is positive for aldosterone synthase (CYP11B2) by immunohistochemistry (IHC), and the other is bilateral idiopathic hyperaldosteronism (IHA), which has image-undetectable CYP11B2-positive cell clusters (termed aldosterone-producing cell clusters, APCC) beneath the adrenal capsule. We and others have previously shown that APCC present in normotensive adrenals and the number is significantly less than IHA adrenals, supporting the disease-causing role of APCC in hyperaldosteronism of IHA. Furthermore, we have also shown that most of APCCs in the IHA and normotensive adrenals harbor somatic mutations in a L-type calcium channel,
CACNA1D
, whereas most of APA has been previously shown to harbor those in a potassium channel,
KCNJ5
. These mutations are reported to increase intracellular calcium levels, resulting in aldosterone overproduction. Here, we seek for the first time to identify the potential role of APCC in non-PA hypertensive patients and to clarify whether these adrenals could be precursor of APA or IHA by performing next-generation sequencing (NGS) on observed APCC.

Method and Results:
We selected fifteen adrenal glands with the evidence of hypertension and/or antihypertensive agents from a cohort of serial Japanese autopsy cases. None of the cases harbored adrenal tumors, but instead nine cases harbored 23 APCCs. We then isolated DNA from each APCC and performed NGS targeting genes mutated in APA. Of observed APCCs, 9 (39%) APCCs harbored somatic mutations in
CACNA1D
.

Interpretations:
These results show that autonomous aldosterone production in a part of patients with essential hypertension is caused by APCCs with aldosterone-driving somatic mutations. In addition, the mutation spectrum observed in this cohort suggests that essential hypertension could potentially develop into IHA through aldosterone overproduction by APCC.
Abstract 099: Carvedilol Selectively Stimulates Beta-arrestin2-dependent Serca2a Activity in Cardiomyocytes to Augment Contractility
Authors: Lymperopoulos Anastasios , Parker Barbara M , Brill Ava , McCrink Katie A , Maning Jennifer , Desimine Victoria L , Wertz Shelby L
Published: December 2018

Background:
Heart failure (HF) is the number one killer in the western world and β-blockers are part of its cornerstone pharmacotherapy. We recently showed that β
1
-adrenergic receptor (AR)-activated βarrestin2, a G protein-coupled receptor (GPCR) adapter protein, promotes Sarco(endo)plasmic reticulum Ca-ATPase (SERCA)2a SUMO (small ubiquitin-like modifier)-ylation & activity, thereby increasing cardiac contractility. Given that carvedilol, unlike other β-blockers, has been shown to activate βarrestins and SERCA2a in the heart, we investigated herein its effects on βarrestin2-dependent SERCA2a modulation and contractility.

Methods:
We studied SERCA2a-βarrestin interactions, SUMOylation & activity in response to therapeutic doses of carvedilol or metoprolol in H9c2 cardiac cells. We also measured contraction amplitude (cell shortening) of neonatal rat ventricular myocytes (NRVMs).

Results:
Carvedilol, unlike metoprolol, acutely induces βarrestin2 (not βarrestin1) interaction with SERCA2a in H9c2 cardiomyocytes, resulting in enhanced SERCA2a SUMOylation. This translates into enhanced SERCA2a activity in the presence of the β
2
AR-selective inverse agonist ICI 118,551 (ICI), indicating an opposing effect of β
2
AR on carvedilol-stimulated, βarrestin2-dependent SERCA2a activation in H9C2 cells. In addition, the amplitude of spontaneous cell shortening of βarrestin2-overexpressing NRVMs is enhanced by carvedilol, especially in the presence of ICI (122+9% of control, ICI only-treated NRVMs; p<0.05, n=10). In contrast, metoprolol alone or with ICI had no effect again (98+6% of control, ICI only-treated NRVMs; p<0.05, n=10).

Conclusions:
Carvedilol uniquely stimulates βarrestin2-mediated SERCA2a SUMOylation/activity through the β
1
AR in cardiac myocytes. This translates into direct positive inotropic effects of this β-blocker, which may be opposed by the cardiac β
2
AR subtype. These findings challenge the conventional wisdom that all β-blockers exert negative inotropy in the heart, complicating their use in human chronic HF treatment, and highlight the particular usefulness of carvedilol for treating this disease, as this drug may have positive inotropic properties on top of its classic reverse remodeling effects.
Abstract 100: Mechanistic Insight into Tannic Acid Mediated Angiotensin Type 1 Receptor and PCSK 9 Downregulation
Authors: Iyer Laxmi , Zhang Yong , Thekkumkara Thomas
Published: December 2018

Deregulated angiotensin type 1 receptor (AT1R) and low-density lipoprotein receptor (LDLR) are implicated in cardiovascular disorders. AT1R is over expressed in hypertension and LDLR is involved in lowering plasma LDL cholesterol. In this study we investigate the role of tannic acid (TA), a hydrolyzable polyphenol, on AT1R and LDLR in rat aortic smooth muscle cells (RASMC) and rat liver epithelial cells (RLEC) respectively. TA (10 μg/ml) treatment exhibited down-regulation of AT1R specific [3H] AngII binding compared to untreated control (3628.32 ± 289.42 vs. 7244.66 ± 40.26 DPM/mg protein,
P
< 0.001). TA mediated AT1R downregulation was dose (0-40 μg/ml) and time dependent (0-24 hours) without significant change in receptor affinity (K

d

1.302 ± 0.152 nM for TA treated vs. K

d

1.391 ± 0.048 nM for untreated). Consistently, TA inhibited mRNA expression (TA 0.023± 0.0012 vs. control 0.051± 0.0045 mRNA fold). Furthermore, TA activated of ERK p42/44, which was inhibited by MEK inhibitor PD98059 (20μM) and an EGFR inhibitor AG1478 (5μM) suggesting a role for EGFR induced ERK p42/44 in AT1R downregulation. A similar effect was observed with EGF (0.5 μg/ml) alone (EGF; 3788.04 ± 206.51 vs. Control; 6549.22 ± 115.48 DPM/mg protein, P < 0.001). LY294002 (10μM), an inhibitor of phosphoinositide 3-kinase (PI3K), inhibited both TA and EGF induced pERK and reversed AT1R downregulation suggesting that TA mediated AT1R downregulation is through EGFR-PI3K-ERK pathway. Consistent with the receptor downregulation significant inhibition of AngII induced intracellular calcium (TA 203.4 ± 7.79nM vs control 408.5 ± 5.33 nM) was observed. Under similar conditions proprotein convertase subtilisin/kexin type 9 (PCSK9), a crucial enzyme responsible for LDLR turn over, was decreased 2.8 fold at the protein level and mRNA levels (reduced 46.78± 2.3%) in RLEC. Alternatively, we observed 3.6 fold increased cell surface expression of LDLR and induced LDL-c uptake in human hepatocyte cells (2677 ± 26.39 untreated vs. 3894 ± 40.1 for TA treated). Our study provides a mean to control hypertension and hypercholesteremia associated cardiovascular disorders.
Abstract 104: DOCA Induced Natriorexigenic and Hypertensive Effects are Blunted by Central Infusion of AT1 Receptor Biased Agonist TRV120027
Authors: Seara Fernando , Nakagawa Pablo , Gomez Javier A , Deng Guorui , Sebag Julien A , Karnik Sadashiva S , Grobe Justin L , Sigmund Curt D
Published: December 2018

TRV120027 (TRV) is biased agonist at the angiotensin AT1 receptor that activates the β-arrestin pathway, causing receptor internalization. AT1 signaling within the hypothalamus is required for the blood pressure, polydipsia, and sodium appetite phenotypes in the deoxycorticosterone acetate (DOCA)-salt model of salt-sensitive hypertension (HTN). We hypothesized that TRV action within the hypothalamus would antagonize the effects of DOCA-salt. Using a novel luciferase-based HiBit tagging assay, we confirmed that TRV decreased AT1 surface density in immortalized mouse hypothalamic N43/5 cells by 35±4% compared to vehicle (n=4, p<0.05), and this was prevented by pretreatment with losartan. TRV (0.05 μg/h, icv n=5-10) or aCSF (n=4-8) were chronically infused into C57BL/6J mice implanted with DOCA (50 mg, sc) pellets, compared with mice undergoing sham surgeries (CTL, n=5-8), and animals were offered chow, water and NaCl
ad libitum
. DOCA increased fluid intake at baseline (CTL 4±0 vs 15±1 mL/day DOCA+aCSF, p<0.05), 0.15 mol/L NaCl (3.8±0 vs 20±2, p<0.05), 0.3 mol/L (4±0 vs 26±2, p<0.05), and 0.45 mol/L (4±0 vs 28±3, p<0.05). DOCA-induced polydipsia was not affected by TRV at baseline (DOCA+aCSF 19±1 vs 15±0 mL/day, p>0.05), 0.15 mol/L NaCl (27±4 vs 20±2 mL/day, p>0.05), 0.3 mol/L (25±4 vs 26±2 mL/day, p>0.05), or 0.45 mol/L (25±3 vs 28±3 mL/day, p>0.05). However, TRV prevented DOCA-induced increase of NaCl preference at NaCl 0.15 mol/L (DOCA+aCSF 29±1 vs 14±2% DOCA+TRV, p<0.05), 0.30 mol/L (29±3 vs 5±1%, p<0.05), and 0.45 mol/L (29±3 vs 5±1%, p<0.05). DOCA+aCSF exhibited increased heart weight / tibia length (CTL 8±0 vs 10±0 g/m DOCA+aCSF, p<0.05), which was prevented by TRV (8±0, p<0.05). In a separate cohort, DOCA-implanted mice were provided only 0.9% NaCl drink for one week, and mean arterial pressure (MAP) was recorded after acute injection of TRV (2 μg icv n=5) or aCSF (n=5). MAP was elevated with DOCA-salt (141±5 mmHg), and at 40 minutes post injection, TRV decreased MAP by 17±4 mmHg (p<0.05). These data underscore the potential for AT1 receptor-biased agonists as tools to treat subtypes of HTN characterized by salt sensitivity and low circulating renin activity.
Abstract 109: Association Between Aortic Stiffness and Lower Retinal Microvascular Blood Flow Revealed by Laser Speckle Flowgraphy: Surrogate Measures of Cerebral Microcirculation in Humans
Authors: Holwerda Seth W , Kardon Randy H , Wetherell Joseph M , Full Jan M , Dubose Lyndsey E , Points Lauren J , Gremaud Allene L , Luehrs Rachel E , Tong Brian , Nellis Julie K , Stroud Amy K , Fiedorowicz Jess G , Pierce Gary L
Published: December 2018

The age-related increase in aortic stiffness increases systolic blood pressure (BP) and flow pulsatility and is associated with target organ damage such as ischemic cerebrovascular disease. However, the mechanistic link between aortic stiffness and cerebrovascular disease remains unclear because techniques available to non-invasively assess dynamic changes in the human brain microcirculation with high spatial and temporal resolution are limited. Given the evidence that disease- and age- related alterations in the retinal microvasculature may mirror changes in the cerebral microvasculature, we assessed resting and dynamic changes in the retinal microcirculation using high resolution laser speckle flowgraphy. We tested the hypothesis that retinal microvascular blood flow at rest and during acute increases in BP would be associated with age-related increases in aortic stiffness (carotid-femoral pulse wave velocity, CFPWV). In 40 healthy subjects (35 ± 2 years; 25-60 years; 18 M/22 F; systolic BP: 123 ± 2 mmHg; diastolic BP: 77 ± 2 mmHg), retinal arterial blood flow was assessed at baseline (following dilation of the left pupil with 0.5% topicamide), and during a sympatho-excitatory stimulus (2-min cold pressor test). At baseline, lower retinal blood flow was significantly correlated with higher CFPWV (R=-0.35, P=0.03) and older age (R=-0.38, P=0.02) independent of mean BP (partial correlation). In response to the CPT, significant increases were observed in mean BP (BL: 92 ± 2 vs. CPT: 106 ± 2 mmHg, P<0.001) and retinal blood flow (BL: 215 ± 6 vs. CPT: 230 ± 8 AU, P<0.001). The relation between higher CFPWV and lower retinal blood flow observed at baseline was maintained during the CPT independent of mean BP (R=-0.37, P=0.03). However, no association was noted between CFPWV and % change in retinal blood flow. These preliminary data suggest that higher aortic stiffness is associated with lower retinal microvascular blood flow at rest and during sympatho-excitation independent of BP. These findings have important implications for age-related ocular degeneration, and lend support for future studies aiming to identify retinal microvascular abnormalities as markers of age-related cerebrovascular pathology.
Abstract 114: Increased Circulating Levels of Mitochondrial N-Formyl Peptides Leads to Vascular Dysfunction and High Blood Pressure in Spontaneously Hypertensive Rats
Authors: Wenceslau Camilla F , McCarthy Cameron G , Webb R. Clinton
Published: December 2018

Aims:
The most powerful signaling pathway that induces actin polymerization and movement in neutrophils is due to formyl peptide receptor (FPR) activation. FPR was originally identified by its ability to bind bacterial N-formyl peptides. Mitochondria carry hallmarks of their bacterial ancestry and one of these hallmarks is that this organelle uses an N-formyl-methionyl-tRNA as an initiator of protein synthesis. Consequently, mitochondrial N-formyl peptides (F-MITs) are recognized by FPR. Recently, we have observed that FPR is expressed in endothelial and vascular smooth muscle cells (VSMC). We hypothesized that increased plasma levels of F-MITs activates FPR and contributes to microvascular damage and hypertension.

Methods:
Male SHR (12-week old) were treated for 10 days with FPR antagonist cocktail: Cyclosporin H (CsH, 0.1 mg/kg/day
i.p.
) and WRW4 (0.1 mg/kg/day
i.p.
) or vehicle. Blood pressure (BP) was measured by telemetry. We also treated Wistar rats (12 week old) with F-MITs (formylated peptide corresponding to the NH2-terminus of mitochondria ND6; 0.02 mg/rat/day
i.p.
) or non-formylated peptides (control) for 10 days. Blood was collected to measure F-MITs. Intrarenal arteries and mesenteric resistance arteries (MRA) were isolated and mounted in a wire myograph. Wistar VSMCs were treated with F-MITs (20 min, 10 μM) or control in the presence or absence of FPR antagonists to evaluate actin polymerization. T-test *p<0.05; n=5-6.

Results and Conclusion:
F-MITs plasma levels were increased in SHR (1.7-fold* vs. Wistar). FPR blockade decreased BP (SHR + vehicle: 151 ± 7.2 vs. SHR+CsH+WRW4: 138 ± 0.4* mmHg) and ameliorated acetylcholine-induced relaxation (Intrarenal; Emax: SHR+vehicle: 10 ± 5 vs. SHR+CsH+WRW4: 28 ± 6* %); (MRA; Emax: SHR+vehicle: 74±10 vs. SHR+CsH+WRW4: 99±0.6* %). F-MITs infusion increased BP in Wistar rats (Control: 133 ± 1.9 vs. F-MITs: 145 ± 4.7* mmHg) and decreased acetylcholine-induced relaxation (EC50; MRA: Control: -8.765 ± 0.05 vs. F-MIT: -8.348 ± 0.08*) (Emax; Intrarenal: Control: 63±4 vs. F-MIT: 46 ± 3* %). F-MITs induced VSMCs actin polymerization (1.8 fold* vs. control) and FPR blockers abolished this result. In conclusion, FMITs play a role in vascular dysfunction and hypertension via FPR activation.
Abstract 118: DNA de novo (de)Methylation in the Kidney Contributes to Salt-Induced Hypertension
Authors: Liu Pengyuan , Liu Yong , Pan Xiaoqing , Li Yingchuan , Usa Kristie , Liang Mingyu
Published: December 2018

Numerous adult diseases involving tissues that consist primarily of non-dividing cells are associated with changes in DNA methylation. It suggests a role for de novo methylation or demethylation of DNA, which is catalyzed by DNA methyltransferase 3 (Dnmt3) and ten-eleven translocases (Tet). However, the contribution of DNA de novo (de)methylation to these diseases remains nearly completely unproven. Broad changes in DNA methylation occurred within days in the renal outer medulla of Dahl SS rats fed a high-salt diet, a classic model of hypertension. Intra-renal administration of anti-Dnmt3a/Tet3 GapmeR’s attenuated high salt-induced hypertension in SS rats (mean arterial pressure 153+/-4 vs. 141+/-2 mmHg, n=9-11, p<0.05). The high salt diet induced differential expression of 1,712 genes in the renal outer medulla. Remarkably, the differential expression of 76% of these genes were prevented by anti-Dnmt3a/Tet3 GapmeR’s. The genes differentially expressed in response to the GapmeR’s were involved in the regulation of metabolism and inflammation and were significantly enriched for genes showing differential methylation in response to the GapmeR’s. These data indicate DNA de novo (de)methylation in the kidney contributes to the development of hypertension in SS rats. The findings should help to shift the paradigm of DNA methylation research in diseases involving non-dividing cells from correlative analysis to functional and mechanistic studies.
Abstract 119: Using Blood Pressure Values from Electronic Medical Records to Identify Hypertension During Pregnancy
Authors: Chen Lu , Easterling Thomas , Cheetham T. Craig , Reynolds Kristi , Avalos Lyndsay , Holt Victoria , Neugebauer Romain , Shortreed Susan M , Akosile Mary , Kamineni Aruna , Walker Rod , Badon Sylvia E , Dublin Sascha
Published: December 2018

Objective:
Hypertension is a major risk factor for poor pregnancy outcomes. Many observational studies have relied on diagnosis codes, particularly from the delivery hospitalization, to identify hypertension in pregnancy. We augmented diagnosis codes with electronic blood pressure (BP) data to improve the identification of pregnant women with hypertension.

Methods:
We studied pregnant women aged 15-49 years enrolled in three Kaiser Permanente health plans who delivered during 2005-2014. Using diagnosis codes, BP values, and antihypertensive medication dispensings, we defined hypertension as: (1) ≥ 2 high BPs (≥ 140/90 mmHg) within 30 days of each other (2highBPs); or (2) ≥ 1 antihypertensive medication fill with ≥1 hypertension diagnosis code from 120 days prior to pregnancy through 20 weeks gestation (chronicHTN); or (3) ≥ 1 high BP, a hypertension diagnosis code, and an antihypertensive fill within 7 days (RapidTx). Among women meeting our study definition, we examined receipt of hypertension diagnosis codes and prevalence of severe hypertension (1+ BP ≥160/110 mmHg).

Results:
Among 553,477 eligible women, 29,933 (5%) met our definition of hypertension, including 26,855 identified via 2highBPs, 5,774 via chronicHTN and 6,198 via RapidTx (not mutually exclusive). Among women meeting our hypertension definition overall, only 64% had 1+ hypertension diagnosis code assigned during pregnancy, and 49% had one at delivery. Among hypertensive women identified via 2highBPs, only 60% (16,057/26,855) had a hypertension diagnosis code in pregnancy and 45% (12,131/ 26,855) at delivery. However, 53% of our hypertensive women (14,972/ 29,933) overall and 56% (14,972/ 26,855) of the 2highBPs women had severe hypertension at some time during pregnancy.

Conclusion:
Incorporating BP values identifies additional pregnant women with hypertension who would have been missed by approaches using diagnosis codes alone. Women identified by our method frequently had severely elevated BP, showing the importance of including these women in future studies of hypertension during pregnancy.
Abstract 121: Endothelial Cullin3 Mutation Causes Vascular Dysfunction, Arterial Stiffening, and Hypertension
Authors: WU Jing , Agbor Larry N , Fang Shi , Hu Chunyan , Liu Xuebo , Mukohda Masashi , Nair Anand , Sigmund Curt D
Published: December 2018

Mutations in CULLIN3 gene (causing in-frame deletion of exon 9) cause human hypertension (HT), which is likely to be driven by a combination of renal tubular and vascular mechanisms. We have recently shown that smooth muscle expression of Cul3Δ9 mutant causes vascular dysfunction and HT via augmented RhoA/Rho-kinase signaling, strongly supporting a vascular role of Cul3 in BP regulation. To test the importance of endothelial Cul3
in vivo
, we bred the conditionally activatable Cul3Δ9 mice with tamoxifen-inducible Tie2-CRE
ERT2
mice. The resultant mice (E-Cul3Δ9) developed nocturnal hypertension (Night time peak systolic BP, E-Cul3Δ9: 138±3 vs Control: 121±4 mmHg, p<0.01) and arterial stiffening (pulse wave velocity, 3.7±0.3 vs 2.7±0.1 m/s, p<0.01). No difference was seen in daytime BP. Nitric oxide synthase (NOS) inhibitor L-NAME induced smaller increases in nocturnal peak SBP and DBP in E-Cul3Δ9 mice (15±1 vs 27±3 mmHg, p<0.01), suggesting their NOS activity is low. Of note, E-Cul3Δ9 mice exhibited impaired endothelial-dependent relaxation in carotid artery (Max ACh relaxation: 69% vs 84%, p<0.05) and cerebral resistance basilar artery (41% vs 77%, p<0.01). No difference in smooth muscle function was observed. To determine the molecular mechanisms, we isolated primary aortic endothelial cells from mice carrying the inducible Cul3Δ9 construct and induced Cul3Δ9 expression
in vitro
using adenovirus carrying Cre recombinase gene. Expression of Cul3Δ9 resulted in marked decreases in wild type Cul3 protein, phosphorylated eNOS, and nitric oxide production. Because protein phosphatase 2A (PP2A) is a known Cul3 substrate which dephosphorylates eNOS, we determined whether impaired eNOS activity was attributable to PP2A. Cul3Δ9-induced impairment of eNOS activity was rescued by a selective PP2A inhibitor Okadaic Acid (4 nM), but not by a Protein Phosphatase 1 inhibitor Tautomycetin (4 nM). These data define a novel regulatory pathway involving Cullin3/PP2A/phospho-eNOS in the endothelium. Selective endothelial expression of Cul3Δ9 partially phenocopies the hypertension observed in Cul3Δ9 patients, suggesting that mutations in Cullin-3 cause human hypertension in part through a vascular mechanism characterized by endothelial dysfunction.
Abstract 123: Effect of Immune Cell-Derived Reactive Oxygen Species on Dahl SS Hypertension
Authors: Abais-Battad Justine M , Lund Hayley , Dasinger John Henry , Fehrenbach Daniel J , Mattson David L
Published: December 2018

Previous studies utilizing the SS

p67
phox–/–

rat have demonstrated the importance of systemic NADPH oxidase NOX2-derived reactive oxygen species (ROS) production in the pathogenesis of Dahl Salt-Sensitive (SS) hypertension and renal damage. Our laboratory has established a key role for renal immune cell infiltration in the development of SS hypertension and observed an enrichment of NOX2 subunits in infiltrating T cells. The contribution of ROS specifically derived from immune cells in SS hypertension remains unknown. To assess the role of ROS in immune cells, SS

p67
phox–/–

rats underwent total body irradiation and received bone marrow transfer from either SS

p67
phox–/–

(p67/p67) or SS (p67/SS) donor rats. p67/SS rats would be capable of producing NOX2-derived ROS only in the transferred SS cells of hematopoietic origin while the p67/p67 control group would be unable to produce NOX2-derived ROS globally. Mean arterial pressure (MAP) was not different between the p67/SS and p67/p67 rats when fed a 0.4% NaCl diet (112.6±1.4 vs 110.7±2.4 mmHg, n=4-5). After 3 weeks of high salt, however, there was a significantly greater increase in MAP in the p67/SS rats (176.1±4.7 mmHg) compared to the p67/p67 control rats (147.9±8.4 mmHg), which was accompanied by a significant increase in albuminuria (230.3±13.9 vs 153.1±14.9 mg/day, p67/SS vs p67/p67, n=5-6). Interestingly, upon analysis of the immune cells in the kidney, no differences were detected in the number of CD45+ total leukocytes, CD11b/c+ monocytes/macrophages, CD3+ T cells, and CD45R+ B cells between the p67/SS and p67/p67 (n=4-5), indicating that the recruitment of immune cells into the kidney during hypertension and renal damage is independent of NOX2-derived ROS production from the renal parenchyma. The respiratory burst response to phorbol 12-myristate 13-acetate stimulus (135 μM) in peritoneal macrophages isolated from p67/SS was 4.0-fold greater compared to nonresponsive p67/p67 cells, and was completely inhibited by superoxide dismutase, demonstrating that the reconstituted bone marrow cells in the p67/SS are capable of producing superoxide. In summary, the production of NOX2-derived ROS from immune cells alone is sufficient for the development of Dahl SS hypertension and renal damage.
Abstract 133: Endothelial-Specific Interference With PPARγ Increases the Susceptibility to Angiotensin II-Induced Endothelial Dysfunction in Adult Offspring Born from AVP-Infused Pregnancies
Authors: Nair Anand R , Mukohda Masashi , Agbor Larry N , Lu Ko-Ting , Wu Jing , Sandgren Jeremy A , Grobe Justin L , Sigmund Curt D
Published: December 2018

Mutations in the ligand-activated transcription factor PPARγ result in hypertension, and synthetic agonists of PPARγ reduce blood pressure. Previously we found that mice expressing dominant-negative (DN) PPARγ driven by an endothelium-specific promoter (E-DN) exhibit vascular dysfunction. Preeclampsia (PE) is a hypertensive disorder of pregnancy which carries cardiovascular risk to offspring. PE is also associated with vascular dysfunction. We hypothesized a role for endothelial PPARγ in the pathogenesis of PE and its sequelae. C57BL/6J dams were bred with E-DN sires, and symptoms of PE were induced by infusion of vasopressin (AVP, 24 ng/hr sc) throughout gestation. Phenotypes of PE were first assessed in pregnant dams and then in adult offspring. Compared to saline infusion (SAL), AVP elevated maternal blood pressure (SBP: 116±3 vs 107±3, p<0.05) at gestational day (GD) 14-15 and urine protein (70±6 vs 27±4 mg/mL, p<0.05) at GD17. Offspring were phenotyped in adulthood. Data were stratified to sex, genotype, and exposure to AVP or SAL. At 20 weeks of age, SBP and vasorelaxation responses to acetylcholine were similar in offspring exposed to PE compared to mice born from SAL pregnancies. Adult offspring were next exposed to a sub-pressor dose of Angiotensin II (ANG; 120ng/kg/hr) for 14 days. Adult male ANG-treated E-DN born to PE pregnancies, but not NT mice exhibited significant impairment in ACh-induced relaxation in carotid artery (% relaxation: 62±10 vs 86±11, p<0.05). Adult female ANG-treated E-DN exposed to PE also exhibited a trend for impaired endothelial function compared to NT controls (% relaxation: 63±9 vs 72±10). Endothelial dysfunction in male ANG-treated E-DN born to PE pregnancies was attenuated by tempol (relaxation improved from 56±9% vs 81±12%, p<0.05), indicative of oxidative stress, and by a Rho-kinase inhibitor (relaxation improved from 73±11% vs 89±12%, p<0.05). This data suggests that interference with endothelial PPARγ in pups born from PE pregnancies increases the risk for endothelial dysfunction upon exposure to a cardiovascular stressor in adulthood. Thus, conditions which impair PPARγ activity, such as obesity and diabetes, may predispose adult offspring born from PE pregnancy to cardiovascular disease.
Abstract 136: Attenuation of Increased Blood Pressure by Chronic Estrogen Supplementation in Female IUGR Offspring at 12 months of Age is Associated With a Shift Towards ACE2
Authors: Davis Gwendolyn K , Newsome Ashley D , Ojeda Norma B , Alexander Barbara T
Published: December 2018

Low birth weight (
LBW
) women exhibit an earlier age at menopause and a two-fold greater prevalence of hypertension in later life. In a model of LBW induced via placental insufficiency in the rat, female LBW rats develop an increase in blood pressure (
BP
) by 12 months of age. This is associated with persistent estrous that occurs 6 months prior to controls, indicative of early reproductive senescence. Although the underlying mechanisms are unknown, a shift in the hormonal milieu in addition to dysregulation of the renin angiotensin system (
RAS
) are potential contributors to the increase in BP that occurs in women after menopause. The vasoconstrictor arm of the RAS is implicated in increased BP; ACE2 counterbalances this shift towards increased BP. We previously reported that renal ACE2 expression is elevated in female LBW rats that are normotensive in young adulthood; an increase that is not present in female LBW rats in later life. Thus, this study tested the hypothesis that chronic 17β-estradiol (
E2
) supplementation would reduce BP and shift the balance of the RAS in female LBW rats. Placebo or E2 pellets (1.5mg 17β-estradiol, 60-day release) were implanted at 12 months of age for 6 weeks followed by measurement of BP in conscious, chronically catheterized animals. Kidneys were collected for molecular analysis. BP was significantly increased in placebo-treated LBW relative to placebo-treated Control (134 vs 122 mmHg, respectively,
p
<0.05); an increase that was abolished by chronic E2 in LBW (116 mmHg,
p
<0.05). Using Real-Time PCR, cortical ACE2 mRNA expression did not differ between Control or LBW rats. Medullary ACE2 mRNA expression was significantly increased in placebo-treated LBW compared to placebo-treated Controls (1-fold increase;
p
<0.05). However, following chronic E2, medullary ACE2 mRNA expression was increased to a greater degree in chronic E2-treated LBW rats, almost two-fold higher, relative to chronic E2-treated Control. Therefore, these results suggest that E2 supplementation attenuates increased BP in female LBW rats in a manner that may involve a shift in the RAS towards ACE2 abundance suggesting that ACE2 via chronic E2 may be beneficial in the treatment of increased BP that occurs after reproductive senescence in LBW.
Abstract P104: Urban And Rural Differences In Exposure And Effects Of Micro Air Particles On Blood Pressure Parameters.
Authors: Nkeh-Chungag Benedicta N , Chungag Anye , Sewani-Rusike Constance R
Published: December 2018

Several studies have shown a relationship between indoor particulate matter and cardiovascular diseases in adults though this relationship is yet to be clarified in children and adolescents. This study purposed to investigate the relationship between indoor air particulate matter and blood pressure in 10-14 year old rural and urban children in Eastern Cape, South Africa. A cross sectional study of 10-14 year old children was carried out in selected rural and urban schools of the Eastern Cape. Anthropometry was performed and blood pressure measured. Indoor air particle counts were determine. Blood pressure parameters and body mass index (BMI) were converted to percentiles for height, sex and age. Prevalence of overweight/obesity were 10.9%/14.0% and hypertension/prehypertension were 12%/20%. Hypertension/prehypertension were more prevalent in rural compared to urban adolescents. All blood parameters were higher in rural adolescents and higher in rural girls compared to boys. In hypertensive/prehypertensive adolescents, systolic blood pressure (SBP) showed a strong correlation with BMI, waist, hip and mid upper arm circumferences (0.50, 0.51, 0.52, 0.53, 0.49; p<0.05 respectively) though SBP was only modestly associated with these variable in normotensive adolescents. Indoor particle counts were higher in rural schools. Blood pressure did not correlate with indoor air particulate matter though there was a weak correlation between PM2.5, 5, 10 and heart rate in rural adolescents. Indoor air particle failed to show any relationship with blood pressure parameters in urban adolescents. Rural adolescents had higher blood pressure and heart rates. Indoor particulate matter may be associated with blood raised pressure via sympathetic activation as suggested by higher heart rates.
Abstract P109: Collecting Duct Renin and Aldosterone Regulate Potassium Homeostasis
Authors: Xu Chuanming , Chen Yanting , Zou Changjiang , Ramkumar Nirupama , Xie Shiying , Wang Fei , Luo Renfei , Yang Tianxin
Published: December 2018

The kalliuric action of the renin-angiotensin-aldosterone system (RAAS) is well established as highlighted by hyperkaliemia side effect of RAAS inhibitors but such action is usually ascribed to systemic RAAS. The present study attempted to address the involvement of intrarenal RAAS in K
+
homeostasis with emphasis on locally generated renin and aldosterone (Aldo) within the collecting duct (CD). In normal C57BL/6 mice, a 1-wk high K
+
(HK) intake (5% KCl in diet) induced parallel increases in renal prorenin (cortex: 298.8%; medulla: 323.8%) assessed by immunoblotting, and urinary prorenin/renin content (URC) (2.5-fold) assessed by ELISA, and urinary renin activity (URA) (1.5-fold), contrasting to suppressed plasma prorenin/renin concentration (by 25.6%) and renin activity (by 61.6%). Following 1-wk HK loading, mice lacking renin in the CD (CD renin KO) had decreased urinary K
+
excretion (by 20.2%) and elevated plasma K
+
level (KO+HK: 4.35 ± 0.14
vs.
Floxed+HK: 3.89 ± 0.04 mM,
P
<0.01), accompanied with a reduction of URC (by 40%), URA (by 58.5%), urinary free and total Aldo excretion (by 50.6% and 36.6%, respectively), and kidney cortical and medullary Aldo (by 33.5% and 49.9%, respectively), without affecting plasma Aldo or renin levels. HK upregulated renal protein expression of Aldo synthase CYP11B2 (cortical: 133.4%, medullary: 221.5%), renal outer medullary K
+
channel (ROMK) (146.4%), calcium-activated potassium channel subunit alpha-1 (α-BK) (160.9%), α-Na
+
-K
+
-ATPase (155.8%), β-ENaC (165.1%), and cleaved-γ-ENaC (1800%), all of which were significantly blunted in CD
renin
KO mice (by 25-75%). We have developed an inducible renal tubule-wide
CYP11B2
KO using the Pax8/LC1 transgenes (termed as RT
CYP11B2
KO). While the homozygous deletion is lethal, the heterozygous mice exhibited normal development and were subjected to 3-day HK. K
+
-loaded RT
CYP11B2
KO mice had decreased urinary K
+
excretion (by 30.5%) and hyperkalemia (KO+HK: 4.63 ± 0.05
vs
. Floxed+HK: 3.98 ± 0.09 mM,
P
<0.01), accompanied with a reduction of urinary free and total

Aldo excretion (by 20.4% and 34.4%, respectively) without affecting plasma Aldo levels. Taken together, these results support a local action of CD renin and Aldo in regulation of K
+
homeostasis.
Abstract P113: Early Life Stress-Induced Increases in Urinary Aldosterone Precedes Renal Dysfunction and Hypertension in Male Mice Fed a High Fat Diet
Authors: Leachman Jacqueline R , Dalmasso Carolina , Xu Xiu , Backus Jason , Cassis Lisa , Loria Analia S
Published: December 2018

We have previously shown that mice subjected to maternal separation and early weaning (MSEW), a model of early life stress, display exacerbated angiotensin II-dependent obesity-induced hypertension and reduced glomerular filtration rate (GFR) after 16 weeks of high fat diet (HF). In this study, we hypothesized that renin-angiotensin aldosterone system (RAAS) activation in MSEW mice fed a HF for only 12 weeks will precede a decline in GFR and heightened hypertension. MSEW was performed by separating the pups from their mother for 4 to 8 hours from postnatal days (PD) 2 to 16. Mice were weaned at PD 17. Control mice remained undisturbed and were weaned at PD 21. We used 16 MSEW and 14 control litters. Eight-week-old mice were fed on a low fat diet (LF) or HF (10 or 60 % fat Kcal) for 12 weeks. Each litter was represented by one male randomly assigned to each diet. After 11 weeks, 24-hr urine was collected to measure proteins, creatinine, aldosterone and electrolytes by Duo ICAP-OES. At week 12, transcutaneous GFR and blood pressure were measured in all mice. Male MSEW and control mice fed a HF displayed similar blood pressure and metabolic parameters, including water intake (3.4±0.3 vs. 3.4±0.2 ml/day), diuresis (1.1±0.1 vs. 1.0±0.2 ml/day), natriuresis (0.10±0.01 vs. 0.09±0.01 mmol/day), proteinuria (3.5±0.7 vs. 2.9±0.5 mg/day) and GFR (0.97±0.04 vs. 0.96±0.03 ml/min/100g BW). However, MSEW increased kaliuresis (0.41±0.07 vs. 0.23±0.02 mmol/day, p<0.05) and urinary aldosterone (25±4 vs. 11±1 ng/g crea, p<0.05). In addition, MSEW mice displayed increased plasma aldosterone (158±32 vs. 87±17 pg/ml, p=0.056) and lower plasma renin concentration compared to controls (2.3±0.4 vs. 3.7±0.4 ng/mL/30’, p<0.05), suggesting elevated circulating angiotensin II. Only MSEW mice showed increased adiposity (36±1 vs. 24±4, %) and a positive correlation between urinary aldosterone and fat mass (2.6±0.9 vs. -0.1± 0.1, p<0.05). These data indicate that MSEW-induced RAAS overactivation precedes the increases in blood pressure and decline in GFR found in HF-fed mice later in life. Thus, increased angiotensin II, most likely due to elevated fat angiotensinogen, may stimulate adipocyte and/or adrenal-derived aldosterone production exacerbating obesity-hypertension in MSEW mice.
Abstract P115: Urinary Angiotensinogen is Increased in Type 1 Diabetes With Mild Albuminuria: Gender Differences
Authors: Sanchez Nadales Alejandro , Ba Aqeel Sheeba Habeeb , Ye Minghao , Wysocki Jan , Rademaker Alfred , Molitch Mark , Batlle Daniel
Published: December 2018

Urinary angiotensinogen (uAOG) can be increased in patients with type 2 diabetes and in hypertension. We wanted to examine uAOG in patients with type 1 diabetes as a possible biomarker of enhanced Renin-Angiotensin system activity within the kidney at an early stage of the disease and study any potential gender differences

Methods:
Urine samples from the Diabetes Control and Complications Trial (DCCT) that were available at the earliest study visit were obtained from NIDDK repository. Cases (N=103) were participants with microalbuminuria at study initiation (30.2-286.5mg/24hrs). Controls (N=103) were patients with type 1 diabetes with normoalbuminuria (<30mg/24hrs). Controls were matched based on age, gender and insulin arm allocation (intensive vs conventional) when the urine was available at the earliest visit (range 0-8). Wilcoxon Rank Sum test was used for non-parametric variables. Gender differences were examined using Mann Whitney test.

Results:
At the study visit there were no significant differences in age, gender, Insulin modality and disease duration. Systolic blood pressure was slightly but significantly higher in cases than in controls (median 116, range 92-180 vs 112, range 90-150mmHg, P=0.005). HbA1c was slightly but significantly higher in cases than controls (median 8.0, range 5.2-14.2 vs 7.8, 5-12.6%, P=0.03). The eGFR was normal in both groups but albumin excretion rate (AER) was increased in the cases as compared to the controls. There was a highly significant difference between cases and controls in terms of uAOG (median 6.6, range 1.0-1070 vs 4.0, range 0.1-65.3ng/mg, P<0.001). This difference persisted after adjusting for GFR, systolic blood pressure and HbA1c (P<0.001). Gender analysis revealed that uAOG was higher in females than males (median 11.7, range 1.0-393 vs 5.4, range 1.26-1070ng/mg, P=0.001)

Conclusion:
We conclude that uAOG is increased in people with type 1 diabetes with normal GFR and microalbuminuria. This increase in uAOG persists after adjustment for blood pressure, GFR and HbA1c. In addition, uAOG is higher in females than males. Gender differences are important for the interpretation of uAOG as a marker of kidney disease in people with type 1 diabetes.
Abstract P118: Tissue Transglutaminase Contributes to Pro-Inflammatory Cytokine LIGHT-Induced Ang II Sensitization
Authors: Liu Chen , Luo Renna , Wang Wei , Peng Zhangzhe , Zhou Zhen , Milewicz Dianna , Johnson Gail , Kellems Rodney , Xia Yang
Published: December 2018

Hypertension is a severe cardiovascular complication with elevated inflammation and pressor sensitivity. Activated enzymatically and transcriptionally by inflammatory cytokines, tissue transglutaminase (TG2), a ubiquitous crosslinking enzyme, is known to stabilize the Ang II receptor AT1 and contribute to hypertension and cardio-renal syndrome.We hypothesized that TG2 play an essential role in the inflammation-induced Ang II sensitization in hypertension via its posttranslational modifications. To test this, we found a significant TG2-dependent increase in renal transglutaminase activity in animals infused with inflammatory cytokine LIGHT/TNFSF14 (4ng/day) for 14 days (PBS=0.5±0.04, LIGHT=0.8±0.03, LIGHT+TG2 inhibitor ERW1041E=0.6±0.05, LIGHT+TG2-/-=0.4±0.01mU/mg protein, n=4-5, p<0.05). Western blot further determined an ~9 fold increase in renal TG2 and ~4 fold in AT1 in animals infused with LIGHT only (n=3, p<0.05). Mutagenesis study on AT1 confirmed the requirement of TG2 modification in LIGHT-induced receptor accumulation. Following this, we identified an ~80% pronouncement in Ang II-induced calcium response in luciferase reporter cells co-treated with LIGHT but not LIGHT plus ERW1041E (n=3, p<0.01). Taken together, our studies may shed light on the intrinsic role of TG2 as the pressor sensitizer downstream of the cytokines. Our results could also be a proof of concept for the use of TG2 inhibitors as novel antihypertensives.
Abstract P119: Hypertension and Renal Dysfunction Are Mitigated by Surgical Renal Denervation in an Adult Rat Model of Polycystic Kidney Disease
Authors: Banek Christopher T , Gauthier Madeline M , Osborn John W
Published: December 2018

Polycystic kidney disease (PKD) is the most common form of inherited chronic kidney disease failure. PKD is also associated with hypertension (HTN) and increased peripheral sympathetic activity (SNA). Recent studies have shown ablation of renal nerves may be effective treatment for varying forms of HTN and renal disease; however, the role and mechanism for renal nerves in PKD-associated renal dysfunction and HTN are unclear. Therefore, this study was conducted to test the hypothesis that renal denervation (RDN) would reduce AP, lower peripheral SNA, and improve renal function in the adult PKD rat. To test this hypothesis, 6-month-old male and female PKD rats underwent bilateral surgical RDN or sham surgery (Total n=18; 9/group; 3M/6F). Cardiovascular parameters were monitored by implanted radiotelemeter. Peripheral sympathetic tone was estimated by depressor response to acute ganglionic blockade (GB). Renal function was assessed by measuring glomerular filtration rate (GFR) by FITC-sinistrin clearance. Two weeks following treatment, rats were euthanized and tissues were collected for further analysis. Data was analyzed by a two-way ANOVA, across sex and treatment groups (α=0.05). Data presented as mean+SEM. At baseline, no differences in mean AP (132±2 vs. 129±3 mmHg), GB response (39±3vs. 42±4 ΔmmHg), nor GFR (0.93±0.06 vs. 0.91±0.04 mL/min/100g) were detected between RDN vs. sham. Two weeks following treatment, AP was lower in RDN (120±2mmHg) vs. sham (128±3mmHg). Similarly, GB was halved in RDN (-16±2 ΔmmHg) compared to sham (-32±3 ΔmmHg). GFR was increased in RDN rats vs. sham (1.18±0.06 vs. 0.84±0.08 mL/min/100g). Lastly, no difference in renal cystic area was detected between RDN and sham (8.27±1.39 vs. 8.74±0.90 %). No effect of sex was detected in any of the reported measurements. Altogether, these data support our initial hypothesis, where RDN effectively lowered AP and peripheral sympathetic tone in the PKD rat. Interestingly, we also observed a substantial increase in GFR in these animals, indicating an improvement in renal function despite no observable effect on cystic area. Of note, RDN was effective in both males and female rats. Further studies using repeated measurements are required to elucidate the order in which these effects occur.