bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract MP33: A20 in the Kidney Epithelium Attenuates Angiotensin II-induced Hypertension by Constraining Renal Tubular NHE3 Expression
Authors: Lu Xiaohan , Ren Jiafa , Wen Yi , Griffiths Robert , Yang Ting , Hammer Gianna , Zhuo Jia , Crowley Steven
Published: October 2024

Whereas the ubiquitin editor A20 limits NF-κB-mediated signaling in myeloid leukocytes, the functions of A20 in renal epithelial cells during hypertension have not been described. We recently found that A20 in the kidney tubule ameliorates hypertension and cardiac hypertrophy induced by chronic angiotensin (Ang) II infusion (500ng/kg/min) in mice. Here we report that following 3 weeks of hypertension, the kidneys of mice with tubular deletion of A20 in the kidney epithelium (“A20 iKKO” =

A20
flox/flox

Pax8-rtTA TetO-Cre
+
) have augmented protein expression of the proximal tubular sodium transporter NHE3 (1.5±0.1 vs. 1.0±0.1;
p
<0.01) but similar levels of ENaC subunit expression in the collecting tubule (
p
=NS for all 3 subunits) vs wild-type (WT) controls. Moreover, kidneys from A20 iKKO mice have upregulated mRNA levels for the T cell chemokine CCL5 (2.4±0.3 vs. 1.0±0.3;
p
<0.01) and higher proportions of effector memory CD8
+
T cells (51.2±2.3 vs. 39.3±2.9 % of CD8
+
;
p
=0.013) compared to WTs despite similar numbers of effector memory T cells in the spleen (
p
=NS). To examine whether A20 in the kidney attenuates hypertension by constraining tubular NHE3 expression, we bred A20 iKKO mice with an

NHE3
flox/flox

line, thereby generating mice with double A20 and NHE3 deletion in the kidney tubule (“Dual KKO”) mice. At baseline, kidneys from Dual KKO mice showed robust reductions in mRNA for A20 (0.5±0.1 vs. 1.0±0.1;
p
=0.022) and NHE3 (0.2±0.05 vs. 1.0±0.15;
p
=0.002) compared to WT littermates. During the first ten days of Ang II infusion, mean arterial pressures (MAPs) in the Dual KKO mice were lower than the MAPs previously recorded in our A20 iKKO cohort (129±3 vs. 152±4 mmHg;
p
<0.01). Moreover, during 3 weeks of Ang II infusion, Dual KKO and WT mice maintained similar MAPs (145±7 vs. 149±13 mmHg;
p
=0.78), and heart weight to body weight ratios (5.9±0.3 vs. 5.5±0.2 mg/g;
p
=0.37) in concomitant measurements. Thus, A20 in the kidney tubule blunts the chronic hypertensive response by curtailing NHE3 expression. Our data further suggest that A20 in the nephron can blunt renal inflammation without altering systemic T cell immunity. Stimulating the A20 signaling pathway in the kidney may thus attenuate NHE3-dependent hypertension without conferring attendant risks of global immunosuppression.
Abstract MP35: Angiotensin II-induced vascular response is enhanced by Extracellular Vesicles
Authors: Wei Jin , Wu Jing , Musante Luca , Liu Ruisheng , Liu Jing , Greig Morgan , Harding Michael , Le Thu , Erdbruegger Uta
Published: October 2024

Intro:
The exact effects of extracellular vesicles (EVs) on renal hemodynamics and blood pressure remain to be determined. In this study, we examined the effect of EVs from hypertensive mice and humans on Angiotensin II (ANG II)-induced vascular response in an
ex vivo
model of isolated and perfused renal afferent arterioles (Af-Arts) and also assessed their effect on renal blood flow (RBF) and systolic blood pressure (SBP) in normotensive mice
in vivo.

Method:
Circulating EVs were collected from mice with ANG II-induced hypertension (n=5) and hypertensive humans (n=2). Differential centrifugation (DCF) (5000G x 30min, followed by 17.000G for 30 minutes) was applied to generate an EV pellet (p20) which was subsequently run through size exclusion chromatography (SEC). EV size and concentration was determined by nanoparticle tracking analysis and spectral flow cytometry. Basic EV characterization was performed with cryo electron microscopy and western blotting of EV and non EV proteins.

Following a 30-minute perfusion with EVs (p20 EV pellet (10
10
/ml) or fraction 1-3 or 4-6 of SEC (doses 1-9
7/ml
)), dose-response curves to ANG II (10
-12
to 10
-6
mol/l) were measured in Af-Arts of C57BL/6 mice. Similar doses of EVs were infused into the penile vein of C57BL/6 mice and RBF and SBP were measured. As EVs might deliver AT1R we explored the final common pathway in vasoconstriction in vessels treated with EVs from hypertensive animals vs controls by performing immunoblotting of myosin light chain II S19 phosphorylation.

Results:
EVs deriving from ANG II-treated mice and hypertensive humans, but not those from normotensive control mice and humans, significantly enhanced the vasoconstrictor response to ANG II in Af-Arts of C57BL/6 mice. The same vasoconstrictor response was observed when Af-Arts were perfused with combined EV preps from DCF and SEC (fraction 1-3 vs fraction 1-6) from hypertensive animals.
In vivo
injection of EVs from ANG II-treated hypertensive mice reduced RBF and increased SBP in C57BL/6 mice compared to EVs from normotensive control mice. Immunoblotting of myosin light chain II S19 phosphorylation was inconclusive.

Conclusion:
These data suggest that EVs from hypertensive animals and humans directly affect ANG II response, renal hemodynamic and blood pressure regulation. Further work is needed to identify the EV effector candidate for enhanced ANG II response and possible effect on ART1 activation.
Abstract P101: Surgical and not medical treatment of Primary Aldosteronism increase skin K
+
levels: pathophysiological and clinical implications
Authors: ROSSI FEDERICO BERNARDO , Torresan Francesca , Caputo Ilaria , Zanin Sofia , Caroccia Brasilina , Bertoldi Giovanni , Kohlscheen Eva , Iacobone Maurizio , Seccia Teresa , rossi gianpaolo
Published: October 2024

Introduction:
Primary aldosteronism (PA), the most common curable salt-dependent form of arterial hypertension, features renal K
+
loss and enhanced Na
+
reabsorption.

Hypothesis:
We assessed the hypothesis that alterations of electrolyte, water, and Tonicity-responsive Enhancer Binding Protein/NFAT5 mRNA content occur in the skin of PA patients and are corrected by surgical cure.

Methods:
We obtained skin biopsies in 80 subjects: 49 consecutive consenting PA patients, optimally treated with a mineralocorticoid receptor antagonist (MRA) before adrenalectomy; 6 patients with essential hypertension, and 25 normotensive controls. We measured Na
+
, K
+
and water content with atomic absorption spectrometry after ashing and NFAT5 mRNA with digital droplet PCR. The PA patients were retested after adrenalectomy.

Results:
We discovered that the skin biopsy specimen dry weight (DW) was higher at surgery than follow-up (p<0.001) and correlated directly with electrolyte and water content (all p<0.01), indicating the need for adjusting electrolyte and water data for DW. Surgical cure of PA markedly increased skin DW-adjusted K
+
(from 1.14±0.1 µg/mg to 2.81±0.27 µg/mg, p<0.001) and water content (from 2.92±1.4 mg/mg to 3.85±0.23 mg/mg, p<0.001), but left DW-adjusted skin Na
+
content unaffected. In the PA patients NFAT5 mRNA copy number was higher (p=0.03) than in the normotensive controls and decreased after surgery (p=0.03).

Conclusions:
In conclusion, surgical cure, but not optimal MRA treatment corrected the prominent skin cell K
+
depletion that occurred despite normal serum K
+
levels. The lack of overt skin Na
+
accumulation in PA patients during MRA treatment could be explained by enhanced skin Na
+
lymphatic drainage due to the documented activation of the skin NFAT5/TonEBP pathway.
Abstract P102: Reducing Testing Burden in Patients Screened for Primary Aldosteronism
Authors: Settelmyer Zachary , Flack John , Carney Annah , Prakash Vivek , hill ashley , Bitner Stephanie , Botchway Albert , King Kristin , Buhnerkempe Michael
Published: October 2024

Introduction:
Primary aldosteronism (PA) is an increasingly recognized secondary cause of hypertension characterized by higher cardiovascular, renal, and metabolic disease rates than blood pressure-matched essential hypertensives. Current literature suggests expanding screening for PA to include the majority, if not all, hypertensive patients. This expansive screening would place considerable burden on patients and the healthcare system. A simple clinical risk prediction model that identifies patients with low risk for PA could reduce this testing burden.

Methods:
We evaluated 111 hypertensive patients in the Southern Illinois University Hypertension Clinic who underwent a screening test and, if positive, had subsequent confirmatory testing for PA. We characterized patients based on routine clinical measures including antihypertensive treatment intensity, blood pressure, serum electrolytes, and body size. Classification trees incorporating 96 patients without missing data points for any considered predictor variable were used to identify hypertensive screen-positive patients who have low to absent risk for PA as determined by confirmatory testing.

Results:
Patients with PA had higher serum aldosterone levels at screening (mean = 22.5 ng/dl ± SD = 11.6 vs. 15.2 ± 4.5, p < 0.001), higher BMI (35.8 kg/m
2
± 9.9 vs. 29.9 ± 5.8, p < 0.001), lower potassium (3.9 mmol/L ± 0.5 vs. 4.1 ± 0.6, p = 0.03), and more intense antihypertensive drug therapy (defined by the World Health Organization defined daily dose [6.1 ± 3.2 vs. 4.4 ± 3.0, p = 0.01]) than hypertensive patients without PA. The optimal classification tree model performed relatively poorly at differentiating those with PA from those without PA (sensitivity = 0.76, specificity = 0.86, positive predictive value = 0.87, and negative predictive value = 0.73). However, the model did reveal a subset of individuals who consistently tested negative for PA. Out of 14 patients with serum aldosterone concentration < 25.5, serum potassium > 3.55, and BMI < 27, none had PA.

Conclusion:
We identified a set of readily available clinical variables for identifying hypertensive patients who screened positive for PA but in whom PA was subsequently proven to be absent. Applying these criteria to our screen-positive hypertensive cohort reduces confirmatory testing by approximately 15%. Study in a larger sample size and external validation of this model is needed prior to recommending it for use in clinical decision-making.
Abstract P103: Assessment of Angiotensin II-induced Calcium Signals in Primary Human Adrenocortical Cells
Authors: Ajjour Hala , Lenzini Livia , Pallafacchina Giorgia , Caroccia Brasilina , ROSSI Gian Paolo
Published: October 2024

Background:
Cytosolic and mitochondrial Ca
2+
concentrations are key signals for triggering the secretion of aldosterone in primary aldosteronism (PA), the most common form of secondary hypertension.

Aim:
To investigate [Ca
2+
]
i
transients and oscillations in human CD56
+
cells isolated from aldosterone producing adenoma (APA) and normal surrounding adrenocortical (AAC) cells.

Methods:
We assessed [Ca
2+
]
i
in CD56
+
APA and APA-adjacent cells (AAC) in basal conditions and after angiotensin II (Ang II) stimulation using Fura-2 dye, live confocal microscopy, and an ad hoc developed custom-made pipeline to analyze [Ca
2+
]
i
signals in terms of frequency and peak parameters as amplitude, full width at half maximum (FWHM) and area under the curve (AUC). As the two-pore domain K
+
TASK- 2 channels are under expressed in APA, we used quinidine (20 μM), 20’ prior to [Ca
2+
]
i
imaging acquisition to examine the effect of the pharmacological inhibition of this channel.

Results:
Resting [Ca
2+
]
i
basal levels were higher in AAC (n=11) (0.75, IQR;0.90-0.56) than in APA (n=11) (0.36 IQR;0.69-0.33), (p < 0.02). We detected spontaneous Ca
2+
activity, during resting conditions, in both cell types without significant difference in peak parameters. Upon 1 nM Ang II simulation, AAC showed elevated FWHM, amplitude, and AUC peak parameters of first Ca
2+
transients. However, APA cells exhibited elevated peak parameters of subsequent Ca
2+
oscillations. Ca
2
oscillations frequency and degree of oscillating cells were significantly higher in APA cells than in AAC. After 24 hours in culture, both AAC and APA cells developed rosette-like cell structure resembling the “glomeruli” of the intact zona glomerulosa clusters. Functional Statistics and clustering analysis showed synchronous behavior of cells belonging to the same cluster without significant differences between AAC and APA.

In AAC, Task-2 Inhibition with quinidine tended to increase spontaneous activity and altered peak’s parameters but did not influence frequency of oscillation.

Conclusion:
Quantitative assessment of [Ca
2+
]
i
signals unveiled significant differences in [Ca
2+
]
i
dynamics between human AAC and APA cells. These results suggest the concurrent modulation of frequency and amplitude of [Ca
2+
]
I
subsequent oscillations as a potential driver of increased rate of aldosterone production.
Abstract P106: Association of long term blood pressure variability and Major Cardiovascular events in patients with primary aldosteronism
Authors: Tulake Simili , Li Nanfang , Luo Qin
Published: October 2024

Objective:
Long-term blood pressure variability (BPV) is associated with an increased risk of major cardiovascular events (MACE), but it is rarely reported in patients with primary aldosteronism (PA). We aim to investigate the association between long-term BPV and cardiovascular events in patients with PA.

Methods:
This was a retrospective cohort study which conducted at the Hypertension Center of People’s Hospital of Xinjiang Uygur Autonomous Region from January 2006 to December 2021. The study included patients with PA aged over 30 years. The primary outcome was the incidence of major cardiovascular events during the follow-up, which extended until June 2023. The BPV index encompassed four parameters: standard deviation (SD), coefficient of variation (CV), average real variability (ARV), and maximum and minimum blood pressure difference (MMD). Multivariate analysis using logistic regression was performed, adjusting for demographic variables such as age, sex, body mass index, as well as clinical characteristics including smoking and alcohol consumption.

Results:
A total of 2,380 patients with PA were ultimately enrolled in this study. Access to clinic blood pressure was ensured at least three times during follow-up visits, with a minimum follow-up interval of 1 month. During a median follow-up of3.6 years, there were 124 cases of MACE. The mean age of the participants was 49.36 years, with men accounting for 1,288 cases. The average blood pressure recorded was 135 mmHg. After adjusting for confounding factors such as age, sex, smoking, alcohol consumption, BMI and cardiovascular risk factors, the systolic BPV (SD, CV, ARV) was found to be significantly associated with MACE (OR= 1.03, CI 1.02-1.05; P<0.05). In the group of mean SBP≥130mmHg, SBP variability had a significant association with MACE (OR: 1.03; CI: 1.01-1.06; P<0.05). However, no significant correlation between SBP variability and MACE in group of mean SBP <130mmHg (P>0 .05).

Conclusion:
Focusing on long-term systolic BP SD, CV, and ARV in patients with primary aldosteronism may help to prevent the occurrence of cardiovascular events.
Abstract P111: Last Recorded Blood Pressure Measurement as a Proxy for Average Blood Pressure Control
Authors: Singh Nausheen , Feinberg Abigail , Barai Nikita , Cluett Jennifer , Fishman Mary , Bryant Kelsey
Published: October 2024

Introduction:
Hypertension is a leading cardiovascular risk factor affecting 46.7% of adults in the United States, yet the definitions of blood pressure (BP) control by society guidelines and quality metrics are not aligned. In 2017, the AHA/ACC guidelines redefined hypertension as BP >130/80 while the National Committee for Quality Assurance (NCQA) continues to define control as BP of <140/90. Further, the AHA/ACC guidelines emphasize the importance of using an average of readings, whereas the NCQA determination of control is based entirely on the last recorded BP measured closest to the end of the calendar year. This analysis investigates the odds of BP control based on average office BPs recorded between January and December 2023 compared with BP control based on the last recorded single BP measurement of 2023.

Hypothesis:
At the population level, odds of BP control based on last recorded BP is a proxy for odds of BP control based on average BPs over the past calendar year.

Methods:
We examined BP control for patients aged 18-85 with hypertension who had ≥3 outpatient measurements between 1/1/2023-12/31/2023 in a New York City health system. The odds of BP control based on average BP compared with last BP at <140/90 and <130/80 thresholds were assessed using logistic regression.

Results:
Office BP measurements were analyzed for 89,130 patients. Of these, 71.2% had controlled hypertension based on NCQA criteria of last measured BP <140/90 versus 68.9% using the average BP <140/90. Using the lower <130/80 threshold, 34.5% patients had controlled hypertension by the last measured BP versus 32.7% using the average BP. When last systolic BP >140 or diastolic BP >90 there was a greater likelihood that the patient had uncontrolled hypertension based on average BP (OR 2.00, 95% CI [1.88-2.12] and 7.43, 95% CI [6.96-7.93] respectively). A similar pattern was observed when the last recorded systolic BP was >130 or diastolic BP >80, (OR: 2.02, 95% CI [1.95-2.08] and 3.99, 95% CI [3.87-4.12] respectively).

Conclusions:
Although a single BP measurement may not be predictive of individual cardiovascular risk, the use of last measured BP to assess control at a population level is a reasonable approach to compare health systems and identify disparities.
Abstract P117: Provider Concordance to the Current 2017 High Blood Pressure (BP) Clinical Practice Guidelines and Patient Blood Pressure Control
Authors: Tedla Yacob , Greenland Philip , Kho Abel , Rothman Russell
Published: October 2024

Introduction:
The AHA/ACC 2017 High BP guidelines introduced key changes, compared to previous guidelines, including a new hypertension definition (≥130/80 mm Hg) and lower medication thresholds for patients at cardiovascular risk. The impact of provider adherence to these guidelines on patient BP control has not been fully explored.

Methods:
We identified 3,749 new stage 1 hypertension patients (7,214 visits) and 11,013 new stage 2 hypertension patients (33,321 visits), aged ≥18, from the Vanderbilt University Medical Center database. We scrutinized outpatient BP levels, prescribed medications, follow-up visits, lab results, demographics, comorbid conditions, and physician notes from January 2018 to December 2022. This data was compared to hypertension treatment criteria developed from the 2017 guidelines that was validated by six hypertension experts. Active primary care patients (with 1 visit per year) were included. Mixed-effects Poisson regression and mixed-effects linear regression examined the relationship between guideline adherence and controlled BP (<130/80 mm Hg) and continuous BP, respectively.

Results:
Among stage 1 hypertension visits, 12.2% were treated with at least one antihypertensive drug according to guidelines, resulting in a 13% increase in BP control and a reduction in systolic BP by 2.02 mm Hg. For stage 2 hypertension visits, 26% received treatment with at least two first-line antihypertensive drug classes, leading to a 45% increase in BP control and reductions in systolic and diastolic BP by 3.79 mm Hg and 1.93 mm Hg, respectively. No significant difference in BP control was observed among stage 2 patients treated with two versus one first-line drug. Adherence to follow-up guidelines resulted in a 24% improvement in BP control for stage 1 patients and stage 2 patients when their follow-up visit occurred within 1 month. Subsequent follow-up visits within 1-month improved BP control by 34% if BP was not controlled previously. Additionally, 42.4% of patients with uncontrolled BP for 1 month received an increase in medication dose or a change/addition of medication class and showed a 7% increase in BP control within next month.

Conclusion:
Provider adherence to recommended antihypertensive drug treatment guidelines (12%-26%), follow-up assessments guidelines (29%-37%), and treatment intensification (42.4%) were poor. However, patients treated according to the guidelines showed significant improved BP control.
Abstract P119: The Heart Failure with Preserved Ejection Fraction Induced By High-Fat Diet and L-NAME in Mice Does not Induce a Renal Damage
Authors: Araos Patricio , Figueroa Stefanny , Arenas Gabriela , Reyes Osorio Javier , Saez Javiera , MOYA JACKELINE , sepulveda Ruth , Rojas Joaquin , Fuenzalida Maria Jose , Gabrielli Luigi , Jalil Jorge , ocaranza Maria Paz , Amador Cristian
Published: October 2024

Heart failure with preserved ejection fraction (HFpEF) is a clinical syndrome, without effective therapy, characterized by diastolic dysfunction, a chronic low-grade inflammation and comorbidities such as hypertension. At the present time, few experimental models of HFpEF represent the hallmarks of patients. Recently, it has been described that the administration of a high-fat diet (HFD) in conjunction with an endothelial nitric oxide synthase inhibitor (L-NAME) can adequately reproduce the pathophysiological features of HFpEF. However, the effects on at renal level in HFpEF models have not been fully described. Our goal was to evaluate the kidney damage in a HFpEF model induced by HFD+L-NAME.

Male C57BL/6N mice (12 weeks old, n=4-6) were treated with control diet or HFD (60% in fat calories) plus L-NAME (0.65 g/L in drinking water) for 15 weeks. We evaluated cardiac function (echocardiographic analysis), systolic blood pressure (SBP, plethysmography), cardiac and renal hypertrophy (morphometry and histology), plasma creatinine, blood urea nitrogen (BUN), plasma neutrophil gelatinase-associated lipocalin (pNGAL), proteinuria, and kidney immune cells infiltration (flow cytometry). All data are presented by mean±standard deviation.

After 15 weeks of HFD+L-NAME we observed diastolic dysfunction (p<0.001), an increased SBP (control=128.0±15.3 vs. HFD+L-NAME=155.7±9.8 mmHg; p<0.01), without changes in ejection fraction and shortening fraction. In addition, we did not observe cardiac or renal hypertrophy, nor apparent histological changes in the renal structure between both groups of animals. No differences were found between groups regarding plasma creatinine, pNGAL or BUN, however, we observed that HFD+L-NAME induced proteinuria in compared to control mice (p<0.05). Finally, when we evaluated the immune cells population in kidney of both groups, we observed no differences in dendritic cells, macrophages, or neutrophils. Nevertheless, the HFD+L-NAME treatment was associated to a significant rising of Ly6G
+
CD11b
-
cells population (CD45
+
MHC-II
-
CD11c
+/-
F4/80
-
) compared to control mice (p<0.05).

In conclusion, our results indicate that there is no renal inflammatory damage after 15 weeks of in HFpEF experimental model. However, to our knowledge, an uncharacterized population of Ly6G cells could be participating at this stage which could trigger a long-term renal damage
.

FONDECYT 1221585 (MPO), 1231604 (JJ), 1231909 (CAA), 11241074 (PA) and FONDAP 15130011(MPO, LG).
Abstract P121: Weight Loss is not a Major Mediator of GLP1-Receptor Agonists’ Risk Reduction in Heart Failure with Preserved Ejection Fraction
Authors: Hartsell Sydney , Wei Guo , Kasera Shalini , Singh Ravinder , Sarwal Amara , Nevers McKenna , Boucher Robert , Beddhu Srinivasan
Published: October 2024

The cardioprotective effects of GLP1-Receptor Agonists (GLP1-RA) and SGLT2 Inhibitors (SGLT2i) are incompletely understood. We compared these two drug classes and their effect on HFpEF hospitalizations and examined the hypothesis that weight loss is a significant mediator of these agents’ HFpEF protection.

Methods:
Using the VA Informatics and Computing Infrastructure, we generated a national cohort of veterans with type 2 diabetes on metformin who initiated SGLT2i, GLP1-RA or Insulin Glargine for the first time from 1/1/18 to 12/31/21. The index date was the date of first outpatient prescription fill. We defined 6-month weight change as change from baseline weight (closest outpatient value prior to index date within 1 year) to 6 months (all averaged outpatient weights from index date to 6 months). Those with prior prescriptions for the study drugs from 1/1/08 and missing baseline or 6-month weights were excluded. HFpEF hospitalizations were defined by a previously-validated natural language processing algorithm using inpatient HF diagnostic codes and cardiac imaging dated closest to and within 180 days of the new code with ejection fraction ≥50%. Follow up was through 3/31/23. We performed mediation analyses to separate the direct effects (independent of 6-month weight change) and indirect effects (mediated through 6-month weight change) of the drug classes on time to HFpEF hospitalizations, adjusted for demographics, comorbidities, baseline labs and other medications, and diabetes severity.

Results:
Overall, 161,405 veterans initiated one of the study drugs and 105,439 had weight data. After excluding 760 veterans with HFpEF hospitalizations in the first 6 months, 104,679 were included in the analysis. Baseline demographics included mean age 64±11 years, 94% male and 75% white. Baseline BMI was 33±6 kg/m
2
. There were 1494 HFpEF hospitalizations/ 256,101 years of follow-up. Figure 1 shows weight over time by study drug class. Table 1 shows mediation analyses results.

Conclusion:
GLP1-RA lowered the risk of HFpEF hospitalizations versus the active comparator insulin glargine. This cardioprotection was not mediated by weight loss. GLP1-RA and SGLT2i show equivalent risk reduction. These agents' direct cardiac and vascular effects deserve further exploration.
Abstract P123: Gender Difference in the In-hospital Mortality for Hypertensive Heart Disease – United States Population-Based Survey Analysis
Authors: Yousuf Muhammad , Almani Muhammad Usman
Published: October 2024

Background:
Females have steeper increases in blood pressure than males throughout life, thus setting the stage for gender based phenotypic and clinical differences in hypertension related pathologies. We set out to determine the in-hospital mortality for the patients primarily admitted for Hypertensive Heart Disease (HHD) in the United States.

Methods:
Data was extracted from the National Inpatient Sample (NIS) 2016 - 2020 Database. We used ICD-10 code I11.9 to identify the patients primarily admitted for HHD. Multivariable Logistic regression was used to estimate Odds Ratio (OR) for in-hospital mortality between the two genders. Multivariable Poisson regression model was performed for yearly trends with year as the independent variable and in-hospital mortality as the dependent variable. In-hospital mortality rate trends between the two genders were compared using a group-year interaction term in a multivariable regression analysis with in-hospital mortality event as the dependent variable. All the outcomes were adjusted for age, insurance status, race, Charlson comorbidity index (CCI) and the hospital characteristics (location, bed-size). The analysis was performed on STATA 16 software.

Results:
We identified 1,877,150 patients with HHD. Mean age of the patient cohort was 70 years, mean CCI was 2.7 and 51% were females. Unadjusted in-hospital mortality rate for males was 1.6% compared to 1.8% for females. When adjusted for cofounders, females had lower risk of in-hospital mortality compared to males (OR: 0.95, 95% CI 0.90 – 0.99). In-hospital mortality rate for either gender did not appreciably change from 2016 – 2020, however, when compared to males the mortality rate for females rose more sharply (Interaction p = 0.038) as shown in
Figure 1.

Conclusion:
In our adjusted population-based survey analysis, short term mortality risk for HHD was lower for females than males, however, during the study period 2016 – 2020 mortality rate for females rose more sharply than for male subjects.
Abstract P125: In an Autocrine Mechanism, Endothelial Progranulin Regulates Vascular Tone and Blood Pressure via AMPK Activation
Authors: Bruder do Nascimento Ariane , Costa Rafael , Bruder Thiago
Published: October 2024

Dysfunctional endothelial cells (ECs) contribute to the development of hypertension (HTN). Progranulin (PGRN) is a secreted protein that plays a major role in the inflammatory response. We recently demonstrated that HTN is associated with increased circulating PGRN, whereas the global lack of PGRN elicits vascular dysfunction and raises blood pressure (BP). However, the cellular origin of PGRN and the molecular mechanisms by which PGRN regulates vascular tone and BP are still to be elucidated. We hypothesized that ECs secrete PGRN, which regulates the vascular phenotype and BP in an autocrine manner. We used mice with tamoxifen-inducible, Cdh5-CreERT2-mediated deletion of PGRN in ECs (PGRN EC-/-) and their counterparts as controls (PGRN EC+/+). The effectiveness of endothelial PGRN depletion was validated using
en face
immunostaining in the aorta and qPCR in ECs from mesenteric arteries (MA). PGRN EC-/- mice exhibited a 2.5-fold reduction in circulating PGRN levels followed by EC dysfunction in MA characterized by reduced maximal response (Emax) and sensitivity (pD2) to acetylcholine [Emax (%): PGRN EC+/+: 99.3 ± 0.7 vs PGRN EC-/-: 95.9 ± 0.5* and pD2: PGRN EC+/+: 8.1 ± 0.2 vs PGRN EC-/-: 7.0 ± 0.3*, *P<0.05]. Supplementation with recombinant PGRN (20 μg/day) normalized endothelial function. Moreover, under baseline conditions, BP (analyzed by telemetry) was slightly higher in PGRN EC-/- mice [MAP (mmHg): PGRN+/+: 110.6 ± 1.6 vs PGRN-/-: 114.3 ± 1.2]. However, treatment with a subpressor dose of angiotensin II (120 ng/kg/day for 2 weeks) only increased BP in PGRN EC-/- mice. Mechanistically, we found that knockdown of PGRN (via siRNA) in cultured mesenteric ECs (MECs) suppressed PGRN secretion, AMPK phosphorylation, and nitric oxide (NO) formation. Conversely, MECs overexpressing PGRN (via lentivirus infection) led to exacerbated PGRN secretion followed by AMPK activation and NO formation, which were blunted by blocking AMPK or EphrinA2 (PGRN receptor) with dorsomorphin (10 μM) or ALWII4127 (5 μM), respectively. Finally, endothelial-specific AMPK-deficient mice exhibited increased BP and endothelial dysfunction in MA; such effects were not affected by PGRN treatment, suggesting that AMPK is a downstream target for PGRN. In summary, our data indicate that ECs are important sources of PGRN, which supports the maintenance of the EC phenotype and BP via EphrinA2, AMPK, and NO formation. This pathway occurs in an autocrine-dependent manner.
Abstract P131: Effect of uncontrolled systolic blood pressure in hypertensive patients as a risk factor for CIN after coronary angiography with preventative management of CIN : single center prospective study
Authors: ki YoungJae
Published: October 2024

Background:
Contrast induced nephropathy(CIN) is associated with poor outcomes like dialysis and prolonged hospitalization and risk of death. Clinical factor for CIN include advanced age, diabetes (DM), female, heart failure(HF) and hypertension(HTN). Many prophylaxis strategies for CIN(normal saline hydration, N-acetylcysteine, NAHCO3) had been studied and their preventive effects had been known to be no difference.

Purpose:
We tried to evaluate the effects of uncontrolled systolic blood pressure(defined as SBP>160mmHg) in hypertensive patients as a risk factor for CIN after coronary angiography(CAG) with various preventative management of CIN (normal saline hydration, N-acetylcysteine, NAHCO3).

Methods:
This study included 41patients who performed CAG combined with various preventative management of CIN in single center. All patients were evaluated with serum creatinine(SCr), creatinine clearance(CrCl) before and after coronary angiography at day0, day1, day2 and day30. Definition of CIN include; SCr increase of ≥0.5 mg/dL, a SCr increase ≥25%, an CrCl decrease of ≥25%, or the composite, occurring 48 to 72 hours after contrast exposure. We also analyzed preexisting renal insufficiency(GFR<60ml/min), DM and HTN, age, sex, anemia, contrast volume and HF which are known risk factors for CIN.

Results:
11 of 41 patients developed CIN. There was no statistical differences between the two groups in the aspect of baseline Cr, age, sex, body weight, contrast volume and the prevalence of known risk factor for CIN (DM, HTN, HF and preexisting renal failure). Mean baseline SBP in the CIN group was higher than non-CIN group(166.3±31.9 vs142.4±25.7, P-value=0.018). Among 23 patients with HTN, 11 patients (47.8%) had uncontrolled SBP. The multivariable logistic regression analysis showed that an uncontrolled SBP (odds ratio [OR] 12.63, 95% confidence interval [CI] 1.83-87.07) was the independent risk factor for CIN.

Conclusions:
CIN associate with variety of short-term and long-term complication after CAG. Hypertension is well known as one of the risk factors for CIN. Even if appropriate preventative management of CIN are taken before and after CAG, uncontrolled SBP in hypertensive patients could increase the risk of CIN.
Abstract P132: Sex Differences in Cerebrovascular Dysfunction in the APP/PS1 TgF344-AD Rat Model of Alzheimer's Disease
Authors: Fang Xing , Zhang Huawei , Ryu Jane , Fan Fan , Roman Richard
Published: October 2024

Alzheimer's Disease (AD) is the primary cause of dementia and is characterized by extracellular beta-amyloid plaques and intracellular hyperphosphorylated tau-containing neurofibrillary tangles in the brain. Emerging evidence suggests that cerebrovascular dysfunction is associated with AD. Although two-thirds of AD patients are female, sex differences in transgenic AD models have been largely ignored. The present study investigated sex differences in cerebrovascular dysfunction and potential mechanisms in AD using a TgF344-AD rat model. The APP/PS1 TgF344-AD (AD) rats serve as a preferred model to study AD since they recapitulate the complete repertoire of AD-like pathologies in humans. We previously longitudinally characterized cerebral hemodynamics in male TgF344 AD rats at 3, 4, 6, and 14 months, and reported that 4-month-old AD males displayed early signs of impaired cerebral blood flow (CBF) autoregulation and reduced functional hyperemia, two months earlier than memory loss, which was co-occurred with the appearance of plaques. These deficits progressed with advancing age. The present study used 13-14 months of female TgF344-AD (AD) and F344 (WT) rats. We found that AD females also displayed impaired learning and memory function in the eight-arm swim and open field tests as seen in males. Amyloid plaques were prominent in the hippocampus and cortex of female AD brains at a similar level in males, while no plaques were observed in WT brains. Functional hyperemic responses to whisker stimulation were markedly impaired in female AD rats. Further, we found evidence of astrocyte activation and microgliosis in female AD brains using immunohistochemistry and western blots. The astrocytes and microglial activation formed clusters around amyloid plaques. The direct contact between astrocytic endfeet and penetrating arterioles was markedly reduced in AD animals. Moreover, the blood-brain barrier (BBB) was disrupted in AD animals as the tight junction marker ZO1 was significantly reduced in AD brains. These results demonstrate cerebrovascular dysfunction contributes to loss of cognitive function in both male and female APP/PS1 TgF344-AD rats and is associated with impaired functional hyperemia, loss of astrocytic endfeet on penetrating arterioles, BBB breakdown, neuroinflammation related to amyloid plaques. They also indicate that the female TgF344-AD rat is appropriate for studying age-related influences of sex hormones on AD.
Abstract P136: Phosphorylation and regulation of URAT1 by insulin and NaCl: a possible mechanism linking insulin resistance and excessive salt intake to hyperuricemia
Authors: Fujii Wataru , Yamazaki Osamu , Kochi Yuta , Shibata Shigeru
Published: October 2024

Objective:
Insulin resistance (IR) is the key environmental factor associated with hyperuricemia; however, the underlying mechanisms are not fully understood. Although several studies suggested that excessive salt intake results in hyperuricemia, the association still remains controversial. Focusing on urate transporter 1 (URAT1), we here investigated the mechanisms by which insulin and salt intake contribute to hyperuricemia.

Methods:
In a large UK Biobank (UKB) dataset involving 377,358 participants, we evaluated the association of TyG index (a surrogate of IR) and habitual salt intake with serum uric acid levels using multivariable regression models. We addressed whether single nucleotide polymorphisms (SNPs) that influence URAT1 expression (eQTL) modify the association of TyG index with serum uric acid levels. We also analyzed underlying mechanisms by using HEK cells expressing hURAT1.

Results:
In UKB dataset, TyG index and habitual salt intake were significantly and independently associated with elevated serum uric acid levels (P < 0.05). From previous studies and imputation accuracy, we extracted an eQTL SNP for
SLC22A12
gene and found that there is a positive interaction between TyG Index and the eQTL SNP. As post-translational modification is the key mechanism that enables adaptation to environmental changes, we evaluated the contribution of phosphorylation by AGC kinases, such as Akt, as possible downstream effectors of insulin signaling and salt-sensing pathway. Western blotting with phospho-Akt substrate motif antibody confirmed URAT1 phosphorylation and identified T350 and T408 as candidate sites. Among them, URAT1 carrying non-phosphorylatable A408 substitution (T408A-URAT1) suppressed glycosylation and plasma membrane translocation, while T350A had no effect. By using kinase screen assay involving 53 AGC and related Ser/Thr kinases, we found that SGK1, PKA, and PKG also phosphorylated URAT1-T408. Insulin and NaCl loading induced Akt and SGK1 expression respectively, which resulted in URAT1 phosphorylation and increased cell-surface membrane expression; this effect was abolished in T408A-URAT1. Finally, we found that URAT1-T408 phosphorylation was also detected in human kidney sections.

Conclusion:
IR and excessive salt intake contribute to hyperuricemia, and the effect is in part mediated by hURAT1. Mechanistically, insulin and NaCl promote membrane trafficking of URAT1 by inducing T408 phosphorylation through Akt and SGK1, respectively.
Abstract P138: Diet Influences the Expression of Renal Sodium-Glucose Cotransporters in Juvenile Male Rats
Authors: Knick Benjamin , Bhullar Kanwar , Deuchler Catherine , Fischer Stephen , Lee Gina , Gigliotti Joseph
Published: October 2024

Poor nutrition and metabolic dysfunction are becoming more prevalent among children, as evident by increasing childhood obesity in recent years. The kidney sodium-glucose cotransporter 2 (SGLT2) is a contributor to overall metabolic homeostasis, with inhibition of SGLT2 significantly reducing circulating glucose and lipids. The study objective was to determine if diet influences the expression of renal glucose transporters in young rats. Weanling male Wistar Kyoto rats received 1) standard laboratory chow, 2) AIN-93G diet, or 3) a high-fat diet (60% kcal, HFD). Each dietary group was then further subdivided to receive either filtered water or 5% (w/v) sucrose solution
ad libitum
. Four weeks later, animals were euthanized and kidneys were collected and processed. mRNA expression of
Sglt1
,
Sglt2
,
Sqstm1
, and
Nlrp3
was determined using gene specific primers and RT-PCR. Renal SGLT2 protein expression was visualized using immunohistochemistry and quantified using Imagej software. Diet significantly influenced the renal expression of both
Sglt1
(
P
=0.005) and
Sglt2
(P=0.046), while beverage did not have a significant effect on either (
P
=0.7 and
P
=0.4, respectively). Rats fed the HFD had more than 30% reduction in
Sglt1
mRNA expression as compared to rats fed chow, and ~20% reduction in
Sglt2
as compared to rats fed AIN. Preliminary data suggest that diet and beverage significantly altered renal protein expression of SGLT2, with rats fed chow having the lowest renal expression that was slightly increased with sugar consumption (1.5% versus 2.7%, respectively). Rats fed AIN had the greatest expression, but was drastically reduced in rats consuming sugar (4% versus 1.2%, respectively). To determine if autophagy and inflammasome activation is involved, the expression of
Sqstm1
and
Nlrp3
was measured by RT-PCR. Similarly, rats fed HFD had the lowest expression of both
Sqstm1
(
P<0.001) and
Nlrp3 (
P=0.03).
Sqstm1
and
Nlrp3
mRNA expression was decreased with sugar consumption in rats fed chow and AIN, whereas
Sqstm1
(
P
=0.01) and
Nlrp3
(
P
<0.001) mRNA expression was increased in rats fed HFD with sugar beverage. Our data suggest that diet significantly influences renal glucose transporter expression in juvenile rats in a way that may be mediated by autophagic impairment. The modulatory effect of sugar consumption on SGLT2 protein,
Sqstm1,
and
Nlrp3
was diet-dependent, highlighting the importance of nutrition to renal physiology within weeks after weaning.
Abstract P139: Exploring the cardiometabolic effects of lipid 12-hydroxyeicosapentaenoic acid through thromboxane receptor partial agonism
Authors: Tomazini Goncalves Tiago , Alnouri Wessan , Offermanns Stefan , Osorio Silveira Leiria Luiz
Published: October 2024

Introduction:
It is well known that lipid mediators secreted by brown adipose tissue (BAT) during cold exposure or beta-adrenergic stimulation can exert beneficial effects on glucose and lipoprotein homeostasis. Recently, Leiria et al. demonstrated that one of these mediators, 12-hydroxyeicosapentaenoic acid (12-HEPE), secreted by BAT, enhances glucose tolerance and insulin sensitivity through the stimulation of a G protein-coupled receptor (GPCR). However, the receptor that 12(S)-HEPE binds to exert its functions was unknown. Therefore, this study aimed to identify the GPCR that 12-HEPE binds to and to investigate whether 12-HEPE affects vascular reactivity.

Methods:
Screening of 70 non-olfactory GPCR candidates was conducted in HEK293 cells, either stimulated or not with 12-HEPE (5µM). Receptor activation was measured using the dynamic mass redistribution (DRM) assay. After identifying the potential receptor, we validated the result by monitoring intracellular Ca2+ levels through a dose-response curve for 12-HEPE. To study the biological effects resulting from this ligand-receptor interaction, thoracic aortic artery rings were isolated from 12-week-old C57BL6 male mice, and their contractile responses were monitored using a myograph in response to cumulative doses ranging from 100pM to 10µM of 12-HEPE.

Results:
Through DRM screening, we found that the thromboxane receptor (TP) exhibited the strongest interaction with 12(S)-HEPE (5µM). To validate the screening, we incubated HEK293 cells with 12-HEPE or U46619 and quantified intracellular calcium mobilization. Our data demonstrate that 12-HEPE is a partial TP agonist, with an EC50 of 314nM compared to 7.2nM for U46619. Furthermore, incubation of the thoracic aorta with 12-HEPE produced dose-dependent increases in arterial constriction (pEC50: 6.129 ± 0.174; Emax: 95.39 ± 15.72), and this effect was completely prevented by the thromboxane antagonist, TPA (1µM).

Conclusions:
Our results suggest that 12-HEPE is an endogenous partial agonist of the TP receptor, and such interaction is necessary for the biological effects of these lipids. Importantly, the vasoconstrictor effect mediated by 12-HEPE may be an important physiological adaptation to prevent body heat loss upon cold exposure, thus allowing for adequate temperature maintenance.
Abstract P146: Sex-Dependent Regulation of the Endothelin System in Multi-End Organ Inflammation in Lupus-Associated Cardiovascular Disease
Authors: McCrorey Marice , Hawkins Kennedy , Oates Jim , Abdul Yasir , Ergul Adviye , Cunningham Melissa , Van Beusecum Justin
Published: October 2024

Lupus is a chronic autoimmune disease characterized by IgG deposition, Toll-like receptor 7 (TLR7) overactivation, and autoantibody production resulting in end organ inflammation. SLE disproportionally affects women, SLE disproportionally affects women to men (9:1 ratio), with a high prevalence in African American women. Highly vascularized organs are severely impacted by lupus, specifically the kidneys and brain. Recent literature has shown that cardiovascular disease (CVD) has significant detrimental impacts on organ crosstalk. Endothelin-1 (ET-1), a potent vasoconstricting peptide, has been shown elevated in patients with CVD and in patients with lupus independent of disease severity. However, the extent to which ET-1 promotes lupus-associated CVD and end organ cross talk remains to be elucidated. Therefore, we hypothesized that TLR7 overactivation in a murine model of SLE-associated CVD would lead to a sex-dependent regulation of the renal and cerebral ET systems. Female and male lupus-prone B6.Nba2 mice were subjected to bi-weekly treatment with the Resiquimod (R848; TLR7/8 agonist; 100ug/30ul) or vehicle control (Acetone) for one month and subsequently studied for 12 weeks. At 16 weeks, the kidneys and brains were collected for histology and ET system (ET-1, ET
A,
and ET
B
) analysis. Exploration of specific end organ damage demonstrates a profound increase in renal and cerebral IgG complex deposition as well as renal macrophages and T cells in both females and males receiving R848. Renal ET-1 levels displayed no time nor treatment-dependent effects in females, while males exhibited a significant time-dependent increase in expression at the 4 and 16-week timepoints. Interestingly, at the receptor level, neither sex demonstrated differences in ET
A
nor ET
B
levels in the kidney. While renal ET-1 level did not change, females demonstrated a significant increase in cerebral ET-1 at the 16-week timepoint, while males trended for increased cerebral ET-1. At the receptor level, females had a trend for increased cerebral ET
A
expression, while there was no change in males, and neither sex displayed a change in ET
B
receptor expression. These findings suggest a potential shift towards a pro-inflammatory state via the cerebral ET-1/ET
A
signaling axis in the R848-treated females. These findings warrant further exploration into the end-organ specific pathophysiological role ET-1/ET
A
signaling in multiorgan crosstalk in lupus-associated CVD.