bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 13: P2X7 Receptor Knockout Attenuates Angiotensin II-Induced Hypertension, Vascular Injury And CD8
+
T Cell Infiltration
Authors: Shokoples Brandon G , Comeau Kevin , Higaki Akinori , Caillon Antoine , Paradis Pierre , Schiffrin Ernesto L
Published: August 2021

Background:
The P2X7 receptor (P2RX7) recognizes damage associated molecule patterns such as adenosine triphosphate (ATP), and triggers the activation of immune cells. Elevated plasma ATP levels have been observed in hypertensive patients, providing a potential mechanism for P2RX7 activation. Additionally, a hypomorphic polymorphism for P2X7 is correlated with a decreased risk for essential hypertension in Chinese post-menopausal women. However, it is unknown whether P2RX7 activation contributes to angiotensin (Ang) II-induced blood pressure (BP) elevation and vascular damage. We hypothesized that
P2rx7
knockout would blunt Ang II-induced BP elevation, vascular injury, and infiltration of activated immune T cells into perivascular adipose tissue (PVAT).

Methods:
Ten-to-12-week-old male C57BL/6J male wild-type (WT) and

P2rx7
-/-

mice were infused or not with Ang II (1000ng/kg/min) for 14 days. BP was determined by telemetry, mesenteric artery function and remodeling using pressurized myography, aortic stiffening by ultrasound and infiltration of activated immune T cells in aortic PVAT by flow cytometry.

Results:
Ang II-infused

P2rx7
-/-

mice display a reduced systolic BP (164±3 vs. 176±2 mm Hg,
P
<0.05) and pulse pressure (37±4 vs. 53±3 mm Hg,
P
<0.001) in comparison to WT mice. Aortic stiffening occurred in WT mice treated with Ang II, demonstrated by an increased pulse wave velocity (7.7±0.7 vs. 5.9±0.3 m/s,
P
<0.05), accompanied by a 3.8-fold increased infiltration of activated CD8
+
T cells in aortic PVAT (60±16 vs 16±3 cells/aortic PVAT,
P
<0.001), which were both absent in

P2rx7
-/-

mice (6.4±1.4 vs 5.5±1.1 m/s and 27±7 vs 16±3 cells/aortic PVAT). In addition, the frequency of IFN-γ producing CD8
+
T cells in the spleen of Ang II-treated WT mice increased (2.6±0.2% vs 1.2±0.2%), which did not occur in

P2rx7
-/-

mice (1.7±0.3% vs 1.7±0.2%). Ang II-infusion induced mesenteric artery endothelial dysfunction in WT mice (61±7 vs 83±4% relaxation response to acetylcholine,
P
<0.05), which was absent in

P2rx7
-/-

mice (89±3 vs 90±3%).

Conclusion:
P2rx7
knockout attenuates Ang II-induced hypertension, vascular injury, and infiltration of activated CD8
+
T cells into aortic PVAT.
Abstract 16: Altered Testicular Macrophage Polarization Is Associated With Reproductive Dysfunction In Hypertensive Mice
Authors: Navaneethabalakrishnan Shobana , Wilcox Brooke K , Goodlett Bethany L , Mitchell Brett M
Published: August 2021

Elevated circulating proinflammatory (M1) and decreased anti-inflammatory (M2) macrophages contribute to hypertension (HTN) and end-organ damage. HTN is associated with reproductive dysfunction in men. However, the impact of HTN on testicular macrophages and inflammation is unknown. We hypothesized that HTN increases M1 and decreases M2 testicular macrophages, which is associated with inflammation and reproductive dysfunction. Male mice were made hypertensive by either providing L-arginine methyl ester hydrochloride (L-NAME) (0.5 mg/mL) in the drinking water for 3 weeks [L-NAME-induced HTN (LHTN)] or L-NAME water for 2 weeks, followed by a 2-week washout period and a subsequent 3-week 4% high salt diet [salt-sensitive hypertension (SSHTN)]. Control (C) mice received tap water and normal diet. Flow cytometry analysis revealed a significant increase in both M1 (C: 15%±1, LHTN: 22%±2; p<0.05) and M2 (C: 10%±1, LHTN: 21%±2; p<0.05) macrophages in testes from LHTN mice. Similarly, testes from SSHTN mice had a significant increase in M1 (C: 17%±1, SSHTN: 28%±2; p<0.05) but had a significant decrease in M2 (C: 14%±1, SSHTN: 7%±1; p<0.05) macrophages. Testes from both hypertension models had a significant increase in gene expression of the proinflammatory cytokines TNFa, IFNg, IL-1b, IL-6, and IL-17. Sperm concentration (C: 8.5±0.7, LHTN: 6.5±0.2, SSHTN: 4.7±0.5; both p<0.05) and the percentage of sperm mitochondrial activity (C: 88%±3, LHTN: 71±5, SSHTN: 64%±3; both p<0.05) were decreased significantly in both hypertension groups. Hypertensive mice presented a significantly increased percentage of sperm with abnormal morphology (C: 5%±1, LHTN: 8%±1, SSHTN: 13%±2; both p<0.05) and damaged acrosome (C: 1.4%±0.2, LHTN: 2.8%±0.2, SSHTN: 4%±0.5; both p<0.05). There was a significant decrease in gene expression of the hormone receptors AR, ERa, and LHR, and the steroidogenic enzymes StAR, 3bHSD, 17bHSD, and CYP17a1 in the testes of LHTN and SSHTN mice. These data demonstrate that HTN alters testicular macrophage polarization which is associated with inflammation and impaired reproductive health. Therapeutic strategies may be developed to improve reproductive health in male hypertensive patients by targeting testicular macrophage imbalance.
Abstract 17: Genetic Deletion Of Angiotensin Ii AT
1a
Receptors Selectively In The Proximal Tubules Of The Kidney Attenuates Two-kidney, One-clip Goldblatt Hypertension In Pt-
Agtr1a
-/-
Mice
Authors: Li Xiao C , Leite Ana , Hassan Rumana , Sato Akemi , Dugas Courtney , Wang Chih-Hong , Sato Ryosuke , Zhuo Jia L
Published: August 2021

The activation of the renin-angiotensin system (RAS) in the clipped kidney plays a critical role in the development of two-kidney, one-clip Goldblatt hypertension (2K1C), but the roles of angiotensin II (Ang II) and AT
1a
receptors in the proximal tubules has not been determined previously. The present study tested the hypothesis that genetic deletion of AT
1a
receptors selectively in the proximal tubules attenuates the development of 2K1C Goldblatt hypertension via AT
1a
receptor-mediated, Na
+
/H
+
exchanger 3 (NHE3)-dependent mechanisms. To test the hypothesis, 2K1C Goldblatt hypertension was induced by placing a silver clip, 0.2 mm internal diameter, on the left renal artery for 4 weeks in adult male wild-type (WT), global AT
1a
receptor knockout (
Agtr1a
-/-
), proximal tubule (PT)-specific
Agtr1a
-/-
(PT-
Agtr1a
-/-
), or PT-
Nhe3
-/-
mice, respectively. In WT mice, systolic blood pressure increased in a time-dependent manner reaching a maximal response by Week 3 (Basal: 112 ± 2 vs. 2K1C: 149 ± 4 mmHg, n=12,
P
<0.01). 2K1C Goldblatt hypertension in WT mice was associated with significant increases in renin mRNA expression in the clipped kidney (Control: 2366 ± 255 vs. Clipped: 3144 ± 569 copies/ng RNA,
P
<0.01) and decreases in renin mRNA expression in the nonclipped kidney (1738 ± 341 copies/ng RNA,
P
<0.05). Plasma Ang II levels were significantly increased in WT mice with 2K1C Goldblatt hypertension (Control: 50.2 ± 7.2 vs. 2K1C: 109.7 ± 17.2 pg/ml,
P
<0.05). Glomerular and tubulointerstitial fibrotic responses were also significantly increased in the clipped kidney (
P
<0.01). In contrast to WT mice, the development of 2K1C hypertension was completely attenuated in
Agtr1a
-/-
(Basal: 88 ± 4 vs. 2K1C: 92 ± 2 mmHg, n=9,
n.s
.), PT-
Agtr1a
-/-
mice (Basal: 101 ± 2 vs. 2K1C: 104 ± 3 mmHg, n=12,
n.s
.) and PT-
Nhe3
-/-
mice (Basal: 103 ± 3 vs. 109 ± 5 mmHg, n=12,
n.s
.). Renin mRNA expression was not different in clipped and nonclipped kidney of
Agtr1a
-/-
mice, but it was decreased in the nonclipped kidney of PT-
Agtr1a
-/-
mice (
P
<0.05). Taken together, these data suggest that genetic deletion of AT
1a
receptors selectively in the proximal tubules attenuates the development of 2K1C Goldblatt hypertension via AT
1a
receptor-mediated, Na
+
/H
+
exchanger 3 (NHE3)-dependent mechanisms.
Abstract 19: Maternal B Cell Depletion Lowers Blood Pressure And Improves Fetal Weights In Offspring In A Rat Model Of Preeclampsia
Authors: Campbell Nathan , Herrock Owen , Solise Dylan , Fitzgerald Sarah , Deer Evangeline , Turner Ty , Cockrell Kathy , Ibrahim Tarek , McCray Morgan , Evans Kymberlee , Ingram Nicole , Amaral Lorena M , Lamarca Babbette A
Published: August 2021

Preeclampsia (PE), new onset hypertension during pregnancy, is the leading cause of death and morbidity for the mother and low birth weight in offspring. PE women have activated B cells producing agonistic autoantibodies to the angiotensin II type I receptor (AT1-AA). We have shown Rituximab (R), used clinically for B cell depletion, lowers mean arterial pressure (MAP), B cells, and AT1-AA in the RUPP rat model of PE. Clinical studies show no untoward effects on offspring of pregnant women maintained on R for treating lymphoma, however, R is not used during PE therefore, effects of maternal B cell depletion on offspring survival and growth in response to placental ischemia is unknown. We hypothesize that R will deplete maternal B cells in RUPP rats without worsening the effect of placental ischemia on pup growth and survival. To test this hypothesis, R (250 mcg/kg) was given on gestation day (GD) 14 via mini-osmotic pump. On GD 19 B cells were measured by flow cytometry, and MAP and pup weights were recorded. A separate group of dams were allowed to deliver, pup weights were recorded within 12 hours and weekly until 16 weeks, and B cells were analyzed. A one-way ANOVA was used for statistical analysis. MAP increased in RUPP 123±2 (n=19, p<0.05) compared to NP controls 101±1 (n=18) and was 106±3 mmHg in RUPP+R (n=8, p<0.05). On GD19, maternal circulating B cells were 16±2 % (n=14) in RUPPs, 8±2 % in NP rats, (n=7, p<0.05), and 5.5±1% gate in RUPP+R (n=5, p<0.05). RUPP male and female offspring were smaller 5.11±0.23 g, 5.19±0.14 g (n=4, n=4) at birth than NP offspring 6.09±0.15 g, 5.87±0.12 g (n=6, p<0.05; n=6, p<0.05) or RUPP+R offspring 5.75±0.24 g, 5.36±0.28 g (n=6, p=0.11; n=6, p=0.67). At 12 weeks, male and female RUPP offspring had elevated circulating B cells (21±3, 20±1 % (n=6; n=9)) compared to NP (1±0.23, 1.6±0.06 % (n=4, p<0.05; n=3, p<0.04)) which was normalized in RUPP+R offspring (0.4±0.1, 0.3±0.03 % gate (n=3, p<0.05; n=8, p<0.05)). At 16 weeks, B cells were comparable in male offspring from NP (0.78±0.09 % (n=10)) and RUPP+R rats (0.80±0.04 % gate (n=3)). Our findings indicate that R lowers maternal circulating B cells and MAP in RUPP rats and improves fetal weight and circulating B cells, indicating that R does not worsen adverse fetal outcomes in response to placental ischemia.
Abstract 23: Body Fluid And Sodium Loss Leads To Aldosterone Hypersecretion And Muscle Catabolism In Hepatocellular Carcinoma Rats.
Authors: Kidoguchi Satoshi , Kitada Kento , Nakano Daisuke , Yokoo Takashi , Nishiyama Akira
Published: August 2021

Background:
The number of cancer survivors coincides with cardiovascular diseases is increasing, therefore, we are promoting the concept of “Onco-Hypertension” to clarify the mechanism linking cancer and blood pressure. In this study, we evaluated body osmolyte and water imbalance in hepatocellular cancer (HCC) model rats.

Methods:
Wistar rats were administered diethylnitrosamine (DEN) (1.5 μg/day, p.o.), a carcinogenic drug, for 8 weeks to establish liver cancer. Three weeks after the completion of DEN administration, we investigated blood pressure, tissue osmolyte and water content, and its association with aldosterone secretion.

Results:
HCC rats significantly reduced blood pressure, skin sodium, potassium, and water content. In the carcass (muscle + bone), dry weight, sodium, potassium, and water content were dramatically reduced without changing bone mass in HCC rats, suggesting that HCC causes muscle wasting to supply osmolyte and water for the dehydrated organs. These osmolytes and water loss were significantly associated with increased urinary aldosterone excretion. Supplementation of 0.25% salt water to drink improved body sodium and water loss and muscle wasting in HCC rats, which were completely suppressed by treatment with spironolactone (75 mg/kg/day, p.o.), a mineralocorticoid receptor (MR) blocker.

Conclusion:
These findings suggest that HCC causes body osmolyte and water loss, which leads to aldosterone hypersecretion and muscle catabolism to compensate for dehydration. A relatively small amount of salt supplement ameliorates the HCC-induced dehydration and muscle wasting via aldosterone/MR-mediated sodium and water restoration.
Abstract 25: Inhibition Of The Agonistic Ang II Type 1 Receptor Autoantibody In A Rat Model Of Preeclampsia Improves Uteroplacental Perfusion And Fetal Growth In Late Gestation
Authors: Ashraf Usman , Campbell Nathan , Waller Jamarius P , Rawls Adam , Solise Dylan , Cockrell Kathy , Bidwell Gene L , Lamarca Babbette A , Alexander Barbara
Published: August 2021

Placenta ischemia, the initiating factor in preeclampsia (PE), is associated with intrauterine growth restriction (IUGR) and increased blood pressure (BP) in offspring. Yet, the only treatment for PE is delivery of the baby and placenta. The Reduced Uterine Perfusion Pressure (RUPP) rat model induced by placental ischemia at gestational day 14 (G14) mimics many facets of human PE including pregnancy-specific hypertension, an increase in the agonistic ANG II Type 1 receptor autoantibody (AT1-AA), IUGR and increased BP in the offspring. Inhibition of AT1-AA using an epitope-binding inhibitory peptide ('n7AAc') attenuates increased BP at gestational day 19 in the RUPP. Yet, whether use of ‘n7aac’ improves fetal growth and mitigates increased BP in the offspring is unknown. Thus, we tested the hypothesis that maternal administration of ‘n7aac’ improves fetal growth by attenuating reduced uterine blood flow and impaired placental remodeling. Sham or RUPP surgery was performed at G14 with administration of vehicle or ‘n7aac’ (144μg/day) via mini osmotic pump until gestational day 20 (G20). At G20 uterine artery resistance index was significantly elevated in vehicle RUPP (0.69±0.02 mm/s n=10) compared to vehicle Sham (0.48±0.02 mm/s n=8) (P<0.0001) and not increased in treated RUPP (0.49±0.02 mm/s n=10) or treated Sham (0.48±0.02 mm/s n=9). Fetal weight was significantly reduced in vehicle RUPP (3.24±0.2 g) compared to vehicle Sham (3.92±0.05 g) (P=0.013) and not decreased in treated RUPP (3.70±0.04 g) or Sham (3.98±0.10 g). Litter size of viable pups at G20 was only reduced in treated RUPP (5.3±1.4) compared to vehicle Sham (11.56±0.7) (P=0.003). Importantly, using in vivo imaging, little to no auto fluorescence of rhodamine-labeled peptide (480 μg/kg/day, n=4) was detectable in the pups at G20. Thus, our results demonstrate that maternal treatment with ‘n7aac’ in the RUPP rat model of PE improve UARI, which is associated with improved fetal weight at G20 in response to placental ischemia. Whether this benefit continues to birth and mitigates increased BP in IUGR offspring is unknown but is the focus of future studies. In conclusion, inhibition of the AT1AA during PE may not only provide benefit to the mother, but may also be associated with benefit in the offspring.
Abstract 27: Unravelling Cell-specific Interactions At The Preeclamptic Maternal-foetal Interface From Early To Late Pregnancy
Authors: Nonn Olivia , Debnath Olivia , Valdes Daniela S , Fischer Cornelius , Secener Kerim , Tiesmeyer Sebastian , Ishaque Naveed , Sallinger Katja , El-Heliebi Amin , Staff Anne , Mueller Dominik N , Dechend Ralf , Gauster Martin , Herse Florian
Published: August 2021

Preeclamptic syndrome arrises in the fetal part of the placenta (villi). In this study, we analyse placental development by single nuclei RNA-sequencing in early and term pregnancy and draw conclusions about pathological processes in preeclampsia (PE) that originate early in gestation. We profiled the transcriptome of 101,067 nuclei obtained from a total of 12 pregnancies, spanning early, term and PE doners. Using unsupervised computational approaches, we identified 12 and 16 different cell types and states in decidua and villi, respectively. Our comprehensively identified catalogue of cell types and states aligns well with the previous single cell studies. We identified different subpopulations of syncytiotrophoblast and GATA3+/GREM2+ trophoblast stem cells (TSC) in villi. Through gestation, gene expression in cell populations from the matrisome or vascular environments show dynamic expression reflecting vascular development associated with spiral artery remodelling and concordant decidual stroma reorganisation. Global differential gene expression analysis shows that trophoblast cell types are most dysregulated in PE. Cell-cell communication analysis revealed important dysregulation between villi and decidual cell types. The secretory signalling characteristic of this trophoblastic disease may be used for early biomarker screening. Overall, this study paves the way to a deeper understanding of the early pathophysiology of PE. Figure 1: Villi (v) and decidua (d) cell clusters from early, late control and preeclampsia (PE) villi and decidua visualised as a UMAP. Datasets were integrated separately for each tissue and merged for embedding.
Abstract 29: Acute Systemic Treatment With Sphingosine -1- Phosphate Receptor 1 Agonist Diminishes Sex-difference In Renal Function Seen In Intrauterine Growth Restricted Mice.
Authors: Bhunu Benjamin , Intapad Suttira
Published: August 2021

IUGR is a risk factor for the early development of cardiorenal diseases in life. We previously showed that our IUGR mouse model exhibits sex differences in blood pressure (BP) and kidney functions as males have elevated BP and impaired kidney function while females are not. Sphingosine-1-Phosphate Receptor 1 (S1PR1) is reported to be involved in developing and progressing several cardiorenal diseases. We found that acute activation of S1PR1 transiently reduced BP in male IUGR; however, the effects of SIPR1 on renal function in IUGR are still unknown. We hypothesize that S1PR1 plays a role in the sex differences of impaired kidney function in IUGR. Here we investigated the acute effects of a specific S1PR1 agonist (SEW2178) on kidney function in IUGR offspring generated through placental insufficiency.

Methods:
C57BL IUGR or control offspring were obtained from the Reduced Uterine Perfusion Pressure (RUPP) or sham surgeries on the 13
th
day of gestation. At 6 months, nephron number was measured by analyzing kidney histology while Glomerular Filtration Rate (GFR) was measured using FITC-Anulin decay after retroorbital injection. Mice were treated with 3mg/kg SEW2871 i.p before GFR measurement.

Results:
Both male and female IUGR had a decrease in nephron number compared to same-sex control (371.8± 28.8 vs 284.0± 11.3, P<0.01, male control vs IUGR) and (333.6±13.3 vs 256± 13.0, P<0.05, female control vs IUGR). Furthermore, IUGR impaired kidney function in male but not female mice as indicated by reduced GFR (8.9 ±0.6 vs 13.9±1.1 μL/min/kg BW, P<0.05, IUGR vs control males) and (12.3±0.9 vs 15.9 ±1.9 L/min/kg BW, control females vs IUGR). Administration of SEW2871 diminished the difference of GFR between male control and IUGR (9.5±0.6 vs 9.7±1.1 μL/min/kg BW, P> 0.05 control vs IUGR). While in females, both IUGR and control had a similar response to SEW2178 (7.4±1.0 vs 10.6 ±1.9 μL/min/kg BW, control vs. IUGR).

Conclusion:
IUGR impairs renal function in male offspring but not in females, indicating sex differences in fetal programming of kidney disease. Male and female IUGR offspring respond differently to acute systemic activation of S1PR1. Further studies are required to investigate the kidney-specific and cell-specific of S1PR1 activation/inhibition.
Abstract 30: Role For Adverse Childhood Experiences And Depression In Preeclampsia
Authors: Myers Monica C , Santillan Mark K , Brandt Debra S , Stroud Amy K , Vignato Julie A , Scroggins Sabrina M , Nuckols Virginia , Pierce Gary L , Santillan Donna A
Published: August 2021

Hypertensive diseases are associated with adverse experiences in childhood as well as depression. In order to determine if these associations were present in women with preeclampsia (PreE), a particularly devastating hypertensive disease in pregnancy, the scores from three questionnaires: Adverse Childhood Experiences (ACE), Edinburgh Postnatal Depression Scale (EPDS), and the Patient Health Questionnaire-9 (PHQ-9) were compared between women with PreE (n=32) and women without PreE (n=46) between 9 and 48 months postpartum (IRB# 201808705). ACE scores are calculated by summing an individual’s affirmative responses to specific adverse experiences during childhood. In our study, the average ACE score of individuals with PreE was higher than that of women without PreE (1.69 vs. 1.02, P=.04). We also divided women into groups based on whether their ACE score was ≤3 or ≥4 due to evidence that individuals who have experienced ≥4 ACEs are at greatest risk for physical and mental health conditions. Among our participants, 80% of women with an ACE score ≥4 (n=10) had PreE while only 35.3% of women with a score ≤3 (n=68) developed the condition (P=0.01). As well, the odds of having PreE were higher in those with ACE scores ≥4, compared with those with scores ≤3 (OR= 7.34; 95% CI = 1.44, 37.33). In a subset of participants, scores were available from EPDS, survey that identifies women who have postpartum depression 6 weeks after birth, and from the PHQ-9, another assessment for depression. Among our participants, the average EPDS score was higher in women with PreE than women without PreE (6.38, n=21 vs. 3.71, n=42 P=0.01), indicating more severe symptoms of postpartum depression in women who also had PreE. In addition, the average PHQ-9 score among women with PreE was higher than that of women without PreE (3.71, n=15 vs 1.86, n=37 P=.02) with a higher score indicating more severe depression. The average PHQ-9 score was also higher in women who had ACE scores ≥4 than women with scores ≤3 (4.00, n=4 vs. 2.27, n=48 P=.01) indicating that women with more adverse childhood events were more likely to experience depression. Together, these findings indicate that PreE may be associated with adverse events during childhood as well as depression in late pregnancy and/or postpartum.
Abstract 31: Esomeprazole Improves Blood Pressure, Intrauterine Growth, Inflammation, And Vascular Function During Placental Ischemia Through Inhibition Of NLRP3
Authors: Cornelius Denise C , Wang Xi , Travis Olivia K , Shields Corbin A , Tardo G A , Giachelli Chelsea , Nutter Christopher W , Cooper Olive G , Williams Jan M
Published: August 2021

Preeclampsia (PE), a multisystem hypertensive disorder of pregnancy is characterized by intrauterine growth restriction (IUGR), inflammation, and vascular dysfunction. NLRP3 inflammasome is a cytoplasmic complex that mediates inflammation and is implicated in CVD. Clinical studies show an association between PE and increased placental NLRP3 expression. We hypothesized that inhibition of NLRP3 using (1) a specific NLRP3 small molecule inhibitor, MCC950 (M9, 20 mg/kg/d, i.p.) or (2) esomeprazole (ESO, 3.5 mg/kg/d, oral), a therapeutic that is safe in pregnancy, would improve MAP, inflammation, IUGR, and vascular dysfunction in the reduced uterine perfusion pressure (RUPP) rat model of placental ischemia. Sham (S) or RUPP surgery was performed in pregnant Sprague Dawley rats on gestation day (GD) 14. A subset of rats from both groups received either vehicle, M9, or ESO on GD14-19 (n=9/group). On GD18, Uterine Artery Resistance Index (UARI) was measured via Doppler Ultrasound. MAP, fetal, and placental weight were measured, and blood and tissues were processed for additional analyses on GD19. MAP (mmHg) was elevated in RUPP (133±1) vs S (108±2; p<0.05). Treatment with M9 or ESO in RUPP decreased MAP (111±1 and 115±3, respectively; p<0.05 vs RUPP). Fetal weight (g) was reduced in RUPP (2.1±0.04) vs S (2.4±0.05), and ESO normalized fetal weight (2.4±0.01; p<0.05 vs RUPP). Placental NLRP3 mRNA expression increased 5-fold in RUPP vs S (p<0.05); and was less than 2 fold in M9 and ESO treated RUPP rats (p<0.05 vs RUPP). Inflammatory T-helper 17 and cytolytic NK cells, evaluated by flow cytometry, were increased in the circulation, placenta, and kidney of RUPP vs S controls. Treatment with M9 or ESO normalized both cell populations in all tissues (p<0.05 vs RUPP). UARI was increased in RUPP (0.71±0.03) versus S (0.56±0.01; p<0.05) and was decreased to 0.48±0.01 in M9 and 0.61±0.03 in ESO-treated RUPPs (p<0.05 vs RUPP). Renal vascular resistance (mmHg/mL/min/g) was increased in RUPP (42±8) vs S (23±4, p<0.05) and was normalized to 26±3 after treatment with M9 (p<0.05 vs RUPP). These data implicate NLRP3 in mediating inflammation and vascular dysfunction to cause maternal HTN and IUGR in RUPP; and identify NLRP3 as a potential target and ESO as a potential therapeutic for PE.
Abstract 34: Diminished Contractile Capability In Cerebral Vascular Smooth Muscle Cells In The TgF344-AD Rat Model Of Alzheimer's Disease
Authors: Fang Xing , Zhang Huawei , Liu Yedan , Wang Shaoxun , Zheng Baoying , Elliott Parker S , Ryu Jane J , Strong Luke B , Crumpler Reece F , Thomas Kirby N , Roman Richard J , Fan Fan
Published: August 2021

We recently reported that cerebral vascular dysfunction leads to impaired autoregulation of cerebral blood flow (CBF), neurovascular coupling (NVC), and blood-brain barrier (BBB) leakage. The present study examined if cerebral vascular dysfunction precedes cognitive impairment in the TgF344-AD (AD) rat model of Alzheimer's disease. In the present study, we confirmed that the AD rats develop learning and memory deficits beginning at 24-week of age using an eight-arm water maze. AD rats (n = 11) took a longer time to escape and displayed more errors than age-matched wildtype (WT) rats (n = 6). We also completed a longitudinal comparison of the myogenic response (MR) of the middle cerebral artery (MCA) and found that the MR was similar in AD and WT rats at 8- to 12-week of age when perfusion pressure was increased from 40 to 180 mmHg. However, the MR was significantly reduced in 16-week old AD rats (n = 6) as the inner diameter of the MCA only decreased by 8.2 ± 2.4% when perfusion pressure was increased from 40 to 180 mmHg compared with 14.5% ± 2.0% in age-matched WT rats (n = 6). The impaired MR of the MCA was exacerbated in AD rats with aging. Autoregulation of CBF AD rats (n = 4)
in vivo
was impaired in the surface and deep cortex at 24-week of age compared to age-matched WT rats (n = 4). Furthermore, we found the contractile capability of the cerebral vascular smooth muscle cells (VSMCs) isolated from AD rats (n = 4) was significantly reduced compared with WT rats (n = 4), detected by the reduction in size of 15.7 ± 0.9% vs. 25.4 ± 1.0% using a collagen gel-based assay kit. These results provide evidence that cerebral VSMC dysfunction, impaired MR, and autoregulation of CBF precede the development of memory and learning deficits in the TgF344-AD rat model. However, the underlying mechanisms for the loss of VSMCs contractility in this AD model overexpressing mutant human amyloid precursor protein (
APPsw
) and presenilin 1 (
PS1ΔE9
) genes remain to be determined. Nevertheless, these results provide novel insight into the vascular contribution to AD.
Abstract 36: High Salt Induces Vascular Damage Through Redox-sensitive Parp/trpm2-induced Calcium Influx And Inflammasome Modulation Without Influencing Blood Pressure
Authors: Alves-lopes Rheure , Neves Karla B , Montezano Augusto C , Mary Sheon , Delles Christian , Touyz Rhian M
Published: August 2021

High salt diet (HSD) has deleterious effects on the vasculature by mechanisms not fully elucidated. We demonstrated tight coupling between Na
+
and Ca
2+
levels in VSMCs, where PARP-regulated TRPM2, a redox-sensitive Ca
2+
channel, plays an important role. Increased [Ca
2+
]i also contributes to inflammasome assembly, which dysregulates vascular function. We hypothesized that HSD induces a pro-oxidant environment that contributes to PARP-induced TRPM2-activation, Ca
2+
influx and inflammasome assembly, leading to vascular damage. WKY rats were treated with 1% HSD (3 weeks). Blood pressure was assessed by tail-cuff methodology, vascular reactivity was assessed in mesenteric arteries and calcium influx, ROS generation, inflammasome and pro-contractile marker in vascular smooth muscle cells (VSMCs) in presence and absence of HS medium (HSM-140mM). HSD did not increase blood pressure (BP), but vascular contractility was exaggerated (Emax(mN): WT 10.20 ± 0.70 vs 15.17 ± 1.74), effect reversed by PARP (Emax: 11.14 ± 1.24) and TRPM2 (8-br) (Emax: 11.45 ± 1.11) inhibitors. In VSMCs, HSM behaves as a pro-oxidant agent (ROS-AU: Control 77.51 ± 2.80
vs
130.04 ± 13.89 HSM), leading to increased [Ca
2+
]i (AUC: Control 25562.45 ± 880.48
vs
30924.8 ± 1263.85 HSM) and activation of myosin light chain (pMLC-AU: Control 99.27 ± 1.01
vs
626.87 ±71.28 HSM), by mechanisms dependent on ROS and PARP/TRPM2 activation. HSM also increased expression of inflammasome components NLRP3 (2
ΔΔCt
: Control 1.20 ± 0.01
vs
1.85 ± 0.19 HSM), ASC (2
ΔΔCt
: Control 1.02 ± 0.01
vs
1.49 ± 0.17 HSM) and Caspase 1 (2
ΔΔCt
: Control 1.06 ± 0.03
vs
2.35 ± 0.46 HSM), which was prevented by ROS scavenger Tiron and PARP inhibitor. In conclusion, HSD-induced vascular hypercontractility involves ROS activation of PARP/TRPM2 signalling. Activation of PARP/TRPM2 was associated with increased [Ca
2+
]i and activation of pro-contractile signaling and inflammasome assembly. Normal BP in HSD-fed rats in the presence of vascular damage suggests that ROS/PARP/TRPM2 signaling induced by salt influences vascular function independently of BP elevation. We identify a novel pathway that underlies salt-induced vascular damage and suggest a potential therapeutic role for PARP/TRPM2 inhibitors in vascular dysfunction.
Abstract 39: Fate Of Renin Cell Progenitors In The Developing Kidney Vasculature
Authors: Martini Alexandre , Gomez Ariel R , Sequeira Lopez Maria Luisa
Published: August 2021

The unique spatial arregement of the kidney arterioles is an essential event for its development. However, the mechanisms that govern this process are still poorly understood. During nephrogenesis, a group of stromal cells expressing the Forkhead Box D1 (
FoxD1
) transcription factor (TF) will give rise to the metanephric progenitors for the mural cells of the kidneys arteries and arterioles. We aim to identify the core TFs involved in the cell fate along the differentiaton pathways of the developing kidney vasculature. Therefore, we generated Foxd1-cre; mTmG mice, whose
Foxd1
derivative cells are labeled with green fluorescent protein (GFP). GFP+ cells were isolated from 5 (P5) or 30 (P30) days old mice kidneys, and processed either for single-cell RNA-Seq (scRNA-Seq) or for single-cell Assay for Transposase-Accessible Chromatin (scATAC-Seq
).
The top5 highly expressed TFs on scRNA-Seq at P5 are:
Tcf21, Zeb2, Meis2, Cebpd and Nme3
(p_adjusted_value(padj)= 0, 3.8E-187, 3.9E-180, 4E-172, 4.1E-172 and 3.2E-154, respectively). They are involved in developmental processes and cell proliferation. At P30, the top5 highly expressed TFs are:
Atf3, klf2, Fos, Nr4a2
and
Junb
(padj= 4.2E-294, 2.1E-200, 3.5E-182, 1.7E-52 and 0.2E-24, respectively). They are implicated with calcium-signaling pathway and inflammation. Additionally, scATAC-Seq identifies regions of accessible chromatin for pontential TFs binding, leading to changes in gene expression content and cell identity. At P30, scATAC-Seq showed differential accessible regions with subsequent putative motif enrichment analysis for the TF N4a2 (padj: 4E-297). This is in accordance with our scRNA-Seq results and might play a role in the
Foxd1
progenitors cell fate decisions. Our results tracks the fate of the Foxd1+ cells during the kidney vasculature assembly and suggest a new transcription factors network that might play a role to orchestrate cell fate decisions during kidney vascular development.
Abstract 47: Circulating Human Sprr Increased Blood Pressure In Female Mice Fed A Low-fat Diet
Authors: Poupeau Audrey , Arthur Gertrude , Nichols Kellea , Yiannikouris Frederique B
Published: August 2021

Elevated plasma soluble prorenin receptor (sPRR) is associated with essential hypertension and obesity-hypertension in men. Additionally, our laboratory previously found that the infusion of mouse sPRR elevates systolic blood pressure (SBP) in high-fat (HF) fed male mice through activation of the sympathetic nervous system but did not elevate SBP in HF-fed female mice. Interestingly, mouse sPRR infusion increased renal and hepatic angiotensinogen (AGT) and plasma renin concentration in female mice fed a low-fat diet. However, whether sPRR-activates the renin angiotensin system (RAS) and increases blood pressure in low-fat fed female mice remains to be investigated. Additionally, little is known concerning the influence of human sPRR on blood pressure in women. Therefore, we developed a humanized mouse model with high circulating human sPRR. Human sPRR-Myc-tag transgenic mice were bred with mice expressing Alb/Cre recombinase to induce human sPRR release in the circulation. Control and Alb-HsPRR female mice were fed a LF-diet for 8 months (n=11/groups). Body weight and body composition were examined and blood pressure assessed by radiotelemetry. Human sPRR-Myc-tag was detected in the liver of Alb-HsPRR female mice and plasma sPRR levels increased by 50-fold (CTL: 3.6±0.5 ng/ml, HsPRR:190.5±24.4 ng/ml; P<0.05), which validated the humanized mouse model. Elevated circulating human sPRR did not change body weight (CTL: 22.2±0.37, HsPRR: 23.0±0.32 g) or fat mass (CTL: 2.5±0.2, HsPRR: 3.1±0.2 g). Liver-derived human sPRR significantly elevated SBP in Alb-HsPRR compared to control female mice (Night SBP: CTL: 130.5±1.2 mmHg; Alb-HsPRR: 135.9±2 mmHg; P<0.05) and acute injection of AngII exacerbated SBP elevation. Interestingly, the decrease in blood pressure mediated by losartan was not different between Alb-HsPRR and control female mice (Night ΔSBP: CTL: -13.11±2.2 mmHg; Alb-HsPRR: -14.8±2.7 mmHg; P>0.05). Plasma AGT and renin activity were similar between Alb-HsPRR and control female mice. Therefore, whether the local RAS or the sympathetic nervous system are involved in human sPRR-mediated increase of SBP remains to be examined. Altogether, our results suggest an important role of circulating human sPRR in blood pressure control in women.
Abstract 51: Ambulatory Blood Pressure Phenotype And Cardiovascular Risk In Youth: The Ship-ahoy Study
Authors: Flynn Joseph T , Khoury Philip , Samuels Joshua A , Lande Marc B , Meyers Kevin , Ferguson Michael A , Urbina Elaine
Published: August 2021

We investigated whether blood pressure (BP) phenotype based on clinic & 24-hour ambulatory BP (ABP) was associated with intermediate markers of cardiovascular disease (CVD) in 374 adolescents enrolled in a study of the relationship of BP to CV risk. Clinic BP was measured by auscultation and categorized using the 2017 AAP guideline. ABP was measured for 24 hours by an oscillometric device and analyzed using the adult ABP wake SBP cut-point (130 mmHg). This created 4 BP phenotype groups: normal BP (n=224), white coat hypertensive (n=48), ambulatory hypertensive (n=57) & masked hypertensive (n=45). Echocardiographic parameters & carotid-femoral pulse wave velocity (PWVcf) were measured to assess CVD risk. Left ventricular mass (LVM) was lowest in the normal BP group, whereas multiple measures of cardiac function and PWVcf were worse in the masked and ambulatory hypertensive groups:

Generalized linear models adjusted for body mass index (BMI) were constructed to examine the associations between BP phenotype and the measured CVD variables. ABP phenotype was an independent predictor of LVM, diastolic and systolic function and PWVcf in the unadjusted model. ABP phenotype remained significantly associated with diastolic function (E/e’, e’/a’), systolic function (ejection fraction) and increased arterial stiffness (PWVcf) after adjustment for BMI percentile (all p<=0.05). We conclude that BP phenotype is an independent predictor of markers of increased CVD risk in adolescents, including impaired cardiac function and increased vascular stiffness. ABP monitoring has an important role in CVD risk assessment in youth.
Abstract 53: Results Of The Nashville Barber-pharmacist Hypertension Management Pilot Study
Authors: Parrish Jarod , Blyler Ciantel A , Okafor Henry E , Chang L. Cindy , Nair Devika , Peek Grayson , Ikizler Talat A , Norris Keith C , Harrison David G , Cavanaugh Kerri
Published: August 2021

Background:
The Los Angeles Barbershop Blood Pressure Study (LABBPS) demonstrated that collaboration between barbers and pharmacists delivering hypertension management could significantly improve blood pressure as well as access to care. It is unknown if this model can be translated to other locations. This single-arm, proof-of-concept pilot study tested the development and implementation of a similar protocol in Nashville, TN (NCT04232124).

Methods:
Between 2019 and 2021, community stakeholders, clinicians, investigators, and the LABBPS team convened to adapt the study design and protocol. Established barbershops with barbers willing to be trained on study procedures were recruited as study sites. Non-Hispanic Black male clients, aged 35-79 years with systolic blood pressure (SBP) > 140 mmHg on two screening days were eligible for participation. Enrolled participants met with a pharmacist for lifestyle and medication management at least once monthly for a six-month period. Barbers measured blood pressure during haircuts for additional monitoring.

Results:
Eight barbershops in business for an average of 20 (± 5) years participated in the trial. Barbers from each shop (range: 1-4) were trained. A total of 419 clients completed screening visit 1, 82 were eligible and 52 (12%) completed visit 2. We enrolled 36, with 30 completing the initial clinical visit and 27 had complete data at 6-months. Participants were on average age 50 (± 10) years, had a body mass index 33 (± 6), 44% were currently smoking, 52% with high school or less education, and 56% reported current primary care. Baseline BP 157.7±17.1/ 95.1±13.9 mmHg improved to 125.7±11.9/75.6±9.5, a change of -32.1±21.6/ -19.5±14.1, respectively. At 6-months 85% of the group had a BP<140/90, 74% BP<135/85, and 67% BP<130/80. At baseline 15% of participants reported health as excellent/very good and this increased to 56% at 6-months (p=0.002). Adverse reactions reported were minimal.

Conclusion:
Implementation of the barber-pharmacist model of hypertension management and care delivery improved BP control among hypertensive Black men in Nashville. Lessons learned regarding adaptation overcoming unforeseen barriers will inform expansion into additional naïve locations for future research.
Abstract 56: Proteome Profile Of Vascular Smooth Muscle Cells During Phenotypic Switching In Human Hypertension
Authors: Camargo Livia L , Mary Sheon , Lilla Sergio , Zanivan Sara , Bulleid Neil , Hartley Richard , Delles Christian , Leiper James , Fuller Will , Montezano Augusto C , Touyz Rhian M
Published: August 2021

Vascular smooth muscle cells (VSMCs) are key players in vascular dysfunction associated with hypertension, where phenotypic switch is a fundamental process. While various transcription factors have been implicated in this process, the proteomic signature associated with phenotypic switching in human hypertension is unknown. Using high fidelity proteomic analysis, we characterized the proteome profile of VSMC in human hypertension. VSMC derived from resistance arteries from normotensive (NT) and hypertensive (HT) subjects were studied. Protein expression and cell migration were assessed by immunoblotting and wound healing assay. VSMC proteins were labelled with isobaric tandem mass tags and identified by liquid chromatography tandem mass spectrometry. The oxidative proteome was assessed using stable isotope-labelled iodoacetamide to target free reduced cysteine thiols. VSMCs from HT subjects exhibit reduced expression of α-SMA (0.05±0.01 vs NT:0.20±0.03, p<0.05), increased expression of the proliferation marker, PCNA (0.162±0.3 vs NT:0.51±0.004, p<0.05), and increased migration (54.68±2.86 vs NT:23.37±8.36, p<0.05). The proteomic analysis identified 207 proteins upregulated in HT subjects (fold change>1.5, p<0.05). There were no changes in protein expression of pathways related to the contractile phenotype (MYH11, CNN1, TAGLN, TPM, CALD1). However, extracellular matrix (ECM) proteins such as COL1A1, COL9A1, COL10A1, FBN1, FBLN1 were increased in cells from HT (fold change>1.5, p<0.05), suggesting a switch to a fibroblast-like phenotype in hypertension. Expression of proteins related to the interferon and IL-1β pathways (IFIT1, IFIT2, IFIT3, MX1, MX2, ABCA1, ABCA2, IL1RAP, CD36, ICAM1) were also increased in cells from HT subjects (fold change>1.5, p<0.05). Considering the importance of oxidative stress in hypertension, we assessed the VSMC oxidative proteome. Results demonstrate that ECM proteins, such as COL11A1 and COL16A1, were highly oxidized in cells from HT (fold change>1.5, p<0.05). Our study provides new insights into the proteomic changes that define the vascular phenotype in hypertension and highlights candidate targets that may drive phenotypic switching associated with vascular injury in hypertension.
Abstract 57: Risk Factors For Severe Reductions In Blood Pressure After Treatment Of Severe Inpatient Hypertension
Authors: Ghazi Lama , Li Fan , Chen Eric , Simonov Michael , Yamamoto Yu , Biswas Aditya , Hanna Jonathan , Shah Tayyab , Peixoto Aldo J , Wilson F. Perry
Published: August 2021

Background:
Incident severe HTN during hospitalization is far more common than admission for HTN, however treatment guidelines are lacking. Severe inpatient HTN is poorly studied, therefore our goal is to characterize inpatients who develop severe HTN and assess BP response to antihypertensive treatment.

Methods:
This is a cohort study of adults admitted for reasons other than HTN and developed severe HTN within a single healthcare system. We defined severe inpatient HTN as the first documentation of BP elevation (>180 systolic or >110 diastolic) at least 1 hour after hospital admission. Treatment was defined as receiving antihypertensive medications within 6 hours of BP elevation. We studied the association between treatment and BP drop ≥30%.

Results:
Among 224,265 hospitalized adults, 23,147 developed severe HTN of which 40% were treated. Compared to inpatients who did not develop severe HTN, those who did were older, more commonly women and Black, and had more comorbidities. Of the treated and untreated patients, 45.5 and 46.4% had a MAP drop ≥30% (p-value= 0.2). Risk factors for severe MAP drop include older age, Black race, HTN, and diabetes. Additionally, treatment vs. no treatment and treatment with intravenous vs. oral medications were associated with greater odds of MAP drop ≥30% (
Table 1
).

Conclusion:
While there was no difference in the proportion of treated and untreated patients with severe MAP reduction, after adjustment for factors independently associated with HTN we found that treatment was associated with severe BP drop. Further research is needed to phenotype inpatients with severe HTN to help establish treatment guidelines.
Abstract 62: Dietary Fibre Attenuates Hypertension By Reducing Macrophage Infiltration Via Gut Microbiota-Derived Metabolite-Sensing Receptors
Authors: R Muralitharan Rikeish , Dinakis Evany , Xu Chudan , Xie Liang , Jama Hamdi , Nakai Michael E , Paterson Madeleine , Salimova Ekaterina , Bitto Natalie , Kaparakis-Liaskos Maria , Barlow Chris , Creek Darren , O'Donnell Joanne , Mackay Charles , Marques Francine
Published: August 2021

High dietary fibre is fermented by the gut microbiota, resulting in the release of metabolites called short-chain fatty acids (SCFAs). Both fibre and SCFAs can reduce high blood pressure (BP) and its associated cardio-renal complications. However, the underlying mechanisms remain unclear. SCFAs can be detected by metabolite-sensing receptors GPR41 and GPR43, highly expressed by immune cells such as macrophages. We hypothesised that dietary fibre attenuates hypertension by modulating renal macrophage infiltration via metabolite-sensing receptors GPR41 and GPR43. To test this, we developed a novel GPR41/GPR43 double knockout (DKO) mice and characterised the cardiovascular and immune phenotype in both sham and angiotensin-II (Ang-II, 0.5mg/kg/day) treated DKO and wild-type (WT) mice (n=7-12 per group). WT Ang-II mice fed a high-fibre diet had significantly lower renal galectin-3 (p=0.0004), a macrophage marker, compared to WT Ang-II mice fed a low-fibre diet. Sham DKO mice on standard chow diet had no difference in BP or heart function but had higher kidney/tibia length index (p=0.049) and renal fibrosis levels compared to WT mice (p=0.004). Moreover, untreated DKO mice had higher numbers of renal macrophages compared to WT mice (p=0.002). Ang-II infusion of DKO mice resulted in higher BP (p<0.0001), renal fibrosis (p=0.007), and mortality (hazard ratio=5.6) compared to WT mice. In the gut, we found significant inflammatory changes, gut barrier integrity disruption, gut microbiota and metabolome changes (all p<0.05). We also found evidence of lipopolysaccharides (LPS) translocation from the gut into the circulation of DKO mice. In conclusion, we show that a high-fibre diet attenuates hypertension by modulating renal macrophages via the gut microbiota-derived metabolite-sensing GPR41 and GPR43 receptors. These receptors can be targeted as a novel treatment for hypertension.
Abstract 63: The Existence Of An Obesity Paradox In Uncontrolled Hypertension: A 1999-2018 National Health And Nutrition Examination Survey Analysis
Authors: Visaria Aayush , Setoguchi Soko
Published: August 2021

Introduction:
An obesity paradox has been described for several CVD conditions (e.g. heart failure), but not CVD risk factors. We aimed to determine whether the association between obesity and uncontrolled BP exhibited an obesity paradox in participants with HTN.

Methods:
We included adults ≥20 years from the 1999-2018 National Health and Nutrition Examination Surveys. Obesity was categorized using BMI (Table). Mean BP was calculated as the average of 3 consecutive BP readings 30 seconds apart. Severe asymptomatic HTN (SAH) was defined as mean SBP≥180 or DBP≥120 mmHg. Overtly uncontrolled BP (OUBP) was defined as SBP≥160-179 or DBP≥100-119. We used survey design-adjusted multivariable logistic regression to determine odds of SAH and OUBP.

Results:
Among 15,042 participants with self-reported HTN (mean age 57, 52% female, 71% White), 7.1% had OUBP and 2.2% had SAH. Participants with BMI ≥30 were more likely to be younger, female, non-Hispanic Black, have dyslipidemia, and report lower sodium intake, and less likely to have CKD or CVD. Those with BMI ≥27.5 had significantly lower adjusted odds of SAH than participants with a normal BMI of 18.5-23, whereas BMI<18.5 was associated with insignificantly increased odds of SAH (Table). Every 1 kg/m
2
increase in BMI was associated with a 4% decrease in odds of SAH. BMI was not significantly associated with OUBP. Addition of waist circumference attenuated, but did not modify, the effect of BMI on SAH or OUBP.

Conclusion:
Even after adjustment for demographic, socioeconomic, and comorbid factors, a BMI above 27.5 kg/m
2
was associated with >40% decreased odds of SAH among participants with self-reported HTN.