bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 021: Role of Central Nervous System Suppressor of Cytokine Signaling 3 (socs3) in Regulating Metabolic and Cardiovascular Function in Obesity
Authors: do Carmo Jussara M , Freeman John N , da Silva Alexandre A , Moak Sydney P , Hall John E
Published: July 2021

Suppressor of cytokine signaling 3 (SOCS3), a negative regulator of leptin signaling, is upregulated in obesity and may contribute to development of leptin resistance. We determined whether deletion of SOCS3 in the entire central nervous system (CNS) would protect mice from adverse metabolic and cardiovascular effects of a high fat diet (HFD). SOCS3
flox/flox
/Nestin-cre mice were generated by breeding SOCS3
flox/flox
with Nestin-Cre mice. Male and female mice with CNS deletion of SOCS3 (SOCS3
flox/flox
/Nestin-Cre, n=5-10) or control (SOCS3
flox/flox
, n=6-10) were fed a HFD plus sucrose from 6 until 22 weeks of age. Mean arterial pressure (MAP) and heart rate (HR) were recorded by telemetry and oxygen consumption (VO
2
) was monitored by indirect calorimetry in 22-week-old mice. Compared to control mice, SOCS3
flox/flox
/Nestin-Cre mice were lighter (male: 34±3 vs. 45±3 and female: 27±1 vs. 37±2 g) and had elevated VO
2
(94±12 vs. 69±6 ml/kg/min) but there were no significant differences in food intake (male: 3.3±0.7 vs. 3.8±0.5 and female: 3.0±0.6 vs. 3.0±0.5 g/day) or plasma glucose (male: 148±8 vs. 198±31 and female 149±12 vs. 164±12 mg/dl). Male SOCS3
flox/flox
/Nestin-Cre mice had similar MAP (115±2 vs. 116±1 mmHg) but higher HR (657±3 vs. 592±3 bpm) compared to control mice. However, female SOCS3
flox/flox
/Nestin-cre mice had higher MAP (121±1 vs. 108±1 mmHg) and HR (655±2 vs. 606±5 bpm) compared to control mice. No significant differences were observed in glucose tolerance in SOCS3
flox/flox
/Nestin-Cre vs. control mice (AUC: 391±36 vs. 429±54 mg/dL x 120 min in males and 459±70 vs. 372±51 mg/dL x 120 min in females). These results indicate that CNS SOCS3 deletion reduced body weight and increased energy expenditure and HR but did not improve glucose tolerance in male or female mice fed a HFD. However, HFD significantly increased BP in female SOCS3
flox/flox
/Nestin-Cre mice compared to control mice fed a HFD, suggesting a sex difference in the role of CNS SOCS3 signaling in BP regulation in obesity. (NHLBI PO1HL51971and NIGMS P20GM104357)
Abstract 041: New Evidence for G Protein-coupled Estrogen Receptor as a Natriuretic Factor
Authors: Gohar Eman Y , Obi Ijeoma E , De Miguel Carmen , Pollock Jennifer S , Pollock David M
Published: July 2021

Premenopausal women have a lower risk of cardiovascular and renal diseases compared to age-matched men. This protection includes fewer salt-dependent complications. We have shown that female rats have a more robust natriuretic capacity compared to males in response to increased dietary salt. The novel estrogen receptor, G protein-coupled estrogen receptor (GPER), is a membrane-bound receptor linked to acute signaling pathways. GPER activation elicits protective effects throughout the cardiovascular system. However, its role in sodium handling is not defined. We hypothesized that activation of GPER in the renal medulla stimulates sodium excretion. In female Sprague Dawley (SD) rats, isosmotic saline was infused into the renal medullary interstitium (500 μl/h) during a 60-80 min equilibration period and 20 min baseline urine collection period. This was followed by infusion of the GPER agonist G1 (50 pmol/kg/min) or vehicle into the renal medulla for two further 20 min periods. Compared with vehicle, G1 significantly increased urinary sodium excretion and urine flow (from 0.5 ± 0.1 to 0.9 ± 0.2 μmol/min and from 5.3±1.1 to 8.3±1.6 μl/min, respectively, n=6, p<0.05). Urinary potassium excretion and mean arterial pressure remained unchanged during the experiments (0.5 ± 0.1 vs. 0.4 ± 0.1 μmol/min and 108.6 ± 3.4 vs. 107.2 ± 5.1 mmHg, respectively, n=6). Immunohistochemical analysis of GPER revealed more predominant staining in both inner and outer renal medulla in female SD rats, compared to males. GPER appears to be predominantly expressed in interstitial cells. These data reveal that renal medullary GPER plays an important role in renal sodium handling and may account for the enhanced ability to handle increased dietary salt in females.

Funded by AHA 15POST25090329 to EYG and P01 HL136267 to DMP
Abstract 045: The Enhanced Pressor Response to AngII in Aged Females is Attenuated by Estrogen Replacement via an AT
2
R-mediated Mechanism
Authors: Barsha Giannie , Mirabito Colafella Katrina M , Gaspari Tracey , Spizzo Iresha , Hilliard Lucinda M , Widdop Robert E , Samuel Chrishan S , Denton Kate M
Published: July 2021

Loss of estrogen (E
2
) following menopause contributes to the sharp rise in cardiovascular risk with age. E
2
is postulated to play a protective role against hypertension and end-organ damage by counterbalancing the pressor actions of the RAS and enhancing the depressor RAS pathways. The aim was to determine whether E
2
replacement in aged females can lower arterial pressure and improve endothelial function via an AT
2
R-mediated mechanism. MAP was measured via telemetry in ovary-intact adult (4-month old), aged (17-month old) and aged+E
2
(3 μg/day sc) FVB/N female mice, which were co-treated with vehicle, AngII (600 ng/kg/min sc) or AngII+PD (PD123319, AT
2
R antagonist; 3 mg/kg/day sc). On day 21 of treatment, endothelium-dependent relaxation in response to acetylcholine was assessed in aortic vessels. Cardiac and renal, tissue fibrosis and gene expression of E
2
receptors and RAS components were also analysed. Basal MAP was lower in E
2
-treated aged mice (90±1 mmHg, n=20) relative to adult (94±1 mmHg, n=10) and aged controls (94±1 mmHg; n=21, both P<0.05). Similar to previous studies, the AngII pressor response was enhanced in aged compared to adult females (Figure). E
2
treatment reduced the AngII pressor response in aged females (Figure). Moreover, the attenuated pressor response observed in the aged E
2
+AngII group was abolished by co-infusion with PD (Figure). Endothelial function was markedly impaired with age (~23% reduced maximal vasodilation, P=0.03) and worsened by AngII infusion (~38%, P=0.001), however E
2
did not significantly improve the response in any group. In conclusion, E
2
replacement may reinstate cardio-protection in aged females via an AT
2
R-mediated mechanism.
Abstract 058: Deoxycorticosterone-induced Hypertension Caused by Fluconazole
Authors: Hakimi Matthew , Krishnan Sandeep , Mason O’Neil , Trabulus Joshua , Zakowski Phillip , McPhaul Michael , Auchus Richard , Victor Ronald G
Published: July 2021

Deoxycorticosterone (DOC), a steroid precursor of aldosterone, constitutes an endogenous mineralocorticoid receptor agonist. Exogenous DOC produces a well-established animal model of mineralocorticoid-induced salt-dependent hypertension (HTN), but, except for rare cases of congenital adrenal hyperplasia with pediatric HTN due to
CYP11
β1
(11
β
hydroxylase, P450c11
β
)
deficiency, DOC is rarely considered a cause of clinical hypertension (HTN). We recently suspected DOC as the novel mechanism of severe low-renin HTN (blood pressure, BP 220/150 mmHg, plasma renin activity 0.21 ng/mL/hour) that developed
de novo
in a 47- year old African American man with previously normal BP (118/74) during treatment of disseminated coccidiodomycosis with high-dose fluconazole, which blocks P450 enzymes in the fungal cell wall. To determine if the fluconazole also inhibited human CYP11β1, the adrenal P450 enzyme that converts DOC to corticosterone, we performed a comprehensive adrenal steroid panel on the patient’s serum using mass spectrometry. During the HTN, his serum [DOC] was 24-times the upper limit of normal (354 vs. <15 ng/dL), while corticosterone was low and aldosterone was undetectable. Both the HTN and the high DOC were caused by fluconazole, because they normalized after treatment ended. We then proved that fluconazole causes dose-dependent inhibition of CYP11β1
in vitro
by transfecting a mammalian (hamster V79) cell line with human
CYP11
β
1
and incubating the cells with DOC. The observed IC50—10 μM— is certainly less than the plasma [fluconazole] during oral high-dose therapy. From these data, we conclude that high-dose fluconazole inhibits DOC conversion by human CYP11β1, causing in this adult patient an acquired form of severe DOC-induced HTN. Given the recent 10-fold increase in coccidiodomycosis cases in the United States and the marked propensity for disseminated disease in African American patients, BP should be monitored carefully during treatment with high-dose fluconazole. Additional case finding studies are warranted.
Abstract 060: Role of Mir-214 in the Regulation of Perivascular Fibrosis in Angiotensin II Induced Hypertension
Authors: Nosalski Ryszard , Siedlinski Mateusz , Nguyen Dinh Cat Aurelie , Skiba Dominik , Mcginnigle Eilidh , Baker Andrew , Denby Laura , Guzik Tomasz J.
Published: July 2021

Objective:
Hypertension (HT) is associated with perivascular inflammation and increased vascular fibrosis. MicroRNAs (miR) are a novel gene expression regulation mechanism and play a pivotal role in a range of pathological processes. The role and mechanism of miR214 in vascular fibrosis is unknown.

Methods:
3-month-old C57BL/6, miR214KO and wild-type littermates were treated with angiotensin II (AngII, 490ng/kg/min; n=6-10) or control buffer for 14 days. PVATs from C57BL/6 animals were analysed using TaqMan_Rodent_microRNA_Arrays. Histological studies, wire myography, lucigenin-enhanced luminometry and cytometrical analysis was conducted, followed by statistical analysis with ANOVA or t-test. Data are expressed as a mean±SEM.

Results:
Out of 381 miRs, 16 were significantly overexpressed in C57BL/6 AngII animals, with only miR214 showing 8-fold induction (p<0.01) after Bonferroni correction. Also, 3-fold elevation of pri-miR-214 was observed. Interestingly, hydralazine treatment prevented both these changes (p<0.01). AngII infusion in miR214 KO animals did not alter blood pressure when compared to WT mice. Mir214 KOs exhibited diminished peri-aortic fibrosis (44779±2491 vs 78805±8696μm, p<0.01), upon AngII hypertension. This was associated with a significantly reduced induction of COL1A1, COL3A1 and TGFβ1 mRNA expression in PVAT and aortas (p<0.05). Vascular studies revealed improved endothelial function (69±10 vs. 22±4%, p<0.01), protection against oxidative stress (66±7 vs 118±19 RLU/sec/mg, p<0.001) and NOX2 mRNA expression (1.9±0.2 vs1.1±0.1, p<0.05) in AngII miR-214-KO aortas, while these parameters were not altered in mesenteric arteries. Recruitment of T cells into aortic PVAT was abolished in KO HT animals in comparison to control group (192±65 vs. 603±164 cell/mg; p<0.05). AngII HT was associated with 4-fold increase of miR-214 expression in the circulating peripheral blood T cells and 2-fold in the spleen. Moreover, AngII infusion increased TNFα mRNA expression in WT T cells (1±0.1 vs 1.6±0, p<0.01) whereas this effect was not seen in miR214 KO T cells (0.9±0.3 vs 0.9±0.1).

Conclusions:
MiR-214 plays a major role in modulation of aortic fibrosis, vascular function, oxidative stress and perivascular inflammation.
Abstract 067: Genetic Complementation Establishes That a K572Q Mutation in Gamma-adducin Plays a Causal Role in Renal Microvascular Dysfunction in FHH and MNS Rats
Authors: Zhang Chao , Ge Ying , Mims Paige N , Fan Fan , Roman Richard J
Published: July 2021

The Fawn-Hooded hypertensive (FHH) rat is a genetic model of hypertension-induced renal disease. However, the causal genes and pathways involved are unclear. We previously reported that the transfer of a small region in Chr. 1 of Brown-Norway (BN) rats which contains 15 genes, including gamma-Adducin (Add3), into the FHH background restores renal microvascular function and attenuates the development of proteinuria in FHH rats. Our further work identified a K572Q mutation in Add3 in FHH rats as a potential candidate variant in the pathogenesis of renal disease. The present study examined the role of Add3 in the impaired myogenic response of the afferent artery (Af-art) and autoregulation of renal blood flow (RBF) using transgenic and KO rats. RBF increased by 21.5 ± 3.0% in SD. Add3 KO rats (n=7) when mean arterial pressure (MAP) was increased from 100 to 150 mmHg. In contrast, RBF only increased by 3.5 ± 0.9% in wildtype SD rats (n=13). The diameters of the renal Af-art decreased by 12.9 ± 0.8% in SD rats when perfusion pressure was increased from 60 to 120 mmHg, but it increased in SD.Add3 KO rats. The myogenic response of the Af-art in FHH rats was markedly impaired and increased by 8 ± 1.2% when the pressure was increased by from 60 to120 mmHg. The myogenic response was restored, and the diameters of the Af-art decreased by 12 ± 0.7% and 7 ± 1.0% in FHH. 1
BN
congenic rats (n=27) and FHH.Add3 transgenic rats that express wt-Add3. RBF increased by 35.1 ± 3.0% when MAP was increased from 100 to 150 mmHg in FHH rats (n=15) versus 7.5 ± 1.7% and 6.0+ 1.3% in FHH.1
BN
or a F1 cross of FHH and FHH.1
BN
rats (n=8) and FHH.Add3 transgenic rats. The myogenic response of Af-art and autoregulation of RBF were also impaired in MNS rats (n=6) that carry the same K572Q mutation in Add3 as FHH rats. These phenotypes were complemented in a F1 cross of FHH and MNS rats (n=7), but the myogenic response and autoregulation of RBF were restored in an F1 cross of FHH and FHH.1
BN
rats with one copy of wt-Add3. These results indicate that the recessive K572Q mutation of Add3 in FHH and MNS rats plays a causal role in renal microvascular dysfunction, which may contribute to the development of chronic kidney disease induced by hypertension in these models.
Abstract 074: Use of Antihypertensive Drugs, Drug Classes, and Combinations in Resistant versus Non-resistant Hypertension, and in True versus White-coat Resistant Hypertension
Authors: De La Sierra Alejandro , Armario Pedro , Oliveras Anna , Banegas Jose R , Gorostidi Manuel , Vinyoles Ernest , Segura Julian , De la Cruz Juan J , Ruilope Luis M
Published: July 2021

We aimed to evaluate the use of specific antihypertensive drugs and drug classes, as well as combinations in patients treated with 3 or more drugs classified as having or not resistant hypertension (RH), controlled or uncontrolled RH and true versus white-coat RH. From the Spanish ABPM Registry, we identified 21238 patients treated with 3 (14264) or more (6974) antihypertensive drugs of different classes. Among patients treated with 3 drugs we compared those with controlled (<140/90 mmHg; No RH) or uncontrolled (RH) office BP. In patients treated with 4 or more drugs we compared controlled versus uncontrolled RH. Moreover in uncontrolled RH patients, we compared those with white-coat (normal ABPM) versus true RH. We evaluated the use of different antihypertensive drug classes, specific antihypertensive drugs inside each class, and types of combinations. Results were adjusted for age, gender, and previous history of cardiovascular disease. With respect to RH treated with 3 drugs, those No RH used more frequently aldosterone antagonists (AA; Relative risk: 1.82; 1.40-2.37. They also used more frequently clorthalidone (CTL) among diuretics (RR: 1.54; 1.24-1.91), amlodipine (AML) among calcium channel blockers (CCB; RR: 1.12; 1.00-1.24) and bisoprolol (BIS) among beta blockers (RR: 1.18; 1.03-1.35). In patients treated with 4 or more drugs, controlled RH was also associated with the use of AA (RR: 1.41; 1.14-1.73) and AML (RR: 1.42; 1.25-1.62). No differences were observed in the type of combination used. When comparing patients with true versus white-coat RH, the latter group used more frequently diuretics (RR: 1.31; 1.16-1.47), CTL (RR: 1.79; 1.48-2.16) among diuretics, and AML (RR: 1.44; 1.32-1.57) among CCB. The triple combination of RAS blockers, CCB, and diuretics was also more frequently used in those with white-coat RH (RR: 1.09; 1.00-1.19). In conclusion, controlled office BP among patients treated with 3 or more drugs is associated with AA, CTL, and AML use. White-coat RH is associated with more diuretic use, especially CTL, AML, and the combination of RAS blockers, CCB, and diuretics. These results support the use of such triple combination, preferably including CTL and AML, and the addition of AA in order to reduce the prevalence of RH and true RH.
Abstract 076: Factors Associated With Home Blood Pressure Monitoring Among US Adults: The National Health and Nutrition Examination Survey (NHANES), 2013-2014
Authors: Marrs Joel C , Anderson Sarah L
Published: July 2021

Introduction:
National prevalence data on home blood pressure monitoring (HBPM) has previously been reported from 2009-2010, but no recent data has been reported.

Methods:
This report is based on national-level, cross-sectional data for noninstitutionalized US adults aged ≥18 years (n = 6,113 participants) from the National Health and Nutrition Examination Survey (NHANES), 2013-2014.

Results:
Overall, 25.5% (1560 of 6113) of the adults engaged in HBPM monitoring in the 2013-2014 survey year. This is an increase from the previous NHANES 2009-2010 analysis reporting a 21.7% (1302 of 6001) rate. The frequency of HBPM increased with higher age, higher education level, having a partner, hypertensive, hypertensive aware, and hypertensive treated. Both SBP and DBP were higher in the HBPM group and were both statistically significant. The groups of hypertensive, hypertensive-aware, and hypertensive-treated patients all showed higher rates of HBPM use than nonuse. The frequency of HBPM monitoring in the categories of less than monthly, monthly, and weekly was reported at rates of 7.8% (476 of 6113), 6.6% (401 of 6113), and 11.2% (683 of 6113), respectively. These reported rates are consistent and slightly increased in the weekly HBPM reported use compared with the NHANEs 2009-2010 analysis (7.2% [432 of 6001], 6.6% [396 of 6001], and 7.9% [474 of 6001), respectively). Adjusting for covariables, those who were aware of, treated for, and had known hypertension were more likely to have a higher frequency of HBPM than the reference: unaware, untreated, and no known hypertension (odds ratio (OR) = 1.98; OR = 2.13; and OR = 1.64, respectively). Individuals with less than a high school diploma and having no partner were less likely to perform HBPM than the reference: high school graduate or greater and having a partner (OR = 0.73; OR = 0.65, respectively).

Conclusions:
Approximately 21.0% (1084 of 6113) of adults engaged in monthly or more frequent HBPM which is an increase from the reported 14.5%(870 of 6001) rate in the 2009-2010 analysis. Having hypertension, being aware of hypertension, and being treated for hypertension were associated with an increased frequency of HBPM. Having no partner and less than a high school diploma were associated with lower frequency of HBPM.
Abstract 088: At
1
Receptor on Glutamatergic Neurons Regulate Autonomic Function Through Modulation of Neuronal Excitability and Sympathetic Outflow
Authors: Xu Jiaxi , Molinas Adrien , Zsombok Andrea , Lazartigues Eric
Published: July 2021

Using a mouse model with Angiotensin II type 1 receptor (AT
1a
R) deletion from the central nervous system neurons, we previously reported attenuated hypertension, improved autonomic function, and importantly, blunted ADAM17 (A Disintegrin And Metalloprotease 17) activation after deoxycorticosterone-salt treatment (DOCA 1mg/g body weight sc + 1% saline po for 3 weeks) compared to their control littermates. Since neuronal AT
1a
R has a pivotal role in ADAM17-mediated ACE2 shedding and the maintenance of neurogenic hypertension, we further investigated the role of central AT
1a
R and ADAM17 in excitatory neurons. New mouse models were generated with deletion of AT
1a
R (AT1G) or ADAM17 (A17G) specifically from glutamatergic neurons. AT1G (n=9), A17G (n=9), and their control littermates (n=10) were implanted with telemetry probes for continuous recording of blood pressure. Following DOCA-salt treatment, both strains showed increased mean arterial pressure, however the pressor responses were significantly lower in both transgenic lines (AT1G: +16 ±3 mmHg, A17G: +21 ±3 mmHg), compared to the controls (+31 ±2 mmHg). Meanwhile, the pulse pressure in the AT1G and A17G mice were not significantly increased compared to their baseline, while it was elevated in the control littermates by around 20 mmHg (
P<0.01
). In addition to a reduced hypertension, AT1G mice exhibited preserved baroreflex sensitivity and autonomic function at the end of the DOCA-salt protocol. The DOCA-salt-induced dysautonomia was also attenuated in A17G mice but still exhibited decreased cardiac parasympathetic tone and increased vascular sympathetic tone, indicating that glutamatergic AT
1
R can modulate autonomic regulation independently of ADAM17. To further dissect this mechanism, patch-clamp recording was performed in AT1G and control mice. Interestingly, lower action potential frequency was recorded in kidney-projecting PVN neurons of AT1G mice (0.43 ±0.06
vs.
2.04 ±0.96 Hz, n=3,
P=0.01
), suggesting that AT
1a
R-knockdown neurons have a lower excitability compared to controls. Altogether, our data provide evidence that AT
1a
R located on glutamatergic neurons regulate autonomic function and the development of hypertension through ADAM17-dependent and independent pathways.
Abstract 096: Comparing Strategies to Improve Systolic Blood Pressure Over 10 Years: A Simulation Study Using the Blood Pressure Control Model
Authors: Bellows Brandon K , Moran Andrew E , Ruiz-Negrón Natalia , Bibbins-Domingo Kirsten , Pletcher Mark , Fontil Valy
Published: July 2021

Background:
Uncontrolled hypertension increases patients’ risk for cardiovascular and kidney disease. This study compared strategies to improve systolic blood pressure (SBP) among 1000 simulated patients with uncontrolled hypertension (SBP >=140 mmHg) from the National Health and Nutrition Examination Survey (NHANES).

Methods:
The Blood Pressure Control Model (BPCM) is a microsimulation, health state transition model that predicts the weekly SBP of patients receiving usual care. In the BPCM, patient SBPs are estimated using office visit frequency, measured SBP accuracy and variability, probability of treatment intensification with uncontrolled SBP, effect of antihypertensive medications, and adherence. BPCM inputs are derived from national survey data, meta-analyses, and other published literature. The effects of usual care on SBP were compared to 10% and 50% increases in global strategies for SBP control (i.e., visit frequency, treatment intensification, and/or adherence) over 10 years. SBP outcomes were validated against published literature values of 44-46% prior to implementation (i.e., usual care) and 74-80% 8-10 years after implementation of aggressive hypertension management programs in large health systems.

Results:
In the simulated NHANES population, the mean (SD) age was 61.1 (14.6), 52% were male, and mean baseline SBP was 153.2 (13.6) mmHg. Under usual care, the BPCM estimated a mean SBP of 140.1 (16.4) mmHg and 49% of patients achieving SBP <140 mmHg after 10 years. Compared to usual care, 50% improvements in global strategies resulted in more rapid reductions in SBP and earlier achievement of SBP control. Simultaneously improving all global strategies by 50% resulted in an estimated mean SBP of 132.4 (15.5) mmHg with 71% achieving control after 10 years.

Conclusions:
Usual care and intervention BPCM predictions are consistent with hypertension control rates observed in contemporary national surveys and the observed results of recent systematic hypertension control improvement programs. These results show the BPCM may be used by health system planners to project the impact of implementing hypertension control strategies.
Abstract 099: Smooth Muscle PPARγ Mutation Causes Salt-sensitive Hypertension
Authors: Wu Jing , Agbor Larry N , Mukohda Masashi , Nair Anand R , Nakagawa Pablo , Grobe Justin L , Sigmund Curt D
Published: July 2021

Abnormal increase in renal salt retention is traditionally believed to be an early pathophysiological event in the causation of salt-sensitive hypertension, whereas increase in systemic vascular resistance (SVR) is a secondary response caused by autoregulation. However, recent studies show that salt-resistant subjects vasodilate and reduce SVR during salt loading, while salt-sensitive humans fail to vasodilate and exhibit salt-induced blood pressure (BP) elevation. Therefore, we tested the hypothesis that primary vascular dysfunction predisposes to salt sensitive hypertension. We used mice with smooth muscle-specific expression of a human hypertension-causing mutation in PPARγ P467L (S-P467L). S-P467L transgenic mice and non-transgenic controls (NT) were fed regular diet (0.4% salt) or high salt diet (4% salt) for 4 weeks. S-P467L mice, but not NT controls, exhibited severe impairment in acetylcholine- and sodium nitroprusside-induced vasorelaxation (31±4.9% S-P467L salt vs. 70±9.5% regular diet, maximal relaxation at 30 μM acetylcholine). This was associated with salt-induced systolic BP elevation in S-P467L mice (142±5 mmHg salt vs 127±2 mmHg regular diet), but not in NT mice (120±2.7 mmHg salt vs 115±4.0 mmHg). These changes were not due to differences in food intake, weight gain or renal sympathetic nerve activity between the two strains. In the 3
rd
week of high salt diet, S-P467L mice and NT controls both had increased water intake by 3-fold compared to those on regular diet; however, S-P467L mice excreted 32% less urine and produced 36% less NO in the kidney as indicated by 24-hour urinary nitrate/nitrite. To assess renal function, mice were subjected to an acute saline challenge (10% body weight, i.p. injection). S-P467L mice exhibited a marked decline in their capacity to excrete this volume/sodium load, indicative of renal dysfunction. Of note, the impaired vasorelaxation in S-P467L occurred as early as day 3 of high salt diet, while renal dysfunction did not develop until day 10, suggesting that vascular dysfunction may serve as an initiation mechanism that reinforces salt-induced hemodynamic changes. These data supports the concept that vascular dysfunction may predispose to renal abnormalities including increased salt sensitivity.
Abstract 102: Loss of Salt Sensing Kinase, SGK1, in T Cells Abrogates Memory T Cell Formation, Hypertension, and End-organ Damage
Authors: Itani Hana A , Pandey Arvind , Norlander Allison E , Madhur Meenakshi S , Harrison David G
Published: July 2021

Accumulating evidence indicates that NaCl can be concentrated in tissues with high salt-intake, age and in the setting of hypertension. Elevated NaCl has been shown to promote T
H
17 cell formation in an SGK1-dependent fashion. We have previously shown that Memory T cells play a major role in the genesis of hypertension. These long-lived cells remain responsive to repeated hypertensive stimuli, such as salt feeding, and can be mobilized to enter the kidney where they release cytokines that promote renal dysfunction. To examine mechanisms by which T cells sense salt and contribute to salt-sensitivity, we tested the hypothesis that SGK1 in T cells is necessary for formation of memory T cells and their activation in salt-sensitive hypertension. To study the role of SKG1 in hypertension, we produced mice with T cell specific deletion of SGK1, SGK1
fl/fl
x tg
CD4cre
mice and used SGK1
fl/fl
mice as controls. To impose repeated episodes of hypertension, we treated these mice with L-NAME (0.5mg/ml) in drinking water for two weeks, allowed a two-week normotensive interval and a then fed high salt (4% NaCl) for three weeks. L-NAME followed by high salt increased memory T cells in the kidney, aorta and bone marrow of SGK1
fl/fl
control mice but not in SGK1
fl/fl
x tg
CD4cre
mice, as identified by the surface marker CD44
hi
. To assess markers of renal injury, we measured albumin in 24-hour urine samples collected at the end of the L-NAME/high salt. L-NAME/high salt caused striking albuminuria in SGK1
fl/fl
mice and was absent in SGK1
fl/fl
x tg
CD4cre
mice. In additional studies, we found that loss of SGK1 in T cells abrogates renal and vascular inflammation and protects against hypertensive renal and vascular injury in the L-NAME/high salt model. Thus, our data provide a potential mechanism by which SGK1 in T cells promotes their development of salt sensitivity and their mediation of renal and vascular dysfunction in hypertension.
Abstract 110: Reduced Expression, Knockout, and Pharmacological Intervention of
Arhgef11
-RhoA Pathway Significantly Attenuates Renal Injury and Blood Pressure
Authors: Johnson Ashley C , Pasco David S , Garrett Michael R
Published: July 2021

Through genetic analysis of the Dahl salt-sensitive (SS) rat, a model of hypertension and chronic kidney disease (CKD), Arhgef11, a Rho guanine nucleotide exchange factor was implicated in kidney injury. Arhgef11, via exchange of GDP for GTP, plays a role in the activation of RhoA signaling cascades through a number of cell stimuli that impact cytoskeletal structure and influence cell-cell contacts and promotes cell transformation. Previously, we demonstrated that reduced
Arhgef11
expression/protein function in an SS-
Arhgef11
SHR
-minimal congenic strain resulted in significantly decreased proteinuria, fibrosis, and improved renal hemodynamics compared to SS-WT, without impacting BP. More recently, an SS-Arhgef11
-/-
knockout rat model was studied. On low-salt (0.3% NaCl), SS-Arhgef11
-/-
animals demonstrated reduced proteinuria and renal injury, with no impact on BP versus SS-WT. In contrast, SS-Arhgef11
-/-
animals on an elevated-salt diet (2% NaCl) demonstrated a significant (p<0.001) attenuation of proteinuria (41±7.9 mg/24 hrs.), along with a substantial reduction in BP (124±2.6 mm Hg) compared to SS-WT (119±15.3 mg/24 hrs. and 151±5.9 mm Hg). This data suggests that reduced expression/loss of
Arhgef11
(on low-salt) similarly leads to renoprotection, whereas the loss of Arhgef11 (vs reduced expression) leads to blunting of salt-induced elevations in BP vs SS-WT. These animal studies, in combination with
in vitro
work, suggest that inhibition of Arhgef11-RhoA could be an effective therapeutic for CKD. Using an
in vitro
cell screen, several hundred natural product compounds were tested for ability to inhibit Arhgef11/RhoA activity and identified 3 compounds. A pilot study done using one compound (sufficient for a one-week study) was very encouraging as animals treated with XTL-019-G7 (20mg/kg/day) exhibited an ~25% reduction in proteinuria compared to vehicle (VEH) treated animals (70±8.5 versus 95±9.6 mg/24hours, p=0.07), demonstrating that
in vivo
testing is feasible and that the compound warrants further testing. In summary, genetic analysis of a model of CKD identified a gene/pathway involved in kidney injury that served as a target to screen natural product derived small molecules, and may ultimately lead to a new treatment for CKD.
Abstract 111: A Kidney Targeted Epoxyeicosatrienoic Acid Analog, EET-F01, Reduces Cisplatin-induced Nephrotoxicity
Authors: Imig John D , Khan Md A , Adebesin Adeniyi M , Falck John R
Published: July 2021

Epoxyeicosatrienoic acid (EET) analogs have exceptional therapeutic potential to combat cardiovascular and kidney diseases. EET analogs combat damage in acute and chronic kidney disease models. Biological actions attributed to EET analogs such as vasodilation, anti-inflammation, anti-apoptosis, and anti-fibrosis are ideally suited to treat kidney diseases. Although EET analogs have performed well in several
in vivo
models, targeted delivery of EET analogs to the kidney can be reasonably expected to reduce the level of drug needed to achieve a therapeutic effect in the kidney and obviate possible side effects. For EET analog kidney-targeted delivery, we conjugated an EET analog to folic acid because there is a high concentration of folate receptors in renal tissue. The EET analog was conjugated to folic acid via a PEG-diamine linker. Next, we compared the kidney targeted EET analog, EET-F01, to a well-studied EET analog, EET-A. EET-A or EET-F01 was infused i.v. (10mg/kg/hr) for 6 hours via the rat jugular vein. Plasma and kidney tissue were collected and EET-A or EET-F01 measured by LC-MS-MS. EET-A plasma level was 1.6 ng/mL, but EET-A was undetectable in the kidney. On the other hand, EET-F01 was 6.5 ng/mL in plasma and 26.7 ng/mL in kidney tissue. These data demonstrate that EET-F01 targets the kidney. Experiments were conducted to compare EET-F01 and EET-A to decrease cisplatin-induced nephrotoxicity. A single injection of cisplatin (7 mg/kg ip) was administered to WKY rats treated with vehicle, EET-A (10 mg/kg ip) or EET-F01 (20 mg/kg or 2 mg/kg ip) for five days. Cisplatin increased BUN (125 ± 11 mg/dL) and NAG (12 ± 4 IU/L) compared to control (36 ± 9 mg/dL and 4 ± 1 IU/L). EET-F01 was as effective as EET-A in decreasing BUN, NAG, and renal histological injury five days following cisplatin administration. Despite it almost 2x-greater molecular weight compared with EET-A, EET-F01 was effective in lowering BUN and NAG at 20 mg/kg/d and at a 10-fold lower dose of 2 mg/kg/d. These data clearly demonstrate that EET-F01 targets the kidney and allows for a lower effective dose. In conclusion, we have developed a kidney targeted EET analog, EET-F01, that demonstrates excellent potential as a therapeutic for kidney diseases.
Abstract 126: Blood Pressure in Childhood Predicts HTN in Adulthood: The International CV Cohorts Consortium
Authors: Urbina Elaine M , Bazzano Lydia , Burns Trudy , Dwyer Terry , Jacobs David , Juonala Markus , Prineas Ronald , Raitakari Olli , Sinaiko Alan , Steinberger Julia , Venn Alison , Woo Jessica
Published: July 2021

The natural history of blood pressure (BP) tracking from childhood to adulthood is not well defined. Using data from the International Childhood CV Cohorts Consortium, consisting of 7 international longitudinal cohorts, participants (N=5042) were evaluated by self-reported adult hypertension (HTN). Correlation between mean of all measures of BP on a subject during childhood (8-11 years) or adolescence (15-18 years) and adult BP (28-31 years) was evaluated with Spearman correlation coefficients. Differences in mean BP, body mass index (BMI) and laboratory values were evaluated by t-tests. Participants at time of self-report of HTN were mean age 48.5 years (81.7% white, 17.4% black; 39% male). Prevalence of HTN (N=1562) was 31%; 3480 were normotensive (NT). Correlations between child and adolescent SBP and DBP were r=0.43, 0.42, respectively, child and young adult r=0.24, 0.23 and adolescent and young adult r=0.41, 0.27 (all p<0.0001). Participants self-reporting HTN were more likely to be non-white (46.8% black, 30.2% white, 29.7% other, p<0.0001). Males were more likely to report HTN (33% vs 28% females, p<0.0001). They had significantly higher SBP and BMI as children and adolescents and also significantly higher DBP and fasting glucose by adolescence and lower HDL and higher TG by young adulthood (all p<0.01). We conclude that adult HTN begins in childhood with higher BP and BMI which appear to be important clinical markers for progressive increases in metabolic risk factors as these individuals age through adolescence and young adulthood.
Abstract 131: Racial Differences in Nocturnal Hypertension and Non-dipping Blood Pressure: The Coronary Artery Risk Development in Young Adults (CARDIA) Study
Authors: Muntner Paul , Booth John N , Thomas Stephen J , Deng Luqin , Schwartz Joseph E , Llyod-Jones Donald , Shikany James M , Lewis Cora E , Shimbo Daichi
Published: July 2021

Risk factors for nocturnal hypertension are more common among blacks compared with whites. We hypothesized nocturnal hypertension and nocturnal non-dipping BP are more common among blacks compared with whites. We analyzed data for 781 participants of the population-based Coronary Artery Risk Development in Young Adults (CARDIA) study who completed ambulatory blood pressure (BP) monitoring (ABPM) in 2015-2016. Awake and sleep periods were defined using actigraphy and self-report. Nocturnal hypertension was defined as mean sleep systolic BP (SBP)/diastolic BP (DBP) ≥ 120/70 mm Hg. Non-dipping SBP and DBP, separately, were defined as a decline in mean sleep BP, relative to mean awake BP < 10%. The mean age of participants was 54.7 years, 21.1% were white women, 38.5% were black women, 16.8% were white men and 23.6% were black men. The prevalence of nocturnal hypertension was 18.2% and 44.5% among white and black women, respectively, and 35.9% and 59.8% among white and black men, respectively. After multivariable adjustment, the prevalence of nocturnal hypertension was higher among black women, white men and black men, each compared with white women (Table). The prevalence of non-dipping SBP was 21.2% and 40.9% among white and black women, respectively, and 19.8% and 37.5% among white and black men, respectively. After multivariable adjustment, non-dipping SBP was more common among black women and black men compared with white women. There were no statistically significant differences in non-dipping DBP across race-gender after multivariable adjustment. Nocturnal hypertension and non-dipping SBP are more common among blacks compared with whites even after adjustment for mean BP.
Abstract 149: Impact of Baseline 10-year Cardiovascular Risk on Benefit and Harm of Intensive Treatment in SPRINT
Authors: Phillips Robert A , Xu Jiaqiong , Peterson Leif E , Diamond Joseph A , Schussheim Adam E
Published: July 2021

We investigated the impact of baseline CV risk on outcomes in SPRINT. Using the ACC/AHA CVD risk algorithm, we stratified the SPRINT population into quartiles of baseline 10-year CV risk. Within each quartile, Cox proportional hazards models were used to examine the effect of intensive treatment vs. standard of care on the SPRINT CV outcomes, all-cause mortality and serious adverse events (SAEs). Number needed to treat (NNT) and number needed to harm (NNH) were calculated for each quartile. There were 9,323 participants with available baseline ACC/AHA 10-year risk scores. In each quartile of risk, the hazard ratio (HR) favored intensive treatment. For CV outcomes, NNT decreased from 91 in the 1
st
quartile to 38 in the 4
th
quartile. For all-cause mortality, NNT decreased from 333 in the 1
st
quartile to 45 in the 4
th
quartile. Although incidence of all SAEs increased with each quartile in both treatment groups (p for trend <0.0001), there was no difference in incidence of SAEs between the treatment groups in each quartile. However, SAEs classified as hypotension were more frequent in the 4
th
quartile for the intensive treatment group (incremental increase 1.8%, NNH = 55) and SAEs classified as acute kidney injury or acute renal failure were significantly more frequent in the 2
nd
, 3
rd
and 4
th
quartiles for the intensively treated group (incremental increase 1.7%, 2.4% and 2.1%; NNH 59, 42 and 48, respectively). Therefore, those with greatest baseline CVD risk got the most benefit from intensive BP treatment but were at greater risk for hypotension and renal injury. This analysis may help providers and patients make decisions regarding the intensity of BP treatment to prevent death and CV events.
Abstract 151: Falls and Syncope Among Treated Hypertensive Individuals With Lower Systolic Blood Pressure
Authors: Sim John J , Zhou Hui , Wei Rong , Tran-Nguyen Jocelyn , Handler Joel , Brettler Jeffrey W , Reynolds Kristi
Published: July 2021

Objective:
To determine whether lower treated blood pressure (BP) was associated with serious fall injuries and syncope among hypertensive patients.

Methods:
A cross sectional study was conducted within Kaiser Permanente Southern California in patients age ≥18 yrs treated for hypertension and with outpatient BP measurements btwn 7/1/2014-6/30/2015. All BP measurements were used to determine mean and minimum (single lowest BP) systolic BP (SBP) and categorized as <110 and ≥110 mmHg. Serious fall injuries and syncope were determined from emergency dept and inpatient encounters.

Results:
Among 477,516 patients (53% women, mean age 65 yrs), 14,724 (3.1%) had mean SBP<110 and 127,581 (26.7%) had minimum SBP<110. Overall, 15,419 (3.2%) experienced a fall (7,458) and/or syncope (8,763) with higher rates occurring in mean SBP<110 (5.4 vs 3.2%) and minimum SBP<110 (5.7 vs 2.3%) compared to SBP≥110. After multivariate adjustment for potential confounders, compared to patients with SBP ≥110, the odds ratios (95%CI) of falls, syncope, and falls or syncope combined were 1.17 (1.04-1.32), 1.79 (1.63-1.96), and 1.52 (1.40-1.63), respectively, for mean SBP <110; the corresponding odds ratios (95% CI) were 1.78 (1.70-1.86), 2.42 (2.31-2.52), and 2.11 (2.04 - 2.18) for minimum SBP<110 vs ≥110. Use of >2 medications, alpha blockers, older age, females sex, dementia, peripheral vascular disease, and congestive heart failure were also associated with falls and syncope.

Conclusion:
Among treated hypertensive patients, SBP <110 mmHg was associated with serious fall injuries and syncope. Low treatment related BP and variability deserve consideration as we manage our hypertension population.
Abstract P100: Bioavailability of Arginine versus Erbumine Formulations of Perindopril in Healthy Subjects
Authors: Bishop Andrew
Published: July 2021

Perindopril is an angiotensin converting-enzyme inhibitor with a proven track record in cardiovascular clinical trials. Traditionally formulated as the
t
-butyl amine (“erbumine”) salt, perindopril’s arginine salt has been FDA-approved in combination with amlodipine besylate. The arginine salt has the advantage of being more stable at higher temperatures or humidity.

To demonstrate bioequivalence (using the standard FDA definition of 90% confidence intervals that are within 80-125% of the values seen with the reference product), the bioavailability of the two perindopril formulations was compared. After screening 121 healthy subjects, 30 (73% male, 38.9±9.5 years of age, 67% African American, BMI of 26.6±2.7 kg/m
2
) signed informed consent, and were studied in a randomized, single-center, single-dose, open-label, 2-period, 2-way cross-over design with a 14-day washout period. After an overnight fast, perindopril was administered orally with 240 mL of water, and blood samples taken at defined time points.

After dose-normalization (the arginine salt has a molecular weight 1.43-fold greater than that of the erbumine salt), all pharmacokinetic parameters for perindopril plasma concentrations were similar between treatments. Mixed model analysis of these parameters demonstrated bioequivalence (e.g., area under the time-plasma concentration curve: 10.2 vs. 10.5 hr•ng/mL/mg, P = 0.54). Very similar results with non-significant differences were also seen for the pharmacokinetics of perindoprilat (perindopril’s active metabolite): 33.3 vs. 38.3 hr•ng/mL/mg, P = 0.17. Two subjects did not complete the perindopril arginine arm of the study: one withdrew consent, and the other experienced a protocol deviation. No subjects experienced a serious adverse event or withdrew due to a treatment-emergent adverse event. Four subjects reported adverse events after perindopril arginine, and two subjects reported neck pain or presyncope after perindopril erbumine.

After dose-normalization, these two formulations of perindopril meet all traditional criteria for AB rating and interchangeability.
Abstract P103: Anti-hypertensive Mechanisms of Action of G
q/11
Inhibitor Ligands
Authors: Meleka Matthew M , Dahlen Shelby , Edwards Alethia J , Medcalf Matthew , Moeller Kevin D , Osei Owusu Patrick
Published: July 2021

Chronic blockade of individual G protein coupled receptors (GPCRs) has proven to be inadequate strategy for managing hypertension partly because the subcellular heterotrimeric G proteins that propagate intracellular signaling can simultaneously couple to several other vasopressor receptors. Whether blood pressure can better be controlled by directly targeting G proteins has not been thoroughly investigated due to paucity of selective, cell-permeable inhibitors. Here, we tested whether chemical inhibition of Gq/11 proteins
in vivo
and
ex vivo
using recently discovered small-molecule inhibitor ligands, YM-254890 (YM), FR-900359 (FR) and WU-07047 (WU) is sufficient to reverse hypertension in mice. Using
ex vivo
vessel reactivity assay, we found that Gq/11 inhibitors markedly reduced vasoconstriction evoked with phenylephrine (PE), vasopressin, endothelin-1, and the thromboxane analog U-46619. Blockade of PE-induced contractility by the Gq/11 inhibitors showed the following rank-order potency: FR
LogIC50
-0.008 ± 0 > YM
LogIC50
-0.49 ± 0 > WU
LogIC50
-64.95 ± 6.4. YM and WU but not FR inhibited PE-induced vasoconstriction through G protein-dependent and independent pathways by blocking L-type calcium channel-mediated Ca
2+
influx. Acute subcutaneous injection of FR and YM (0.3 mg/kg, s.c.) in normotensive and
N
ω
-Nitro-L-arginine methyl ester (L-NAME) hypertension mice elicited marked hypotension, which was more severe (ΔSBP = -25 ± 2.7 vs. ΔSBP = -21 ± 2.2 mmHg) and long lasting (FR
t1/2
≅ 12 hr vs. YM
t1/2
≅ 4 hr) after the injection of FR relative to YM. In DOCA-salt hypertension mice, chronic injection of FR (0.3 mg/kg, s.c., daily for seven days) reversed hypertension (vehicle SBP: 149 ± 5 vs. FR SBP: 117 ± 7 mmHg) and sustained blood pressure reduction several days after terminating the injection regimen (DOCA SBP: 141 ± 2 vs. SBP 5 days post FR: 128 ± 5 mmHg). Our results together support the hypothesis that increased Gq/11 activity in blood pressure-regulating organs is involved in the pathogenesis of hypertension, and that direct systemic blockade of Gq/11 reverses hypertension. The findings provide clear evidence for targeting Gq/11 in the cardiovascular system as an effective therapy for treating hypertension.
Abstract P114: White Coat Effect is Common Among Stroke Survivors
Authors: Morell Adriana , Okpala Munachi , Savitz Sean , Sharrief Anjail Z
Published: July 2021

Background and Purpose:
Among stroke survivors, uncontrolled hypertension is a major risk factor for recurrent stroke. Blood pressure (BP) medication titration often relies on office BP measures, which may be inaccurate due to the white coat effect (WCE). We sought to determine the prevalence of the WCE in stroke survivors and to determine whether clinical and demographic factors were associated with WCE.

Methods:
We followed ischemic and hemorrhagic stroke and transient ischemic attack patients with prior hypertension presenting to our stroke clinic for a BP study. Sitting BP was obtained by a medical assistant using an office automated BP machine (OABP). Patients also underwent BP measurement using BPtru, an automated machine that measures and averages five BPs with the patient alone in a room. BPtru approximate BPs obtained by the gold standard ambulatory blood pressure machine. Systolic BP (SBP) obtained by BPtru was subtracted from that obtained by OABP. WCE was defined as SBP difference ≥ 10mmHg. Uncontrolled BP was defined as SBP ≥ 135 mmHg by BPtru or ≥ 140 mmHg by OABP. We used student t-tests (continuous) and chi-squared or Fischer’s exact tests (categorical) for univariate analyses.

Results:
Of 94 patients, mean age was 60 (SD 12), 60.6% were male, 26.6% were Non-Hispanic White, 46.8% were Black, and 23.4% were Hispanic. Systolic OABP was 13.2 mmHg (SD 19.3) higher than BPtru SBP (student t-test; p <0.001). WCE was present in 58.5 % of participants and BP was misclassified as uncontrolled in 21.2%. In univariate analyses, age (p = 0.14), sex (p = 0.78), race (0.07), stroke type (0.92), body mass index (p = 0.65), and tobacco use (p = 0.35) were not significantly associated with presence of WCE. The presence of normal SBP by OABP was associated with a decreased likelihood of WCE (p = 0.006).

Conclusions:
Among hypertensive stroke patients following in a clinic, WCE was highly prevalent and one-fifth of patients were misclassified as uncontrolled. Neither race nor other previously described predictors of WCE were associated with WCE in this study. Our findings suggest that in patients with elevated office BP, findings should be confirmed with an automated machine like BPtru in order to avoid over-titration of medication or incorrect assessment of BP control.