bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 025: Role Of The C-C Motif Chemokine Ligand 5 (CCL5) And Its Receptor, C-C Motif Chemokine Receptor 5 (CCR5) In The Genesis Of Aldosterone-induced Hypertension, Vascular Dysfunction, And End-organ Damage
Authors: Costa Rafael , Alves Juliano , Santos Débora , Awata Wanessa , Singh Shubhnita , Bruder-Nascimento Ariane , Ho Jacqueline , Bruder Thiago
Published: October 2023

Background:
Hyperaldosteronism (HA) is the most common cause of endocrine hypertension. CCL5 is a pro-inflammatory chemokine acting primarily via C-C Motif Chemokine Receptor 5 (CCR5). We hypothesize that HA leads to hypertension, vascular injury, and renal demage via CCL5/CCR5 and Nox1-dependent mechanisms.

Methods:
10-12-weeks-old male wild type (CCR5+/+) and CCR5 receptor knockout (CCR5-/-) mice were infused with aldosterone for 14 days (600ug/Kg/day, osmotic mini-pump). Endothelial function was studied in aortae and blood pressure was analyzed by radiotelemetry. Mouse endothelial cells (MEC) were used to examine the molecular mechanisms.

Results:
HA increased circulating CCL5 levels [Vehicle: 7.41±1.6 vs. HA: 13.2±1.4 (pg/mL)] and reduced acetylcholine (ACH)-induced vasodilation (% relaxation) in CCR5+/+, which was abolished by inhibiting Nox1 (NOXA1ds, 10 μM) [Vehicle: 91.2±2.1 vs. HA: 42.7±3.9 vs. HA+NOXA1ds: 89.7±1.9]. To understand if CCL5 induces endothelial dysfunction via CCR5 and Nox1, aortae were incubated with CCL5 (100 ng/mL for 24 h) with vehicle or CCR5 antagonist (Maraviroc, 40 μM) or NOXA1ds and endothelial function and Nox1 expression were analyzed. CCL5 treatment induced endothelial dysfunction [ACH response, Vehicle: 90.1±1.9 vs. CCL5: 31.9±2.7] and increased Nox1 expression, which were blunted by Maraviroc or NOXA1ds, indicating that CCL5 affects endothelial function dependent on CCR5 and Nox1. We found that CCR5-/- mice are protected from HA-induced endothelial dysfunction, hypertension [mean arterial pressure (mmHg) - CCR5+/+: 154.8±3.7 vs. CCR5-/-: 104.5±2.9], and renal damage (analyzed by NGAL, Synaptopodin, and Colagen III gene expressions). On mechanistic levels, we found that CCL5 decreases nitric oxide production, leads to NF-κB activation, induces oxidative stress (elevated superoxide anion and H2O2) and ICAM and VCAM expressions in MEC, which were associated with higher macrophage adhesion, such effects were prevented by Maraviroc.

Conclusion:
These findings reveal a novel signaling pathway in HA-associated vascular and renal injuries and hypertension, CCL5 activates CCR5 stimulating NF-κB and Nox1 activities, and place CCR5 as a pharmacological target for cardiovascular outcomes in HA.
Abstract 026: Three-dimensional Visualization And Evaluation Of Differences In Renin Expression In Kidneys Exposed In Vivo To Either Low Or High Arterial Pressure
Authors: Yamaguchi Manako , Yamaguchi Hiroki , Medrano Silvia , Tainaka Kazuki , Sequeira Lopez Maria Luisa S , Gomez Ariel R
Published: October 2023

Background:
Renin cells are essential for the regulation of blood pressure (BP). In adult mammals, renin cells are present in small numbers in the juxtaglomerular area, but when homeostasis is threatened, such as by hypotension, additional cells along the renal arterioles are recruited to synthesize and release renin. However, there have been no studies to date that have shown changes in the distribution of renin cells in response to changes in BP by three-dimensional (3D) imaging.

Objective:
Establish a method to visualize and measure renin cells, and comparison of renin expression differences in kidneys exposed to hypo- or hypertension in 3D.

Methods:
We used our new mouse model (

Ren1
c-tdTomato

) that expresses renin and the fluorescent tdTomato under the control of the endogenous renin gene environment. To test whether we could detect differences in the distribution of renin in response to changes in BP, we subjected

Ren1
c-tdTomato

mice to aortic coarctation at 3 months of life. The right and left kidneys (RK, LK) are exposed to high and low BPs, respectively. We harvested kidneys after 72 hours and performed tissue-clearing. We used light sheet microscopy for imaging and the Imaris software for 3D reconstruction and data analysis. We defined the 3D renin cell index (3D-RI) as total volume of renin cells (μm
3
) divided by total number of glomeruli. We extracted 10 cubes of 500 μm square each from the L and R renal cortex, calculated their individual 3D-RI and compared them.

Results:
With tissue-clearing-based 3D imaging, renin cells were clearly visualized, allowing observation of their distribution and appearance at a one-cell resolution. In the LK exposed to hypotension, several afferent arterioles were identified where renin cells extended close to their branching point. Further, the average value of the 3D-RI was significantly higher in LK (21460 ±6303) than in RK (12838 ± 4632; p = 0.003).

Conclusion:
Using our new

Ren1
c-tdTomato

mouse and 3D imaging technology we detected differences in renin cells appearance and distribution in kidneys exposed to hypo- or hypertension. Combined with spatial single cell multiomics, 3D visualization of renin cells will open new avenues for understanding how vascular cells acquire their positional identity in health and disease.
Abstract 033: Innate Lymphoid Cells Mediate Hypertension In T Cell Deficient Hosts
Authors: Chen Wei , Xiao Liang , Mohammed Marwa , Patrick David , Smart Duncan , Mingfang Ao , Perrotta Sara , Carnevale Lorenzo , Pallante Fabio , Lembo Giuseppi , Carnivale Daniela , Harrison David
Published: October 2023

T cells have been shown to contribute to hypertension, however, hypertension occurs in settings of T cell deficiency, such as human immunodeficiency virus infection and following chemotherapy. We studied two colonies of RAG-1
-/-
mice with disparate responses to angiotensin II (Ang II) infusion. Ang II increased both systolic and diastolic pressure in RAG-1
-/-
mice from Jackson labs (RAG-1
-/-US
) but had no effect in a colony of RAG-1
-/-
mice housed at our Pozzilli facility (RAG-1
-/-POZ
) for the past 14 years (SBPs 169 ± 2 vs 114 ± 3 mmHg, p < 0.003). While both lacked T and B cells, the RAG-1
-/-US
mice exhibited a striking expansion of natural killer and both type 1 and type 3 innate lymphoid cells, while the RAG-1
-/-POZ
mice did not. In RAG-1
-/-US
mice, NK cells and ILC1s produced IFNγ, while ILC3s produced IL-17A. Immunoclearing of IL-17A, but not IFNγ prevented the hypertension in RAG-1
-/-US
mice. There were differences in global gene expression and DNA methylation status in NK1.1
+
/NKp46
+
cells between these two colonies and notably β2-AdR and IL-10 expression was significantly lower in RAG-1
-/-US
mice. Knockdown of the β2-AdR using intra-splenic infusion of siRNA in RAG-1
-/-POZ
mice led to a hypertensive response resembling that observed in wild type and RAG-1
-/-US
mice. We also found striking differences in DNA methylation patterns in innate lymphoid cells of RAG-1
-/-US
vs RAG-1
-/-POZ
mice and treatment with the DNA methyltransferase inhibitor 5-azacytidine prevented hypertension in RAG-1
-/-US
mice. Single cell sequencing showed that 5-azacytidine also reduced splenic populations of NK and ILC3 cells and reduced the presence of the transcription factor RORγt, which defines ILC3s. Ang II infusion caused only a transient elevation of blood pressure in NSG/MHCI/II
-/-
mice that lack T cells and innate lymphoid cells but led to sustained hypertension following adoptive transfer of either T cells or NK1.1
+
/NKp46
+
cells to these animals. We conclude that DNA methylation likely modulates expansion of NK and innate lymphoid cells. Either T cells or innate lymphoid cells can contribute to hypertension, and the latter can compensate by serving as a source of cytokines like IL-17A when T cells are deficient.
Abstract 038: Adipocyte-specific BBSome Deficiency Impacts The Sympathetic Nervous System And Baroreceptor Reflex Sensitivity
Authors: Zhao Yuying , Guo Deng-Fu , Morgan Donald , Rahmouni Kamal
Published: October 2023

The BBSome, a protein complex of 8 Bardet-Biedl syndrome (BBS) proteins including BBS1, has emerged as an important regulator of metabolic, cardiovascular, and autonomic functions. Patients with
Bbs
gene mutations display severe obesity, type II diabetes and hypertension. Previous studies demonstrated that neuronal BBSome contributes to metabolic homeostasis and blood pressure regulation, however the importance of adipocyte BBSome in controlling glucose homeostasis and blood pressure remains poorly understood. To address this, we generated mice lacking the BBSome specifically in adipocytes through selective deletion of the
Bbs1
gene (Adipo
Cre
/Bbs1
fl/fl
) by crossing the adiponectin
Cre
(Adipo
Cre
) male mice crossing with female Bbs1 floxed (Bbs1
fl/fl
) mice. Adipocyte-specific Cre recombinase was validated by further crossing the Adipo
Cre
mice with td-Tomato reporter mice.
Bbs1
gene deletion in adipocytes was confirmed by measuring
Bbs1
gene expression in adipose tissue using qRT-PCR. To induce obesity, mice were fed a high fat high sucrose diet (HFHSD) for 16 weeks starting at 4 weeks of age. Interestingly, the weight gain evoked by HFHSD was comparable between male and female Adipo
Cre
/Bbs1
fl/fl
mice and littermate controls. Similarly, Adipo
Cre
/Bbs1
fl/fl
mice displayed unaltered blood glucose as indicated by glucose and insulin tolerance tests. Using radiotelemetry, we found no significant difference in systolic, diastolic, and mean arterial pressure and heart rate between control and Adipo
Cre
/Bbs1
fl/fl
mice fed HFHSD. Surprisingly, a significant increase in renal sympathetic nerve activity was observed in Adipo
Cre
/Bbs1
fl/fl
mice (Control: 101.1 ± 16.23 spikes/sec, Adipo
Cre
/Bbs1
fl/fl
: 133.2 ± 32.15 spikes/sec,
p
< 0.05). Next, we measured baroreceptor reflex sensitivity by analyzing the responses to sodium nitroprusside (SNP) and phenylephrine (PE). The slope analysis showed that Adipo
Cre
/Bbs1
fl/fl
mice were less sensitive to SNP (Control: -2.56 ± 0.82, Adipo
Cre
/Bbs1
fl/fl
: -1.746± 0.29,
p
< 0.05), demonstrating altered baroreflex. Taken together, our data implicate the adipocyte BBSome is a regulator of sympathetic nervous activation and baroreflex sensitivity.
Abstract 042: Therapeutic Administration Of L-lysine Leads To The Attenuation Of Salt-sensitive Hypertension
Authors: Dissanayake Lashodya V , Levchenko Vladislav , Palygin Oleg , Staruschenko Alexander
Published: October 2023

Our previous studies using a multilayered omics approach showed a deficiency of lysine metabolites in Dahl Salt-sensitive (SS) rats fed a high-salt (HS) diet. Consequent studies attempted to rewire these metabolic changes by treating the SS rats with lysine in drinking water while on an HS diet. Rats treated with lysine revealed that the treatment significantly attenuated the development of SS hypertension (HTN) and decreased kidney damage via increased diuresis, accelerated lysine metabolism, and inhibited tubular albumin uptake. Our current study hypothesizes that lysine administration will also be beneficial when blood pressure is already elevated. For this purpose, we used a therapeutic approach where we fed 8-week-old male rats with an HS diet for 10 days (at which point they develop moderate kidney damage (by historic data) and a mean arterial pressure (MAP) of ~140 mmHg, N=13) and consequently put on either L-lysine (17 mg/ml drinking water supplement) or vehicle treatment for 11 more days. MAP was continuously measured using radiotelemetry. At the end of the protocol, the L-lysine-treated group showed a significant decrease in MAP compared to the vehicle group (endpoint 145±5 vs. 176±8 mmHg, N=6-7). Next, using targeted metabolomics, we measured 42 amino acids and their derivatives in the kidney, plasma, and urine at the endpoint of these rats to determine the influence of the treatment on amino acid metabolism. Lysine was upregulated in the treatment group in the kidney, plasma, and urinary excretion compared to the control (log
2
FC & p-value: 0.24, 0.01; 1.22, 3.37E-06; 6.6, 0.04; N=4-5 per group). Lysine degradation product, alpha-Aminoadipic-acid was also increased by 1.56-fold and 1.86-fold in the kidney and plasma, respectively (p-value:0.03 & 0.006). Additionally, 13 other amino acids or derivatives significantly changed in the kidney, plasma, or urine. Our current data suggest that L-lysine administration in hypertensive subjects can have therapeutic effects on the progression and magnitude of SS HTN. As shown by our previous study, it is likely the mechanisms behind this beneficial effect lie in the increased lysine metabolism, which can affect critical downstream metabolic pathways such as the TCA cycle and Fatty Acid metabolism.
Abstract 045: High Fat Diet Increases Blood Pressure In Sprague Dawley Rats And Induces Salt Sensitive Hypertension: Role Of NKCC2 In The Thick Ascending Limb (TAL)
Authors: Granados Pineda Jessica , Haque Mohammed Z , Maskey Dipak , Liao Tang-Dong , Eaton Andrew J , Ortiz Pablo A
Published: October 2023

The Western Diet is characterized by elevated added sugars, fat, and salt and is known to promote hypertension. Chronic high fat intake induces obesity in rodents but the effects on blood pressure are variable between rat/mouse strains. The early effects of a high fat diet, prior to the onset of obesity, in kidney function, signaling and NaCl reabsorption have been poorly studied. In the kidneys, the thick ascending limb (TAL) absorbs 25% of filtered NaCl through the apical Na/K/2Cl cotransporter NKCC2. NKCC2 is activated by phosphorylation at residues Thr
96,101
, or Ser
126
or by increasing its trafficking to the apical surface. The effects of a high fat diet on NKCC2 activity and blood pressure have been poorly studied in the Sprague Dawley (SD) rat, a widely used rodent model. We hypothesized that a high fat diet (60%kCal saturated fat, HF) activates NKCC2 and induces salt sensitive hypertension. We first tested the effects HF on blood pressure (BP). SD rats (Charles Rivers) were trained for tail cuff SBP and placed on HF or control diet for 4 weeks. After 2 weeks, HF diet increased SBP by 9±4 mmHg and at 4 weeks by 13±4 mmHg (p<0.05). We then added high salt diet (4% Na, HS, n=7) which further increased SBP by 19±5 mmHg in HF rats reaching 151±5 mmHg after 4 weeks. SBP did not change in rats fed HS or HF alone (HS: 121±4, HF: 132±4 mmHg, p<0.05 vs HF+HS). In isolated TALs, 2 weeks of HF did not change total NKCC2 expression or P-Thr
96,101
, but enhanced P-Ser
126
NKCC2 (78±26%, p<0.05) and increased surface/intracellular NKCC2 ratio by 22±8% (p<0.03). After 8 weeks of HF diet, surface to intracellular NKCC2 was higher in HF (+54±9% from HS, p<0.05) and HF+HS rats (+40±18% of HS) compared to those fed HS alone. Body weight gain was similar in groups after 2 weeks, but after 8 weeks, rats on HF or HF+HS gained more weight than HS alone (p<0.05). Blood glucose was not different between groups. Our data suggest that 2 weeks of a HF diet induces early signaling in TALs, prior to obesity, that may contribute to increase BP or induce salt sensitivity in normal rats.
Abstract 056: Pooled, 6-month Renal Safety And Blood Pressure Reductions Using The Symplicity Spyral Radiofrequency Renal Denervation Catheter
Authors: Secemsky Eric , Mahfoud Felix , Mancia Giuseppe , Townsend Raymond R , Kandzari David E , Whitbourn Robert , Hettrick Doug , Böhm Michael
Published: October 2023

Background:
Renal denervation (RDN) targets the sympathetic nervous system to lower blood pressure (BP). Impact on long-term renal function after RDN is less well known. We report 6-month renal safety and BP reductions after RDN from the largest database, using the latest generation, multi-electrode, radiofrequency (RF) Symplicity Spyral™ catheter.

Methods:
A broad spectrum of patients with hypertension were enrolled in the SPYRAL HTN Global Clinical Program (4 major studies; SPYRAL First-In-Man, SPYRAL HTN-OFF MED, -ON MED and Global SYMPLICITY Registry [GSR]). GSR is an all-comer registry for patients with uncontrolled hypertension, including chronic kidney disease (CKD), whereas patients enrolled in the SPYRAL HTN-OFF and -ON MED trials required to have OSBP ≥150 and <180 mmHg and ODBP ≥90 mmHg. OFF MED patients were required to have a drug washout period prior to treatment and did not take drugs for the first 3 months. ON MED patients were on a stable 1-3 antihypertensive drug regimen for 6 months.

All were treated with RF RDN using the Spyral catheter. We pooled reductions in office and 24-h ambulatory BP, safety outcomes include changes in the estimated glomerular filtration rate (eGFR) and renal artery stenosis incidence (>70%, confirmed by angiography).

Results:
As of December 2022, 1,284 patients have received RF RDN with the Spyral catheter. Patients at baseline were 58±12yrs old, 37.6% female, 29.1% had type 2 diabetes, eGFR was 77.7±23.9mL/min/1.73m2, 20.7% had CKD stage 3 or higher, OSBP was 166±21mmHg and ASBP was 153±16mmHg. 6 months after RDN, OSBP changed by -14.8±22.1 (n=940; p<0.001) and ASBP changed by -8.9±16.0 (n=678; p<0.001). Significant diastolic BP reductions were also observed. eGFR changed by -2.0mL/min/1.73m2. There was no incidence of renal artery stenosis (>70%).

Conclusion:
In this large, pooled population of patients receiving RF RDN using the Spyral catheter, there were significant and consistent BP reductions through 6 months. There was no incidence of renal artery stenosis, and eGFR change was consistent with the natural progression of the aging hypertensive population. These data highlight the safety and efficacy of RF RDN as an adjunctive therapy to antihypertensive drugs to treat high BP.
Abstract 059: (Pro)renin Receptor Ablation In Tyrosine Hydroxylase Neurons In The Paraventricular Nucleus Of The Hypothalamus Improves Insulin Sensitivity In Mice Fed A High-fat Diet
Authors: Pan Shiyue , Worker Caleb , Gayban Ariana , Feng Earley Yumei
Published: October 2023

We recently reported that neuron-specific (pro)renin receptor (PRR) deletion reduces blood pressure (BP), fasting blood glucose (BG) and improves glucose tolerance in high-fat diet (HFD)-treated mice. To investigate underlying mechanisms, we focused on tyrosine hydroxylase (TH) neurons in the hypothalamic paraventricular nucleus (TH
PVN
neurons), a largely GABAergic population that expresses PRR and projects to autonomic centers important for BP and BG regulation. The PRR was ablated specifically in the TH-positive neurons in the PVN using Cre-LoxP approach. Radio telemetric BP recording, fasting blood glucose (FBG) and frequently sampled intravenous glucose tolerance test (FSIVGTT) were performed following 6-week HFD (60% calories from fat). HFD induced elevation of BP within the first week (
p
< 0.05) and maintained elevated during 6-week protocol; however, there was no difference in BP between TH
PVN
PRRKO and WT control mice in both sexes following HFD. In male mice, the controls had higher FBG when compared with the TH
PVN
PRRKO mice (144.1 ± 5.74 vs. 121.9 ± 4.34 mg/dl,
p
< 0.05) at the end of the 6-week HFD treatment. Interestingly, PRR ablation in the TH
PVN
neurons did not reduce FBG in female mice. The TH
PVN
PRRKO mice showed improved glucose handling during FSIVGTT (AUC: 3481 ± 247.5 vs. 4263 ± 153.9,
p
< 0.05), while there was no difference in insulin secretion (AUC: 1782 ± 339.2 vs. 2324 ± 495.6,
p
> 0.05) between the two groups suggesting that TH
PVN
PRR ablation attenuates HFD-induced glucose intolerance through either an increase in insulin sensitivity or an insulin-independent pathway. Detected by western blot, the glucose transporter 4, an insulin-dependent glucose transporter, was higher on the plasma membrane of skeletal muscle in the TH
PVN
PRRKO mice (1.9 ± 0.2 vs. 1.0 ± 0.1,
p
< 0.05) compared with the controls. The hepatic mRNA expression of glucose-6-phosphatase, a gluconeogenic gene transcriptional negative-regulated by insulin, was lower in the TH
PVN
PRRKO (0.60 ± 0.07 vs. 0.95 ± 0.10,
p
< 0.05) compared with the control mice, suggesting improvement of insulin sensitivity in the TH
PVN
PRRKO mice. In conclusion, PRR ablation in TH
PVN
neurons attenuates FBG and glucose tolerance through improvement of insulin sensitivity without impacting BP.
Abstract 070: Deletion Of A Non-coding Genomic Segment Orthologous To The Human Rs1173771 Haplotype Region Attenuates Hypertension In Dahl Salt-sensitive Rats
Authors: Xue Hong , Liu Pengyuan , Vanden Avond Mark , Grzybowski Michael , Wang Jingli , Ying Rong , Westhoff Justin , Widlansky Michael E , Cowley Allen W , Geurts Aron M , Liang Mingyu
Published: October 2023

Most SNPs associated by GWAS with blood pressure (BP) phenotypes are located in noncoding regions of DNA. Investigating the physiological effects of these SNPs and their underlying mechanisms will advance our understanding of how genetic and epigenetic mechanisms regulate BP. The BP-associated SNP rs1173771 is in linkage disequilibrium (LD) with 10 other SNPs (r
2
>0.80) forming a haplotype spanning 17.5 kb of noncoding DNA. We explored the effects of the rs1173771 LD orthologous region on BP on the genetic background of Dahl salt-sensitive (SS) rats and the physiological and molecular mechanisms involved.

A ~29.5 kb noncoding segment on rat chromosome 5 orthologous to the human rs1173771 LD region was identified using LiftOver. The model SS-rs1173771LD
-/-
was generated using CRISPR-Cas9 to delete this region in SS rats. BP was not significantly different between SS-rs1173771LD
-/-
rats and wildtype (WT) littermates on a 0.4% NaCl diet. After 14 days of a 4% NaCl diet, 24-hour mean arterial pressure (MAP) and systolic blood pressure (SBP) of SS-rs1173771LD
-/-
male rats were significantly lower compared with WT (149.1 ± 2.7 vs 155.9 ± 2.5 mmHg for MAP and 169.6 ± 4.3 vs 179.0 ± 3.0 mmHg for SBP). No difference in diastolic blood pressure was observed. No significant difference of BP was observed between genotypes in females. The transcription start sites of human natriuretic peptide receptor 3 (NPR3) and rat Npr3 are located ~129-kbp and ~80-kbp from the human rs1173771 LD region and the rat orthologous region, respectively. RNA-seq indicated that Npr3 expression was upregulated in the mesenteric artery in SS-rs1173771LD
-/-
rats. Vasodilation of mesenteric arteries in response to C-type natriuretic peptide was significantly enhanced in SS-rs1173771LD
-/-
male rats compared with WT littermates. A selective NPR-C antagonist abolished vasodilation in both groups, suggesting the vasorelaxation was NPR-C dependent.

In conclusion, deletion of the rs1173771 LD orthologous region increased mesenteric artery vasorelaxation and attenuated salt-induced hypertension in male SS rats. Our findings provided evidence that evolutionarily conserved noncoding DNA regions located far from protein-coding genes can influence BP
in vivo.
Abstract 103: Combined Dapagliflozine And Eplerenone Treatment Improve Cardiorenal Function In Rats With Chronic Kidney Disease.
Authors: SOULIE Matthieu , Manon Durand , STEPHAN Yohan , Lima Posada Ixchel Quetzaliztli , Palacios-Ramirez Roberto , Lopez-Andres Natalia , Mulder paul , Jaisser Frederic
Published: October 2023

Background:
Patients with chronic kidney disease (CKD) present higher risk of cardiovascular (CV) complications as well as renal function decline. The DAPA-CKD trial showed reduced CV events in diabetic and non-diabetic CKD patients treated with Dapagliflozine (DAPA). Mineralocorticoid Receptor Antagonists (MRAs) have shown similar beneficial effects in diabetic CKD patients (FIDELIO-CKD & FIGARO-CKD trials) but impact in non-diabetic CKD patients is unkown. The aim of this study was to compare DAPA and Eplerenone (EPLE), alone or in combination on renal and CV functions in a non-diabetic rat CKD model.

Methods:
CKD was induced in Sprague Dawley rats by 5/6 nephrectomy. Rats were treated with DAPA (10 mg/kg/day PO), EPLE (100 mg/kg/day PO) or the combination during 3 months.

Results:
After 3 months kidney fibrosis is decreased in the 3 treated groups compared to untreated CKD. No changes of creatinine clearance were observed while albuminuria increased in the DAPA groups with no significant effects of EPLE. The fractional shortening and the cardiac output were not modified by the treatments (not shown). Cardiac hemodynamic was improved with reduced LV end diastolic pressure (LVEDP) in the CKD rats treated with EPLE or a combination of EPLE and DAPA while LV end diastolic pressure volume relationship (LVEDPVR) was decrease by both DAPA and EPLE and further decreased by DAPA + EPLE. Decreased cardiac perfusion was prevented with EPLE alone or with the combination.

Conclusion:
The use of DAPA + EPLE appears to be more effective than DAPA or EPLE alone on diastolic function in non-diabetic CKD rats while cardiac perfusion is improved by EPLE only.
Abstract 112: Effects Of Intensive Blood Pressure Treatment On Orthostatic Hypertension: An Individual-level Meta-analysis
Authors: Juraschek Stephen P , Hu Jiun-Ruey , Cluett Jennifer , Mita Carol , Lipsitz Lewis , Appel Lawrence J , Beckett Nigel , Davis Barry R , Holman Rury , Miller Edgar R , Mukamal Ken J , Peters Ruth , Staessen Jan A , Taylor Addison A , Wright Jackson T , Cushman William C
Published: October 2023

Background:
Orthostatic hypertension (OHTN) is associated with cardiovascular disease (CVD). We previously demonstrated how more intensive blood pressure (BP) treatment reduced orthostatic hypotension (OH) via a net increase in BP upon standing. Whether this effect also increased OHTN was not examined.

Methods:
We used pooled data from an individual participant data meta-analysis of 9 randomized BP pharmacologic treatment trials with seated and standing BP (search conducted through May 13, 2022). OHTN was defined as an increase in SBP ≥20 or DBP ≥10 mm Hg after changing from sitting to standing. Effects were examined overall and by baseline characteristics.

Results:
Of 31,124 participants with 315,497 standing BP assessments, 9% had OH, and 20% had OHTN at baseline. Trial effects were similar (

I
2

= 38.0%). During follow-up, 17% of those assigned more intensive treatment had OHTN, while 19% of those assigned less intensive treatment had OHTN. Intensive BP treatment significantly lowered OHTN risk (OR 0.93; 95% CI: 0.90, 0.96) (
Figure
). Effects were significantly greater among non-Black adults (OR 0.86 vs 0.97,
P
-interaction =0.003) and adults without diabetes (OR 0.88 vs 0.96;
P
-interaction =0.047). Effects did not differ by age≥75 years, sex, baseline seated BP≥130/≥80 mm Hg, obesity, stage 3 kidney disease, stroke, cardiovascular disease, standing SBP ≥140 mm Hg, or pre-randomization OHTN (
P
-interactions ≥0.05).

Conclusion:
In this pooled cohort of adults with hypertension or higher CVD risk, OHTN was more common than OH and more intensive BP treatment reduced OHTN risk. The relationship between this effect with respect to CVD should be a focus of subsequent research.
Abstract 122: Cerebral Artery Function And Stiffness Is Dependent On
Apoe
Genotype And Ovariectomy Status In Mice
Authors: Kehmeier Mackenzie N , Cannon Audrey T , Khurana Aleena , Cullen Abigail , Alkayed Nabil J , Walker Ashley E
Published: October 2023

The major risk factors for late-onset Alzheimer’s Disease (LOAD) include old age, female sex and the
APOE4
allele. Post-menopausal females with an
APOE4
allele have higher rates of LOAD compared with age-matched males with an
APOE4
allele, suggesting an interaction between
APOE
genotype and estrogen deficiency. Importantly, cerebrovascular dysfunction is a contributor to LOAD risk. However, it is unknown if
APOE
genotype and estrogen deficiency interact to impact cerebrovascular function.Young female homozygous
APOE3
and
APOE4
mice (n=4-6 per group; ~6 months old) fed a high-fat, high-cholesterol diet were intact (sham) or ovariectomized (OVX). We assessed endothelium-dependent dilation to increasing doses of acetylcholine (ACh) in pressurized posterior cerebral arteries (PCAs). There was an interaction effect of
APOE
genotype and OVX status for endothelium-dependent dilation (p=0.03), such that
APOE3
-Sham mice had a greater endothelium-dependent dilation compared to the
APOE3
-OVX group (70±9.1% vs. 39±7.6%, p=0.003). These differences were mediated by nitric oxide as indicated by the absence of dilation in the presence of N omega-Nitro-L-arginine methyl ester hydrocholoride (L-NAME, Sham: 3.7±3.3% vs OVX: 4.1±4.0%, p= 0.99). In contrast,
APOE4
-OVX had preserved endothelium-dependent dilation compared with
APOE4
-Sham (49.9±14.8% vs 56.1±9.2%, p=0.8). Endothelium-independent dilation was assessed by the response to increasing doses of sodium nitroprusside and did not differ among groups (all p>0.05). Passive stiffness was measured in isolated PCAs after incubation in a calcium-free solution. For the elastic modulus at low pressures, there was an interaction effect of
APOE
genotype and OVX status (p=0.03), with
APOE4
-OVX having a higher elastic modulus compared to
APOE4
-Sham (0.93±0.3 vs. 0.47±0.1, p=0.03). The elastic modulus was similar in
APOE3
groups (sham: 0.64±0.01, OVX: 0.62±0.09, p=0.99). Overall, we found that estrogen deficiency impacts cerebral artery endothelial function in
APOE3
, but not APO
E4
mice, while estrogen deficiency impacts cerebral artery stiffness in
APOE4
, but not
APOE3
mice. These results shed light on the impact of
APOE
genotype on the cerebral vasculature, suggesting a potential mechanism for LOAD.
Abstract 123: Partial Endothelial Cell
In Vivo
Reprogramming Reverses The Hypercontractility To Phenylephrine In Resistance Arteries Isolated From Males But Not From Females BPH/2J Hypertensive Mice
Authors: Pernomian Laena , Waigi Emily W , Townsend Paul , Wilson Emily W , Pujhari Sujit , McCarthy Cam , Wenceslau Camilla F
Published: October 2023

The Schlager hypertensive (BPH/2J) mouse strain is a genetic model of spontaneoushypertension. Endothelial dysfunction is a hallmark characteristic of hypertension,including endothelial phenotypic transition. We hypothesized that treating endothelialcells with the transcription factors Oct4, Sox2 and Klf4, will lead to partial reprogramming,reduce non-functional phenotype changes, and prevent vascular dysfunction. Mouseendothelial cells were transduced with a lentiviral vector containing VE-cadherin-Oct4-Sox2-Klf4-eGFP (LVTF), control lentivirus (LVCO), or phosphate buffer saline (PBS).After 72h, cell migration and proliferation, Ki67 marker, and the endothelial progenitormarkers (CD31, CD133 and CD34) were evaluated. Male or female BPH/2J mice (33-42-week-old) and their age-matched controls BPN/3J (normotensive) received a singleinjection of LVCO or LVTF (100μL) via the tail vein and were evaluated after 10 days.Systolic blood pressure (SBP) was measured directly by catheterization of the left carotidartery. Mesenteric resistance arteries were isolated and mounted on wire myographs toevaluate phenylephrine-induced contraction. Data were analyzed using One-way orTwo-way ANOVA, with Bonferroni correction (
p
<0.05). Lentivirus-transduced endothelialcells had eGFP fluorescence (LVCO: 45±2%, LVTF:19±1%, n=5), and LVTF increasedOct4 (LVTF: 5.6±0.7%
vs.
LVCO: 1.8±0.2%
vs.
PBS: 1.5±0.4%, n=5) and CD133 (LVTF:11.7±0.8%
vs.
LVCO: 7.6±0.3%
vs.
PBS: 6.5±0.5%, n=5), but not CD31, CD34 or Ki67.Cell migration was lower in LVTF (71±2%) than LVCO cells (90±2%, n=5). Male andfemale BPH/2J treated with LVCO had higher SBP (male: 103±3mmHg, female:95±4mmHg, n=2-3) compared to BPN/3J mice (male: 88±1mmHg, female: 80±4mmHg,n=2-6), and LVTF treatment tended to decrease SBP in male BPH/2J (86±2mmHg, n=2,
p
=0.0548) but not in female mice (90±4mmHg, n=3). Phenylephrine-induced contractionwas reduced in arteries from male BPH/2J treated with LVTF (6.77±0.90mN, n=3), butnot in female mice (11.01±1.45mN, n=3) compared to LVCO (male: 12.23±0.15mN;female: 10.77±0.92mN; n=2-3). These data reveal that partial endothelial cellreprogramming
in vivo
is a new strategy to prevent endothelial dysfunction in malehypertensive mice.1
Abstract 125: Elevated Bacterial Fatty Acid Metabolism And Fecal Calorific Values Are Associated With Reduced Diet-induced Adiposity And Weight Gain In Mice
Authors: Alviter Plata Adriana , Martyniuk Chris , Gadient Jennifer , Zubcevic Jasenka
Published: October 2023

Introduction:
Activation of the sympathetic nervous system (SNS) leads to exaggerated immune responses in cardiometabolic conditions. We previously showed that ablation of beta-adrenergic receptors (ADRB) on the bone marrow (BM) hematopoietic cells reduces blood pressure, dampens the immune responses, and modifies gut microbiota, suggesting multisystem host-microbiota interactions. To investigate the role of the microbiota in these complex interactions, we tested the effects of high fat diet (HFD) on the gut microbiota and adiposity in BM mouse chimera characterized by reduced SNS-immune signaling.

Methods:
Male C5BL/6 wild type (WT) mice were irradiated and reconstituted with BM cells from either the C57BL/6 WT controls or the ADRB 1/2 knock-out mice (KD) to generate BM chimera. Following recovery, mice were fed with a control diet (CNT) or HFD for two weeks
ad libitum
. Food intake and body weights were measured weekly, and body fat (NMR) and visceral fat pad weights were measured at endpoint. At endpoint, 16S sequencing and metagenomic analyses (QIIME2, PiCRUST2) were performed in cecal samples, and residual fecal calorific values were determined using a simultaneous thermogravimetric analyzer differential calorimeter (TA Instruments).

Results:
No change in food intake was observed in either group. However, significant increases in body weight, total body fat, and visceral fat weight were observed in the WT compared to KD chimera on HFD (
p<0.05)
. HFD elevated gut bacterial Shannon α-diversity
(p< 0.05)
in the WT but not the KD chimera, and we observed a decrease in
g_Faecalibacterium
and
p_Actinobacteria
in WT, and an increase in
p_Bacteroidetes
and
o_Bacteroidales
in the KD mice on HDF. Enrichment in fatty acid metabolic pathways and increased fecal calorific values were observed in the KD compared to WT chimera on HFD (
p<0.05)
.

Conclusion:
Reduced SNS-immune signaling is protective against diet-induced weight gain and body fat accumulation. This may in part be mediated by the gut microbiota, reflecting in increased bacterial fatty acid metabolism and elevated residual fecal calorific values, suggesting reduced fat absorption. We propose that selective enrichment of gut bacteria is a potential therapeutic target in diet-induces obesity.
Abstract 127: Time Restricted Feeding In Diet Induced Obesity Reduces Kidney Cd8
+
T Cells
Authors: Edell Claudia , Colson Jackson , Pati Paramita , Pollock David M , De Miguel Carmen , Pollock Jennifer
Published: October 2023

Diet induced obesity (DIO) is one of the leading risk factors for chronic kidney disease. Previous studies identified a role for pro-inflammatory T cells in mediating DIO-driven renal damage. We found that time restricted feeding (TRF; food available only in 12-hr dark phase) during the final 2 weeks of a 20-week DIO model (45% fat) significantly improves renal damage compared to DIO
ad libitum
fed male mice. DIO+TRF also significantly reduced total T cells in the kidney adjacent to vasa recta in the outer medulla compared to DIO
ad libitum
mice assessed at the start of the dark phase or Zeitgeber Time (ZT) 12. We hypothesized that DIO+TRF decreases renal and circulating pro-inflammatory CD4
+
and CD8
+
T cells compared to DIO
ad libitum
in a time-of-day dependent manner
.
Using flow cytometry, we found that DIO significantly increases total renal T cells compared to normal diet (ND; 10% fat) at ZT13-15 (dark phase) only. DIO+TRF significantly decreases total renal T cells compared to DIO in the dark phase (n=7-14; 2-way ANOVA, Interaction: p=0.016, Effect of Time of Day: p=0.010, Effect of Diet: p=0.001). Our data showed no significant changes in renal T cell numbers in the light phase. Compared to ND, DIO increased renal CD8
+
T cells at ZT13-15 and DIO+TRF normalized renal CD8
+
T cells (n=7-14; 2-way ANOVA, Interaction: p=0.007, Effect of Diet: p=0.004). DIO and DIO+TRF did not alter renal CD4
+
T cells at either ZT1-3 or ZT13-15 compared to ND. We further examined the kidney CD8
+
T cell phenotype with DIO and TRF. We also found no significant changes in CD8
+
T cell activation markers, perforin and granzyme B, CD8
+
T cell development marker EOMES, or CD8
+
T cell pro-inflammatory cytokines, IFNγ and TNFα. We assessed circulating CD8
+
and CD4
+
T cells via flow cytometry to analyze systemic effects of TRF. DIO+TRF led to a diurnal rhythm of CD8
+
T cells with increased circulating CD8
+
T cells in the dark phase compared to the light phase that was not observed in the ND or DIO groups (n=7-17; 2-way ANOVA, Interaction: p=0.012). No significant changes were observed in circulating CD8
+
T cell activation markers or cytokines. DIO and TRF had no significant effect on numbers of circulating CD4
+
T cells. We conclude that DIO+TRF influences rhythms of circulating and renal CD8
+
T cells, but not CD4
+
T cells.
Abstract 129: Ngal Is A Novel Target In Hypertension By Modulating The Ncc-mediated Renal Na Balance
Authors: Jaisser Frederic , Bonnard Benjamin , Shimosawa Tatsuo , Xhaard Constance , rossignol patrick
Published: October 2023

Background:
The expression of neutrophil gelatinase-associated lipocalin (NGAL/lcn2) is directly modulated by mineralocorticoid receptor activation but its role in blood pressure control is unclear.

Methods:
a potential relationship between NGAL plasma levels, systolic blood pressure and urinary Na excretion was assessed in the STANISLAS cohort. The specific role of NGAL/lcn2 in salt-sensitive hypertension was studied using lcn2-depleted mice (lcn2 KO) fed with low Na diet (0Na).

Results:
we show that NGAL plasma levels positively correlate with systolic blood pressure (SBP), whereas they negatively correlate with urinary Na excretion in subjects of the STANISLAS cohort. Prolonged feeding of lcn2 KO mice with a 0Na diet induced lower SBP than that of the control group (WT), suggesting a role for NGAL/lcn2 in Na-balance homeostasis. Short-term or prolonged 0Na increased Na-Cl cotransporter (NCC) phosphorylation in the cortex of WT mice, which was prevented in lcn2 KO mice. Recombinant mouse lcn2 injections in lcn2 KO mice induced NCC phosphorylation in the kidney cortex, associated with decreased urinary Na excretion.
Ex-vivo
experiments using kidney slices from lcn2 KO mice showed increased NCC phosphorylation by recombinant murine lcn2. In addition, recombinant murine lcn2 induced activation of CamK2β phosphorylation in lcn2 KO mice and in kidney slices, providing an underlying mechanism involved in lcn2-induced NCC phosphorylation. Indeed, the inhibition of CamK2β prevented NCC phosphorylation induced by recombinant lcn2 in kidney slices.

Conclusion:
we highlight a novel role of NGAL/lcn2 as a modulator of the activity of the renal sodium transporter NCC affecting salt-sensitive blood pressure
Abstract 132: Proteomic Profiles Of Human Arterioles Isolated From Fresh Adipose Tissue Or Following Overnight Storage
Authors: Pandey Rajan , Roberts Michelle L , Wang Jingli , Pereckas Michaela , Jensen David , Greene Andrew S , Widlansky Michael E , Liang Mingyu
Published: October 2023

Resistance arteries, or arterioles, are key determinants of the total peripheral vascular resistance, which, in turn, is a key determinant of arterial blood pressure (BP). However, the amount of protein available from one isolated human arteriole may be less than 5 μg, making proteomic analysis challenging. In addition, obtaining human arterioles requires manual dissection of unfrozen clinical specimens. This limits its feasibility, especially for powerful multi-center clinical studies in which clinical specimens need to be shipped overnight to a research lab for arteriole isolation. We performed a study to address low input, test overnight tissue storage, and develop a reference human arteriolar proteomic profile. We found that, in tandem mass tag proteomic analysis, the use of a booster channel consisting of endothelial and vascular smooth muscle cells (1:5 ratio) increased the number of proteins detected in a human arteriole segment with FDR < 0.01 from 1,051 to more than 3,000. We collected adipose tissues from three human subjects and isolated two arterioles from each fresh aliquot of the tissue or after 24h of cold storage of unfrozen aliquots in MACS tissue solution. The correlation coefficient of proteomic profile was similar (p=0.6) between replicate arterioles isolated freshly, following the cold storage, or before and after the cold storage. We built a human arteriolar proteomic profile consisting of 3,836 proteins based on the analysis of 12 arteriole samples from the three subjects. The average arteriolar protein had a mean copy number of 2.76х10
6
per cell using the histone proteomic ruler, which is based on the fact that the mass of DNA per cell is approximately equal to the protein mass of histones. Transgelin was the most abundant protein detected. We curated a set of BP-relevant human genes, which encode 1,945 proteins. Of these BP-relevant proteins, 476 (12.5%) were detected in the arteriolar proteome, which was a significant overrepresentation (p<0.05, Chi squared test). These findings demonstrate that proteomic analysis is feasible with arterioles isolated from human adipose tissue following cold overnight storage and provide a reference human arteriolar proteome profile highly valuable for studies of arteriole-related traits.
Abstract 133: Urine Derived Human Renal Proximal Tubule Cells Isolated From Inverse Salt Sensitive Participants Have Higher C-src Kinase Activity And Aminopeptidase N Expression
Authors: Gildea John J , Xu Peng , Felder Robin
Published: October 2023

We have previously published that 11% of individuals in a salt sensitivity clinical trial are determined to be Inverse Salt Sensitive (ISS), and are associated with SNPs in the Dopamine D2 Receptor (D
2
R) that are shown to have decrease D
2
R expression in urine derived human renal proximal tubule cells (uRPTC) compared to salt resistant (SR) and salt sensitive (SS) individuals. We also previously have shown that ISS uRPTCs have increased expression of aminopeptidase N (APN), the enzyme capable of converting AngIII to AngIV. Inhibition of this activity in ISS may be a therapeutic strategy for ISS individuals. We previously reported that a nanobody designed to inhibit human APN activity added to cells reverses a number of adverse cell physiologic responses to low sodium conditions in ISS derived uRPTCs. Here we show evidence that the increased expression of APN in ISS uRPTCs is due to increased constitutive C-SRC activity. Three stable ISS uRPTCs, three SR uRPTCs and three SS uRPTCs were grown and lysed for analysis by Western blotting. The human homolog of the V-Src Avian Sarcoma viral oncogene tyrosine kinase (C-Src) activation state was measure using a Y-416 C-Src tyrosine phosphorylation specific antibody. There is a 266.1±21.7% increase in ISS Y416 p-Src vs SR (p<0.01 ISS vs SR, N=3) and no difference in SS vs SR. Plasma membrane localized APN was measured by In-cell Western and the increase in APN expression between ISS and SR was completely blocked by incubating cells for 4 hours with the C-Src inhibitor PP2 (ISS 49.1±8.8% increase over SR vs -14.6±11.9% with PP2, p<0.01, N=4 per group). Similarly lithium chloride (10 mM 4hrs), and D
2
R overexpression normalized the differential expression of D
2
R in ISS and completely blocked the increased APN expression found in ISS. In summary, low D
2
R expression in ISS leads to constitutively active C-Src and increased APN expression, and re-expression of D
2
R or inhibition of C-Src normalizes expression of APN. Future studies will determine if the tyrosine phosphorylation site in the intracellular domain of APN is a direct target of C-Src.
Abstract 139: Hypertension Control In The Afro-dpp Program, A Virtual Cardiometabolic Health Intervention Among African Immigrants
Authors: Ogungbe Oluwabunmi V , Turkson-Ocran Ruth-Alma , HINNEH THOMAS , Owusu Loretta , Spaulding Erin , Mensah Jasmine , Gbaba Serina , Sinyan Aminata , Assani-Uva Adeline , Commodore-Mensah Yvonne
Published: October 2023

Background:
Black people and immigrants in the US are disproportionately affected by poor cardiometabolic health.

Aim:
To test the effectiveness of a virtual, culturally tailored lifestyle intervention with remote blood pressure (BP) monitoring on BP control among African immigrants.

Methods:
In this pilot cluster-randomized trial (NCT05144737), two churches were randomly assigned to first intervention or delayed intervention groups. African immigrants aged 25-75 years with a smartphone and ≥ 2 of the following were enrolled: body-mass index (BMI) ≥ 25 kg/m
2
, HbA1c 5.7-6.5%, systolic BP (SBP) ≥ 140 mmHg or diastolic BP (DBP) ≥ 90 mmHg. Intervention involved lifestyle coaching using the diabetes prevention program curriculum, adapted to African immigrants, Bluetooth-enabled BP monitor synced to a mobile app (Sphygmo
TM
), allowing for remote BP readings. The primary outcome was 6-month change in BP.

Results:
A total of 60 participants, 30 per site, were recruited; mean age 50.6(±11.9). At baseline, mean SBP was 144.5 mmHg in the first intervention group and 148.3 mmHg in the delayed intervention group. At 6 months, there was 11.32 mmHg SBP, 9.73 mmHg DBP reduction in the first intervention group and 9.31 mmHg SBP, 5.34 mmHg DBP reduction in the delayed intervention group (
Figure
); mean SBP reduction was 4.35 mm Hg greater in the first intervention group (95% CI, -13.56, 4.87). Additional BP control (< 130/80 mmHg) was achieved in 22% of participants in the first intervention group, and 10% in the delayed intervention group.

Conclusions:
Among African immigrants at risk for cardiovascular disease, a culturally tailored, lifestyle intervention led to BP reduction.
Abstract P114: Patient Preferences And Experiences With Cuffless Wrist Versus Upper Arm Cuff Blood Pressure Measurements In The

CO

ntinual Vs.

O

ccasiona

L

B

lood

P

ressure Monitoring (COOL-BP) Study
Authors: Unlu Ozan , Lee Simin , Gabovitch Daniel , Perruchoud David , Almeida Tiago , Collins Emma , Figueroa Christian , Ruggiero Ryan , Chin Nicholas , Cannon Christopher P , Scirica Benjamin , Fisher Naomi D
Published: October 2023

Background:
Cuffless BP devices can potentially enhance hypertension management and remote BP monitoring. Patient experience with these devices is a key driver of their acceptability and uptake.

Methods:
We implemented a remote hypertension program within the Mass General Brigham health system using traditional home BP monitoring to adjust medications. Selected patients in this program were enrolled in an ancillary study where BP was also measured via a cuffless wrist monitor (Aktiia SA, Switzerland). We analyzed patient experience surveys of all patients who successfully used each device for at least one full week in the program and completed surveys at 30, 60, and 180 days after enrollment. Final (180-day) surveys were analyzed.

Results:
Of 35 patients who used both devices, 34 completed all 3 surveys. Thirty-one patients (91%) indicated they preferred using the cuffless wrist monitor over the upper arm BP cuff, 1 (3%) preferred the BP cuff, and 2 (6%) had no preference. Top reasons for preference of the bracelet included ease of use, convenience, and number of readings (Figure). Most patients (27/34, 79%) used the cuffless device either daily or most days of the week, and 23 (68%) found it easy or very easy to use. Twenty-eight (82%) patients believed the bracelet could help manage their hypertension; 32 (94%) indicated that direct transmission of bracelet BP to their providers would be valuable.

Conclusion:
Among patients who used both traditional home BP monitoring and a cuffless continual BP monitoring bracelet in a remote hypertension management program, the vast majority found the cuffless device easier to use and more convenient, and preferred it over traditional BP cuffs.