bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 024: Early Gestation Estrogen Supplementation In The Preeclamptic-like BPH/5 Mouse Model: Effects On Uteroplacental Expression Of Sex Steroid Hormone Receptors
Authors: Gomes Viviane C , Woods Ashley K , Gilbert Bryce M , Crissman Kassandra R , Landry Camille A , Sones Jennifer L
Published: October 2023

Preeclampsia (PE), a hypertensive disorder of pregnancy, is a leading cause of maternal/fetal morbidity and mortality worldwide. Defective placentation is a major event in PE pathogenesis, albeit underlying molecular mechanisms remain poorly understood. Estradiol (E2) deficiency has been reported in multiple PE cohorts and correlated with impaired placentation and disease severity. Early gestation uteroplacental dysregulation of sex steroid hormone receptors (SSHr) may further impact trophoblast cell differentiation, angiogenesis, and placental perfusion. The BPH/5 mouse spontaneously develop impaired trophoblast cell invasion, poor placentation, and PE-like clinical signs. Adult BPH/5 females display hypoestrogenism and hyperandrogenism pre-pregnancy, premature and depressed E2 surge during early gestation, and delayed embryonic development and decidualization. We aimed to elucidate expression of estrogen (
Esr1
and
Gper30
) and androgen receptors (
Ar
) in BPH/5 embryonic implantation sites (eIS) during nidation (e4.5) and decidualization (e7.5). We hypothesized that SSHr would be dysregulated in natural (NAT) BPH/5 eIS vs. normotensive C57 controls. Furthermore, early gestation E2 supplementation (E2-S), a protocol recently shown to improve BPH/5 placental expansion and umbilical blood flow, was expected to normalize eIS SSHr. In NAT BPH/5 and C57 pregnancies, eIS were harvested at e4.5 and e7.5 (n=5-8/group). Early gestation E2-S consisted of ovariectomy in BPH/5 an C57 females at e2.5, subcutaneous administration of 25 ng E2, and daily subcutaneous injections of 1 ng of progesterone until eIS collection 2 days post-E2 treatment (n=8-9/group). Gene expression was assessed via RT-PCR, and Student’s T-tests were used for pairwise comparisons.
Esr1, Gper30,
and
Ar
were 2- to 4-fold higher in NAT BPH/5 vs. NAT C57 eIS at e4.5 and e7.5 (p<0.01). After E2-S,
Esr1
was downregulated in BPH/5 eIS (p<0.01), while
Gper30
and
Ar
remained elevated in E2-S BPH/5 vs. E2-S C57 (p<0.05). In conclusion, eIS
Esr1, Gper30,
and
Ar
upregulation accompanies the abnormal E2 profile in BPH/5 early gestation. However, only
Esr1
upregulation was altered by early gestation E2-S. Further investigation of
Esr1
role in PE defective placentation is warranted.
Abstract 027: Chronic Unpredictable Stress Affects Mitochondrial Function In The Hypothalamic-Pituitary-Adrenal Axis And Impairs Vascular Reactivity In Male And Female Mice
Authors: dos Passos Junior Rinaldo R , Priviero Fernanda , dos Anjos Moraes Raiana , Arishe Olufunke , Wilczynski Stephanie , Snyder Ashley , Crockett Alexia , Frambres Noelle , Stegmann Kyle , Churillo Amelia , Wood Susan K , Spinale Francis G , Ryan Michael J , Hollis Fiona , Webb R Clinton C
Published: October 2023

Chronic stress is associated with hypertension but the underlying mechanisms linking the interplay between the changes in the hypothalamic-pituitary-adrenal axis and vascular dysfunction remain unclear. We hypothesize that chronic unpredictable stress (CUS) induces mitochondrial dysfunction in the HPA axis and in the vasculature, associated with GasderminD pore formation in the mitochondrial membrane. Adult male and female C57Bl6/J mice were exposed to 28 consecutive days of CUS or handling (CTL). Anxiety-like behavior, coping behavior, and systolic blood pressure (SBP) were assessed. Mitochondrial function was assessed using high-resolution respirometry in the hypothalamus, amygdala, and adrenal tissues. The vascular reactivity of mesenteric resistance arteries (MRA), pudendal arteries (PA), and aorta were assessed in response to phenylephrine or acetylcholine. GasderminD protein expression was measured in the MRA. Data are presented as mean ± S.E.M and significance was set at p<0.05. In both sexes, CUS increased SBP by 12.1±4.9%. CUS decreased body weight gain by 53% (p=0.04) and increased anxiety-like behavior (p=0.01) in males (vs CTL) but did not affect females. CUS decreased latency to immobility in the forced swim test by 46% (p=0.0001) in both sexes. Sex-dependent decreases in mitochondrial respiration were observed as follows: in the amygdala of males (20%, p=0.05) and the hypothalamus (20%, p=0.004) and adrenal glands (17%, p=0.06) of females. In the vasculature, CUS led to endothelial dysfunction observed by reduced potency of ACh in the MRA (pEC
50
CUS 6.17±0.09 vs CTL 6.81±0.11; p=0.003) and PA (pEC
50
CUS 6.31 ±0.07 vs CTL 6.65±0.09; p=0.008) of male mice, as well as in the MRA of females (pEC
50
CUS 5.94±0.30 vs CTL 6.98±0.15; p=0.041). Contractile responses to PE were significantly reduced in the aorta of CUS males only (E
MAX
CTL: 4.70±0.48 mN vs CUS 3.01± 0.23 mN; p=0.007). The expression of gasderminD was increased in the MRA of female mice only. CUS increased SBP in males and females, induced sex-specific changes in the mitochondrial function in key areas of the brain, and caused vascular dysfunction, suggesting that behavioral, brain, and vascular adaptations to CUS are sex-dependent.
Abstract 030: Perinatal Paradigms Of Opioid Use To Investigate Cardiovascular Disease (CVD) Risk In Adult Offspring
Authors: Ahmed Nermin , Dalmasso Carolina , Loria Analia S
Published: October 2023

Adversity and dysfunctional households during childhood have been shown to increase systolic blood pressure and overall CVD risk in the long term. A common contributor to dysfunctional households is the parental drug abuse, which transplacental effects acts as a stressor to the developing offspring. Opioid use disorder (OUD) among pregnant women is an understudied area related to the opioid epidemic, causing a 5-fold increase in neonatal opioid withdrawal syndrome (NOWS) incidence. In former studies, we subjected female rats to two models of maternal opioid exposure 1) daily escalating doses of morphine from gestational day 1-19, or 2) fentanyl self-administration during preconception and gestation. Female and male opioid-exposed offspring are smaller throughout the lifespan than vehicle (VEH)-exposed counterparts, displaying NOWS-like somatic withdrawal signs and catch-up growth. Also, in utero opioid-exposed adult offspring displayed sympathetic activation and increased CVD risk factors. Perinatal exposure to opioids has been shown to dysregulate the endogenous opioid peptides (EOP) and alter behavioral outcomes later in life, but the effects on blood pressure regulation remain understudied. Therefore, this study aimed to investigate the expression of proenkephalin (PENK) in brain regions known to control the sympathetic outflow (PVN, RVLM, NTS) and the pressor response to Leu-Enkephalin (100 ug/kg i.p) in adult offspring exposed to VEH or opioids in utero. PENK protein expression in RVLM showed a ~50% reduction in opioid-exposed offspring vs. VEH (p<0.05). As expected, the pressor response to Leu-ENK in conscious VEH-exposed rats showed a ~30% increase in mean arterial pressure from baseline, while the pressor response was blunted in opioid-exposed offspring (12.2±3.7 vs. -4.5±2.1 delta mmHg, n=4-5, p<0.05). As EOP exert inhibitory effects on adrenergic signaling, our data indicate that perinatal exposure to opioids may induce a permanent downregulation of EOP in the central nervous system. Similar to what we reported in vasculature and heart, reduced PENK may contribute to increasing the sympathetic tone and desensitize acute pressor responses to EOP, suggesting that optimal EOP levels may be critical for blood pressure regulation.
Abstract 046: Association Of Cancer With The Risk Of Developing Hypertension
Authors: Nagasawa Hajime , Kaneko Hidehiro , Suzuki Yuta , Okada Akira , Fujiu Katsuhito , Takeda Norifumi , Morita Hiroyuki , Nishiyama Akira , Yano Yuichiro , Node Koichi , Yasunaga Hideo , Komuro Issei
Published: October 2023

Background and Aims:
Although the importance of hypertension in patients with cancer is widely recognized, little is known about the risk of developing hypertension in patients with a history of cancer.

Methods:
This retrospective observational cohort study analyzed data from the JMDC Claims Database between 2005 and 2022, including 78,162 patients with a history of cancer and 3,692,654 individuals without cancer. The primary endpoint was the incidence of hypertension.

Results:
At the initial health check-up, the median systolic and diastolic blood pressure were 116 (IQR:106-126) and 71 (IQR:64-80) mmHg. The median age of the study participants was 43 years (IQR, 35-51 years), and 2,118,101 (56.2%) were men. During a mean follow-up period of 1,208 ± 966 days, 311,197 participants developed hypertension. Compared to individuals without a history of cancer, the cumulative incidence of hypertension was higher in individuals with a history of cancer (log-rank test, P<0.001). The incidence of hypertension was 364.6 (95% CI 357.0-372.2) per 10000 person-years among those with a history of cancer and 247.2 (95% CI 246.3-248.1) per 10000 person-years in those without cancer. Individuals with a history of cancer had an elevated risk of developing hypertension according to multivariable Cox regression analyses (HR 1.17, 95% CI 1.15-1.20). Both cancer patients requiring active chemotherapy (HR 2.01, 95% CI 1.85-2.20) and those who did not require active chemotherapy (HR 1.14, 95% CI 1.12-1.17) had an increased risk of hypertension. A multitude of sensitivity analyses confirmed the robustness of the relationship between cancer and incident hypertension. Patients with certain types of cancer were found to have a higher risk of developing hypertension than those without cancer, with varying risks dependent on the types of cancer.

Conclusions:
Our analysis of a nationwide epidemiological dataset revealed that individuals with a history of cancer have a higher risk of developing hypertension, and that this finding applies to both cancer patients who require active chemotherapy and those who do not. The results of this study confirm the importance of the novel scientific field of "onco-hypertension”.
Abstract 048: Clinical Impact Of 3 Versus 5-minute Delay And 30 Versus 60-second Intervals On Automated Office Blood Pressure Measurements
Authors: Lynn-Green Erika , ishak anthony , Cluett Jennifer , Turkson-Ocran Ruth-Alma N , Mukamal Ken J , Cohen Marc L , Li Jonathan , Juraschek Stephen P
Published: October 2023

Background:
Guidelines advise automated office blood pressure (AOBP) with an initial 5-minute delay and multiple measurements at least 60 seconds apart. Recent studies suggest that AOBP may be accurate with shorter delays or intervals, but evidence in clinical settings is limited.

Objective:
To evaluate shorter delays and intervals for AOBP accuracy compared to awake-time ambulatory BP measurement (ABPM).

Methods:
Patients referred to one hypertension (HTN) center underwent ABPM and one of four non-randomized, unattended AOBP protocols: a 3 or 5-minute delay with a 30 or 60-second interval, i.e., 3m/30s/30s, 3m/60s/60s, 5m/30s/30s and 5m/60s/60s. HTN was defined as SBP≥140 or DBP≥90 mmHg. We compared differences in mean BP and HTN classification between average AOBP and awake-time ABPM by t-tests and Fisher’s exact test.

Results:
Among 195 participants (mean 59±16 years, 63% women, 25% Black), overall AOBP average was 133.9±15.7/78.5±11.2. Differences between average AOBP and awake ABPM were: 1.3±14.8 mmHg (5m/60s/60s;
P
for difference=0.46), 7.0±16.7 mmHg (3m/60s/60s;
P
=0.004), 6.7±12.1 mmHg (5m/30s/30s;
P
<0.001), and 5.0±12.9 mmHg (3m/30s/30s;
P
=0.06) (
Figure
). The proportion with HTN was statistically equivalent to ABPM for the 60s arms, but significantly different for the 30s arms (both
P
-values for difference = 0.01).

Conclusion:
In this quality-improvement study, 5m/60s/60s was most similar to the awake-time ABPM average. While shorter protocols may improve AOBP adoption in clinical practice, the potential loss in accuracy should be evaluated in a larger, randomized trial.
Abstract 054: Role Of The Immunoproteasome In Hypertension-associated Immune Activation
Authors: de la Visitacion Nestor , Chen Wei , Krishnan Jaya , Van Beusecum Justin P , Amarnath Venkataraman , Hennen Elizabeth M , Zhao Shilin , Saleem Mohammad , Ao Mingfang , Harrison David G , Patrick David M
Published: October 2023

Background:
Inflammation is essential for the development of hypertension (HTN), however the mechanisms underlying this process remain unclear. The immunoproteasome processes intracellular peptides for display within class I major histocompatibility complexes (MHC-I). LMP7 is the chymotrypsin subunit of the immunoproteasome. Isolevuglandins (isoLGs) are reactive oxygen species (ROS) that drive immune activation and contribute to the development of HTN.

Methods:
IsoLG adduct-MHC-I interaction was measured by fluorescence resonance energy transfer. Mice with LoxP sites flanking exons 1 and 2 of
Psmb8
, the gene encoding the immunoproteasome subunit LMP7, were generated and crossed to mice expressing dendritic cell (DC) and endothelial cell (EC) specific cre-recombinase. Mice were treated with angiotensin II as a model of HTN and blood pressure was measured by radiotelemetry. Tissue inflammation was measured by flow cytometry.
In vitro
, DCs, ECs, and B8 fibroblasts were used to study IsoLG adduct processing. Mouse aortic ECs (MAECs) were studied in models of isoLG-mediated T cell proliferation, gene expression, and antigen presentation.

Results:
Overexpression of immunoproteasome subunits augments isoLG-adduct MHC-I presentation. Oxidative stress induces IsoLG adduct MHC-I presentation by ECs and DCs. EC-specific IsoLG adduct MHC-I presentation contributes to hypertensive T-cell proliferation. Conditional deletion of LMP7 in DCs attenuated both tissue inflammation and HTN (142.3 ± 13.3 mmHg in

Psmb8
fl/fl

mice compared to 127.1 ± 9.5 mmHg in

Psmb8
fl/fl
/CD11c-Cre

mice, N = 5-6, p=0.0363). Similarly, EC-specific LMP7 loss-of function attenuated inflammation and HTN (142.9 ± 3.4 mm Hg in

Psmb8
fl/fl

mice compared to 119.2 ± 2.6 mm Hg in

Psmb8
fl/fl
/vecadherin-Cre

mice, N = 5, p= 0.0094). IsoLGs increase LMP7 expression and STING activation in MAECs.

Conclusions:
These studies define a critical role of the immunoproteasome in the regulation of immune cell activation and tissue infiltration in HTN. Moreover, they define a mechanism of isoLG adduct MHC-I presentation as a driver of this process. Finally, these studies define a critical role of antigen presentation in T cell activation, proliferation, and migration in HTN.
Abstract 063: Pregnancy Protects Aged G Protein-coupled Estrogen Receptor 1 Knock-out Mice Against Hypertension
Authors: Singh Ravneet , Almutlaq Rawan N , Nasci Victoria L , Gohar Eman Y
Published: October 2023

Hypertension, a major cause of premature death worldwide, is more prevalent in postmenopausal women compared to premenopausal women. Data implicate a protective effect of G protein-coupled estrogen receptor 1 (GPER1) in cardiovascular diseases. A recent study showed that previous pregnancy lowers blood pressure in hyperandrogenemic rats. We hypothesized that pregnancy protects against hypertension elicited in aged female mice lacking GPER1. 16-20 month-old GPER1 wild-type (WT) and knock-out (KO) mice with or without a history of former pregnancies were implanted with telemeters for blood pressure recording and placed in metabolic cages for 24h urine collection. Plasma, adrenal glands and kidneys were collected. GPER1 deletion increased mean arterial pressure in virgin mice (125±3 vs. 109±2 mmHg; n=3,7; p=0.0005), and previous pregnancies (PP) eliminated this genotypic effect (106±1 mmHg; n=6; p≤0.0001). Aldosterone plays a critical role in blood pressure regulation, we measured its urinary excretion and revealed lower levels in PP KO mice compared to virgin KO mice (4.8±0.8 vs. 13.0±3.0 pg/day; n=6; p=0.0099). PP KO mice elicited greater levels of circulating estradiol levels compared to virgin KO mice (42.8±5.2 vs. 29.5±3.4 pg/mL; n=8,6; p=0.0451). To explore the role of adrenal estrogen receptors (ER) in regulating aldosterone production, we assessed the adrenal expression of ER and CYP112B, the gene coding for aldosterone synthase. No difference was observed in ERα, however ERβ expression was higher in PP KO mice compared to virgin KO mice (2.4±0.3 vs. 1.1±0.1 fold change; n=6,5; p=0.0359). GPER1 deletion increased CYP112B expression in virgin mice (2.0±0.4 vs. 1.0±0.2 fold change; n=7,5; p=0.0265), and this was eliminated with PP (0.9±0.2 fold change; n=5; p=0.0260). Aldosterone regulates renal epithelial sodium channel (ENaC), so we assessed its subunits’ expression. Renal ENaCα and ENaCβ increases with the deletion of GPER1 in the virgin group, and this effect was lost with PP. ENaCγ withheld the same trend, but it did not reach significance. Overal, our data suggest that GPER1 and former pregnancy protect from hypertension in aged females at least partially via regulating aldosterone and ENaC signaling.
Abstract 066: Decoy Peptide Attenuates The Syndrome Of preeclampsia In A Rodent Model
Authors: Kundysek Waverly B , Pantho Ahmed , Afroze Syeda , Vora Niraj , Kuehl Thomas , Zawieja David C , Uddin Mohammad N
Published: October 2023

Background:
Preeclampsia (preE) is a serious complication of pregnancy manifested by high blood pressure, proteinuria, and edema. In a previous study, we demonstrated that circulating levels of soluble (pro)renin receptor ((P)RR) were elevated at delivery in patients with preE, and that both plasma and placental (pro)renin were elevated in preE patients and in a rat model of preE. Decapeptide based on this handle-region sequence (handle-region peptides or HRP) can block binding of (pro)renin to (P)RR.

Objective:
The goal of this study is to develop and characterize HRP as an innovative treatment for preE.

Methods and Materials:
The human extravillous cytotrophoblast (CTB) cell line Sw-71 used in these studies was derived from first trimester chorionic villous tissue. (P)RR (2 nM) was incubated in 1.0 mL medium with the receptor-expressing CTB cells for 0, 6, 12, 18, or 24 hours, followed by renin activity assay to determine the percent binding and activation of (P)RR. Female Sprague-Dawley rats (200-250 g, first-time pregnant with confirmation by vaginal plug, Harlan) are randomly assigned to three groups (n=8 per group): 1) NP: normal pregnant rats; 2) PDS: pregnant animals injected initially with 12.5 mg of DOCA in a depot form intraperitoneally, followed by a weekly injection of 6.5 mg, and whose drinking water was replaced with 0.9% saline; 3) PDS-HRP: rats administered DOCA and saline as for Group 2, and also given HRP (10 mg/kg, i.p.) daily from day 10 through day 20 of pregnancy.

Results:
(Pro)renin was non-proteolytically activated by binding to (P)RR on the cell membrane of CTB cells, while HRP inhibited binding and attenuated activation of (P)RR. The HRP decoy peptide normalizes blood pressure, proteinuria, and birth numbers in DOCA model of preE. When HRP was administered following the onset of hypertension, the BP, proteinuria, and number of pups were normalized to control normal pregnancy.

Conclusions:
HRP inhibits the binding of (pro)renin to (P)RR. This study in animal model of preE suggests HRP blockade of (P)RR can attenuate the disease. (P)RR and (pro)renin levels are elevated in preE patients and in their placenta, as well as in a rat model of preE. This data suggests HRP as a potential therapeutic for intervention of RAS signaling in preE.
Abstract 073: Blood Pressure Trajectories Among Young Veterans: Differences By Sex, Race, And Ethnicity
Authors: Chang Tiffany E , Gaffey Allison E , Buta Eugenia , brandt cynthia , Lichtman Judith H
Published: October 2023

Background:
Veteran status (~6% of U.S. population) is an independent risk factor for cardiovascular disease (CVD). High blood pressure (BP) increases CVD risk for older Veterans. Less is known about BP levels and trajectories for younger Veterans.

Objective:
Assess sex- and racial/ethnic differences in longitudinal BP trajectories for young Veterans accessing the Veterans Affairs (VA) healthcare system.

Methods:
We used national 2001-2016 Veterans Health Administration data for Veterans who served in conflicts in Afghanistan and Iraq. Systolic (SBP) and diastolic (DBP) BP were collected from the first to most recent outpatient visit. Linear mixed effects models estimated annual rate of change in BP. Models adjusted for sociodemographic, behavioral, and clinical factors included a time*sex, time*race/ethnicity interaction term.

Results:
We included 764,652 Veterans (13% women; median age 29.5 years) with 14,788,864 BP measurements over a mean of 6.1 years. There was a significant interaction by sex and race/ethnicity (
P
<0.0001). In sex-stratified models, women vs. men had lower initial SBP (113.3 vs 120.9) and DBP (70.1 vs 73.0), but women had a markedly rapid BP increase over time (SBP rate of change= 0.36 vs 0.10 mmHg/year; DBP = 0.46 vs 0.31;
Figure
). For women, Non-Hispanic (NH) Black women had the highest initial BP and annual rate of change (SBP/DBP: 0.46/0.50).

Conclusion:
Younger Veterans’ have normal BP when ending military service, but there are sex, racial, and ethnic differences in BP patterns over time. NH Black women Veterans show the greatest rate of BP increase. Targeted interventions may be needed to reduce disparities in trajectories to reduce future CVD risk.
Abstract 084: Smooth Muscle Cullin-3 Regulates The Renal Baroreceptor Mechanism Via Lamin AC
Authors: Golosova Daria , Kumar Gaurav , Lu Ko-Ting , Brozoski Daniel T , Muskus Veitia Patricia , Nakagawa Pablo , Sigmund Curt D
Published: October 2023

Cullin-3 (CUL3) is a critical component of the CUL3-Ring-Ligase (CRL) ubiquitin ligase complex. Patients carrying mutations in CUL3 exhibit early onset of hypertension (HTN). We demonstrated that conditional ablation of CUL3 in vascular smooth muscle cells (VSMCs) results in vascular dysfunction, arterial stiffness, and severe HTN in mice. We hypothesized that 1) conditional deletion of CUL3 in VSMCs (S-CUL3KO) results in severe HTN via the renin-angiotensin system (RAS)-dependent mechanism, and 2) S-CUL3KO mice exhibit an impaired baroreceptor mechanism regulating renin. Mice carrying a conditional allele of CUL3 were bred with mice expressing a tamoxifen-inducible CRE-recombinase driven by a smooth muscle promoter (ISM-CRE, control). S-CUL3KO mice exhibited elevated systolic blood pressure (SBP) 3 weeks after tamoxifen administration (S-CUL3KO: 164±1 vs. ISM-CRE: 124±4 mmHg, p<0.05). To test the first hypothesis captopril or candesartan (10 mg/kg/day and 100 mg/kg/day in drinking water, respectively) were administered to S-CUL3KO once severe HTN was already established. Captopril or candesartan treatment for 1 week reversed elevated SBP to 117±4 mmHg (p<0.05) and 127±2 mmHg (p<0.05), respectively, in S-CUL3KO. Despite the severity of HTN in S-CUL3KO, the level of renin mRNA in the kidney was surprisingly unaltered (p>0.05) when compared to the normotensive control mice. The protein expression of lamin AC, a crucial component of the nuclear mechanotransducer controlling renin expression, was significantly increased in S-CUL3KO kidneys (0.11±0.02 vs. 0.05±0.01 normalized RFU, p<0.05). This suggests lamin AC may be a target protein of CUL3-ubiquitin ligase complex. Supporting this concept, treatment of A10 smooth muscle cells with MLN4914, an inhibitor of the Cullin pathway, results in a 2-fold increase in lamin AC expression (0.007±0.001 vs. 0.0154±0.0004, normalized RFU, p<0.05). Furthermore, co-immunoprecipitation of CUL3 in HEK293 cells immunoprecipitated lamin AC. Our findings support the hypothesis that 1) CUL3 regulates RAS via the renal baroreceptor mechanism and 2) lamin AC is a potential target of the CRL3-mediated regulation. Understanding CUL3 target proteins might help the development of new strategies to mitigate HTN.
Abstract 088: Modulatory Effects Of The Gut Microbiota-derived Short Chain Fatty Acids On Heart Rate And Vascular Tone In Larval Zebrafish
Authors: Sree Kumar Hemaa , Wisner Alexander , Schiefer Isaac T , Zubcevic Jasenka
Published: October 2023

Introduction:
Short chain fatty acids (SCFAs), such as acetic, propionic, and butyric acid, areproduced by fermentation of dietary fibers by the gut bacteria. Deregulation in production andhandling of SCFAs is associated with cardiovascular dysfunction in humans and rodents.However, limited studies have investigated the effect of SCFAs on cardiovascular parameters inthe zebrafish (ZF), an increasingly popular model in cardiovascular and gut microbiota research.

Methods:
Bacterial 16s sequencing and GC-MS were performed in adult male and female Tg(NFkB:EGFP) ZF fecal samples for analysis of the gut microbiota and SCFAs respectively. Inanaesthetized ZF larvae (7-9 dpf; MS-222, 0.3mM), measurements of heart rate (HR, bpm) andvascular tone (width of dorsal aorta, pixels) were performed every 10 minutes for an hour usinglive imaging (Nikon, 4x magnification) following administration of physiologic concentrations ofacetate (27.4 μM), butyrate (2.2 μM) and propionate (1.7 μM; pH=7.5) via immersion.

Results:
Relative proportions of SCFAs in the ZF were similar to those previously reported in rats andhumans, with highest levels of acetic acid (27.43 μM) followed by butyric (2.19 μM) and propionic(1.65 μM) acid. At the phylum level, the observed abundances of Proteobacteria, Firmicutes, andFusobacteria were similar to those reported in rodents and humans. Administration of selectSCFAs resulted in a ~20% decrease in HR at all time points following administration of butyrate(P<0.0001), and at 40-60min following administration of acetate (P<0.05), with no effect ofpropionate. In addition, a ~25% decrease in the width of the dorsal aorta was observed at 60 minfollowing administration of butyrate (P<0.05), with no effect of acetate or propionate.

Conclusion:
We report a similar composition and abundances of the gut microbiota and SCFAs between ZF,rodents and humans. In addition, both butyrate and acetate produced a bradycardia in the ZF,while the observed butyrate-dependent decrease in the vascular tone may be compensatory tobradycardia. Our findings demonstrate the potential for ZF as a model to study host-microbiotainteractions in regulation of cardiovascular function.
Abstract 090: Using Evidence-Based Resources And Unique Self-Measured Blood Pressure Program Designs To Improve Blood Pressure Control
Authors: Smith Alison , Wagner Ashley , Brown Leslie , Cardoso Celicia , Dadkhah Mahsa , Decuir Crystal , Fiegel Sondi , Harrison Nekeisha , Mooney Mark V. , Pena David , Pillai Jan , Ramirez Sonia , Royer Amy , Tate Jodi , Sanchez Eduardo J
Published: October 2023

Background:
The AHA National Hypertension Control Initiative (NHCI) is a $32M, 3-year grant from the Health Resources and Services Administration (HRSA) and the Office of Minority Health (OMH) to provide training and technical assistance (T/TA) to 350 HRSA-funded community health centers (CHCs) with blood pressure (BP) control rates below 58.9% in 2019.

Hypothesis:
NHCI can improve BP control particularly among Black, Hispanic, American Indian/Alaskan Native (AI/AN) persons through systematic, culturally sensitive, and evidence-based interventions, including use of self-measured blood pressure (SMBP).

Methods:
AHA’s T/TA included a training curriculum, evidence-based tools from Target: BP™ and the AMA MAP™ framework, and new tools, including a BP Measurement Policy and a BP Treatment Algorithm. CHCs used these resources to change clinical care and to implement SMBP though multidisciplinary and culturally relevant approaches that reflect the organizational capacities and characteristics of their patients. For example, 3 CHCs successfully implemented SMBP programs using different designs and achieved notable improvements in BP control (defined by the performance measure <140/90 mm Hg). SWLA in Louisiana established a team-based care model that emphasizes standardized processes, including an SMBP onboarding protocol and a BP treatment algorithm. Canyonlands in Arizona coupled systematic in-clinic practices with AI/AN community health workers (CHW) to reach patients from the Navajo Nation in a culturally relevant manor. Via Care in California developed a collaborative practice model with their clinical pharmacists and clinicians for systematic medication management, SMBP and lifestyle education, and team-based visits.

Results:
SWLA increased BP control from 44% to 69% in 2022, Canyonlands from 53% in 2021 to 70% in 2023 in AI/AN patients that partnered with a CHW, and Via Care from 55% in 2021 to 74% in 2022. So far, the highest BP control rate percentage change was for CHCs that participated with NHCI and had a 5.1% year-over-year increase compared to 3.8% at all HRSA centers from 2020 to 2021.

Conclusion:
Various SMBP program designs with systematic processes, team-based care models and culturally sensitive approaches can improve BP control.
Abstract 095: Sex-specific Mechanisms Of Obesity-associated Hypertension Are Not Dependent On Ovarian-derived Sex Hormones
Authors: Barris Candee , Kress Taylor , Antonova Galina , Faulkner Jessica L , Belin De Chantemele Eric J
Published: October 2023

Traditionally, female sex hormones were thought to protect premenopausal women from cardiovascular disease, yet recent studies indicate that obesity abolishes these protective effects and predisposes women to vascular dysfunction and hypertension. Previously, our laboratory demonstrated that hypertension in obesity involves the adipokine leptin via sex-specific mechanisms: leptin-mediated sympatho-activation in males and leptin-mediated aldosterone production in females. Novel data gathered from discarded human adrenals indicate that obesity increased aldosterone synthase expression in women. However, whether female sex hormones modulate this sex difference remains unknown. We hypothesized that the loss of ovarian-derived sex hormones induced by ovariectomy (OVX) will alter the mechanism by which obese female mice develop hypertension and impair vascular function. Obese agouti yellow (Ay) mice and lean controls (a/a) underwent OVX or sham (S) surgery. Radiotelemeters were implanted for blood pressure (BP) monitoring at baseline and in response to leptin receptor (LepR) blockade. Mice were euthanized and mesenteric arteries isolated to measure vascular function via wire myography. Obesity increased BP (a/a
S
: 109±0.9 vs. Ay
S
: 119±2.5 mmHg, p<0.05) which was not further elevated by OVX (a/a
OVX
: 103±3.5 vs. Ay
OVX
: 116±7.5 mmHg). LepR blockade decreased MAP in obese OVX and S mice (Ay
S
: 112±3.5 vs. Ay
OVX
: 111±4.2 mmHg), indicating that BP elevation remains leptin-mediated in the absence of female sex hormones. Obesity impaired endothelial-dependent relaxation to acetylcholine (a/a
S
: 94±6.6, a/a
OVX
: 95±4.6, Ay
S
: 82±10, Ay
OVX
: 71±13%) but OVX did not cause further impairment. OVX did not alter adrenal aldosterone synthase expression nor plasma aldosterone levels. Human adrenocortical cells (HAC15) treated with combination of estradiol ± progesterone exhibited no increased aldosterone synthase expression. These results indicate that loss of ovarian-derived sex hormones does not alter the mechanisms for hypertension in obese female mice. This data suggests that female sex hormones likely play a limited role modulating the sex-specific mechanisms for hypertension in obesity.
Abstract 096: Effects Of Chronic Hypertension And Sex On Spatial Reference And Working Memory
Authors: Reulbach Jamie C , Baumoel Emily , Johnson Abbie C
Published: October 2023

Chronic hypertension is a primary risk factor of vascular dementia. The hippocampus is a brain region critical to learning and memory and is highly susceptible to hypoxia/ischemia. Here, we investigated how chronic hypertension affects two different types of memory function, and whether one sex is more susceptible to hypertension-induced memory impairment. Memory function in adult male and female SHR (spontaneously hypertensive rats) and normotensive Wistar rats was tested using two behavioral tasks (n=6/group). The radial arm maze (RAM) tested reference memory that is largely hippocampal-dependent by measuring the time it took to retrieve rewards from three baited arms during daily 10 min sessions. A continuous y maze task was used to calculate alternation index and determine working memory that is cortico-hippocampal-dependent. Data are mean±SEM and comparisons made via a two-way ANOVA with a post-hoc Tukey test. In the RAM task, female rats had steeper learning curves than males as they completed the task in less time with each session (Fig 1). There was a significant interaction between sex and hypertension in the y maze task (F
(1,20)
=13.26, p=0.002). Working memory was impaired in female SHR, with a lower alternation index (index=0.60±0.03) than Wistar rats (index=0.74±0.03). However, male SHR that had a higher alternation index of 0.75±0.02 compared to 0.58±0.6 in male Wistar rats. Overall, females had stronger reference memory than males regardless of hypertension. Working memory was differentially affected in male and female SHR, suggesting sex differences in the susceptibility of different brain regions and types of memory to hypertension-induced dysfunction.
Abstract 097: Sexually Dimorphic Transcriptional Phenotypes In Tubular Epithelial Cells Of The Rat Kidney
Authors: Kramer Benjamin K , Zhang Ronghao , Gonzalez-Vicente Agustin
Published: October 2023

Sexual dimorphism is a key biological variable in different organ systems. Although a growing body of evidence at single cell-resolution highlights sex differences in the kidneys of humans and mice, information regarding rats is limited. We integrated publicly available datasets to construct a single-cell RNA sequencing atlas of the rat kidney, incorporating data from 3 females and 3 males. Transcriptomes from the tubular epithelial cell clusters, exhibited strong correlation with those from rat-kidney microdissected tubule segments and single-cell RNA sequencing of human kidney biopsies. We hypothesize that transcriptomes of rat tubular epithelial cells present sexual dimorphism. To test this hypothesis, we identified genes that were differentially expressed between sexes (DEGs: log2FC >= 1; adjusted-p <= 0.05) in proximal tubules (PT), thick ascending limbs (TAL), distal convoluted tubules (DCT), principal cells (PC) and intercalated cells A and B (IC-A and IC-B, respectively). Gene enrichments of PT DEGs were conducted using WikiPathways. The number of genes positively enriched in males was as follows: 43 PT, 9 TAL, 16 DCT, 23 PC, 12 I-CA and 19 I-CB. While females presented 44 PT, 4 TAL, 4 DCT, 3 PC, 1 IC-A and 3 IC-B. Four transcripts were upregulated in all male segments, 1) glutathione peroxidase 3 (
Gpx3
), 2) lysozyme 2 (
Lyz2
), 3) locus A1 (
RT1-A1
) which enables beta-2-microglobulin and peptide binding activity, and 4)
AABR07060872.1
an Ig-like domain-containing protein. Only one long non-coding RNA (
AABR07039356.2
) was upregulated in all female segments. The three most enriched pathways in male PT were: 1) Proximal tubule transport (WP4917; p < 1x10
-14
), 2) Glycolysis and gluconeogenesis (WP534; p < 2x10
-12
) and 3) Metabolic reprograming in colon cancer (WP4290; p < 8x10
-9
). In females, the most enriched pathways were: 1) Trans-sulfuration and one-carbon metabolism (WP2525; p < 6x10
-9
), 2) One-carbon metabolism related pathways (WP3940; p < 1x10
-8
) and 3) Amino acid metabolism (WP3925; p < 7x10
-8
). Our results show the presence of sex-related transcriptional differences across the rat nephron, but predominantly in PT. These differences impact key metabolic functions of PT such amino acids and carbohydrates metabolism and transport processes.
Abstract 098: Collecting Duct-derived (pro)renin Receptor Contributes To 2-kidney, 1-clip-induced Renovascular Hypertension And Ischemic Nephropathy In Mice
Authors: Fu Ziwei , Kaewsaro Kannaree , Hamby Jacob A , Kimball Alex , Yang Tianxin
Published: October 2023

The 2-kidney, 1-clip (2K1C) Goldblatt model features overactivation of the systemic renin-angiotensin-aldosterone system (RAAS) due to increased renin release from juxtaglomerular cells. However, no prior study functionally evaluates possible involvement of the intrarenal RAAS in this model. Within the kidney, (pro)renin receptor (PRR) is predominantly expressed in the collecting duct (CD) implicated in regulation of the intrarenal RAAS under physio-pathological conditions. In the present study, we employed a mouse model of CD-specific deletion of PRR (CD PRR KO) to examine the role of CD PRR in pathogenesis of 2K1C-induced renovascular hypertension and ischemic nephropathy and to further explore the underlying mechanism. Floxed and CD PRR KO mice were subjected to sham-operation or 2K1C procedure, followed by analysis of blood pressure, renal injury, and indices of the RAAS, as well as the expression of subunits of ENaC. Clipping-induced hypertension and renal injury were both attenuated in CD PRR KO mice as compared with floxed controls (MAP on day24: Floxed/2K1C 148.2±3.9 mm Hg vs. CD PRR KO/2K1C 136.9±2.5 mm Hg, n=6, p < 0.05) (urinary albumin/creatinine: Floxed/2K1C 67.5±6.4 mg/g vs. CD PRR KO/2K1C 30.9±3.5 mg/g, n=6, p < 0.01). Of note, the reductions of clipping-induced protein abundance of fibronectin and α-SMA in CD PRR KO mice were 70% and 55%, respectively. Similarly, clipping induced paralleled increases in renal mRNA expression of IL-1β, TNF-α, MCP-1 and TGF-β1 in floxed mice, which were all blunted in CD PRR KO mice. The protective phenotype of the null mice was paralleled with suppressed intrarenal renin and aldosterone (Aldo) levels. In contrast, clipping-induced enhancement of circulating renin and Aldo remained unchanged between the genotypes. Moreover, renal medullary α-ENaC mRNA and protein expression was elevated by clipping in floxed mice, which was respectively blunted 52% and 54% in CD PRR KO mice. Together, these results suggest that the activation of CD PRR stimulates components of the intrarenal RAAS and renal medullary α-ENaC, which result in increased tubular sodium reabsorption and thus contribute to 2K1C-induced renovascular hypertension and ischemic nephropathy.
Abstract 101: Similar Cd28
+
Delta T Effector Memory Cell Subpopulations Are Present In Murine Perivascular Tissues Of Angiotensin II-infused Male Mice And Subcutaneous White Adipose Tissue Of Obese Women
Authors: Paradis Pierre , Berillo Olga , Comeau Kevin , Caillon Antoine , Leclerc Séverine , Shokoples Brandon , Mahmoud Ahmad , Andelfinger Gregor , Schiffrin Ernesto L
Published: October 2023

Introduction:
The innate-like γδ T cells play a role in angiotensin II (AngII)-induced hypertension, mesenteric artery injury and T cell activation in mesenteric vessels/perivascular adipose tissue (MV/PVAT) in mice. Blood γδ T cells frequency is associated with systolic blood pressure in humans. Obesity was associated with small artery injury in subcutaneous white adipose tissue (WAT) and is a risk factor for hypertension in humans. We hypothesized that single cell RNA sequencing (scRNA-seq) in MV/PVAT of AngII-infused vs control mice and in subcutaneous WAT of obese vs lean humans will reveal γδ T cell subpopulations involved in hypertension, vascular injury and T cell activation.

Methods:
Male C57BL/6J mice were infused SC or not with 490 ng/kg/min AngII for 14 days (n=3). Hypertension was confirmed by tail cuff blood pressure measurement. MV/PVAT were collected, and T cells were isolated and used for scRNA-seq. γδ T cell subpopulations were studied by subclustering T cells expressing the T cell receptor δ constant chain. Cell subpopulations were validated by flow cytometry. Subcutaneous WAT CD45
+
cell ScRNA-seq data of 3 lean and 3 obese women were obtained from GEO NCBI database (GSE155960) and γδ T cell subpopulations analyzed as above.

Results:
ScRNA-seq identified 6 murine and 9 human δ T cell subclusters. AngII increased
Cd28
high
(% of T cells: 3.6±0.9 vs 1.4±0.2) and
Trdv4
+
δ T effector memory (EM) cells (7.3±1.3 vs 3.4±1.7) and lowered resting δ T cells (1.0±0.2 vs 4.2±1.5) in mice.
Cd28
high
δ T
EM
and resting δ T cell study by flow cytometry using unique marker pairs, CD28
+
SELL
-
and CD28
-
SELL
+
, showed that AngII increased CD28
+
SELL
-
(cell number: 376±48 vs 220±40) and decreased CD28
-
SELL
+
δ T cells (23±4 vs 45±4) in MV/PVAT.
Cd28
/
CD28
subclustering of δ T cells identified 4 human and murine cell populations and showed that AngII increased
Cd28
+
γδ natural killer T
EM
(% of T cells: 1.9±0.3 vs 0.5±0.2) and

Cd28
high

δ T
EM
cell subpopulations (1.6±0.5 vs 0.2±0.1) in mice, and that

CD28
+

δ T
EM-1
cell subpopulation tended to be increased in obese vs lean women.

Conclusion:
ScRNA-seq revealed similar δ T
EM
cell subpopulations expressing CD28 in murine MV/PVAT and human subcutaneous WAT that could play a role in hypertension, vascular injury and T cell activation.
Abstract 104: Assessment Of Brain Venous System In Hypertension
Authors: Glodzik Lidia , Biskaduros Adrienne , Butler Tracy , Li Yi , Chiang Gloria , Tanzi Emily , Harvey Patrick , de Leon Mony
Published: October 2023

With an increasing recognition of the contribution of venous collagenosis to cerebral small vessel disease there is a growing interest in the imaging of brain veins. Some studies showed increased prominence of cerebral veins in animal models of hypertension (HTN) and increased venous visibility in patients with white matter lesions using susceptibility weighted imaging (SWI). Since SWI uses deoxygenated blood as an intrinsic contrast agent, reduced cerebral blood flow (CBF) could result in increased oxygen extraction and thus greater vein prominence. However, others reported that fewer veins visible on SWI correlated with more brain atrophy. We examined whether the number of deep medullary veins (DMVs) was related to HTN and CBF, in a group of cognitively healthy subjects from a brain aging research center (n=351, 220 women, mean age 69.4 ± 7.1). DMVs were assessed by visual inspection on SWI on the level of lateral ventricles, and the number was defined as the average count of both hemispheres. CBF was assessed using arterial spin labeling. The number of DMVs did not differ (p=0.17 after adjusting for age, sex and acquisition type) between normotensive subjects (n=194, <140/90, and no antihypertensive treatment), subjects with controlled HTN (n=74, <140/90, and antihypertensive treatment), and subjects with uncontrolled blood pressure (n=72, ≥ 140/90). It was, however, numerically the lowest in the uncontrolled group (22.8 ± standard error (SE) 1.1, 25.6 ± 1.1 for controlled, and 24.4 ± 0.7 for normotensive individuals). In the entire group there was a negative correlation between CBF and DMVs: r=-0.23, p<0.001. The correlation became progressively stronger across HTN spectrum: r=-0.16, p=0.03 for normotensive subjects, r=-.23, p=0.03 for the controlled group and r=-.038, p=0.001 for the uncontrolled HTN group. At the same time there was a linear trend (p=0.06 after adjusting for age and sex) for CBF decrease from 59.5 ml/100mg/min ± SE 0.4 in normotensive subjects, 58.3±0.7 in controlled and 58.0±0.7 in the uncontrolled group. A greater reliance on oxygen extraction due diminishing perfusion across HTN spectrum may obscure structural changes in cerebral veins previously reported in subjects with vascular disease.
Abstract 109: Geriatric Unhoused Patients Presenting To A Free Clinic In Miami- Dade Had Significantly Higher Rates Of Stage 1 And 2 Hypertension And Are At Significantly Increased Risk For Untreated Hypertension Compared To Age- Matched General Population
Authors: Morgan Orly , Seshadri Suhas , Moore Alana , Bergholz Daniel , Henderson Armen , Laban Joshua
Published: October 2023

Introduction:
People over age 50 comprise 30% of people experiencing homelessness (PEH) in the United States and are predicted to increase exponentially in the next ten years. Few studies have documented PEH hypertension (HTN) prevalence, and treatment protocols rarely test their efficacy specifically when treating older PEH. The few geriatric PEH HTN studies that have been conducted are based out of homeless shelters, with even less known about the HTN rates of people experiencing unsheltered homelessness (PEUH) who constitute one-third of PEH nationally. Our objective is to assess prevalence of elevated blood pressure (BP) and HTN treatment rates among geriatric PEUH presenting at a free clinic in Miami-Dade County.

Methods:
Clinical information and BP measurements of 105 PEUH over 50 were documented as part of care by a street outreach medical team. Clinical information was recorded in a REDCap electronic database. De-identified data was compared to the Centers for Disease Control National Health and Nutrition Examination Survey 2017-2019 pre-pandemic cohort (CDC NHANES) and analyzed in R studio version 4.3.2.

Results:
PEUH were at significantly increased relative risk (RR) for Stage 1 HTN (RR 3.97, Confidence Interval (CI) 2.68-5.89, p<0.0001) when compared to age- matched NHANES dataset. After 10-year age cohort stratification the small PEH sample size (N=11) lacked enough power to reach significance. PEUH were also found to be at significantly increased risk of Stage 2 HTN (RR 6.17, CI 4.93-7.73, p<0.0001). Here findings remained significant after 10-year age cohort stratification (N=41). NHANES reports medication use by 20-year age groups: 50.8% of people aged 40-59, and 49.4% of people over 60 with diagnosed HTN were on BP control medication. PEUH were at significantly increased risk of untreated HTN (RR 1.67, CI 1.50-1.87, p<0.0001). Only 15.9% were prescribed medication.

Conclusion:
Increased risk of elevated BP and untreated HTN among geriatric PEUH likely reflect barriers of access to chronic disease management. There are likely also unidentified confounding factors preventing proper care for this vulnerable population. Further investigation should critically assess gaps in the care to devise protocols for improving treatment outcomes.
Abstract 113: Aldosterone Synthase Inhibition With Lorundrostat For The Treatment Of Uncontrolled Hypertension: The Target-HTN Randomized Clinical Trial
Authors: Laffin Luke J , Rodman David , Luther James M , Vaidya Anand , Weir Matthew R , Nissen Steven E
Published: October 2023

Importance:
Excess aldosterone production contributes to hypertension in both classical hyperaldosteronism and obesity-associated hypertension.

Objective:
To compare the safety and efficacy of lorundrostat, an aldosterone synthase inhibitor, with placebo, and characterize dose-dependent safety and efficacy.

Design, Setting, and Participants:
Randomized, placebo-controlled, dose-ranging, double-blind clinical trial among adults with uncontrolled hypertension taking 2 or more antihypertensive medications. A first cohort of participants with suppressed plasma renin (plasma renin activity (PRA) of less than 1.0 ng/mL/h) and elevated plasma aldosterone (greater than 1.0 ng/dL) were enrolled (n=163), with subsequent enrollment of participants with PRA greater than 1.0 ng/mL/h (n=37).

Interventions:
Participants were randomized to placebo or one of five doses of lorundrostat (12.5, 50, or 100 mg once daily, or 12.5 or 25 mg twice daily.)

Results:
Following 8 weeks of treatment in participants with suppressed PRA, dose-dependent changes in office systolic BP were observed (figure). The primary endpoint, placebo-subtracted change in modeled systolic BP was -9.6 mmHg (90% confidence interval [CI], -15.8 to -3.4, p=0.01) for the 50 mg once daily dose and -7.8 mmHg (90% CI, -14.1 to -1.5, p=0.04) for 100 mg daily. Six participants had increases in serum potassium above 6.0 mmol/L that were reversed with dose reduction or drug discontinuation.

Conclusions and Relevance:
Among individuals with uncontrolled hypertension, use of lorundrostat was effective at lowering blood pressure compared with placebo.