bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P199: Mas Receptor Deficiency Regulates Obesity-Hypertension and Cardiac Function in Female and Male Mice
Authors: Shoemaker Robin C , Wang Yu , Thatcher Sean , Cassis Lisa
Published: June 2021

Angiotensin-1-7 (Ang-(1-7)) counteracts angiotensin II through effects at Mas receptors (MasR). We demonstrated that sexual dimorphism of obesity-hypertension was associated with dysregulated production of Ang-(1-7). However, the role of MasR in sexual dimorphism of obesity-hypertension has not been examined. MasR deficient mice have also been reported to exhibit deficits in cardiac function. In this study, we hypothesized that deficiency of the MasR would differentially regulate obesity-hypertension in male
versus
(
vs
) female mice. In addition, we quantified effects of MasR deficiency on cardiac function in obese male mice. Male and female

MasR
+/+

and

-/-

mice were fed a low fat (LF, 10%kcal) or high fat (HF, 60% kcal) diet for 16 weeks, and blood pressure was quantified by radiotelemetry. As demonstrated previously, male

MasR
+/+

mice (24 hr diastolic blood pressure, DBP: LF, 90 ± 3; HF, 96 ± 2 mmHg; P<0.05), but not females (LF, 85 ± 1; HF, 85 ± 2 mmHg), developed hypertension in response to HF feeding. MasR deficiency converted female HF-fed mice to an obesity-hypertension phenotype (DBP: 92 ± 1 mmHg; P<0.05). Surprisingly, male HF-fed

MasR
-/-

mice exhibited reduced DBP compared to HF-fed

MasR
+/+

males (90 ± 1
vs
96 ± 2 mmHg; P<0.05). To define mechanisms for reductions in DBP of HF-fed male

MasR
-/-

mice, we performed cardiac magnetic resonance (CMR) imaging in both genotypes at 1 month of HF feeding.

MasR
-/-

mice had significantly reduced ejection fraction (EF) compared to

MasR
+/+

mice at baseline (51.4 ± 2.5
vs
59.3 ± 2.1%; P<0.05) and after one month of HF-feeding (49.8 ± 2.4
vs
52.6 ± 1.9%; P<0.05). Further, CMR imaging demonstrated a thickening of the ventricle wall in

MasR
-/-

mice with 1 month of HF-feeding.

MasR
+/+

, but not

MasR
-/-

mice, exhibited diet-induced reductions in EF (by 16%; P<0.05) at 1 month of HF feeding, which were reversed by infusion of Ang-(1-7). These results demonstrate that MasR contributes to sexual dimorphism of obesity-hypertension. Ang-(1-7) protects females from obesity-hypertension through the MasR. In contrast, reductions in DBP in obese male mice with MasR deficiency may arise from deficits in cardiac function. These results suggest that MasR agonists may be effective therapies for obesity-associated cardiovascular conditions.
Abstract P200: Role of Macula Densa Neuronal Nitric Oxide Synthase in Control of Renin Release and Blood Pressure Recovery Following Hemorrhagic Shock
Authors: Wang Lei , Wang Shaohui , Buggs Jacentha , Zhang Jie , Wei Jin , Zhang Gensheng , Liu Ruisheng
Published: June 2021

Renin is a rate limiting factor for generation of angiotensin II, which is essential for blood pressure regulation. The role of macula densa nitric oxide (NO) in renin release is not conclusive. The goal of this study was to elucidate the role of macula densa neuronal NO synthase (NOS1) in control of renin release in response to sodium challenges and hemorrhagic shock, as well as in blood pressure recovery after hemorrhagic shock.

C57BL/6L mice and macula densa specific NOS1 knockout (MD-NOS1KO) mice were given 10 days of low (0.1% NaCl), normal (0.4% NaCl) and high (1.4% NaCl) sodium diet. Hemorrhagic shock was induced by withdrawing 0.4 ml whole blood from the right retro-orbital sinus. Mean arterial pressure (MAP) in conscious mice was monitored by radio-telemetry system. Plasma renin concentration (PRC) was determined by radioimmunoassay.

Low sodium diet stimulated PCR by 29% (from 685 ± 32 to 883 ± 112 ng/ml/hr) in WT mice and by 16% (from 652 ± 24 to 756 ± 124 ng/ml/hr) in the MD-NOS1KO mice (n=5/group, p<0.01 vs WT). PCR was not significantly different between the WT and MD-NOS1KO mice fed a normal or high salt diet. As shown in Fig1A, following removal of 0.4ml of blood, MAP dropped to about 40mmHg in the WT mice and 35mmHg in the MD-NOS1KO mice. MAP recovered faster in WT mice than the MD-NOS1KO mice. In Fig1B, PRC increased over 200% of the basal value in WT mice, but only increased about 26% in the MD-NOS1KO mice (n=4/group, p<0.01 vs WT).

We conclude that NOS1 in the macula densa facilitates renin release. Lack of macula densa NO generation impairs blood pressure recovery, which may be mediated by limiting renin release during hemorrhagic shock.
Abstract P206: Increased Short-term Dietary Sodium Intake Does Not Increase Blood Pressure or Measures of Aortic Stiffness in Young Healthy Adults
Authors: Rosenblum Hannah , Katz Stuart
Published: June 2021

Background:
Sodium (Na) sensitivity is defined as the short-term increase in blood pressure (BP) after increased Na intake. The impact of Na sensitivity on measures of aortic stiffness has not been previously characterized in non-hypertensives. We tested the feasibility of an oral Na loading protocol to characterize the effects of increased dietary Na on blood pressure and measures of aortic stiffness in health young adults.

Methods:
In an open-label repeated measures pilot study, we enrolled 40 healthy adults age 21-30 years (18 females). Subjects were evaluated before and after 7 days of oral Na loading (12g (205 mmol) of salt tablets/day). BP (mmHg, cuff method) was measured in supine, seated and standing positions. Radial artery tonometry (Sphygmacor) was used to assess measures of aortic stiffness: aortic BP (mmHg), aortic pulse pressure (mmHg), aortic augmentation index (%) and adjusted Tr, time of return of the reflected pressure wave as an index of aortic pulse wave velocity (m/s). Serum aldosterone, urine Na and potassium (K) were measured.

Results:
Nine subjects dropped out due to GI intolerance of the salt pills. In the remaining 31 subjects, Na loading did not change BP in any position, aortic BP, aortic pulse pressure, or Tr (Table: mean±SD for seated R arm BP and measures of aortic stiffness). There was no measurable aortic augmentation in the cohort. Na loading decreased serum aldosterone (9.1±1.5 to 5.2±0.8 ng/dl, p=0.02), and increased urine Na:K ratio (1.8±0.2 to 3.5±0.5, p=0.002).

Conclusion:
Oral Na loading was associated with suppression of the renin-angiotensin aldosterone system, did not increase BP, or measures of aortic stiffness in healthy young adults.
Abstract P219: Exercise Training Normalizes Vascular Changes in Aging Hypertension Involving microRNAs Profile and Target Genes
Authors: Fernandes Tiago , Roque Fernanda R , Neves Vander J , Penteado João L , Silveira André C , Mesquita Suliana , Jordão Camila P , Souza Rodrigo , Rossoni Luciana V , Oliveira Edilamar M
Published: June 2021

MiRNAs profile and target genes involved in hypertension-associated vascular changes were evaluated. In addition, we checked the therapeutic role of exercise training (ET) on these parameters. Spontaneously hypertensive rats (SHR) aged 6 months and their controls Wistar Kyoto (WKY) were divided into 4 groups: SHR, trained SHR (SHR-T), WKY and trained WKY (WKY-T). Swimming ET consisted of 60 min of duration, 1x/day/10 weeks, with 4% caudal body weight workload. SHR showed an increased in systolic blood pressure (207 ± 5.5 mmHg) compared to WKY (133 ± 3.9 mmHg) analyzed by tail-cuff system, with no changes in baseline heart rate. We observed a reduction in VO
2
peak (WKY: 62 ± 1.5; SHR: 53 ± 2.5 mL.kg
-1
.min
-1
) accompanied by soleus muscle atrophy (fiber type I - WKY: 4039 ± 195; SHR: 2658 ± 53; type IIa - WKY: 2903 ± 182, SHR: 2050 ± 68; Intermediate - WKY: 2663 ± 136, SHR: 1967 ± 95 μm
2
) in SHR. Vascular function of femoral artery was similar in SHR and WKY, however, wall-to-lumen ratio was increased in femoral artery (WKY: 0.17±0.01, SHR: 0.27±0.01 a.u.) and muscular arteriole (WKY: 0.54±0.02, SHR: 1.14±0.03 a.u.) accompanied by capillary rarefaction (WKY: 1.2 ± 0.05, SHR: 0.6 ± 0.03 capillary-to-fiber ratio) in soleus muscle from SHR vs. WKY. In contrast, ET promoted reduction in blood pressure and resting bradycardia in trained animals. ET corrected the VO
2
peak reduction, muscle wasting, microvascular remodeling in SHR-T toward control levels. ET downregulated 8 miRNAs (-96, -205, -182, -146b-5p, -140, -328a, -665, -1) and upregulated 3 miRNAs (-499, -208b and -99b) in SHR-T when compared to SHR. Bioinformatics study for functional analysis of the predicted targets genes for the 11 miRNAs restored by ET demonstrated enrichment of different signaling pathways including cell death (p value <0.001; Fold enrichment 16.78) and vascular development (p value < 0.001; Fold enrichment 10.62). Thus, targets involved in angiogenesis and vascular integrity by the VEGF/VEGFR2/AKT/eNOS/Bcl-2 pathway were impaired in hypertension and corrected by ET. The results support the hypothesis that the structural changes arising from the progression of hypertension may be regulated by a set of miRNAs and target genes; and ET participates in restoring the vascular remodeling.
Abstract P224: Endothelin-1 Overexpression Preserves Endothelial Function in Mice with Vascular Smooth Muscle Cell-specific Deletion of PPAR-gamma
Authors: Ouerd Sofiane , Idris-Khodja Noureddine , Trindade Michelle , Gornitsky Jordan , Rehman Asia , Barhoumi Tlili , Offermanns Stefan , Gonzalez Frank J , Neves Mario F , Paradis Pierre , Schiffrin Ernesto L
Published: June 2021

Objective:
Peroxisome proliferator-activated receptor γ (PPARγ) agonists reduce blood pressure (BP) and vascular injury in hypertensive rodents.
Pparγ
inactivation in vascular smooth muscle cells (VSMC) using a tamoxifen inducible Cre-Lox system enhanced angiotensin II-induced vascular damage. Transgenic mice overexpressing endothelin (ET)-1 in the endothelium (eET-1) exhibit endothelial dysfunction, increased oxidative stress and inflammation. We hypothesized that inactivation of the
Ppar
gene in VSMC (sm
Pparγ
-/-
) would exaggerate ET-1-induced vascular damage.

Methods and Results:
Eleven-week-old male control, eET-1, sm
Pparγ
-/-
and eET-1/sm
Pparγ
-/-
mice were treated with tamoxifen (1 mg/kg/day, s.c.) for 5 days and sacrificed 4 weeks later. Systolic BP determined by telemetry was higher in eET-1 (123±5 vs 109±2 mm Hg,
P
<0.05) and unaffected by sm
Pparγ
inactivation. Mesenteric artery (MA) vasorelaxation to acetylcholine was impaired only in sm
Pparγ
-/-
(E
max
: 52.0±6.7 vs 82.2±4.9%,
P
<0.05). MA reactive oxygen species levels were increased 1.7±0.3-fold in sm

Pparγ
-/-

(
P
<0.05) and further increased in eET-1/sm

Pparγ
-/-

(2.5±0.3-fold,
P
<0.05). MA perivascular fat monocyte/macrophage infiltration was higher in eET-1 and sm

Pparγ
-/-

(331±34 and 326±49 vs 140±8 cells/mm
2
,
P
<0.05), and further increased in eET-1/sm

Pparγ
-/-

(557±77,
P
<0.05). The spleen fraction of CD11b
+
cells was increased in sm

Pparγ
-/-

(1.1±0.1 vs 0.47±0.1%,
P
<0.05) and further increased in eET-1/sm

Pparγ
-/-

(1.8±0.2%,
P
<0.05). The spleen fraction of Ly-6C
hi
monocytes was increased in eET-1 and sm

Pparγ
-/-

(24±3 and 27±4 vs 14±1%,
P
<0.05) but not in eET-1/sm

Pparγ
-/-

. The spleen fraction of T regulatory cells was increased in sm

Pparγ
-/-

(13±2 vs 9±1%,
P
<0.05) and decreased in eET-1 (7±1%,
P
<0.05), which was further decreased by eET-1/sm

Pparγ
-/-

(3±1%,
P
<0.05).

Conclusions:
These results suggest that increased ET-1 paradoxically preserves endothelial function in mice with inactivated VSMC
Pparγ
despite enhanced oxidative stress. Flow cytometry data indicate that infiltrating monocyte/macrophages in these mice might be anti-inflammatory.
Abstract P225: Nox1 or Nox4 Deletion Prevents Type-1 Diabetes-induced Endothelial Dysfunction
Authors: Ouerd Sofiane , Idris-Khodja Noureddine , Trindade Michelle , Coelho Suellen C , Neves Mario F , Jandeleit-Dahm Karin A , Paradis Pierre , Schiffrin Ernesto L
Published: June 2021

Objective:
The prognosis of type-1 diabetes is in part related to the increased risk of vascular complications such as atherosclerosis. Overproduction of reactive oxygen species by NADPH oxidase (NOX) is believed to play an important role in diabetes-related vascular injury. NOX1 may play a role in the macrovascular disease, whereas NOX4 may have protective actions. Nevertheless, their role in diabetic vascular injury is less well understood. We hypothesized that deletion of
Nox1
would prevent diabetes-induced endothelial dysfunction and vascular remodeling of small arteries whereas
Nox4
would exaggerate vascular injury in atherosclerosis-prone apolipoprotein knockout (

Apoe
-/-

) mice.

Methods:
Diabetes was induced by streptozotocin IP injections (STZ, 55 mg/kg/day) for 5 days in 6-week-old male

Apoe
-/-

mice,

Apoe
-/-

mice deficient in
Nox1
(

Apoe
-/-

/

Nox1
y/-

) and
Nox4
(

Apoe
-/-

/

Nox4
-/-

). Mice were studied 14 weeks later. Endothelial function and vascular remodeling were assessed in mesenteric arteries (MA) using pressurized myography.

Results:

Apoe
-/-

mice presented a maximal endothelium-dependent vasodilatory response (E
max
) to acetylcholine of 48±8%, which was further decreased by diabetes to 20±6%. In contrast, endothelium-dependent relaxations to acetylcholine were 1.5-fold higher in diabetic

Apoe
-/-

/

Nox1
y/-

and

Apoe
-/-

/

Nox4
-/-

mice compared to non-diabetic

Apoe
-/-

mice (E
max
: 72±9 and 70±7 vs 48±8%). Diabetes decreased MA stiffness in

Apoe
-/-

mice, as indicated by a rightward displacement of the stress-strain curves (strain at 140 mm Hg: 1.02±0.04 vs 0.75±0.04), which was blunted by
Nox1
or
Nox4
knockout (strain at 140 mm Hg: 0.77±0.02 and 0.81±0.02). MA media/lumen was unaltered by diabetes. Knockout of
Nox4
but not
Nox1
increased MA media/lumen 1.4-fold in diabetic

Apoe
-/-

mice (4.1±0.4 and 3.5±0.2 vs 2.9±0.2%).

Conclusions:
These results suggest that NOX1 and NOX4 play a pathophysiological role in diabetes-induced endothelial dysfunction and contribute to potentially maladaptive changes in vascular stiffness. NOX4 seems to have dual actions on the vasculature, as it is also protective against vascular remodeling of small arteries in type 1 diabetes.
Abstract P228: Protective Role of TRPM7-kinase Against Vascular Dysfunction and Fibrosis Induced by Aldosterone and Salt
Authors: Rios Francisco J , Hood Katie Y , Harvey Adam , Neves Karla B , Nosalski Ryszard , Anyfanti Panagiota , Camargo Livia L , Montezano Augusto C , Touyz Rhian M
Published: June 2021

TRPM7 is a cationic ion channel and with a serine/threonine kinase important for cellular Mg
2+
homeostasis. We recently showed that TRPM7-kinase plays a role in aldosterone-mediated vascular effects and inflammation. Here we explored the role of TRPM7-kinase in cardiac fibrosis and vascular function in aldosterone-induced hypertension in mice. Wild-type (WT) or heterozygote TRPM7-kinase domain (TRPM7+/-) were treated with infused aldosterone (600 μg/Kg/day) and NaCl 1% in drinking water (aldo/salt) for 4 weeks. Blood pressure (BP) was evaluated by tail-cuff. Vessel function was investigated in mesenteric arteries by wire and pressure myography. Protein expression was assessed in cardiac tissue by western-blot and histology. Aldo/salt increased BP in TRPM7+/- and WT to similar levels (137mmHg vs control 118mmHg). Mesenteric arteries from untreated TRPM7+/- mice were more sensitive to relaxation induced by acetylcholine (LogEC50: 7.6±0.1 vs 7.1±0.2, TRPM7+/- and WT, respectively), effects that were reduced by Aldo/salt treatment (LogEC50: 7.2±0.1). Phenylephrine-contraction and sodium nitroprusside-relaxation curves were similar among groups. Pressure myography showed that in WT, aldo/salt increase the diameter (26%) and cross-sectional area (40%), resulting in hypertrophic outward remodelling, whereas in TRPM7+/-, the treatment decreased the diameter (16%) and increase the wall/lumen ration (82%), resulting in eutrophic inward remodelling. Hearts from TRPM7+/- presented decreased expression of annexin-1, which is a target protein of TRPM7-kinase, that was further decreased by aldo-salt. Hearts from untreated TRPM7+/- mice had increased fibrotic markers: plasma galectin-3 (2.5ng/mL) vs WT (1.4ng/mL) and protein expression for fibronectin (2.4-fold) and TGFβ (2-fold), and the aging marker p-P66Sch (47%) which were similar to WT-aldo/salt. Aldo/salt induced higher collagen expression in TRPM7+/- than in WT animals (15%), as observed by picrosirius red staining. Our findings provide some insights into aldosterone signalling through TRPM7-kinase and suggest that this chanzyme may have protective actions, which when downregulated, promotes vascular remodelling and cardiac fibrosis in aldosterone-induced hypertension.
Abstract P234: CTMP Negatively Regulates Pathological Cardiac Hypertrophy via Akt-dependent Manner
Authors: Li Hongliang , Deng Ke-Qiong , Zhang Xiao-Jing
Published: June 2021

Pathological cardiac hypertrophy which represents a leading cause of morbidity and mortality worldwide is a pathological process related to multifactorial and multiple molecules and regulated by numerous signaling pathways. Deregulation of AKT signaling is important in cardiac hypertrophy and cardiac dysfunction, but the underlying mechanism is not fully understood. In this study, we identified carboxy-terminal modulator protein (CTMP), an endogenous AKT inhibitor, as a key regulator of cardiac hypertrophy in response to pressure overload. Our results demonstrated that CTMP levels were downregulated by about 40% in aortic banding (AB)–induced hypertrophied mice hearts and 50% in failing human hearts compared to their controls respectively. Mice overexpressing CTMP specifically in the heart were resistant to AB-induced cardiac hypertrophy, whereas cardiac-specific conditional CTMP-knockout mice exhibited an aggravated phenotype induced by pressure overload. Additionally, gain-or-loss of function experiments mediated by adenovirus demonstrated that CTMP also prevented an angiotensin II–induced hypertrophic response in isolated cardiomyocytes
in vitro
. Mechanistically, we discovered that AKT signaling was significantly activated in AB-treated WT hearts, which was blocked by cardiac overexpression of CTMP, whereas being enhanced by loss of CTMP in response to chronic pressure overload and agonist stimulation. Moreover, rescue-experiments revealed that inhibition of AKT activation through LY294002 ameliorated the cardiac abnormalities in CTMP-knockout mice after AB. Taken together, our present study provides both
in vitro
and
in vivo
evidences that CTMP functions as a novel negative regulator factor of pathological cardiac hypertrophy. The underlying mechanisms responsible for CTMP-elicited effects are dependent on the inhibition of AKT signaling. The above-mentioned findings also expand our knowledge of the mechanisms of cardiac hypertrophy and provide potential therapeutic targets for pathological cardiac hypertrophy and heart failure.
Abstract P257: Taking Fresh Aim at Improving Blood Pressure Control
Authors: Rakotz Michael K , Hasan Omar , Sanchez Eduardo , Wozniak Greg
Published: June 2021

Nearly 70 million adults in the United States have hypertension, and only about 50% of those adults have a blood pressure reading below 140/90 mm Hg. Target: BP is a national initiative co-led by the American Heart Association and American Medical Association aimed at improving blood pressure control nationally to reduce the number of Americans who suffer the consequences of high blood pressure including myocardial infarctions, heart failure, strokes, and chronic kidney disease. Target: BP will equip physician practices and health care systems with the resources and technical assistance to achieve a minimum 70% blood pressure control rate with a target of reaching 80% or higher. Target: BP is a multi-faceted, evidence-based, data-driven quality improvement initiative that leverages the AMAs membership and strategic focus on improving health outcomes and the AHAs success in disseminating large scale improvement initiatives like Get With the Guidelines - Stroke, an effective cerebrovascular quality improvement and recognition program. This initiative offers participating healthcare systems, ambulatory clinical practices and individual clinicians the following: A mass communication campaign to raise public awareness about the importance of controlling high blood pressure A quality improvement program and relevant tools to help clinicians and care teams implement the latest hypertension guidelines and improve management and treatment of patients with hypertension, including check lists, treatment algorithms, protocols and fact sheets Opportunities to contribute data for tracking blood pressure control rates and benchmarking against other participants regionally and nationally Opportunities to be recognized formally for achieving a 70% blood pressure control rate, improving blood pressure control, reducing therapeutic inertia and using self-measured blood pressure monitoring Access to a national advisory group of experts in improving blood pressure control Since its initiation in November 2015, over 250 healthcare systems and clinics have committed to participate in Target: BP.
Abstract P281: Alamandine-induced Vasorelaxation is Selectively Increased in Sp-shr
Authors: Leao Nadia , Etelvino Gisele , Santos Robson
Published: June 2021

The renin angiotensin system is implicated in hypertension and cardiovascular diseases. It’s actions are dependent of counter-regulatory modulation of its vasopressor and vasodepressor axes. Recently, our laboratory described and characterized a new component the RAS, alamandine and its receptor, the MrgD. Alamandine can be formed from angiotensin A through the action ACE2 or from angiotensin-(1-7), by a still unknown decarboxylase. Among the actions of alamandine, a endothelium-dependent vasorelaxation in aortic rings of mice and rats has been described. The aim of this study was to investigate the vasorelaxing effect of alamandine in aortic rings of hypertensive rats. The vasorelaxing effect of alamandine was tested in aortic rings taken from SP-SHR, Wistar and SHR, pre-contracted with phenylephrine (0.1 μM) with and without enthotelium or in the presence of L-Name (100 μM ) or Indomethacin ( 10 μM). In aortic rings from SP-SHR alamandine produced a pronounced dose-dependent relaxation when compared with Wistar (Emax= 80± 6,0 vs 45 ± 4). This response was diminished in the presence of L-Name (Emax= 39 ± 7 in SP-SHR and 1,0 ± 3 in Wistar). Indomethacin also attenuated the vasorelaxation produced by alamandine in aortic rings from SP-SHR (Emax = 40 ± 4,0), while a smaller effect was observed in Wistar rats (Emax= 31 ± 4). The vasorelaxing effect of alamandine in SP-SHR was abolished in endothelium desnude rings (Emax= -9 ± 4). When we compare two models of animals with hypertension, SHR-SP and SHR the vasorelaxing effect of alamandine was also more pronounced in SP-SHR ( Emax = 80± 6,0 and 38 ± 4 respectively).Taken together these results suggest that the vasorelaxing effect of alamandine is selectively increased in SP-SHR. The mechanism of it effect in SP-SHR appears to involve NO and prostaglandins release.
Abstract P282: Use of a CRISPR/Cas9 System for Specification of the Renin Cell Phenotype
Authors: Medrano Silvia , Brown Robin Isadora , Martinez Maria F , Sequeira-Lopez Maria Luisa S , Gomez R A
Published: June 2021

Renin is a key enzyme/hormone that controls blood pressure and fluid-electrolyte homeostasis. Renin transcription is subject to complex developmental, physiological, and pathological regulation. We have identified the cAMP pathway and associated epigenetic marks as crucial regulators of renin expression. However, those studies involved the use of cAMP activators or analogues or histone deacetylase inhibitors that alter the transcriptome and epigenome of the whole cell without exclusively targeting the renin locus.

The CRISPR/Cas9 Type II bacterial immune system has been modified for genome editing in eukaryotes. This system consists of a CRISPR-associated endonuclease (Cas9) and single guide RNA (sgRNA) to target specific genomic areas. Modifications to the Cas9 enzyme have allowed the use of CRISPR to regulate gene expression by targeted modification of epigenetic marks.

In this study we tested whether renin expression can be induced by a nuclease-null dCas9 protein fused to the catalytic core of the acetyltransferase p300 (dCas9p300) and sgRNAs directed to the renin promoter and enhancer.

We used cultured arteriolar smooth muscle cells of the renin lineage that constitutively express CFP (a renin lineage marker) and YFP only when the renin gene is turned on. Cells were transfected with a dCas9p300 and sgRNA expression vectors and analyzed for YFP expression 48h after transfection.

We tested four sgRNAs targeting the renin enhancer and five sgRNAs targeting the renin promoter either individually or in combination. We found few YFP+ cells when all 4 enhancer sgRNAs or 5 promoter sgRNAs were simultaneously used. The highest YFP expression (32±9 cells/well) was observed when two enhancer sgRNAs (at positions -2,757 and -2,631) and three promoter sgRNAs (at positions -719, -50 and -25) were simultaneously added to the cells. We did not find any YFP+ cells when the dCas9p300 plasmid was transfected without sgRNAs or when cells were transfected with a control plasmid.

Our data support targeted acetylation as a causal mechanism of renin transactivation. CRISPR/Cas9 provides a tool to study the regulation of renin expression by targeting epigenetic marks in the promoter and enhancer of renin.
Abstract P283: Effects of Ovarian Estrogens on Local RAS Activity in Cardiomyocytes and Non-cardiomyocytes
Authors: Sun Xuming , Ahmad Sarfaraz , Lin Marina , Zapata-Sudo Gisele , Cheng Cheping , Varagic Jasmina , Ferrario Carlos , Wang Hao , Groban Leanne
Published: June 2021

The relatively low efficacy of ACE-Is in the treatment of heart failure (HF) in women after estrogen (E2) loss may be due to their inability to reach the intracellular sites at which Ang II is generated and/or the existence of cell-specific mechanisms in which ACE is not the essential processing pathway for Ang II formation. We compared the metabolic pathway for Ang II formation in freshly isolated cardiomyocytes (CMs) and non-cardiomyocytes (NCMs) extracted from hearts of gonadal-intact and ovariectomized (OVX) adult WKY and SHR rats. Circulating levels of angiotensinogen (AOGEN) were higher in WKY (958 ± 47 pg/mL) vs. SHR (SHR: 626 ± 40 pg/mL; P<0.05 strain effect) and E2 loss augmented this effect in WKY (WKY OVX: 1,169 ± 66 pg/mL vs. SHR OVX: 625 ± 41 pg/mL). Correspondingly, plasma Ang II levels were higher in WKY vs. SHR (strain effect: WKY: 62 ± 6 pg/mL vs. SHR: 42 ± 9 pg/mL), independent of OVX. Chymase activity was nearly 40-fold higher in NCMs compared to CMs, and for the NCMs, activities were highest in cells from WKY vs. SHR and OVX vs. intact rats (P<0.05 strain and E2 effects, respectively) (Figure).

Neither strain nor gonad status influenced the lower ACE activity found in both NCMs and CMs. In contrast, ACE2 activity in CMs and NCMs was higher in cells from WKY vs. SHR (P<0.05 strain effect), independent of E2 status. We conclude that NCMs from WKY and SHR express significantly higher levels of chymase, ACE, and ACE2 activities. E2 loss leads to selective changes in the activity of chymase, but not ACE, in NCMs. The significance of these novel findings is that targeted cell-specific chymase rather than ACE inhibition may have a greater benefit in the management of HF in women after menopause.
Abstract P284: Angiotensin II Upregulates Cytochrome-450 4A Expression in Rat Kidney Through Type 1 Receptor
Authors: Wang Wanting , Rong Rong , Ito Osamu , Ogawa Yoshiko , Muroya Yoshikazu , Kohzuki Masahiro
Published: June 2021

20-hydroxyeicosatetraenoic acids (20-HETE) is a cytochrome P-450 (CYP) 4A-dependent metabolite of arachidonic acid and regulates vascular tone and renal tubular function. Previous studies showed that angiotensin II (Ang II) stimulated the renal CYP activity and 20-HETE production through the Ang II type 1 (AT1) receptor and that the Ang II-increased the 20-HETE was linked to the Ang II type 2(AT2) receptor. Thus, the study was designed to clarify the role of Ang II in CYP4A isoforms expression in the rat kidney. Male Sprague-Dawley rats were infused Ang II at low dose (AL, 0.17mg/kg/min, sc) and high dose (AH, 0.70mg/kg/day, sc) by using osmotic mini pump, with or without AT1 receptor blocker candesartan (1 and 3mg/kg/day, po) for 1 week. The protein expression of CYP4A isoforms, AT1 receptor and AT2 receptor in the renal cortex, outer medulla, and inner medulla was examined by immunoblot analysis. The mRNA expression of CYP4A isoforms was examined by reverse transcription and polymerase chain reaction (RT-PCR). Ang II at high dose increased systolic blood pressure (control, 109±2; AH, 164±8 mmHg, p<0.01), creatinine (control, 0.24±0.00; AH, 0.29±0.01 mg/dl, p<0.01) and urinary albumin excretion (control, 20.3±5.9; AH, 2398.6±303.6 μg/mg creatinine, p<0.01). In the control group, the CYP4A1, 4A2, and 4A8 proteins were highly expressed in the renal cortex, lowly expressed in the outer medulla, barely detected in the inner medulla. The AT1 receptor was expressed in kidney sections; highly in the outer and inner medulla, the AT2 receptor was only detected in the outer medulla. Ang II dose-dependently increased all CYP4A isoform proteins in the renal cortex and outer medulla (CYP4A1, 24% and 222%; CYP4A2, by 51% and 258%; CYP4A8, by 52% and 550%, p<0.05). Ang II also increased all CYP4A isoform mRNAs in the renal cortex and outer medulla. The candesartan treatment dose-dependently inhibited the Ang II-increased blood pressure, creatinine, urinary albumin excretion and CYP4A isoform expressions. These results indicated that Ang II increases CYP4A isoform expressions in the kidney through AT1 receptor. The Ang II-upregulated CYP4A expressions may play an important role in hypertension and renal function.
Abstract P295: Endothelial Sodium Channel Activation Promotes Cardiac Stiffness in Obese Female Mice
Authors: Habibi Javad , Aroor Annayya R , Ma Lixin , Jia Guanghong , Whaley-Connell Adam , Garro Mona , Barron Brady J , DeMarco Vincent G , Sowers James R
Published: June 2021

Cardiac diastolic dysfunction (DD) and diastolic heart failure is increasing in concert with obesity and aging population in the United States. In obese and diabetic women, DD is more common than in their male counterparts. This disproportionate increase in DD in obese females may partly explain their loss of sex-related cardiovascular (CV) disease protection. Recent studies have suggested a role for endothelial sodium channel (ENaC) activation in promotion of endothelial stiffness and suppression of flow- (nitric oxide) mediated vasodilation. Moreover, increased mineralocorticoid receptor (MR) activation mediated endothelial stiffness is promoted, in part, by ENaC activation. In this regard, we have recently reported increased plasma aldosterone levels, aortic and cardiac stiffness, and cardiac and vascular MR expression in female mice fed a high fat and high fructose diet (western diet [WD]). This increase in CV stiffness was prevented by very low dose MR antagonism. Accordingly, we hypothesized that inhibition of MR-mediated ENaC activation by using a very low dose of the ENaC inhibitor, amiloride would prevent cardiac stiffening (DD) in WD-fed female mice. Four week old C57BL6/J mice were fed a WD containing high fat (46%), sucrose (17.5%), and high fructose corn syrup (17.5%) with or without a very low dose of amiloride (1mg/kg/day) for 16 weeks. Amiloride significantly attenuated WD-induced impairment of cardiac relaxation in vivo as measured by high resolution magnetic resonance imaging (MRI) as well as cardiac interstitial fibrosis as measured by immunohistochemistry by picrosirius red staining. Moreover, amiloride prevented the development of DD in obese female mice without having effects on blood pressure. These observations support a role for ENaC activation in diet-induced cardiac stiffening (DD) in obese females.
Abstract P298: Finerenone Protects Against the Acute and Chronic Consequences of Renal Ischemia/reperfusion Injury
Authors: Barrera-Chimal Jonatan , Le Mercier Alan , El-Moghrabi Soumaya , Kolkhof Peter , Jaisser Frederic
Published: June 2021

Introduction:
One of the most common causes of acute kidney injury (AKI) is renal ischemia/reperfusion (IR). Mineralocorticoid receptor (MR) antagonism has shown beneficial effects against renal IR consequences. The potential benefit of novel non-steroidal MR antagonists such as finerenone has not been explored.

Objective:
Evaluate the efficacy of finerenone to prevent the acute and chronic consequences of ischemic AKI.

Methods:
For the acute study (24 hours), 18 rats were divided in: sham, rats subjected to bilateral renal ischemia of 25 min and rats that received three doses of finerenone at -48 h, -24 h and -1 h before the ischemia. For the chronic study (4 months), 21 rats were divided in: sham, rats with 45 min of bilateral ischemia and rats treated with Finerenone at day -2, -1 and 1h before IR. The left kidney was used for histology and the right kidney for molecular analysis.

Results:
After 24 h of reperfusion, the untreated IR rats presented a 3-fold increase in plasma creatinine, accompanied by 40% of tubules presenting cell detachment and casts. Kim-1 and NGAL mRNA levels were induced by 30-fold. In contrast, the rats that received finerenone presented normal creatinine and significantly fewer injured tubules (11%) and a less pronounced induction of kim-1 and NGAL (8-fold). After 4 months, the untreated IR rats developed chronic kidney disease (CKD), evidenced by kidney dysfunction, increased proteinuria (121.6 vs. 14.3 mg/24h in sham) and renal vascular resistance (16.8 vs. 11.4 mmHg/mL in sham). Tubular dilation, extensive tubule-interstitial fibrosis and an increase in kidney TGF-β and Collagen-I mRNA levels also characterized CKD. The transition from AKI to CKD was fully prevented by finerenone administration at the time of IR.

Conclusion:
Altogether, our data shows that finerenone is able to prevent AKI induced by IR as well as the chronic and progressive deterioration of kidney function and structure.
Abstract P300: Cardiac Ly6C
high
Monocyte Accumulation and Remodeling Depend on the Mineralocorticoid Receptor in Endothelial Cells
Authors: Lother Achim , Hübner Aurelia , Hilgendorf Ingo , Schnick Tilman , Moser Martin , Bode Christoph , Hein Lutz
Published: June 2021

Introduction:
Inflammation is a key driver for the development of cardiac fibrosis and diastolic dysfunction. Aldosterone promotes the expression of adhesion molecules and vascular inflammation. Thus, the goal of the present study was to examine the significance of endothelial MR for pressure overload induced cardiac inflammation and remodeling.

Methods and results:
Mice with endothelial cell-specific deletion of the mineralocorticoid receptor (MR
Cdh5Cre
) were generated using the Cre/loxP system. MR
Cdh5Cre
and Cre-negative littermates (MR
wildtype
) underwent transverse aortic constriction (TAC, n=5-7 per group).

After two weeks of pressure overload echocardiography revealed diastolic dysfunction in MR
wildtype
(mitral valve E acceleration time TAC 15.7 ± 0.5 vs. sham 12.8 ± 0.4 ms, P<0.05) but not in MR
Cdh5Cre
mice (TAC 11.2 ± 0.6 vs. sham 12.2 ± 0.9 ms, n.s.).

Cardiac hypertrophy (ventricle weight 143.2 ± 5.2 vs. MR
wildtype
167.3 ± 6.7 mg, P<0.001) and interstitial fibrosis (sirius red stained area 8.2 ± 4.7 vs. MR
wildtype
13.5 ± 4.5 %, P<0.05) following TAC were attenuated in MR
Cdh5Cre
mice. mRNA expression of atrial natriuretic peptide (
Nppa
, 2429 ± 1230 vs. MR
wildtype
7051 ± 3182 copies/10
4
copies
Rps29
, P<0.01) or the fibrosis marker gene collagen 1a1 (
Col1a1
, 256 ± 89 vs. MR
wildtype
432 ± 165 copies/10
4
copies
Rps29
, P<0.05) as determined by qRT-PCR confirmed these findings.

Cardiac leukocytes were quantitatively analyzed by fluorescence assisted cell sorting using specific antibodies. Numbers of CD45
+
leukocytes were similarly increased after TAC in the hearts of both genotypes (MR
Cdh5Cre
3840 ± 443 vs. MR
wildtype
4051 ± 385 /mg tissue, n.s.). Subtype analysis revealed a shift towards CD45
+
CD11b
+
F4/80
low
Ly6C
high
monocytes vs. CD45
+
CD11b
+
F4/80
high
Ly6C
low
macrophages in the heart of MR
wildtype
(TAC 20 ± 6 vs. sham 4 ± 1 % of CD45
+
CD11b
+
, P<0.05) but not of MR
Cdh5Cre
mice (TAC 6 ± 2 vs. sham 3 ± 1 % of CD45
+
CD11b
+
, n.s.).

Conclusion:
MR deletion from endothelial cells ameliorates left ventricular remodeling and diastolic dysfunction after pressure overload. The protective effect of endothelial MR deletion is associated with a shift towards less pro-inflammatory Ly6C
high
monocytes and more reparative Ly6C
low
macrophages.
Abstract P308: Pilot Study: 24 Hour and Daytime Blood Pressure Lowering Effect of Melatonin in Younger and Older Non-hypertensive Adults
Authors: Cheng Cynthia , Gooneratne Nalaka S , Payton Colleen , Keith Scott , Hafycz John
Published: June 2021

The study objective was to study the effect of melatonin on blood pressure (BP) in both younger and older normotensive and prehypertensive individuals.

Preliminary data available at the time of abstract submission are presented for 12 participants (10 ≤ 50 years old; two ≥ 60). free of known hypertension or diabetes. All subjects had office and 24 hour ambulatory BP measured before and after dosing of 9 mg controlled release melatonin for six weeks.

Subjects are described in
Table 1
. Full data analysis for all 30 pilot subjects, including 12 older and 10 control subjects, will be available for the presentation.

Table 2
shows results comparing baseline and follow-up BP. Mean 24 hour SBP/DBP, as well as ambulatory daytime SBP/DBP, were significantly lower following melatonin administration.

In conclusion, these findings suggest possible future utility of melatonin for lowering BP in both younger and older non-hypertensive individuals.
Abstract P318: miR-155 Induces Inflammation by Polarizing M1 Macrophages in a TLR3-induced Preeclamptic Mouse Model
Authors: Chiasson Valorie L , Bounds Kelsey R , Charles Derek , Dusio Giuseppina , Tobin Richard P , Newell Rogers M. Karen , Chatterjee Piyali
Published: June 2021

Placental microRNA (miRNA) expression is known to be dysregulated during preeclampsia (PE), a hypertensive pregnancy disorder but the role it plays in the pathogenesis of PE is currently unknown. We have previously demonstrated that placental miR-155 expression is upregulated (P-PIC: 3.15 fold, p<0.05 vs. controls) in a TLR3-induced PE mouse model similar to PE patients. In addition, poly I:C (PIC) treatment significantly increased systolic blood pressure (SBP) at gestational day 17 in P-PIC WT (147±4.5 mmHg) compared to P WT mice (103±2.7 mmHg), but did not have any effect in P-PIC miR-155 KO mice (101±2 mmHg). In PE there is an increase in the number of total and activated macrophages. M1 macrophages display the capacity to shift T cell responses toward a TH1 while M2 macrophages promote a TH2 response. We hypothesized that TLR3-induced upregulation of miR-155 contributes to hypertension in part by polarizing the macrophages to a M1 phenotype and these effects will be attenuated in P-PIC miR-155 KO mice. Placental flow cytometry analyses demonstrate that administration of poly I:C induced CD45(+) CD11b(+) macrophages in P-PIC WT mice which is attenuated in P-PIC miR-155 KO mice. In addition, placental classical ‘M1’ macrophages CD45(+) CD11b(+) CD86(+) CD206(-) were increased and alternate ‘M2’ macrophages CD45(+) CD11b(+) CD206(+) CD86(-) were decreased in P-PIC WT mice compared to controls. Interestingly, administration of poly I:C did not change M1 macrophages but increased M2 macrophages in P-PIC miR-155 KO mice. P-PIC WT mice exhibited increased placental expression of additional M1 markers (NOS2, IFNg) and decreased expression of M2 markers (Arg1, IL-10) compared to controls by qRTPCR but not in P-PIC miR-155 KO mice. The above observations are also consistent with our splenic flow cytometry and qRTPCR studies. Based on our results, miR-155 activation induces inflammation in part by increasing M1 macrophages thus contributing to hypertension. Targeting the innate immune system by inhibition of monocyte/macrophages may have beneficial cardiovascular effects in women with PE.
Abstract P322: Vasopressin Release is Enhanced Before the Development of Preeclampsia in Humans Despite Exaggerated Suppression of Plasma Osmolality
Authors: Santillan Mark K , Scroggins Sabrina M , Ray Alyssa T , Witcher Phillip C , Sandgren Jeremy A , Linggonegoro Danny W , Pierce Gary L , Santillan Donna A , Grobe Justin L
Published: June 2021

Plasma osmolality (Osm) suppression is of critical importance to maintain appropriate blood volume to perfuse the uterus during pregnancy. Osm is reduced starting at the fifth week of gestation via increased arginine vasopressin (AVP) secretion. This increased secretion is maintained via a decrease in the AVP/osmotic release threshold. We previously demonstrated that pregnant women who develop preeclampsia (PreE) exhibit exaggerated AVP secretion as early as the 6th week of gestation via measurement of copeptin, the stable C-terminal fragment of AVP. It is unclear whether AVP secretion is elevated before the onset of PreE due to osmotic or non-osmotic stimuli. We tested the hypothesis that elevated AVP secretion before PreE may be associated with elevated Osm (a strong stimulant of AVP secretion). Plasma and clinical data from pregnant women were obtained from the University of Iowa Maternal-Fetal Tissue Bank (IRB#200910784). Osm was measured using the freezing-point suppression technique. Osm was assessed in non-pregnant women (n=109), pregnant women who later developed PreE (n=12 for 7-12 weeks, n=9 for 16-24 weeks), and maternal and gestational age matched controls (n=25 for 6-13 weeks, n=15 for 14-27 weeks). As expected, Osm was decreased in control pregnancies (non-pregnant 291±1 vs pregnant 286±1 mOsm/kg, p<0.05). Contrary to our hypothesis, the Osm decrease was exaggerated in women who would later develop PreE (1st trimester: PreE 279±4 vs control 287±3, and 2
nd
trimester: PreE 277±4 vs control 285±3 mOsm/kg; effect of PreE p<0.05, gestational age p=NS, interaction p=NS) even after controlling for age, BMI, diabetes, chronic hypertension, history of preeclampsia, and gravida (model p<0.05). Despite suppressed Osm, plasma copeptin was elevated in the PreE group at all timepoints (p<0.05). These data support the conclusion that long before the development of clinical symptoms of PreE, the rate of secretion of AVP is inappropriately increased despite maintenance of normal osmotic-regulating actions of AVP. This effect must be the result of increased non-osmotic stimuli for AVP, and a suppression of the AVP/osmotic release threshold beyond that observed in control pregnancies.
Abstract P328: Reduced Salt Taste Sensitivity Leads to Increase of Daily Salt Intake
Authors: Isomura Minoru , Nabika Toru
Published: June 2021

Objective:
It is empirically aware that inter-individual differences exist in sensitivity of salt taste. It is assumed that person with reduced sensitivity to salt taste may consume a higher amount of salt than those with a normal sensitivity of salt. The objective of this study is to reveal the relationship between the sensitivity of salt taste and salt intake.

Design and method:
In this study, we defined salt taste sensitivity by the threshold of the sense of salt taste. Therefore, we defined as reduced sensitivity when individuals were unable to sense salt taste in the concentration that majority of individuals could. Japanese residents who came to annual health checkups were recruited to this study. A series of salt-impregnated taste strips were used to measure salt taste intensity. Each taste strip was impregnated with sodium chloride at concentrations of 0, 0.6, 0.8, 1.0, 1.2, 1.4 and 1.6 g/cm
2
. Daily sodium intake was estimated from sodium excretion in the spot urine using Tanaka’s equation.

Results:
Among 1,207 participants, 979 (81.1%) individuals were considered to bear normal sensitivity because they were able to detect salt taste by taste strip of 0.6 or 0.8 g/cm
2
. However, 67 (5.6%) individuals were categorized as reduced sensitivity group because they were able to detect it only by the taste strip with the highest amount of salt, or unable to detect salt taste. Daily salt intake of reduced sensitivity group was significantly higher than those in normal sensitivity group (10.3g vs 9.7g, p<0.04). No significant difference was identified in blood presser. Salt taste sensitivity decreased by age, history of smoking and presence of diabetes mellitus.

Conclusions:
Our study revealed the sensitivity of salt taste is associated with daily salt intake, even adjusted by gender and age. Strict reduction of salt is recommended especially for Smokers or persons with diabetes mellitus, not only because low BP recommendation but also they might pretend to consume salt because of reduced sensitivity of salt taste.
Abstract P332: Role of T-Lymphocytes in the Long-Term Blood Pressure and Renal Disease Effects of Acute Renal Ischemia-Reperfusion Injury
Authors: Lopez Bernardo , Petrova Galina , Abais-Battad Justine M , Lund Hayley , Fehrenbach Daniel , Mattson David L
Published: June 2021

Epidemiological data indicates that acute kidney injury (AKI) is an independent risk factor for the development of hypertension and chronic kidney disease in patients. Previous studies demonstrated that rats develop sodium-dependent hypertension and kidney damage following experimental AKI induced by a renal ischemia-reperfusion (IR) insult; furthermore, these high salt deleterious effects could be blunted by administration of immunosuppressive agents. The present study was performed on Dahl SS (SS) rats and SS rats with a null mutation in the CD247 gene (SS-CD247) leading to depletion of T-lymphocytes in order to specifically examine the role of T cells in this response (n=5-6 rats/group). As assessed by serum creatinine (SCr) levels, no difference was observed in the initial response to IR injury between SS and SS-CD247: SCr increased from 0.44±0.03 to 2.16±0.32 mg/dl in SS rats 24 hours after an initial 30 minute period of renal ischemia and returned to control levels after 8 days of recovery. Moreover, no differences were noted in mean arterial pressure (MAP) or albumin excretion rate (UAlb) between SS and SS-CD247 after 43 days of recovery from IR injury while the rats were maintained on a low salt (0.4% NaCl) diet. When the rats were fed a 4.0% NaCl diet for two weeks, MAP and UAlb significantly increased in the sham SS to 178±9 mmHg and 189±25 mg/day, respectively; values significantly greater than observed in the sham SS-CD247 rats (148±2 mmHg and 87±17 mg/day). As expected, the SS rats recovered from IR injury demonstrated an exaggerated increase in MAP (peaking at 183±2 mmHg) and UAlb (275±54 mg/day) in response to high salt. There was no difference in the number of total CD3+ lymphocytes in the kidneys of IR and sham SS after high salt, though the ratio of CD4+/CD8+ T cells was increased in the IR group. Compared to sham CD247, an exaggerated elevation of MAP (157±9 mmHg) and UAlb (210±32 mg/day) was also observed in the SS-CD247 rats recovered from IR injury, demonstrating enhanced responsiveness following IR injury in animals lacking T cells. These data indicate that T lymphocytes amplify salt-sensitive hypertension and renal damage, but other mechanisms also mediate the salt-sensitive hypertension and renal damage that occurs in animals recovered from IR injury.