bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 068: Diabetic CD4+T Cells Causes Increased Glucose, Hypertension, Renal Injury, And Decreased Vascular Endothelial Cell Function In A Novel Rat Model Of Diabetes During Pregnancy
Authors: Deer Evangeline M , Williams Jan M , Wilson Mariah , Herrock Owen , Campbell Nathan , Amaral Lorena , Lamarca Babbette B
Published: October 2023

Gestational diabetes mellitus (GDM) is a complication of pregnancy that is characterized by glucose intolerance, insulin resistance, and beta-islet cell dysfunction. Additionally, increased levels of maternal glucose, insulin, inflammatory mediators, abnormal placental vascular function, and reduced renal function are associated with the pathogenesis of GDM, however, we don’t know the importance of placental factors or inflammatory cells causing HTN or other pathophysiological factors associated with GDM. We hypothesize placental CD4+ T cells from pregnant diabetic patients (GDM) play a role in the pathophysiology of GDM by stimulating antibodies leading to β-cell destruction causing characteristics of diabetes during pregnancy. CD4+ T cells were magnetically separated from placentas from GDM patients and injected into normal pregnant athymic nude rats on Gestation Day (GD) 12. On GD19, blood pressure (MAP), blood, tissues, and urine were collected and fasting glucose was measured in GDM CD4+ T cell recipients and pregnant nude control rats. In another set of animals, a glucose tolerance test (GTT) was performed. A student’s t-test was used for statistical analysis. On GD19, MAP increased to 121±2 mmHg (n=8, p<0.05) in GDM T cell recipients compared to normal pregnant (NP) athymic nude control rats (105±3 mmHg, n=8). Blood glucose levels were elevated in GDM CD4+ T cells recipients (201±46 mg/dl, n=8, p<0.05) compared to NP athymic nude controls (82±9 mg/dl, n=5). After performing a GTT, glucose tolerance was impaired in recipients of GDM CD4+ T cells (p<0.05) compared to NP nude controls. Moreover, sera from GDM recipient rats increased endothelial cell mtROS (71±1% Gated, n=7, p<0.05) compared to sera from NP controls (54±4% Gated, n=6). Protein excretion increased in GDM CD4+ T cells recipients (8±0.6 mg/day, n=5, p<0.05) compared to NP nude controls (1±0.1 mg/day, n=4). Collectively, these data indicate that adoptive transfer of GDM CD4+ T cells may result in GDM phenotype in pregnant women by causing impaired glucose utilization and decreased vascular endothelial cell and renal function resulting in hypertension during pregnancy. Thereby, the data introduces a new model to investigate mechanisms and new therapies for diabetes during pregnancy.
Abstract 069: Klf2, A Transcriptional Component Of The Renin Cell Baroreceptor
Authors: Yamaguchi Hiroki , Smith Jason P , Li Minghong , Watanabe Hirofumi , Medrano Silvia , Sequeira Lopez Maria Luisa S , Gomez Roberto A
Published: October 2023

Background:
Renin regulates blood pressure (BP) and fluid homeostasis. Renin cells harbor a pressure sensor, the renin cell baroreceptor that controls renin synthesis and release. We recently discovered that the renin cell baroreceptor is a nuclear mechanotransducer within the renin cells that transmits external pressures to chromatin and regulates
Ren1
expression. However, which transcription factors (TF) regulate renin expression during pressure sensing remains unclear.

Objective:
Define the responsible TFs for
Ren1
as part of the renin cell baroreceptor.

Methods:
Forkhead box protein D1 (FoxD1) expressing stromal cells are the progenitors for juxtaglomerular cells (JG), mesangial cells (MC), pericytes (PC), and vascular smooth muscle cells (VSMC) of the renal arteriole. We generated

FoxD1-GC; R26R
TdTomato

mice, in which all FoxD1
+
cells and their descendants are labeled with tdTomato reporter. We applied a surgical model of aortic coarctation (AoCo) between the roots of the left and right renal arteries and sorted tdTomato
+
cells from the cortices of the left kidney (LK, low pressure) and the right kidney (RK, high pressure). Then, we performed single-cell RNA-seq to identify candidate TFs activated at low pressure. Candidates were then validated and deleted in the renin cells of mice exposed to hypotensive conditions.

Results:
1) By single-cell RNA-seq, FoxD1
+
cells were clustered into JG, VSMC, MC and PC, and other cell types. Differentially expressed gene (DEG) analysis in LK compared to RK showed that
Ren1
was the highest DEG in JG. In VSMC, DEG includes
Ren1
and Kruppel-like factor 2 (
Klf2)
, which showed the highest Log
2
fold change (0.36) among the major TFs. 2) We generated

Klf2
fl/fl
; Ren1
dCre

(
Klf2
cKO) mice, and found a 55.1% decrease of kidney cortices
Ren1
mRNA expression by qPCR (n = 5, P=0.022) and reduced plasma renin levels by ELISA (40.1 ± 9.6 vs 15.1 ± 7.3 ng/mL, n = 12, P<0.001) compared to wild type mice.
Klf2
cKO mice treated with captopril-added water (0.5 g/L) and a low Na
+
diet for 7 days displayed reduced renin immunostaining in the JG and along the afferent arterioles.

Conclusion:
We identified Klf2 as a key TF for
Ren1
as a part of the renin cell baroreceptor. Experiments for
Klf2
cKO mice with AoCo surgery are ongoing.
Abstract 072: Sex Differences In The Relationship Between The Age Of Hypertension Diagnosis And Brain Structure In Midlife And Older Population: New Insights From The Uk Biobank
Authors: Kaur Amanpreet
Published: October 2023

Background:
Hypertension is an established leading risk factor for morbidity and mortality from vascular diseases across the lifespan. Early adulthood to midlife high blood pressure has been linked to later reduced brain volume and white matter hyperintensities, both of which are hallmarks of cerebral small vessel disease (CSVD). However, whether the association of age at diagnosis of hypertension with CSVD-related brain atrophy differs between females and males is not fully understood. Therefore, our objective was to assess for sex differences in the association between the age at diagnosis of hypertension and CSVD-related brain structural changes.

Method:
We used data from the UK Biobank. We selected participants with a known age of diagnosis of hypertension who had also obtained a brain MRI between 2014 and 2016 (n = 9,410) and stratified by age of diagnosis of hypertension (<35 years, 35-44 years, 45-54 years, 55-64 years, ≥65 years old) and sex. Using propensity score matching a control participant with MRI data but no hypertension was chosen at random. Generalized linear models were used to determine changes in brain structure as a function of age of hypertension diagnosis and sex while controlling for vascular risk factors and demographic covariates.

Results:
There was a trend for lower brain grey matter volume (adjusted for head size) in males diagnosed with hypertension at a younger age of diagnosis (45-54 years) (β –8448.5 (95%CI, [–2876.7 to –5662.6]mm
3
), (55-64 years) (β –7977.0 (95%CI, [–3283.0 to –5630.0 ]mm
3
) as well as lower volume of white matter (<35 years) (β –25788.3 (95%CI, [– 1965.7 to –13877.0 ]mm
3
), but not in females. The volume of white matter hyperintensities was greater in both males (β (95%CI, 1488.1 [1149.2 to 1827]mm
3
) and females with hypertension (1700.1 [1351.9 to 2048.3]mm
3
) than those without hypertension; more so in females (P
interaction
=0.03).

Discussion:
Hypertension diagnosed in mid-life was associated with smaller brain grey matter and white matter volumes in males. Even though both males and females with hypertension had a higher volume of white matter hyperintensities, hypertensive females had the highest volume. Therefore, sex-specific analyses are necessary to uncover effect of hypertension related to CSVD.
Abstract 074: Discovery Of Post-Translationally Modified Self-peptides Responsible For T-cell Autoimmunity In Hypertension
Authors: Bloodworth Nathaniel , Chen Wei , Patrick David M , Ao Mingfang , Moretti Rocco , Meiler Jens , Davies Sean S , Harrison David G
Published: October 2023

Introduction:
Isolevuglandins (isoLGs) are lipid peroxidation products that covalently bond lysine residues, forming self-antigens that drive T cell inflammation in hypertension. We created a library of isoLG-adducted self-antigens and tested their ability to activate T cells and induce hypertension.

Methods and Results:
Using the Rosetta protein modeling software we developed a computational pipeline estimating isoLG-modified peptide binding for class I major histocompatilibty complex (MHC-I), creating a screening library of peptide candidates. We pulsed dendritic cells (DCs) with candidate peptides before co-culture with T cells from hypertensive mice stained with CFSE cell tracker dye (Fig 1A). Conjugating an MHC-I probe with candidate peptides showed 11 isoLG adducted peptides were recognized by CD8
+
T cells in the aortas of hypertensive mice (Fig 1B), of which 6 also caused T cell proliferation (Fig 1C-D). We adoptively transferred DCs pulsed with these 6 peptides before treatment with a sub-pressor dose of angiotensin II. 4 of the isoLG-adducted peptides induced hypertension
in vivo
, while unadducted peptides did not (Fig 1E).

Conclusion:
Our work identifies specific self-peptides that, when modified by isoLG, cause T cell activation and hypertension.
Abstract 077: Protein Biomarkers Of Hypertension
Authors: Aggarwal Mohit , Lee Dong Heon , Huan Tianxiao , Jenna Mcneil , Courchesne Paul , Joehanes Roby , Dupuis Josee , O'Connor George , Levy Daniel
Published: October 2023

Background:
Hypertension is a major modifiable riskfactor for cardiovascular disease. The discovery of novel protein biomarkers ofhypertension can provide information on disease pathophysiology and highlightpromising therapeutic targets.

Methods:
We measured plasma levels of 2925proteins in 740 participants (mean age 51.7 ±11.3 years; 46% women) in the Framingham HeartStudy using the Olink Explore proximity extension assay. Linear mixed modelswere used to explore the associations of plasma proteins with systolic anddiastolic blood pressure (BP) and prevalent hypertension. We definedhypertension as having elevated BP (systolic ≥ 140 mmHg or diastolic ≥90 mmHg) or current use of antihypertensive medication. We adjusted all themodels for age, sex, batch effects, eGFR (estimated glomerular filtration rate),pack-years of smoking, and lung function status (normal/obstructive/restrictive). Sensitivity analysis was performed by excluding individuals treated withantihypertensive medication. Using overrepresentation analysis, we annotatedthe proteins associated with hypertension. We adjusted for multiple tests using the Benjamini-Hochberg method and a false discovery rate of < 0.05 wasused to define statistical significance.

Results:
We observed differential expression of 526proteins (465 upregulated, 61 downregulated) in participants with prevalenthypertension (N=257) vs. normal BP (N= 483). Systolic BP was associated with571 proteins and diastolic BP was associated with 662 proteins. We observed 507protein associations with systolic BP and 626 protein associations withdiastolic BP after exclusion of treated individuals (N=177). Functionalannotation of proteins associated with hypertension showed enrichment of genesassociated with fatty acid transport, regulation of MAPK cascade, response tointerleukin-1 and inflammatory response.

Discussion:
This study elucidates protein signaturesassociated with BP and hypertension. Hypertension is enriched in pathwaysrelated to fatty acid transport and inflammatory response. These results mayprovide information on the pathogenesis ofhypertension and highlight promising therapeutic targets.
Abstract 078: Epigenetic Chronotherapy For Hypertension: Sleep-time Supplementation Of 1,3-butanediol Confers Better Protection Against Hypertension Via Enhanced Histone β-hydroxybutyrylation
Authors: Aryal Sachin , Chakraborty Saroj , Mandal Juthika , Mell Blair , Issa Ryan , Manandhar Ishan , Joe Bina
Published: October 2023

The leading global contributor to cardiovascular disease and early death is hypertension. Previously, we demonstrated that the ketone body, β-hydroxybutyrate (BHB), is a potent antihypertensive metabolite. BHB is newly recognized to epigenetically modify histones by β-hydroxybutyrylation (BHBylation). As it is known that BHB is predominantly synthesized when there is a lack of food and blood pressure (BP) is lower while animals are sleeping, we hypothesized that an increased histone BHBylation occurs during sleep-time to promote ketosis and lower hypertension. To test this hypothesis, we examined the epigenetic effect of enhancing systemic BHB on BP of the Dahl Salt-Sensitive (S) rat.

Methods:
Male S rats that were six weeks old and fed on a high salt diet were divided into two groups. They were gavaged daily for nine weeks at two different time points, 2 pm/sleep-time (n=6) and 2 am/awake-time (n=5) with 5g/kg/day of a precursor of BHB, 1, 3-butanediol (1,3-BD). BP was monitored by radiotelemetry until euthanasia when tissues were harvested for further analyses.

Results:
The sleep-time chronotherapy group had significantly lower mean arterial BP over the course of twenty-four hours than the awake-time chronotherapy group (130±4 mmHg vs. 138±8 mmHg, p<0.0001). Renal histone3lysine9 (H3K9) BHBylation examined by western blotting was elevated in the sleep-time compared to the awake-time chronotherapy group. Ketosis, as monitored by the expression of the lipid metabolism genes,
Hmgcs2, Acaa1b, Cyp2d4, and Cyp2e1,
was significantly higher in the sleep-time compared to the awake-time chronotherapy group. Further, the expression of renal injury and inflammation markers KIM1, MCP1, IL-18, IL-16, and TGF-β were significantly lowered in the sleep-time compared to the awake-time chronotherapy group.

Conclusion:
This is the first study to demonstrate that enhanced renal epigenetic histone BHBylation is mechanistically linked to better antihypertensive outcomes during sleep-time compared to awake-time chronotherapy with 1,3-BD.
Abstract 079: Multi-omics Analyses Identify Galectin-1 As A Potential Novel Mediator Of Heart Failure With Preserved Ejection Fraction In Mice And Humans
Authors: Wassenaar Jean W , Smart Charles D , Fehrenbach Daniel J , Dungan Matthew M , Amanda C Doran , Shuey Megan , Madhur Meena S
Published: October 2023

Heart failure with preserved ejection fraction (HFpEF) is a clinical syndrome where patients present with symptoms of volume overload despite having a normal systolic function. HFpEF is associated with elevation of inflammatory biomarkers, but the precise pathways leading to cardiac dysfunction is not known. In a deoxycorticosterone acetate (DOCA)-salt model of hypertension and HFpEF, we performed CITE-Seq analysis on total leukocytes isolated from the heart, which revealed significant differential gene expression primarily in the myeloid population. We took the top 20 genes that were differentially expressed between myeloid cells from DOCA-salt vs sham treated mice and performed a genetic analysis called PrediXcan in Vanderbilt’s DNA databank, BioVU. Of these, the gene encoding galectin 1,
Lgals1
, had the lowest p-value (0.02) and highest beta coefficient (0.32) corresponding to an odds ratio for HFpEF of 1.38. The role of galectin-1 in heart failure is unknown. We hypothesized that
Lgals1
exacerbates HFpEF. We found that siRNA knockdown of
Lgals1
in bone marrow derived macrophages stimulated with lipoprotein saccharide and IFNγ; expressed significantly more inflammatory genes including IL-1α; (p<0.0001), IL-6 (p=0.0219), iNOS (p=0.043), and TNF-α; (p=0.0031) using a 2-way ANOVA followed by Sidak’s multiple comparison test. Next, we explored the role of
Lgals1
in the pathophysiology of HFpEF using
Lgals1-/-
mice. Surprisingly,
Lgals1-/-
mice did not show a significant difference in HFpEF phenotype (blood pressure, exercise tolerance, echocardiography, heart weight, or pulmonary congestion) after DOCA-salt treatment compared to littermate
Lgals1+/+
controls in either males or females (p > 0.05 using Student’s t-test, n=10-12). One hypothesis for this negative result is that
Lgals1
is ubiquitously expressed, and a total body knockout may have compensatory mechanisms. We subsequently created
Lgals1
floxed mice and ongoing studies will determine the effect of macrophage specific deletion of galectin-1 on HFpEF. In conclusion, while multi-omics approaches in mice and humans identify galectin-1 as a novel mediator of HFpEF, further studies are needed to determine its role as a potential therapeutic target.
Abstract 083: Neurovascular And Cognitive Dysfunction In Middle-Aged Mice With High Blood Pressure Variability
Authors: Mendiola Perenkita J , Oherron Philip , Xie Kun , Brands Michael W , Coleman Kathleen , Patterson Rachel , Folk Cameron , Di Stefano Valeria , Filosa Jessica A
Published: October 2023

Blood pressure variability (BPV) is a novel risk factor for cognitive decline. This study tested the hypothesis that BPV disrupts cellular function and communication at the neurovascular complex (vascular, glia, and neuron) contributing to BPV-induced neurovascular dysfunction. A main goal of the study was to establish a murine BPV model and examine how this risk factor impacts key functional outputs of the neurovascular complex. Middle-aged C57BL6 mice were implanted a cranial window and injected GCaMP viral vectors to monitor Ca
2+
activity in cortical neurons or astrocytes. DSI telemetry was used to monitor cardiovascular variables simultaneously (
in vivo
) with arteriole diameter and astrocyte/neuronal Ca
2+
responses via two-photon imaging. In addition, mice were implanted a programmable pump and infused 6-8 pulses/day of saline (Control) or angiotensin II (Ang II, 18 μg/day) for 20-25 days. Ang II infusions evoked transient BP increases (Δ 23±2 mmHg, mean arterial pressure (MAP, P=0.0001, n=14)), and increased BPV (P=0.04, n=12) measured by the average real variability. The 24-hour MAP was not different compared to Controls, allowing for the investigation of BPV-evoked changes in neurovascular function in the absence of hypertension. BPV mice displayed cognitive decline (NOR, P=0.006, n=10). Myogenic responses were lost in parenchymal arterioles of BPV mice but preserved in Controls (P<0.01 n=6). In addition to impaired vascular function, during Ang II-induced BP increases, astrocytes of BPV mice exhibited greater Ca
2+
event frequency (P<0.0001, n=4-6), but of lesser amplitude and reduced area (P=0.02, n=4-6). Furthermore, the neurovascular coupling response, evoked via whisker stimulation, was blunted in mice with high BPV (P<0.0001, n=6-10). Relevant to inflammation, microglial cells from BPV mice had elevated Iba1 (P=0.03, n=4-5) and CD68+ (P=0.003), an inflammatory marker. These findings show that BPV, in the absence of hypertension, result in a loss of cerebral autoregulation, a shift in spontaneous astrocyte Ca
2+
dynamics during blood pressure fluctuations, and increased microglia reactivity. Future mechanistic studies are needed to determine how BPV contributes to cognitive decline.
Abstract 086: Examining The Role Of Bacterial Extracellular Vesicles In Host Blood Pressure Regulation
Authors: Wagle Rishi , Shi Feiya , Ayyaswamy Sriram , Durgan David J
Published: October 2023

We have demonstrated that disruption of the normal gut microbiota, termed gut dysbiosis, is an underlying cause of hypertension (HT). However, the mechanisms by which the microbiota influences host blood pressure (BP) are largely unknown. We hypothesized that extracellular vesicles (EVs) released by gut microbiota gain access to the circulation, deliver cargo to host tissues involved in BP regulation, inducing inflammation and ultimately HT. Bacterial EVs (bEVs) were isolated from cecal content of spontaneously hypertensive stroke prone rats (SHRSP) or Wistar-Kyoto rats (WKY) by size exclusion chromatography. BEV cargo including small RNAs, LPS, and 16s rRNA, was assessed. To directly examine the role of bEVs on BP, purified WKY and SHRP bEVs were transplanted to recipient rats of the same and opposite strain by oral gavage (1.5x10
10
EVs/rat) every other day for 4 weeks. The inflammatory status of ileum and brain were assessed by flow cytometry. Global analysis of small RNA cargo of bEVs was found to be significantly different between WKY and SHRSP (n=6, p=0.001). BEVs from SHRSP contained significantly greater LPS as compared to those from WKY (49 vs 38 EU/ml, n=6, p=0.01). EVs isolated from plasma of both strains contained bacterial 16s rRNA, demonstrating bEVs gain access to host circulation. Finally, as compared to WKY rats receiving WKY bEVs, WKY rats transplanted with SHRSP bEVs had 1) significant elevations in BP at 4 weeks (176 mmHg vs 150 mmHg, n=6, p<0.01), 2) significant reduction of T regulatory cells in ileum (49% vs 58%, n=6, p<0.05) and brain (9% vs 18%, n=6, p<0.05), and 3) significant increases in macrophages in ileum (39% vs 6%, n=6, p<0.001) and microglia in brain (24% vs 8%, n=6, p<0.05). SHRSP rats transplanted with WKY bEVs had significantly lower BP at 4 weeks (160 mmHg vs 175 mmHg, n=6, p<0.05), reduced macrophages in ileum (5% vs 42%, n=6, p<0.001) and microglia in brain (5% vs 29%, n=6, p<0.01), as compared to SHRSP receiving SHRSP bEVs. In conclusion, BEVs originating from the dysbiotic gut of hypertensive SHRSP rats carry distinct cargo, reach the host circulation, and are involved in host gut- and neuroinflammation and HT. These studies identify bEVs as a previously unrecognized mechanism for microbiota-host interaction involved in BP regulation.
Abstract 087: Genetically Engineered Gut Microbiota For Delivery Of Angiotensin Converting Enzyme 2 Attenuates Hypertension
Authors: Manandhar Ishan , Mei Xue , Mell Blair , Aryal Sachin , Tummala Ramakumar , Zubcevic Jasenka , Lai Khanh , Yang Tao , Li Qiuhong , Joe Bina
Published: October 2023

Engineered gut microbiota represents a new frontier in medicine, in part, serving as a vehicle for the delivery of therapeutic biologics to treat a range of host conditions. Since gut microbiota plays a significant role in blood pressure (BP) regulation, using engineered gut microbiota for treatment of hypertension is a promising novel prospect. In this study, we hypothesized that genetically engineered
Lactobacilli
that biosynthesize human ACE2 (Lacto-hACE2) target AngII in the gut to lower BP. To test this hypothesis, a novel
Ace2
knockout (KO) rat (r
Ace2
-/-
for female rats, r
Ace2
-/y
for male rats) was generated using the CRISPR/Cas9 technology. Age and sex-matched
Ace2
KO rats were surgically implanted with radiotelemetry transmitters to monitor BP and administered either Lacto-hACE2 (n=8 male; n=8 female) or wild-type
Lactobacillus
(Lacto-Control, n=7 male, n=8 female) for 3 weeks. BP was monitored by radiotelemetry. ACE2 activity and AngII levels were compared in both sexes. As expected, compared to non-founder control rats,
Ace2
KO rats of both sexes had a significantly higher mean arterial pressure (MAP) (females: control
vs.
r

Ace2
-/-

, 132±2 mmHg
vs
167±10 mmHg, p<0.0001; males: control
vs.
r

Ace2
-/y

, 116±2 mmHg
vs
132±6 mmHg, p<0.0001). Moreover, both renal and colonic Ang II levels were elevated in the
Ace2
KO rats (Renal: females-control
vs.
rAce
2
-/-
, p<0.05; males-control
vs.
r
Ace2
-/y
, p<0.0001; Colon: females-control
vs.
r
Ace2
-/-
, p<0.001; males-control
vs
r
Ace2
-/y
, p<0.05). However, treatment with Lacto-hACE2 showed a sex-specific lowering of BP in females (MAP, control
vs.
treated, 158±7 mmHg
vs
150±7 mmHg, p<0.01), which was not observed in males. The treatment effect was largely due to a significant decrease in diastolic BP (control
vs
. treated, 145±8 mmHg
vs
133±9 mmHg, p<0.001). Interestingly, only colonic AngII, but not renal AngII, was lowered in the treated rats. In conclusion, this study is the first to demonstrate the therapeutic benefit of a commensal microbe engineered to deliver human ACE2 to the colon to lower BP in
Ace2
KO rats. Our study is the first to highlight the potential of targeting the colon with engineered bacteria to develop microbiome medicine as the treatment of hypertension.
Abstract 089: Periodontitis Accelerates The Onset Of Arterial Hypertension In SHR, While The Electrical Activation Of The Carotid Sinus Nerve Delays The Beginning Of The Increase In Blood Pressure.
Authors: Buzinari Tereza C , Castania Jaci A , Salvador Sergio L , Fazan Rubens , Salgado Helio C
Published: October 2023

Background:
We demonstrated that electrical stimulation of thecarotid sinus nerve (CSN) has a protective effect on the development ofPeriodontitis (PER). However, whether PER accelerates the onset ofhypertension in SHR is unknown. If this is true, it could be expected that theactivation of the CSN would retard the onset of hypertension.

Objective:
To evaluate whether PER accelerates the onset of hypertension inSHR; and whether the electrical stimulation of the CSN in SHR would delay theonset of this outcome.

Methods:
Three weeks old male SHRs were implanted with electrodes aroundthe CSN for daily electrical stimulation (3V; 1 ms; 30Hz; 10 minutes per day, fortwo weeks), combined with bilateral ligation of the lower first molar with a silksuture for PER induction. Besides, PER induction was combined with theadministration, p.o., of
Porphyromonas gingivalis
(strain W83) injected threetimes a week. The femoral artery of a five-week-old SHR was cannulated, and 24hours later, with the animals awakened, the systolic (SAP), diastolic (DAP), andmean (MAP) arterial pressure were recorded. Control SHR (SHAM) was notexposed to PER.

Results:
Five weeks old control SHR (SHAM) did not have installed arterialhypertension. However, when the SHR were submitted to PER, they alreadyexhibited a higher arterial pressure at five weeks, when compared to the SHAMgroup (SAP PER 150±4
vs.
SHAM 124±3 mmHg, n=8, p≤0.001; DAP PER 103±5
vs.
SHAM 85±5 mmHg, n=8, p≤0.05; MAP PER 119±4
vs.
SHAM 98±4 mmHg,n=8, p ≤0.01). Nevertheless, the stimulation of the CSN prevented this increasein blood pressure in the five weeks old SHR submitted to PER (PAS SHR PERstim. 130±4
vs.
SHAM 124±3 mmHg; PAD SHR PER stim. 84±3
vs.
SHAM 85±5mmHg; PAM SHR PER stim. 99±4
vs.
SHAM 98±4 mmHg, n=8, p>0, 05).

Conclusion:
PER accelerated the onset of arterial hypertension in SHR withinfive weeks. However, the activation of the baroreceptor and chemoreceptor reflexes through the electrical stimulation of the CSN in SHR exposed to PERdelayed the onset of arterial hypertension.
Abstract 092: All-cause And Cardiovascular Mortality In White Coat Hypertension: An Updated Systematic Review And Meta-analysis
Authors: Cohen Jordana , Cohen Debbie L , Townsend Raymond R
Published: October 2023

Background:
Evidence is inconsistent regarding the risk of white coat hypertension. Our goal was to perform an updated meta-analysis evaluating the mortality risk of untreated white coat hypertension (WCH) and treated white coat effect (WCE).

Methods:
We searched PubMed and Embase from inception to May 2021. Studies with at least three years of follow-up after ambulatory or home blood pressure monitoring and which evaluated all-cause mortality and/or cardiovascular mortality were included. Studies were excluded if participants with WCH and WCE were combined in all analyses, given hypothesized differences in risk across these groups. Studies were evaluated for quality, including use of validated devices and adequate adjustment for confounders. Results were contrasted to our prior meta-analysis, which included the Spanish Registry results prior to retraction. Pooled hazard ratios (HR) were estimated using random effects meta-analysis with profile likelihood estimation of variance.

Results:
A total of 7 studies were included, comprising 19,546 participants with untreated WCH or treated WCE followed over a mean duration of 5 to 16 years. Similar to the prior meta-analysis, there was no significant association of treated WCE with all-cause mortality (HR 0.93, 95% CI 0.83-1.27) compared to controlled hypertension. However, unlike our prior meta-analysis that demonstrated an elevated risk of untreated WCH, the current meta-analysis showed that untreated WCH was associated with a similar risk of all-cause (HR 1.08, 95% CI 0.91-1.36) and cardiovascular mortality (HR 1.06, 95% CI 0.68-4.83) compared to normotension. Influence analyses (i.e., leaving each study out) revealed that the recently republished Spanish Registry results drove the diminished risk of untreated WCH (all-cause mortality HR 1.19, 95% 1.02-1.41 after excluding the new Spanish Registry study).

Conclusions:
Treated WCE has no relationship with all-cause mortality. Untreated WCH may be associated with slightly elevated risk of all-cause and cardiovascular mortality, though the risk is small in magnitude and inconsistent across studies. These findings reinforce the critical importance of out-of-office blood pressure monitoring for the diagnosis and management of hypertension.
Abstract 093: Does Ambulatory Blood Pressure Variability Predicts Target Organ Damage In Youth?
The Ship Ahoy Study
Authors: Urbina Elaine M , Hanevold Coral , Falkner Bonita , Ferguson Michael , Lande Marc B , Meyers Kevin , Mitsnefes Mark , Samuels Joshua A , Flynn Joseph T
Published: October 2023

Altered BP and HR variability are associated with abnormalities of cardiac structure and function in adults. Our aim was to determine if BP and HR variability increase prediction of subclinical cardiac changes in youth after adjustment for mean ambulatory BP (ABP) measures. We measured anthropometrics and performed echocardiography for left ventricular mass index (LVMI), LV systolic strain and LV diastolic function (e’/a’). ABP was measured with an oscillometric device and ABP mean percentiles were calculated per pediatric guidelines to adjust mean ABP for differences in age-, height- and sex. ABP and HR coefficients of variation (CV) were calculated as SD/mean. HR dipping was day mean HR - night mean HR. General linear models were used to determine if ABP and HR CV measures remained significant determinants of cardiac measures in models containing mean ABP levels, age, sex, race/ethnicity, BMIz, HR. Mean age was 15.6 ± 1.7 years (63% white, 59% male) with mean casual BP 122.7/71.6 mmHg ±12.3/10.5, mean daytime ABP 124.2/72.0 ± 11.3/7.7 mmHg. In linear models, DBP day CV, DBP nite CV and HR dip remained significant determinants of LVMI even after adjustment for covariates and ABP percentile (R
2
0.36, p
<
0.0001). HR nite CV was a determinant of e’/a’ after adjustments (R
2
0.16, p
<
0.0001). No BP or HR variability parameters enter the model for LV systolic strain. When stratified by ABP phenotype, participants with white coat HTN (WCH) had higher 24-hour BP, HR variability and HR dipping compared to other phenotypes. Measures of BP and HR variability which may represent altered autonomic tone, may affect LVM and diastolic function in youth especially those with WCH.
Abstract 094: Sex-specific Effects Of Sleep Apnea And Medication-treated Depression On Left Ventricular Mass Index And 3D Left Ventricular Global Longitudinal Strain In The CARDIA Study
Authors: Campeanu Ion , Rossi Noreen F , Komnenov Dragana
Published: October 2023

Background:
Cardiovascular disease (CVD) is the leading cause of death and disability worldwide. CVD risk increases with age; however, sex differences are known to significantly affect progression and survival. We designed this study to assess CVD-related risks associated with depression, sleep apnea, and depression comorbid with sleep apnea, in a biracial cohort of middle-aged men and women. We assessed outcomes such as left ventricular mass index (LVMI)-an index of left ventricular hypertrophy, and 3D left ventricular global longitudinal strain (3D LVGLS), an index of global myocardial function.

Methods:
Data were obtained from the Coronary Artery Risk Development in Young Adults (CARDIA) Study from black and white men and women (n=2778) who attended visit 9 (year 30 of follow-up) echocardiography scan. Demographics and diagnoses of depression and sleep apnea were obtained from visit 8 (year 25 of follow-up). Binary logistic regression was used to asses factors associated with high blood pressure (BP > 130/80 mmHg) and heart failure with reduced ejection fraction (HFrEF). Linear regression analyses were used to assess the strength of the following predictors: race, age, depression diagnosis and medication use for depression on LVMI and 3D LVGLS.

Results:
In men, depression alone and comorbid with sleep apnea were associated with higher risk of BP >130/80mmHg (OR = 1.47, p = 0.036 and OR = 2.739, p = 0.006, respectively). In both men and women (median age 55), black race was significantly associated with LVMI (p = 0.008 and p <0.001, respectively). Additional predictors of LVMI found in women were age (p = 0.019) and current medication use for depression (p = 0.006). Black race and current use of medications for depression were significantly associated with 3D LVGLS in both men (p <0.001 and p = 0.029, respectively) and women (p < 0.001 and p = 0.007, respectively).

Conclusion:
Our results suggest that in middle-aged women, but not men, medication-treated depression is a significant predictor of LVMI. The sex differences were not observed for 3D LVGLS. These results suggest that the severity of depression (i.e. necessitating medication use) may be an important predictor of CVD, especially in women. However, a direct effect of these medications on CVD may occur.
Abstract 100: Activation Of The Skin Renin-angiotensin System Contributes To The Development Of Hypertension
Authors: Taguchi Shinya , Azushima Kengo , Kitada Kento , Morisawa Norihiko , Wakui Hiromichi , Nishiyama Akira , Tamura Kouichi
Published: October 2023

Recent studies suggest skin vasoconstriction and sodium (Na
+
) accumulation are associated with hypertension. The skin has a localized renin-angiotensin system (RAS), a key regulator of blood pressure (BP), but its functional role remains unclear. This study aims to investigate the role of skin RAS in hypertension. Skin tissue from 32 individuals, including patients with hypertension, was used to evaluate the association between skin RAS component expression and office BP. Among the RAS components, skin expression of ATRAP (type-1 angiotensin II (Ang II) receptor (AT1R)-associated protein), which selectively inhibits pathological AT1R signaling, was inversely correlated with systolic and diastolic BP (R
2
= 0.12,
P
< 0.05; R
2
= 0.20,
P
< 0.05, respectively). To investigate the causal relationship between skin ATRAP and hypertension, mice lacking ATRAP in skin keratinocytes (KO: K14
Cre
;ATRAP
flox
) were generated. Telemetry analysis showed that Ang II (500 ng/kg/min)-induced hypertension and related cardiac hypertrophy was exacerbated in KO mice compared to control mice (mean BP, 123.3 ± 4.7 vs. 132.6 ± 3.1 mmHg, n = 5,
P
< 0.05; heart/body weight, 4.7 ± 0.4 vs. 5.2 ± 0.7, n = 11-12,
P
< 0.05). KO mice showed increased skin expression of angiotensinogen and AT1R (
P
< 0.05 for each; n = 11-12), while those in the kidneys and heart were similar in both groups, suggesting skin-specific enhancement of tissue RAS activity. AT1R blocker treatment eliminated exaggerated Ang II-induced hypertension and enhancement of skin RAS activity in KO mice. Sympathetic nerve activity, plasma volume, and skin Na
+
content did not differ between the groups. In KO mice, urine volume per water intake increased (
P
< 0.001), despite similar body weight changes in both groups, suggesting decreased extra-renal water loss. Consistent with these findings, skin blood flow (22.6 ± 3.8 vs. 16.5 ± 4.5 AU,
P
< 0.001) and transepidermal water loss (9.0 ± 1.9 vs. 6.5 ± 1.9 g/m
2
/h,
P
< 0.01) were decreased in KO mice. These differences, including exaggerated Ang II-induced hypertension, were eliminated by skin vasodilation via body temperature elevation. In conclusion, enhanced skin RAS activity contributes to BP elevation via skin vasoconstriction. Skin RAS may be a novel target for treating hypertension.
Abstract 102: Targeted Disruption Of Ctcf Binding Near The
Ren
Gene Alters Local Gene Expression In Response To Dietary Salt
Authors: Vanden Avond Mark , Grzybowski Michael , Kurth Terry , Greene Andrew , Liang Mingyu , Cowley Allen , Geurts Aron M
Published: October 2023

Blood pressure regulation and hypertension risk may be impacted by changes in chromatin conformation (CC) caused by common genetic variations. CCCTCF-binding factor (CTCF) is a DNA architecture protein that regulates CC and thereby gene expression. We hypothesized that disruption of local CC surrounding the renin gene can alter local gene expression and influence blood pressure changes to dietary salt. Three mutant rat models were created using CRISPR-Cas9 in the Dahl salt-sensitive rat with mutations in one or more CTCF motifs surrounding the renin gene. Disruption of CTCF binding was confirmed at specific mutated sites in all three models. Plasma renin activity (PRA) was measured and RNA-seq analysis was performed in multiple tissues of wild-type (WT) and mutant rats fed a 0.1%, 0.4% or 4.0% NaCl diet for four days. While PRA in WT males responded as expected (17.52±0.81, 12.87±1.00, and 7.13±0.19 ng/mL; p<0.05 for 0.1% and 4.0% vs. 0.4%) on these diets respectively, males from two of three mutant strains failed to increase their PRA on 0.1% NaCl diet. Females were unaffected. At the transcriptional level in kidney cortex, one mutant strain failed to increase renin mRNA expression on 0.1% NaCl diet, while another did not decrease expression on 4.0% NaCl diet. Other local genes
Kiss1
and
Snrpe
were also significantly disrupted in kidney cortex. Renin expression was also disrupted in liver and adrenal gland in two of three mutant strains. Blood pressure and urinary protein excretion were measured in CTCF mutant and WT littermates over the course of 28 days at baseline (3 days, 0.4% NaCl diet), followed by seven days on 0.1% NaCl, then 21 days 4.0% NaCl diet. While no significant effects were observed between WT and mutant males or females on 0.4% or 0.1% NaCl diet, mean arterial pressure response to 4.0% NaCl diet was significantly attenuated in females of one of three mutant strains on days 16, 17 and 20 (169.6±7.09 vs. 144.2±2.69 mmHg at Day 20; p<0.005). No changes in urinary albumin or protein excretion in mutants of either sex were detected. Collectively, these data suggest that disruption of CTCF binding around the renin gene alters PRA and local gene expression in multiple tissues in response to dietary salt.
Abstract 107: Longitudinal Associations Of Dietary Fructose And Psychological Stress With Vascular Aging Index And Incident Cardiovascular Disease Among Us Black And White Adults In The CARDIA Study
Authors: Osborne Meaghan , Bernard Alexa , Falkowski Emily , Peterson Deni , Vavilikolanu Anusha , Komnenov Dragana
Published: October 2023

Background:
Cardiovascular diseases (CVD) is the leading cause of death and disability worldwide. CVD risk increases exponentially with age, largely due to pathophysiological changes within the vasculature. As per AHA’s Life’s Essential 8, modifiable lifestyle factors such as diet and psychological stress are important predictors of CVD. We designed this study to assess how dietary behaviors, specifically sucrose, fructose and sodium, and endured psychological stress in young adult men and women impact the vascular aging index (VAI) and CVD risk by mid-life.

Methods:
All data were obtained from the Coronary Artery Risk Development in Young Adults (CARDIA) Study. We used the data from black and white men and women (n=2656) who had undergone carotid artery ultrasound Doppler scans at visit 7 (year 20 of follow-up) and used these measures to calculate the VAI, as shown previously. Baseline demographics, dietary data and stress/depression measures (CES-D scores) were obtained from visits 1 (at enrollment, year 0) and 7 and compared (men vs. women) with Kruskal-Wallis and Pearson’s chi-square tests. Regression analyses were used to assess the predictive value of these variables on VAI. Cox regression analyses were conducted to assess the risk of fatal and non-fatal CVD, hypertensive CVD, and stroke.

Results:
Significant associations were found for fructose intake (expressed as percent total calories, P=0.042) and BP >30/80 mmHg (P=0.025) at baseline with VAI in men but not women. Conversely, CES-D scores at baseline were significantly associated with VAI in women (P=0.014) but not men. On Cox regression analyses, baseline BMI was significantly associated with the incidence of any CVD (HR=1.065, P <0.001), hypertensive CVD (HR=1.083, P<0.001) and stroke (HR=1.068, P=0.002) 30 years later. Dietary fructose intake recorded in mid-life was significantly associated with the incidence of any CVD (HR=1.041, P=0.044), and having BP >130/80 mmHg at baseline was associated with stroke (HR=1.996, P=0.029). No sex differences were noted in the Cox regression analyses.

Conclusion:
Our results suggest that in young adult black and white men but not women BMI and fructose consumption, but not sodium are associated with increased VAI and incidence of CVD by mid-life.
Abstract 108: Rejuvenation Potential Of Extracellular Vesicles From Renal Distal Tubular Cells - A Promising Strategy To Prevent Aging-associated Hypertension And Renal Function Impairment
Authors: Aung Lynn-Htet-Htet , Sun Zhongjie
Published: October 2023

Ageing is characterized by progressive multi-organ functional decline and disrupted homeostasis adaptation. Hypertension and kidney damage become increasingly prevalent as individuals age. Given its high metabolic activity and constant exposure to reactive oxygen species, the kidney is particularly susceptible to age-related deterioration. In our study, aged mice exhibited a notable decline in glomerular filtration rate (GFR) and urinary excretion of sodium, potassium, and calcium, alongside elevated urinary creatinine levels, indicating impaired renal function associated with ageing. Histologically, aged kidneys showed glomerular collapse, thickening of the glomerular basement membrane, and increased glomerular sclerosis. Cellular senescence plays a critical role in renal ageing. Senescent cells display an array of changes in gene and protein expression and thus also altered the contents packaged into extracellular vesicles (EVs), secreted into the extracellular milieu. The protein and RNA composition of EVs varied depending on the age and pathological status of the originating parent cells. Our data showed an increased expression of EV biogenesis protein HRS (hepatocyte growth factor-regulated tyrosine kinase substrate) in aged DCT cells (aDCTs). Consistently, with an increase in EV secretion in aDCTs. Administration of young DCT (yDCT)-derived EVs into aged kidneys resulted in a significant improvement of GFR and urinary ion excretion, accompanied by a reduction in age-associated elevation of systolic blood pressure (SBP). Conversely, when administering aDCT-derived EVs to young mice, we saw a significant decrease in GFR and an increase in SBP, indicating a detrimental effect of aDCT-EVs. Proteomic analysis revealed distinct protein expression profiles between yDCT-EVs and aDCT-EVs. These differentially expressed proteins are highly enriched in ageing regulatory pathways, underscoring the crucial role of DCT-EVs in kidney rejuvenation. Intriguingly, we identified a subset of proteins exclusively present in either yDCT-EVs or aDCT-EVs. Our future research will focus on identifying the key components of DCT-EVs responsible for driving kidney rejuvenation.
Abstract 115: An Action Plan To Improve Biochemically Detected Non-adherence To Antihypertensive Treatment Results In A Short-term Reduction In Arterial Stiffness And A Long-term Reduction In Albuminuria (the ATHAN Clinical Trial)
Authors: Oliveras Anna , Vázquez Susana , Camps Carme , Fontdevila Laia , Illana Francisco J , Galceran Isabel , Andolza Laura , Vinyoles Ernest , Hernandez Juanjo , Crespo Marta
Published: October 2023

We previously shown that a specific 3-months(m) intervention improved therapeutic adherence in patients with non-controlled hypertension (HT) decreases blood pressure (BP). We aim to evaluate if there is also a decrease in HT-mediated organ damage (HMOD) at short- and long-term. We conducted a prospective, randomized clinical trial in a HT Unit. Consecutively attended patients with 24h-BP >or=130/80 mmHg despite receiving at least 2 antihypertensive drugs and with therapeutic non-adherence confirmed by determination of antihypertensives in urine (n=47) were randomized (1:1) to receive a specific 3m program to improve adherence (INT=intervention) or routine follow-up (C=control). Antihypertensive treatment was not changed. Prior to randomization, and at 3m and 12m, the presence of antihypertensives in urine and albuminuria were determined. BP and pulse wave velocity (PWV) were evaluated by 24h- ambulatory monitoring in all periods. After 3m, all patients received usual care. At 3m, the adjusted BP differences (mean; 95% CI) from baseline (INT vs. C) were -14.3 mmHg (-5.1 to -23.5), p=0.003, and -10, 0 mmHg (-4.0 to -16.0), p=0.002, for 24h-SBP and 24h-DBP, respectively. The INT group maintained the decrease in BP at 12m. vs baseline: mean change (95% CI) of 24h-SBP = -16.1 mmHg (-22.8 to -9.4), p <0.001 and 24h-DBP change = -7.8 (-12, 8 to -2.7), p=0.002. In the INT group, the percentage of undetected antihypertensive drugs decreased from baseline (median [interquartile range]): 40% [20-100], 0% [0-25], and 18.3% [0-40] at baseline, 3m and 12m, respectively; Z= -3.426 (p<0.001) and Z= -3.055 (p=0.002) for changes at 3m and 12m, respectively. PWV decreased at 3m only in the INT group: mean difference (95% CI) -0.402 m/sec (-0.054 to -0.749), p=0.025, but not at 12m. Albuminuria (median [IQR]) decreased in the INT group at 12m: 27.1 mg/g [11.1-116.2] and 22.4 mg/g [6.4-91.9] at baseline and 12 m, respectively (Z=-2.207, p=0.027). In conclusion, urine biochemical detection of therapeutic non-adherence and a specific 3m-intervention to improve adherence lead to better BP control, which maintains in the long term. In addition, there is a decrease in arterial stiffness in the short term and a regression of albuminuria in the long term, thus reducing the HMOD.