bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 54: The kidney-OVLT axis in renovascular hypertension: Individual and combined effects of afferent renal denervation and OVLT lesion in 2K1C rats.
Authors: Ruiz Lauar Mariana , Collister John , Evans Louise , Osborn John
Published: October 2024

We have recently reported that afferent renal denervation (ARDN) markedly attenuates the pathogenesis of hypertension in the 2 kidney 1 clip (2K1C) model in the rat. This was associated with a reduction in neurogenic pressor activity (NPA). We have also recently shown that lesion of the organum vasculosum of the lamina terminalis (OVLT), a forebrain circumventricular organ, had nearly identical effects in 2K1C rats as ARDN. Together these findings suggest that a neural pathway from the kidney to the OVLT plays an important role in 2K1C hypertension. Based on these findings we hypothesized that the combined effect of ARDN and lesion of the OVLT (OVLTx) on arterial pressure in 2K1C rats would not be greater than the individual treatments.

Male Sprague-Dawley rats were randomly selected to receive either OVLT lesion (x) or sham-OVLTx. One week later, rats were instrumented with telemeters to measure mean arterial pressure (MAP). One week later, renal artery stenosis was created by placing a clip on the left renal artery and then, rats were subjected to ARDN by periaxonal capsaicin or sham treatment of the clipped kidney creating 4 experimental groups: sham+sham (n=6), sham+ARDN (n=6), OVLTx+sham (n=5), OVLTx+ARDN (n=5). MAP was measured daily for 28 days, and at the end of the protocol, NPA was assessed by measuring the peak MAP response to ganglionic blockade.

MAP was similar in all 4 groups at the beginning of the study averaging 108±3 mmHg. MAP increased to 180±10 mmHg in the sham+sham group compared to 130±13mmHg in sham+ARDN, 142±18mmHg in OVLTx+sham rats and 158±14mmHg in OVLTx+ARDN rats (p<0.001). NPA in Sham+sham rats was significantly higher (-44 ± 8 mmHg) compared to sham+ARDN (-18 ± 3 mmHg), OVLTx+sham (-21 ± 2 mmHg) and OVLTx+ARDN (-19 ± 4 mmHg) rats (p<0.05). Notably, the combined effects of OVLTx+ARDN on MAP and NPA were not significantly different from either OVLTx (p=1.00 and p=0.819) or ARDN (p=0.113 and p= 0.878) alone.

In summary, ARDN and OVLTx attenuated hypertension and NPA similarly in 2K1C-HTN rats but the combination of these treatments did not produce any additional effect on MAP or NPA. These findings are consistent with the hypothesis that afferent renal nerves and the OVLT are linked in a common neural pathway required for the development of hypertension following renal artery stenosis.
Abstract 55: Mouse Offspring Exposed to Preeclampsia/Eclampsia-like Symptoms Exhibit Changes in Regional Cerebral Perfusion at 2 months of age
Authors: Saffold Karen , Tall Andrea , Pryor Tyranny , Shao Qingmei , Jones- Muhammad Maria , Stancil Kennedi , Warrington Junie
Published: October 2024

Children born to mothers with preeclampsia, a hypertensive pregnancy disorder, or eclampsia, new-onset seizures during pregnancy, are more likely to develop learning and memory deficits and are more susceptible to neurovascular diseases compared to those born from normal pregnancies. The contributing mechanisms are unknown. The objective of this study was to determine whether mice exposed to reduced uteroplacental perfusion (RUPP), to mimic preeclampsia with or without pentylenetetrazol (PTZ) injection (to induce seizures and model eclampsia), results in regional cerebral perfusion changes at 2 months of age. On gestational day (GD) 13.5, pregnant C57BL/6 mice underwent sham or RUPP surgery followed by injection or no treatment with PTZ (40 mg/kg) on GD 18.5. At 2 months of age, cerebral perfusion was measured using Laser Speckle Imaging, and averaged to obtain mean data per sex, per litter (n = 3-6 average datapoints per group). Perfusion data were normalized to the area of each region of interest. Data were analyzed using Two-Way ANOVA. Our findings indicate a reduction in cerebral perfusion in offspring exposed indirectly to RUPP specifically of the right parietal cortex (p = 0.013), superior sagittal sinus (p = 0.003), cerebellum (p = 0.003) and whole brain (p = 0.021). We also observed an overall significant reduction in perfusion, after indirect PTZ exposure, in the right parietal cortex (p = 0.002), transverse sinus (p = 0.003), and cerebellum (p < 0.001); and increase in prefrontal cortex perfusion (p = 0.001). Further observation showed an overall difference in hematocrit (p = 0.001), SPO2 (p < 0.001), and cerebellum water content (p = 0.007) after indirect PTZ exposure. There was no difference in perfusion of the left parietal cortex, body weight, organ weight (except brain, where RUPP exposure led to overall increased brain weight; p = 0.030). Together, our results are consistent with the hypothesis that exposure to preeclampsia/eclampsia conditions leads to changes in offspring cerebrovascular function, with a net reduction in perfusion. Reduced cerebral perfusion is generally associated with reduced cognitive function. Whether this occurs along with changes in capillary density, or is exacerbated with age are areas of active investigation.
Abstract 56: High Blood Pressure Is Associated with Lower Brain Volume and Cortical Thickness in Healthy Young Adults
Authors: Won Junyeon , Ashley John , Cardim Danilo , Vongpatanasin Wanpen , Zhang Rong
Published: October 2024

Background:
High blood pressure (BP) in middle-aged and older adults is associated with lower brain volume and cortical thickness assessed with structural MRI. However, little evidence is available in young adults.

Objective:
We investigated the associations of high BP with brain volumes and cortical thickness in healthy young adults. We hypothesized that high BP is associated with lower brain volume and cortical thickness in young adults.

Methods:
This cross-sectional study included 1095 young adults (54% women, 22-37 years) from the Human Connectome Project (HCP) who self-reported not having a history of hypertension or taking antihypertensive medications. Brachial systolic (SBP) and diastolic BP (DBP) were measured with semi-automatic or manual sphygmomanometer during study visits. Structural MRI was used to measure gray matter (GM) and white matter (WM) volume and mean cortical thickness. Associations of BP and hypertension stage with total and regional brain volumes and cortical thickness were analyzed using linear regression and analysis of covariance (ANCOVA) after adjusting for age, sex, education years, body mass index (BMI), smoking, alcohol consumption history, zygosity, and total intracranial volume.

Results:
SBP and DBP were (mean ± SD) 123.6 ± 14.2 and 76.5 ± 10.6 mmHg, respectively (n = 1095). High DBP was associated with lower total GM (
p
= 0.012), cortical GM (
p
= 0.004), subcortical GM (
p
= 0.012), and total WM volumes (
p
= 0.031). There were no associations of SBP and hypertension stage with total brain volume and cortical thickness. High SBP and DBP were associated with lower regional cortical thickness in the parietal lobe and lingual gyrus, and lower regional cortical volume in the postcentral gyrus, inferior parietal lobe, lateral occipital gyrus, and middle temporal gyrus.

Conclusion:
Associations of high BP, particularly DBP, with lower total and regional brain volume and cortical thickness were observed in healthy young adults. These findings suggest the importance of prevention or control of high BP early in life for brain health.
Abstract 57: Macrolides for KCNJ5–mutated Aldosterone-Producing Adenoma (MAPA): a Study for Personalized Management of Primary Aldosteronism
Authors: ROSSI Gian Paolo , Seccia Teresa , Caroccia Brasilina , Bressan Alessandro , Rossitto Giacomo , Byiringiro Clement , Ceolotto Giulio , Briones Ana
Published: October 2024

Background:
Somatic mutations in the KCNJ5 potassium channel involve up to 70% of the patients presenting with primary aldosteronism (PA) caused by an aldosterone-producing adenoma (APA). Macrolides corrected the altered function of the two common mutations G151R and L168R,
in vitro
suggesting their potential usefulness for personalized management of patients with KCNJ5-mutated APA. Hence, we investigated if the macrolide roxithromycin that restored the KCNJ5 channel function could lower plasma aldosterone and blood pressure (BP) in KCNJ5-mutated APA patients.

Methods and Design:
In a within-patient study we assessed changes of plasma aldosterone, active renin, cortisol and BP induced by an oral roxithromycin (150-300 mg) challenge, in consenting consecutive hypertensive patients. In parallel, we investigated the vascular and BP effects of macrolides
ex vivo
and
in vivo
in mice.

Results:
We recruited 425 consecutive patients: 19 had G151R - or L168R - mutated APA, 27 KCNJ5 wild-type APA, and the rest had no identifiable cause of hypertension. Roxithromycin lowered the plasma aldosterone concentration (p<0.001), not BP, in the G151R- and L168R-mutated APA patients, but not in the wild-type. However, an aldosterone-independent BP decrease was seen in the much larger cohort of hypertensive patients without PA. In mice experiments, studies showed that this was accounted far by a significant anti-hypertensive effect of macrolides that involved an endothelium-NO-dependent vasodilatory action.

Conclusions:
The roxithromycin-induced fall of plasma aldosterone concentration might be a marker of the presence of KCNJ5-mutated APA. Moreover, our studies unveiled an anti-hypertensive effect of macrolides that involves endothelium- and NO-dependent vasodilation.
Abstract 58: The 5-HT7 receptor contributes to increased hindquarter blood flow caused by skeletal muscle contraction
Authors: Krieger-Burke Teresa , Garver Hannah , Yoder Elise , Watts Stephanie , Fink Gregory
Published: October 2024

Exogenous serotonin (5-hydroxytryptamine, 5-HT) causes a reproducible decrease in blood pressure in normotensive and hypertensive rats. This is associated with dilation of arterioles in skeletal muscle. The 5-HT7 receptor KO rat strain allowed us to test the role of this receptor in both responses as well as the effects of endogenous 5-HT -- acting at 5-HT7 receptors -- on normal circulatory regulation. First, we hypothesized that the 5-HT7 receptor KO rat would have a higher hindquarter (HQ) vascular resistance (HQVR) than wildtype (WT) because of loss of this receptor. In rats anesthetized with isoflurane, instrumented with arterial catheters and flow probes on the terminal aorta, male KO rats had significantly higher resting HQVR [mm Hg/ml/min; KO (16.0±2.0) vs WT (10.8±0.6.0), p = 0.04] and lower HQ blood flow than normal Sprague-Dawley (SD) rats. No differences in resting HQVR or HQ flow were observed in female rats. Intravenous infusion of 5-HT (50 ug/kg/min) reduced systemic blood pressure and HQVR; these parameters were significantly attenuated (~56%) in male KO rats versus WT and SD animals. In females, falls in systemic blood pressure and HQVR were absent in both KO and WT rats. Second, as 5-HT has been implicated in the pathophysiology of heart failure (HF), a left anterior descending artery ligation model of HF was used to investigate the potential role of 5-HT7 regulation of HQ flow in HF-associated exercise intolerance. The ability of hindlimb muscle stimulation to increase blood flow using transdermal neuromuscular electrical stimulation was tested (NMES). M and F KO rats showed a reduced ability to increase HQ flow during muscle contraction vs WT (% over basal: M WT = 118±18 vs KO =14.6±7.1; F WT= 101±12 vs KO = 7.6±6) rats. This was observed in both intact and HF rats (in which flow changes were already impaired). Acute administration of the 5-HT7 receptor antagonist SB269970 abolished the ability of NMES to increase HQ flow in normal SD rats. Importantly, the microcirculatory capacity, visualized by Lectin-594, was not significantly different in the gastrocnemius muscle of WT (arbitrary fluorescence units/area: 946±40) vs KO (914±69). These data support that the 5-HT7 receptor, likely activated by endogenous 5-HT, is an important regulator of skeletal muscle vascular resistance. Thus, 5-HT may have a larger role in normal and pathophysiological regulation of the circulation than has previously been appreciated.
Abstract 61: Protective role of caspase-1 inhibition in experimental model of neonatal hyperoxia-induced cardio pulmonary dysfunction in adults born preterm.
Authors: Ballengee Sydne , Tian Runxia , Jamal Aden , Machi Jacqueline , Schmidt Augusto , Rodrigues Claudia , Young Karen , Wu Shu , Benny Merline
Published: October 2024

Background:
Adults born preterm are at an increased risk of systemic hypertension and cardiopulmonary morbidities. However, little is known of the underlying mechanisms connecting neonatal hyperoxia (O
2
) exposure to these long-term cardiovascular consequences in adults born preterm. Pro-inflammatory caspase-1 mediates the activation of cytokines IL-1β, and pore-forming protein Gasdermin-D.

Objective:
Test the hypothesis that caspase-1 inhibition prevents long-term cardiopulmonary consequences of preterm birth.

Design/Methods:
Newborn mice (n=60) exposed to normoxia (RA) or hyperoxia (O
2
) from postnatal day (P) 1 to 14 were randomly assigned to receive daily intraperitoneal (IP) injections of caspase-1 inhibitor (VX-765, 50 ug/Kg) or DMSO as placebo. The mouse pups recovered in RA into adulthood until 3 months of age. The effect of caspase-1 inhibition on Blood pressure (BP), exercise endurance, cardiac function, systemic vascular stiffness, pulmonary vascular remodeling was assessed at 3 months.

Results:
Adult rats exposed to neonatal O
2
had significantly increased BP associated with vascular stiffness, reduced exercise capacity, cardiac hypertrophy, and cardiovascular inflammation. Caspase-1 inhibition prevented systemic hypertension, systemic vascular stiffness, improved exercise tolerance and decreased cardiac hypertrophy (Fig 1A-D) in adult mice exposed to neonatal O
2
. Moreover, caspase -1 inhibition improved pulmonary vascular remodeling and pulmonary hypertension in these mice (Fig 2A-E). Additionally, western blot analysis demonstrated that early supplementation of VX-765 in O
2
exposed mice led to reduced cardiac proinflammatory markers GSDMD and IL-1β in adulthood (Fig 3A-B).

Conclusion:
Neonatal O
2
exposure predisposes to cardiopulmonary morbidities in adults born preterm. Inhibition of caspase-1 reduces cardiovascular inflammation and cardiopulmonary dysfunction in mice exposed to hyperoxia during the neonatal period. The functional and molecular impact of the use of caspase-1 suggests that it can be used as a novel preventive strategy for long-term cardiopulmonary dysfunction due to neonatal O
2
exposure in preterm survivors.
Abstract 63: Acetylation of Mitochondrial Cyclophilin D Increases vascular Oxidative Stress, Induces Glycolitic Switch, Promotes Endothelial Dysfunction and Hypertension
Authors: Dikalov Sergey , Fehrenbach Daniel , Mayorov Vladimir , Panov Alexander , Ao Mingfang , Lantier Louise , Amarnath Venkataraman , Lopez Marcos , Billings Frederic , Sack Michael , Dikalova Anna
Published: October 2024

Background:
Nearly half of adults have hypertension, a major risk factor for cardiovascular disease. Mitochondrial hyperacetylation is linked to hypertension, but the role of acetylation of specific proteins is not clear. We hypothesized that acetylation of mitochondrial Cyclophilin D (CypD) at K166 contributes to endothelial dysfunction and hypertension.

Methods:
To test this hypothesis, we studied CypD acetylation in patients with essential hypertension, defined a pathogenic role of CypD acetylation in deacetylation mimetic CypD-K166R mutant mice and endothelial specific general control of amino acid synthesis 5 like 1 (GCN5L1) deficient mice using an angiotensin II model of hypertension.

Results:
Arterioles from hypertensive patients had 280% higher CypD acetylation coupled with reduced Sirt3 and increased GCN5L1 levels. GCN5L1 regulates mitochondrial protein acetylation and promotes CypD acetylation which is counteracted by mitochondrial deacetylase Sirt3. In human aortic endothelial cells GCN5L1 depletion prevents superoxide overproduction. Deacetylation mimetic CypD-K166R mice were protected from vascular oxidative stress, endothelial dysfunction and angiotensin II-induced hypertension. Angiotensin II-induced hypertension increased mitochondrial GCN5L1 and reduced Sirt3 levels resulting in 250% increase in GCN5L1/Sirt3 ratio leading to imbalance between GCN5L1-mediated acetylation and Sirt3-mediated deacetylation and promoting CypD acetylation. Treatment with mitochondria-targeted scavenger of cytotoxic isolevuglandins, mito2HOBA, normalized GCN5L1/Sirt3 ratio, reduced CypD acetylation, and attenuated hypertension. The role of mitochondrial acetyltransferase GCN5L1 in endothelial function was tested in endothelial specific GCN5L1 knockout mice. Depletion of endothelial GCN5L1 prevented angiotensin II-induced mitochondrial oxidative stress, reduced maladaptive switch of vascular metabolism to glycolysis, prevented inactivation of endothelial nitric oxide, preserved endothelial-dependent relaxation, and attenuated hypertension.

Conclusions:
These data support the pathogenic role of CypD acetylation in endothelial dysfunction and hypertension. We suggest that targeting cytotoxic mitochondrial isolevuglandins and GCN5L1 reduces CypD acetylation which may be beneficial in cardiovascular disease.
Abstract 70: Cellular senescence as a driver of preeclampsia
Authors: Suvakov Sonja , Gojkovic Tamara , Hatamova Jennet , Dokic Vladimir , Gavrilovici Paul , Garovic Vesna
Published: October 2024

Background:
Preeclampsia (PE) is a severe hypertensive disorder of pregnancy of unknown pathophysiology. We previously have reported the presence of senescence in kidney and fat tissue of PE women, as well as accelerated epigenetic aging, which prompted us to postulate that cellular senescence may drive development of PE.

Methods:
We prospectively collected sera from pregnant women throughout pregnancy, and using multiplex Luminex analysis, measured 50 markers of the senescence-associated secretory phenotype (SASP). Comparisons were made between women who developed PE (n=10) vs. normotensive pregnancies (n=10). We then injected human PE serum into the IL-10KO mouse to induce PE and assess whether PE is associated with development of senescence. We measured plasma p16, a circulating indicator of cellular senescence, before and after inducing PE. We also compared plasma p16 and expressions of senescence-associated genes in the hearts and aortas of regular pregnant (PG, no PE serum injection) and PE mice (n=4).

Results:
There was a significant increase in several SASP markers in sera of women who developed PE, beginning in the first trimester, through delivery. Notably, eotaxin, MCP1, GROb, MIF, MMP2, TNFRII, IGFBP2 were significantly increased in PE women compared to their normotensive peers throughout all trimesters. Mice injected with PE serum exhibited all features of PE, as reported earlier. When measuring plasma p16 before and after PE serum injection, animals had higher values upon inducing PE. Plasma samples of PE mice demonstrated higher p16 concentrations compared to the PG group. The aortas of PE mice exhibited increased expressions of senescence associated genes p16, il6, and mcp1, whereas in the hearts, p16, Il1a and il8 were upregulated.

Conclusion:
The SASP is present very early in human pregnancy, even before PE has developed. Injection of SASP-enriched PE serum into IL-10KO mice induces cellular senescence in different tissues resulting in a PE phenotype. These results suggest that senescence may be an underlying mechanism of PE.
Abstract 71: Analysis of First Morning Urine Transcriptomes in Normotensive and Hypertensive Patients Identify Upregulated Inflammatory and Signaling Pathways Associated with Hypertension
Authors: Umanath Kausik , Sohaney Ryann , Atchison Douglas , ABRAHAM EMMY , Meng Ze , She Ruicong , Adrianto Indra , Levin Albert , Wu Andrew , Ortiz Pablo
Published: October 2024

Introduction:
Hypertension (HTN) remains poorly controlled at the population level. The kidney plays a prominent role in HTN, but the molecular mechanisms are unclear. We enrolled a cohort of patients with HTN, and controls, to study new molecular signatures in the transcriptome of cells shed in first morning urine samples.

Methods:
We collected demographics, 24-h and first-morning urine collections from 80 non-diabetic patients aged >40 with HTN and 20 controls (no HTN). Descriptive variables were summarized. Gene expression was assessed after Next Gen sequencing of mRNA obtained from cells pulled down from first-morning urine and assessed for differentially expressed genes (DEG) (<1 Log2 fold-, p<0.05 FDR) by the Wilcoxon Rank-Sum test. Pathway analysis for DEGs was performed with an in-house Python script that highlights genes associated with cardiovascular diseases including HTN, CKD, type 2 diabetes, inflammation, surface proteins (GPCR, transporters, etc), and the kinome.

Results:
Cohort characteristics are shown in Table 1. After QC, full transcriptomes were analyzed for 11 control and 34 HTN subjects. 20,800 transcripts were identified (including known genes, ORFs and LncRNAs) and compared between groups. 916 DEGs (FDR p<0.05) were identified: 911 genes up- and 5 down-regulated. In HTN subjects, there was overrepresentation of genes in inflammatory signaling pathways (12.5% of all DEGs) including those in IFN-alpha, IFN-gamma, Interleukin (IL)9, IL17 and IL18, and innate immunity. 15 genes, associated with HTN, and 45, associated with type-2 diabetes, at the genome-wide level were up-regulated in the HTN cohort including (LINC00592, ADRA1D, SLC39A12, WNT3A, RXFP2, DEFB136, PDE3A, OPRM1, RIBC2, GRIN2A, CHEK2, HOXA3, CCDC149, COL4A2). Transcripts for major apical nephron NaCl transporters were not significantly different, however, SLC12A5 (K/Cl cotransporter KCC2) and the voltage gated K channels (KCNE1, KCNE2) were up-regulated in HTN patients.

Conclusions:
We conclude that analysis of transcriptomes of cells shed in first morning urine of control and HTN patients allows for the identification of DEG. These data align well with our previous finding indicating that renal inflammation is associated with difficult-to-control hypertension, and as shown here, are also associated with HTN in general. This method allows for discovery of new inflammatory pathways that could render additional treatments for HTN or CKD.
Abstract 73: Extracellular vesicles-derived miR-125b-5p/miR-125a-5p promotes hypertension by regulating angiotensin-converting enzyme 2
Authors: Tarhriz Vahideh , MohammadiNasr Mina , Xia Huijing , Lazartigues Eric
Published: October 2024

Therapeutic targeting of non-coding RNAs, particularly microRNA (miRNA), presents an appealing approach in the treatment of chronic diseases such as resistant hypertension. Recently, miRNA have emerged as promising targets for therapeutic intervention, capable of regulating critical genes to provide a suitable solution to the lack of efficacy of current treatments. Our comprehensive miRNA profiling of plasma extracellular vesicles (EV) from DOCA-salt hypertensive mice of both sexes, compared to sham controls, identified two miRNA implicated in salt-sensitive hypertension, namely miR-125b-5p and miR-125a-5p. Our bioinformatic analysis suggests that these miRNA could be involved in angiotensin-converting enzyme 2 (ACE2) downregulation in hypertension. The present study intended to validate ACE2 as a target of miR-125. Quantitative RT-PCR and capillary western analysis of hypothalami from C57BL/6J DOCA+Salt hypertensive male and female mice following 3-weeks of DOCA+Salt treatment (n=5-6 per group, 3-month old), detected a significant decrease in ACE2 mRNA (males: 0.34±0.03
vs.
1.02±0.02; females: 0.65±0.08
vs.
1.01±0.01; P<0.05) and protein (males: 0.17±0.02
vs.
0.31±0.05; females: 0.16±0.03
vs.
0.37±0.07; P<0.05) expression for both sexes. Further investigation in bEND.3 cells treated with different concentrations of Ang-II (0, 10, 100 nM) for 12, 24, and 48 hours, demonstrated a dose-dependent negative correlation between miR-125b-5p (r≈-0.9978) or miR-125a-5p (r≈-1.002) and ACE2 gene expression, independently of time. When treated with miR-125 mimics the cells demonstrated a consistent downregulation of ACE2 protein levels (miR-125b-5p: 0.6±0.1 and miR-125a-5p: 2.0±0.2
vs.
control: 2.8±0.3). Conversely, no significant change in ACE2 protein levels was observed with the respective miRNA inhibitors. Using a firefly/Renilla luciferase assay, we further identified ACE2 as a validated primary target for these miRNA. While miR-125b-5p indirectly modulates the expression of the
Ace2
gene by targeting important transcription factors like Sp1 and STAT3, our data further elucidate that miR-125b-5p and/or miR-125a-5p directly reduce ACE2 levels by targeting ACE2 3’UTR mRNA. Additional work is warranted to investigate these specific regulatory mechanisms based on cell specificity, physiological conditions, and sex differences.
Abstract 75: CTCF and Rad21 Mediated Regulation of Gene Expression in Human-Induced Pluripotent Stem Cells-derived Endothelial Cells
Authors: Ray Atrayee , Pandey Rajan , Stelloh Cary , Liu Pengyuan , Liu Yong , Kwitek Anne , Geurts Aron , Cowley Allen , Greene Andrew , Liang Mingyu , Rao Sridhar
Published: October 2024

Blood pressure (BP)-relevant non-coding SNPs identified by GWAS may influence the expression of distal protein-coding genes. Our previous study detected 195 SNPs (out of 26585) in CTCF-binding sites. CTCF and Cohesin are known to modulate cell-type-specific gene expression from a distance by forming chromatin loops. We hypothesized that both may regulate gene expression in BP-relevant cell types. To test this, we depleted CTCF and Rad21 (Cohesin subunit) in human induced pluripotent stem cells-derived endothelial cells (iEC) and performed genome-wide transcriptomic analysis.

We detected 366 and 4184 differentially expressed genes (DEGs) in CTCF and Rad21 depleted iEC (CTCF-/Rad21-KD-iEC) compared to iEC control, respectively. Among them, 111 DEGs are common between the CTCF-KD and Rad21-KD-iEC, though majority showed opposite regulatory effects (Figure 1A). We found 5619 DEGs in the CTCF-KD vs Rad21-KD-iEC (Figure 1B). Pathway analysis detected elevated cell cycle (G2M checkpoint), DNA replication (E2F targets), and repair pathways in Rad21-KD compared to the CTCF-KD-iEC. Rad21 plays an important role in chromosome partitioning and repair, so its depletion may cause dysregulation of these pathways. While the CTCF-KD-iEC showed enrichment of the Epithelial-to-mesenchymal transition (EMT), inflammatory (IFN-gamma, IFN-alpha), and hypoxia pathways (Figure 1C). As CTCF maintains the chromosome boundaries, CTCF-KD may affect chromatin compactness and trigger abnormal gene expression related to these pathways.

In conclusion, our study detected different effects of depletion of CTCF and Rad21 on gene expression in a BP-relevant cell type. The findings provide a basis to explore the role of chromatin looping in BP-relevant gene regulation.
Abstract 76: CASC15 lncRNA as a Mediator of Vascular Senescence in the Renin-Angiotensin System
Authors: Cunha Warlley , Martin Maria , Ahmed Ibrahim , Gomez Delphine , Espinosa-Diez Cristina
Published: October 2024

The renin-angiotensin system (RAS) regulates vascular function and maintains blood pressure homeostasis. Angiotensin-II (Ang-II) and angiotensin-1-7 (Ang-1-7) have opposing effects on vascular health. Ang-II promotes vascular senescence and pathophysiological remodeling by inducing oxidative stress, inflammation, vascular hypertrophy, and endothelial dysfunction. In contrast, Ang-1-7, known for its vasodilatory, anti-inflammatory, and antioxidative properties, counteracts these detrimental effects, enhancing endothelial function, reducing oxidative stress, and inhibiting inflammation.

Our laboratory has identified the long non-coding RNA CASC15 as a critical regulatory element within the RAS. CASC15 expression is significantly repressed by Ang-II in aortic vascular smooth muscle cells (SMCs), showing a 59.25% decrease compared to the vehicle control (n=10, p=0.000333). Loss of CASC15 induces cellular senescence, increasing senescent cells from a mean of 14.09% (control inhibitor, n=9, SEM=4.824) to 57.79% (CASC15 inhibitor, n=9, SEM=6.420, p<0.0001), and leads to vascular hypertrophy, with cell area increasing from a mean of 46.89% (control inhibitor, n=8, SEM=9.229) to 221.2% (CASC15 inhibitor, n=9, SEM=67.53, p=0.0300).

We hypothesize that CASC15 repression by Ang-II (1 µM) contributes to vascular senescence and increased DNA damage, while Ang-1-7 (1 µM) preserves CASC15 expression and mitigates these effects. Pro-senescent stressors like hydrogen peroxide and doxorubicin also repress CASC15 expression in SMCs. Using RT-qPCR, we observed decreases in CASC15 expression by 29.56% (hydrogen peroxide vs. vehicle, n=4, p=0.0790) and 44.09% (doxorubicin vs. vehicle, n=3, p=0.0733). Ang-1-7 alone did not significantly change CASC15 expression, but combined with Ang-II, it showed potential recovery from 40% repression to 94% recovery. Further research is needed. Immunofluorescence against gamma-H2AX revealed increased DNA damage in SMCs lacking CASC15, with positive foci increasing from 26.27% (control inhibitor, n=4, SEM=1.459) to 63.71% (CASC15 inhibitor, n=4, SEM=1.806, p=0.0009).

In conclusion, CASC15 is a crucial regulator within the RAS. Its repression by Ang-II promotes vascular senescence, hypertrophy, and DNA damage, effects countered by Ang-1-7. Targeting CASC15 could offer new strategies for mitigating vascular aging and improving vascular health, particularly in chronic kidney disease.
Abstract MP02: Inhibition of RORγt Ameliorates Angiotensin II-Induced Blood-Brain Barrier Disruption and Microglia Activation by Reducing Interleukin-17A Production in Rats
Authors: Yu Yang , Xue Baojian , Bassuk Alexander , Johnson Alan , Wei Shunguang
Published: October 2024

Inflammation-induced blood-brain barrier (BBB) disruption and microglial activation in the autonomic and cardiovascular brain centers contribute largely to sympathetic excitation and hypertension. We recently reported that interleukin (IL)-17A, a major mediator of tissue inflammation, promotes BBB disruption, neuroinflammation, and sympathetic activation in angiotensin (ANG) II-induced hypertension and myocardial infarction-induced heart failure. The nuclear receptor retinoid-related orphan receptor γt (RORγt) has been identified as a master transcription factor regulating the differentiation of T helper 17 cells, the primary producer of IL-17A. The present study sought to determine whether inhibition of RORγt to suppress IL-17A production can ameliorate BBB disruption and attenuate microglia activation in ANG II-induced hypertension model. SD rats were treated with a 2-week subcutaneous (SC) infusion of ANG II (150 ng/kg/min) via mini-pumps, combined with daily SC injections of RORγt inhibitor digoxin or vehicle (VEH). Compared to the VEH control, IL-17A levels in plasma and cerebrospinal fluid were significantly (*P<0.05) elevated in ANG II+VEH rats and were reduced in ANG II + digoxin rats. In ANG II+VEH rats, BBB permeability, measured by fluorescein isothiocyanate-dextran (FITC) 10 kDa, was elevated (6.82±0.61%* vs. 2.36±0.28% FITC10 extravasation), along with increased mRNA levels of caveolin-1 (a transcytosis marker, 2.26±0.39* vs. 1.07±0.17) and decreased mRNA levels of mfsd2a (a BBB-specific lipid transporter and inhibitor of transcytosis, 0.55±0.04* vs. 1.06±0.09) in the hypothalamic paraventricular nucleus (PVN). However, BBB permeability and mRNA levels of caveolin-1 and mfsd2a in the PVN were reversed in ANG II + digoxin rats. Immunostaining revealed that both caveolin-1 and mfsd2a were primarily located in the endothelial BBB within the PVN. Microglia activation, assessed by the number of branches (26±2* vs. 37±4) and total branch length (184±15* vs. 250±23), as well as the mRNA levels of tumor necrosis factor-α (2.78±0.45* vs. 1.04±0.12) and IL-1β (2.57±0.53* vs. 1.03±0.14) in the PVN were augmented in ANG II+VEH rats but were attenuated (40-58%*) in ANG II + digoxin rats. Additionally, ANG II-induced hypertension and sympathetic tone were attenuated by digoxin. These data suggest that inhibiting RORγt diminishes BBB disruption, microglial activation and sympathetic excitation induced by ANG II, likely via reducing IL-17A production.
Abstract MP08: Long-Term Impact of Bariatric Surgery on Hypertension Control and Remission: A Meta-Analysis of Randomized Controlled Trials
Authors: Sebastian Sneha Annie , Shah Yash , Krishnamoorthy Geetha
Published: October 2024

Background:
Bariatric surgery is widely recognized as a highly effective intervention for addressing obesity and its associated metabolic complications. However, there is a paucity of evidence from randomized controlled trials (RCTs) with primary endpoints focusing on its impact on hypertension.

Objective:
We aimed to synthesize findings from pivotal RCTs until May 2024 to evaluate the long-term effects of bariatric surgery on hypertension in obese individuals compared to medical or lifestyle management.

Methods:
We analyzed data using RevMan 5.4 with a random effects model, employing the inverse variance method to report outcomes as risk ratios (RR) or weighted mean differences (WMD) with 95% confidence intervals (CI). Our study protocol is registered in PROSPERO (CRD42024527369).

Results:
The study comprised 18 RCTs involving 1386 obese patients (62.7% women) with a mean BMI of 38 kg/m
2
and an average follow-up duration of 1 to 5 years. On pooled analysis, there was a statistically significant difference in the rate of hypertension remission between patients who underwent bariatric surgery versus patients on medical/lifestyle management with an RR of 2.77 (95% CI: 1.26 to 6.10, p = 0.01). Additionally, a substantial reduction in the use of antihypertensive medications while maintaining controlled blood pressure was observed (RR: 7.10; 95% CI: 4.38 to 11.51, p < 0.00001), with no heterogeneity. Bariatric surgery also significantly improved systolic blood pressure control (WMD: -3.67; 95% CI: -5.53 to -1.80, p = 0.0001).

Conclusion:
Bariatric surgery emerges as a durable solution for obesity-related hypertension, offering remission and control of hypertension while reducing the dependence on antihypertensive medications.
Abstract MP11: Evaluation of a Remote Patient Monitoring Hypertension Program with Telehealth Pharmacist Management in Nephrology Clinic Patients
Authors: Bitton Neria , Krumich Christine , Webster LeeAnn K , Andersen Stacy , Quattrocchi John , Gendy Fady , Namjouyan Kamran , Bucaloiu Ion , Jones Thomas , Bali Atul , Fry Emily , Chang Alexander
Published: October 2024

Introduction:
Remote patient monitoring (RPM) and physician-pharmacist collaborative models have potential in improving hypertension control. Herein, we report the effectiveness of a community based, self-measured blood pressure (SMBP) RPM program on BP control and healthcare utilization in a largely rural health system.

Methods:
A multidisciplinary team developed a self-measured blood pressure (SMBP) RPM program in central and northeast PA using a virtual platform with bluetooth-connected BP devices targeting patients with BP ≥140/90 mmHg seen in nephrology clinics. From 3/2022 to 9/2022, high BP notifications were transmitted to nephrologists. After 9/2022, BP notifications were transmitted first to pharmacists, who co-managed BP through collaborative practice agreements via telehealth. We captured pharmacist interventions by reviewing electronic health record documentation. Changes in SMBP over time and the effect of pharmacist interventions were analyzed using mixed effects models. All-cause hospitalizations and emergency room (ER) visits were compared in 6-month pre/post-enrollment periods.

Results:
A total of 205 patients (mean age 62.3y, clinic BP 146/84, 3.1 BP classes, 48% female, 53% CKD) were enrolled into the program and had ≥ 6 months follow-up data. SMBP control <140/90 improved from 30% during week 1 to 67% at 6 months and 74% at 12 months. Overall, systolic SMBP improved over the first 6 months with stronger effect on patients with higher baseline BP (baseline BP ≥150/100 mmHg: -3.3 mmHg/month (95% CI: -3.5, -3.1); baseline 140-149/90-99: -2.4 (95% CI: -2.6, -2.1) mmHg/month; baseline <140/90: -0.6 (-0.9, -0.4) mmHg/month). Pharmacist telehealth encounters were documented in 65% of patients; 46% had a BP medication adjustment; 37% had new BP medication classes, including 9% mineralocorticoid receptor antagonist. Pharmacist medication intervention was associated with greater decline in systolic BP over time (-1.3 mmHg/month; 95% CI: -1.6, -1.1). Comparing the 6 months pre vs. post-enrollment, patients experienced a reduction in hospitalizations (6.0% vs. 0%; Exact Mcnemar p=0.0005) and no difference in ER visits (14.9% vs. 12.9%; p=0.6).

Conclusion:
Participation in a nephrologist-pharmacist SMBP RPM program was associated with improvement in BP control and reduced hospitalizations. Future efforts are underway to refine delivery of this program for the largest impact on improving BP control.
Abstract MP12: Reproducibility of White Coat Hypertension and White Coat Uncontrolled Hypertension Among U.S. Adults
Authors: Giacona John , Rugovac Adzona , Kotamraju Manasa , Sundaram Venkatraghavan , Gates Jay , Vazquez Miguel , Vongpatanasin Wanpen
Published: October 2024

White coat hypertension (WCH) and white coat uncontrolled hypertension (WUCH) are phenomenon characterized by uncontrolled office blood pressures (BP) in the presence of normal out-of-office BPs among untreated and treated individuals, respectively. Although prior studies have investigated the reproducibility of WCH and WUCH, they were mainly limited to non-U.S. adults.

We assessed white coat status in 272 (75% treated) adults with elevated office BP(≥130/80 mmHg) who had home BP (HBP) measurements for three consecutive months without initiation or intensification of antihypertensive medications during the three-month time interval of interest. HBP measurements were included only when number of readings were ≥12/month. WCH and WUCH was defined as HBP≤130/80 mmHg in untreated and treated adults, respectfully. The reproducibility was assessed using the Kappa coefficient (K) with 95% confidence intervals (CI).

Mean age was 76±5 years (57% women, 12% Black adults). WCH was found in 39 of the 68 untreated adults with elevated office BP. WCH adults had an average home BP of 118/72±8/5 mmHg after the initial month of HBP monitoring (HBPM). As shown in
Figure 1
, 87% and 85% had WCH status confirmed on repeat assessment after an additional one- and two-month interval of HBPM (K=0.54 and 0.48, respectfully). WUCH was found in 92 of the 204 treated adults with elevated office BP and had an average home BP of 120/71±5/5 mmHg. Of those, 86% and 84% had WUCH confirmed on repeat assessment after an additional one- and two-month interval of HBPM (K=0.65 and 0.52, respectfully).

In conclusion, WCH and WUCH phenotypes when assessed by home blood pressure monitoring exhibited fair-to-moderate reproducibility among U.S. adults.
Abstract MP14: Identifying markers of Concentric Arterial and Arteriolar Hypertrophy (CAAH) Using Multiomics.
Authors: Almeida Lucas , Medrano Silvia , Smith Jason , Yamaguchi Manako , Gomez Ariel , Sequeira Maria Luisa
Published: October 2024

Experimental or spontaneous mutations of any of the renin-angiotensin system (RAS) genes or treatment with RAS inhibitors in mammals lead to kidney
concentric arterial and arteriolar hypertrophy (CAAH).
Despite the prevalent use of RAS inhibitors in children and adults, our knowledge of this disease is very limited.

We assessed the hypothesis that mouse models of CAAH will produce identifiable and reproducible markers of early and progressive disease.

We generated murine models that tracks the lineage of renin-producing cells with deletion (

Ren1
c-/-
;Ren1
cCre
;R26R
mTmG

, Ren1cKO) or an intact renin gene (

Ren1
c+/-
;Ren1
cCre
;R26R
mTmG

, control). Kidney cells expressing GFP and tdTomato were isolated by FACS and processed for scRNA-seq. Plasma and urine were collected for proteomics, metabolomics and lipidomics. To study kidney function and structure, we measured transdermal glomerular filtration rate (tGFR) and performed comprehensive three-dimensional (CUBIC) and immunohistochemical analyses.

Ren1cKO mice presented with a decrease in the GFR (1.47±0.02
vs
0.40±0.06 mL/min/100g b.w (
n=
6;
P
<0.05)). They also had polydipsia (3.9±0.05
vs
9.6±0.09 mL/24h (
n=
6;
P
<0.05), polyuria (2.4±0.07
vs
5.9±0.09 mL/24h (
n=
6;
P
<0.05) and marked reduction in urinary osmolality (1384±92.09
vs
588±68.04 mOsm/kg (
n=
6;
P
<0.05). The wall thickness of the afferent arterioles in Ren1cKO mice was significantly increased when compared to controls (4.54 ± 0.17 μm
vs
11.57 ± 0.91 μm,
P
<0.05). The hypertrophic arterioles were covered along their lengths with cells positive for renin. Plasma metabolites showed a significant increase in fatty acids and phosphatidylcholines (
P
<0.05) in Ren1cKO mice. Collagens (
P
<0.05), lymphocyte antigens (
P
<0.05), Spink 1 (
P
<0.05), and cadherin 13 were differentially abundant with cadherin 13 exclusively detected in Ren1cKO mice at 1 and 6 months. Our scRNA-seq data also found Spink 1 (
P
Holm-adj
.=0.00) and Cadherin 13 (
P
Holm-adj
.=5.49X10
3
) significantly increased in renin null cells, corroborating the proteomic findings.

1) CAAH involves smooth muscle cell transdifferentiation to renin-expressing cells that exhibit a mesenchymal phenotype characterized by pro-inflammatory pathways, and deregulation of lipid-metabolic homeostasis. Further, the exclusive expression of Spink and Cadherin 13 suggest their involvement in the pathogenesis of these severe, and silent arteriolar disease. Those molecules may serve to mark the initiation and progression of CAAH.
Abstract MP23: Arm Size Coverage of Popular Over-the-Counter Blood Pressure Devices and Implications in US Adults
Authors: Kaur Eileen , Rayani Asma , Brady Tammy , Matsushita Kuni
Published: October 2024

Background:
Recent findings show the impact of an inappropriate cuff size on blood pressure (BP) reading. Despite the emphasis on the value of home BP monitoring, little is known about the coverage of arm circumference in popular over-the-counter (OTC) devices.

Aims:
To evaluate the coverage of arm circumference in popular BP devices and quantify the proportion of U.S. adults not eligible for using those devices due to arm size.

Methods:
We searched a large online retailer, Amazon, for ranks, ratings, and number of sold devices and purchased ten of the most popular OTC devices. We recorded data on the cuff sizes and corresponding aim circumference ranges offered in those devices. Subsequently, using data from the National Health and Nutrition Examination Survey 2015-2020 (13,826 adults), we quantified the number of US adults not eligible to use those popular devices due to arm circumference.

Results:
Nine devices offered cuff size covering arm circumference from 22-42 cm, and one from 22-40 cm (
Table
). We found that ~22 million US adults (6.7% of US adults) could not use these OTC devices since their arm circumferences were <22 cm (1.0 million [0.3%]) or >42 cm (21.4 million [6.4%]). The proportion of adults not covered by these devices was higher in Blacks (11.7%) than in other racial/ethnic groups (≤6.5%).

Conclusions:
Despite the emphasis on the importance of home BP monitoring, over 20 million US adults may not be eligible to use popular BP devices due to their arm circumference, and Blacks are more susceptible to this issue than other racial/ethnic groups. Manufacturers of OTC blood pressure devices should address this unrecognized disparity for quality and equitable hypertension diagnosis and management integrating home BP monitoring.
Abstract MP32: Megalin and lysosomal disruption in the kidney epithelial cells of high salt diet fed rats: protective role of angiotensin II type 2 receptor
Authors: Kulkarni Kalyani , Patel Sanket , Hussain Tahir
Published: October 2024

Earlier we have reported that 2-days high salt diet (HSD) feeding caused proteinuria in obese rats and the AT
2
R agonist treatment prevented proteinuria. Since HSD feeding for 2 days and the AT
2
R activation did not affect blood pressure, GFR, glomerular structure, and inflammation, the mechanism responsible for proteinuria remains unknown. Megalin is a proximal tubule epithelial cells endocytic receptor responsible for filtered protein reabsorption. The absorbed proteins are directed to lysosomes for degradation. Lysosomes is a nutrient sensing subcellular compartment and a critical part of the endocytic machinery and cellular trafficking.
We hypothesized that high salt intake caused lysosomal stress, which negatively impacted megalin-protein cellular trafficking and thus caused proteinuria.
Therefore, we evaluated the markers of lysosomal stress and megalin expression in the proximal tubules of HSD fed obese Zucker rats. Urinary excretion of N-acetyl-glucosaminidase (NAG), which is a marker of lysosomal injury and oxidative stress, was significantly increased in HSD fed rats compared with normal salt diet (NSD: 16.6 vs HSD: 40.5 RFU/hr/µg Cr). This increase was associated with an increase in another oxidative stress marker H
2
O
2
in the urine (NSD: 0.37 vs HSD: 1.72 ΔRFU/min). Confocal analysis revealed that megalin was localized in the lysosomes in HSD fed rats, as megalin was co-localized with the lysosomal marker LAMP-1 in the proximal tubules. Moreover, in HSD fed rats there was a remarkable increase (2-fold) in the lysosomal size (fig) and number per tubule (NSD: 18 vs HSD: 48). The AT
2
R agonist C21 (0.3 mg/Kg/day, i.p.) treatment of HSD-fed rats restored surface megalin expression, reduced urinary NAG and H
2
O
2
and the lysosomal number and size. Our
in vitro
studies performed in OK cells (proximal tubule epithelial cells) treated with high NaCl (100 mM) without/with the AT
2
R agonist C21 (1µM) also revealed that compared to NSD, high NaCl reduced megalin surface expression and C21 treatment prevented it. We conclude that high salt intake causes lysosomal stress. Particularly, large lysosomal size potentially is affecting endocytic machinery, including impairing megalin-protein dissociation and megalin recycling, thus causing proteinuria.