bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P194: Metabolic effects of GDF15 and Activin A on trophoblasts
Authors: Hartmann Sunhild , Zucker Theresa , Botha Stefan , Schmidt-Ullrich Ruth , Secener Kerim , Stern Christina , Huppertz Berthold , Forstner Desiree , Geisberger Sabrina , Herse Florian , Gauster Martin , Ishaque Naveed , Mueller Dominik , Dechend Ralf , Nonn Olivia
Published: October 2024

Preeclampsia (PE), a pregnancy-related complication, is a major cause of maternal and fetal morbidity and mortality. The pathophysiology is not fully elucidated.

Activin A and GDF15 are part of the TGFβ superfamily and markers for trophoblast differentiation. Both are senescence associated secretory phenotype (SASP) factors that have been associated with cardiovascular disease. Mitochondrial dysfunction is associated with SASP and both have been described in PE. Serum levels of GDF15 and Activin A, were elevated in the maternal blood prior to the onset of PE and found to be elevated in placental tissue of women with PE.

BeWo cells were metabolically reprogrammed to a physiological glucose-environment by cultivating them for 10-15 passages in a cell culture medium with 5.5 mM glucose as opposed to the standard high-glucose environment at 17.1mM. Next, BeWo cells were differentiated into multinucleated syncytiotrophoblasts with Forskolin, and with DMSO as solvent control to model cytotrophoblasts. Subsequently, these two trophoblast differentiation states were treated with GDF15 (20 ng/ml) and Activin A (20ng/ml) and the cell metabolic profile was analyzed.

Additionally, a single cell human trophoblast organoid transcriptomics dataset from Shannon et al. was investigated for expression patterns of
GDF15
and
INHBA
(Activin A) together with the overall metabolic profile of trophoblast cells.

Treating BeWo cells with Activin A and GDF15 leads to an increase in mitochondrial activity, shown by elevated ATP levels in syncytiotrophoblast and an increase in mitochondrial membrane potential in cytotrophoblasts. Additionally, the oxygen consumption rate is significantly higher in syncytiotrophoblasts after Activin A and GDF15 treatment.

In human trophoblast organoids, syncytiotrophoblasts showed the lowest metabolic activity measured by the expression of genes related to glycolysis and mitochondrial respiratory chain. Furthermore,
GDF15
and
INHBA
(Activin A) were strongly expressed in syncytiotrophoblasts.

Together, Activin A and GDF15 stimulation increases the metabolic activity in BeWo cells. As preeclampsia is associated with mitochondrial dysfunction as well as with senescence, our data indicate a potential role of Activin A and GDF15 in preeclampsia development.
Abstract P196: Association between Preeclampsia and Risk of Arrhythmias and Stroke: A Meta-analysis of 3,871,787 women patients.
Authors: Jaiswal Vikash , Jaiswal Akash , Hanif Muhammad , Tibon Deborah , Rajak Kripa , Tummala Nayanika , Biswas Monodeep
Published: October 2024

Background:
Preeclampsia, a hypertensive disorder of pregnancy has been known to be associated with adverse outcomes including cardiovascular complications. However, data on the association between preeclampsia and the incidence of arrhythmias are sparse with limited literature to date.

Objective:
We sought to evaluate the association between preeclampsia and the risk of arrhythmias.

Methods:
We performed a systematic literature search on PubMed, Scopus, and Clinicaltrial.gov for relevant studies from inception until April 2024 without any language restrictions. Adjusted Hazard ratios (aHR) and 95% confidence intervals (CI) were pooled using a random-effect model.

Results:
A total of 4 studies with 3,871,787 women patients (86,452 women in the preeclampsia group and 3,785,245 women in non preeclampsia) were included in the analysis. Pooled analysis showed that women with preeclampsia were having a higher incidence of arrhythmias (aHR, 1.71 (95%CI:1.24-2.18), P<0.001), and stroke (aHR, 1.72(95%CI: 1.25-2.19), P<0.001) when compared with the non preeclampsia group of women.

Conclusion:
Our study shows that women with Preeclampsia were having a higher incidence of arrhythmias and stroke. Proper and timely cardiovascular assessment may benefit from morbidity and mortality.
Abstract P197: Identification and Characterization of Rho-related BTB Domain Containing 1 (RhoBTB1)-Cullin 3 (CUL3) Proteins in Placenta
Authors: Tvina Alina , Kumar Gaurav , Lu Ko-Ting , McIntosh Jennifer , Sigmund Curt
Published: October 2024

Preeclampsia is life threatening condition affecting about 10% of all pregnancies. A potential mechanism is via malfunctioning syncytiotrophoblasts in the placenta. RhoBTB1 is an adapter protein for the CUL3 E3 ubiquitin ligase which delivers protein substrates for proteasomal degradation. An analysis of gene expression databases revealed that the highest level of RhoBTB1 expression is in the placenta. RNAscope in situ hybridization data reveals that RhoBTB1 expression is specifically localized to syncytiotrophoblasts. In blood vessels, RhoBTB1 controls the state of nitric oxide mediated vasodilation because it acts as a substrate adaptor for Phosphodiesterase 5. Thus, we wanted to identify potential protein targets of RhoBTB1 in the placenta to assess if RhoBTB1 plays a role in placental function. To simplify the first identification of RhoBTB1 binding proteins, we employed
HTR-8
/SVneo (HTR-8) cells as a model of placental trophoblast cells. We biotinylated the HTR-8 proteome utilizing ascorbate peroxidase (APEX2) proximity labelling system followed by mass spectrometry to identify RhoBTB1 targets. We employed a fusion protein consisting of the C-terminal half of RhoBTB1 (B1B2C), the region required for the binding of other protein substrates, and a control lacking the most C-terminal region (B1B2). We next chose proteins with binding to B1B2C > B1B2 of 2-fold or greater that were expressed in the placenta based on the Human Protein Atlas. Rho GTPase Activating Protein 29 (ARHGAP29), which controls the regulation of small GTP binding proteins, and S-phase Kinase associated Protein 2 (SKP2), a regulator of cell cycle progression was studied further. SKP2 is interesting because it exhibits a tissue specific tropism for placental expression with its highest expression in extravillous trophoblasts, cytotrophoblasts, and syncytiotrophoblasts. Expression of ARHGAP29 and SKP2 in HTR-8 cells were markedly upregulated by MLN4924 suggesting they are regulated by the CUL pathway. They were also modestly upregulated after partial inhibition of RhoBTB1 by siRNA suggesting they may be RhoBTB1-CUL3 targets. Studies are ongoing to validate the physical interaction of ARHGAP29 and SKP2 with RhoBTB1 by co-immunoprecipitation. We are also using RNAscope to assess the co-expression of RhoBTB1, ARHGAP29 and SKP2 in the human placenta from normal and preeclamptic pregnancies.
Abstract P205: Single-nucleus Sequencing Reveals Endothelial Cell-specific Dysregulation in the Kidney in Multiple Models of Hypertension
Authors: Qiu Qiongzi , Liu Yong , Xue Hong , Liu Jing , Therani Bhavika , Usa Kristie , Huang Jing , Greene Andrew , Rao Sridhar , Geurts Aron , Cowley Allen , Liu Pengyuan , Liang Mingyu
Published: October 2024

The kidney plays a crucial role in the onset and progression of hypertension. To investigate the roles of different renal cell types in hypertension, we applied single-nucleus sequencing and spatial transcriptome sequencing to kidney samples from three well-established hypertension models: C57BL/6 mice treated with Ang II, the Dahl salt-sensitive (SS) rat on high-salt diets, and the spontaneously hypertensive rat (SHR), along with their respective controls including Sprague-Dawley (SD) and Wistar Kyoto (WKY) rats. Our dataset comprised 179,637 nuclei from 24 samples across 12 experimental conditions, identifying 17 major cell types including various epithelial cells, endothelial cells, vascular smooth muscle cells, fibroblasts, and immune cells. At baseline, differentially expressed genes between the SS and SD groups were specifically enriched in endothelial cell pathways related to vascular development. This included expression deficiencies in genes involved in angiogenesis in SS rats, such as Arhgap24 and Nox4. Compensatory expression changes in these genes were observed in the SS rats after a high-salt diet and in 26-week-old SHR, but not in their respective controls. These genes were also associated with multiple blood pressure-associated single-nucleotide polymorphisms (SNPs). Furthermore, changes in endothelial cell communication were most prominent upon stimuli, with impaired PTPRM (Protein Tyrosine Phosphatase Receptor Type M) signaling being a robust feature in hypertension models, involving angiogenic endothelial cells in the SS model and proliferating capillary endothelial cells in the SHR model. Our study highlights the critical role of renal endothelial cell angiogenic dysfunction in the pathogenesis of hypertension and suggests a link between specific renal endothelial cell subtype dysregulation and hypertension development.
Abstract P214: Targeted disruption of CTCF binding in the Dahl salt-sensitive rat reveals
Renin
transcriptional dynamics
Authors: Vanden Avond Mark , Grzybowski Michael , Greene Andrew , Cowley Allen , Liang Mingyu , Geurts Aron
Published: October 2024

Changes in chromatin conformation caused by common genetic variations may impact blood pressure regulation and hypertension risk. A major function of CCCTCF-binding factor (CTCF) is to bind to DNA to regulate and maintain chromatin conformation, and thereby gene expression. Cellular expression of CTCF is critical for proper
Renin
expression and the
Renin
gene is surrounded by multiple CTCF-binding motifs
.
In the Dahl salt-sensitive (SS) rat, CRISPR-Cas9 was utilized to create three models harboring mutations to the CTCF motif surrounding the
Renin
gene. CTCF binding was confirmed to be disrupted in all three models. The mutants and wild-type (WT) littermates were then placed on a 0.1%, 4.0% or maintained on a 0.4% NaCl diet for four days, and
Renin
mRNA and plasma renin activity (PRA) was examined. mRNA analysis showed no differences in
Renin
expression on any diet for any model compared to WT. Interestingly, two of our male mutant models failed to increase their PRA on the 0.1% NaCl diet (CTCF1=13.87±0.86; CTCF2=13.95±0.59; p<0.05 compared to WT=17.52±0.81 on 0.1% NaCl). We hypothesized that
Renin
dynamic transcriptional response to salt depletion is delayed in the mutant models compared to WT. To test this, we have utilized an ex vivo kidney slice culture approach. Kidneys were excised from SS rats fed a 4.0% NaCl diet for 1-2 days, sliced to ~1-3 mms, and cultured for 24 hrs.
Renin
expression was evaluated at 0, 1, 2, 4, 8, 12, 16, 20 and 24 hrs of incubation in basal media.
Renin
expression was elevated in SS rats at 8 hrs (3.82±0.38 compared to 0 hr = 0.93±0.32) and continued to increase until 12 hrs (7.05±1.87) and reduced at 16 hrs (1.42±0.70). CTCF1 mutant
Renin
expression was significantly reduced at 12 hrs (3.77±0.58 compared to WT=7.05±1.87), while CTCF2 mutants showed no differences in
Renin
expression compared to WT. Thus, we established an ex vivo approach for monitoring the dynamic changes in
Renin
expression by culturing kidney slices. This method could be used to evaluate other stimuli, such as therapeutics.
Abstract P219: Risk factors for Accelerated Vascular Aging in Resistant Hypertension: Should we screen our patients earlier?
Authors: Bag Soumyadeep , Sehar Alina , Rathi Hinal , Qamar Aleeza , Patel Aesha , Roth Kaehler , Fanning John , Dudenbostel Tanja
Published: October 2024

Background:
There is an increase in Cardiovascular disease(CVD) mortality rates among young adults worldwide. Patients with resistant hypertension (RHTN) are at particularly high risk for CVD compared to those with more easily controlled hypertension (HTN). RHTN is defined as blood pressure above goal despite using ≥3 antihypertensive medications. The role of accelerated vascular aging (AVA) in this group has not been well investigated. Our study aims to evaluate risk factors for AVA and its role in RHTN.

Methods:
We conducted a cross-sectional analysis of a large cohort of 965 black and white patients with RHTN referred to the RHTN Clinic at the University of Alabama at Birmingham. Patients with HTN served as a control group.

Results:
The most compelling data was seen in young patients (age 41-55 yrs). Those with RHTN had significantly higher arterial stiffness than patients with HTN (pulse pressure: 63.3±17.9 vs 51.2 ±11.7 mm Hg, p=<0.001) earlier onset of HTN (34.2 ±7.3 vs 39.4±10.3 yrs, p=0.001) and longer duration (15.5±9.6 vs 10.7±9.0 yrs, p=0.001), higher BMI (35.4±8.3 vs 31.9±6.6 kg/m2), higher Aldosterone Renin Ratio (12.4±13.2 vs 7.9±12.9, p=0.026), and 24-hour urinary sodium excretion (217.2±104.6 vs 182.8±77.2 mmol, p=0.036).

Conclusion:
Younger patients with RHTN, when compared with HTN, seem to have AVA as evidenced by higher prevalance of obesity, arterial stiffness, overactivation of the renin-angiotensin-aldosterone system, and higher salt intake. Earlier screening, starting at the age of 35 yrs, may help detect AVA quicker and prevent complications associated with RHTN in younger adults
Abstract P224: RbFox2 is a Rho-related BTB Domain Containing 1 (RhoBTB1)-Cullin 3 (CUL3) Pathway Target that Regulates the Actin Cytoskeleton
Authors: Kumar Gaurav , Golosova Daria , Chaihongsa Nisita , Vazirabad Ibrahim , Muskus Veitia Patricia , Wackman Kelsey , Lu Ko-Ting , Brozoski Daniel , Sigmund Curt
Published: October 2024

Blood pressure (BP) is controlled by an array of cellular pathways. Our prior research revealed that endothelial nitric oxide (NO)-dependent vessel tone and BP are regulated through the RhoBTB1-CUL3 mediated proteasomal degradation of Phosphodiesterase 5 (PDE5). RhoBTB1 is an adaptor protein which delivers protein substrates to the CUL3 E3 ubiquitin ligase for proteasomal degradation. However, the range of protein targets of RhoBTB1 remains unknown. We employed the ascorbate peroxidase (APEX2) proximity labelling system coupled with mass spectroscopy to identify RhoBTB1 targets in A7r5 smooth muscle cells. This analysis identified nearly 75 proteins which putatively bind to the C terminal half of RhoBTB1 (known as B1B2C), the region we previously identified was essential for adaptor function. One of these proteins was RNA binding motif protein 9, known as RBM9 and RbFox2, a key regulator of splicing events. Subsequent co-immunoprecipitation (Co-IP) assays validated the interaction of RbFox2 with RhoBTB1 and its B1B2C domains. Expression of Rbfox2 was increased in response to treatment of A7r5 cells with either MG132, an inhibitor of the proteasome, and MLN4924, an inhibitor of the CUL family. Similarly, RbFox2 expression was increased in CRISPR-Cas9 generated CUL3-deficient HEK293 cells, in aorta isolated from SMC-specific CUL3-deficient mice, and in A7r5 cells expressing an siRNA targeting RhoBTB1. Ang-II treatment of mice, which raised blood pressure by 40 mmHg, also increased RbFox2 expression, presumably through down regulation of RhoBTB1. Targeting RbFox2 by siRNA in A7r5 cells resulted in a decrease in actin fiber formation detected by phalloidin staining. In summary, our findings suggest that RbFox2 regulation may occur through the RhoBTB1-CUL3 proteasomal pathway and may potentially play a mechanistic role in actin cytoskeleton organization and thus arterial stiffness associated with hypertension.
Abstract P226: Sex-Dependent Effects of Perivascular Adipose Tissue on Arterial Stiffness in Hypertensive
Authors: McClintock Dillon , Bush Sydney , Sather Lisa , Tykocki Nathan , Lauver Adam , Watts Stephanie , Roccabianca Sara
Published: October 2024

Dillon McClintock, Department of Mechanical Engineering, Michigan State University, East Lansing, MI

Sydney Bush, Department of Mechanical Engineering, Michigan State University, East Lansing, MI

Lisa Sather, Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI

Nathan Tykocki, Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI

Adam Lauver, Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI

Stephanie W Watts, Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI

Sara Roccabianca, Department of Mechanical Engineering, Michigan State University, East Lansing, MI

Perivascular adipose tissue (PVAT) plays a crucial role in arterial mechanics by significantly reducing arterial stiffness in the thoracic aorta in normotensive rats. This study aims to test the hypothesis that impact of PVAT on arterial stiffness is sex-dependent in an adiposity-driven hypertension rat model. Thoracic aorta from male and female Dahl Salt-Sensitive rats were fed control (10% fat-CD) and high-fat (60% fat-HFD) diets over 16 weeks. Uniaxial mechanical tests were then conducted on aorta samples with PVAT (+PVAT) and without PVAT (-PVAT). In -PVAT samples, arterial wall stiffness at 20% stretch (mimicking
in vivo
conditions), was higher in males compared to females on a HFD but not on a CD. HFD rats also had higher systolic blood pressures, but the increase was significant only in females. +PVAT samples’ stiffness was reduced by a 95% factor in all groups as compared to -PVAT samples. Furthermore, when including PVAT, the sex difference in the HF diet group was absent. These results suggest that PVAT has a beneficial effect that is preserved in this adiposity-driven hypertension animal model after 16 weeks on diet. Furthermore, remodeling of the arterial wall is more pronounced in male versus female rats and the inclusion of PVAT dampens this sex difference.
Abstract P227: Potential roles of vascular glucose transporter 1 in development of abdominal aortic aneurysm induced by angiotensin II
Authors: Nakayama Yuki , Torimoto Keiichi , Rizzo Victor , Eguchi Satoru
Published: October 2024

It has been reported that pharmacological inhibition of glycolysis attenuated abdominal aortic aneurysm (AAA) development in mice. Glucose transporters (GLUTs) incorporate glucose as a first step of glycolysis. GLUT1 over-expression in vascular smooth muscle cell (VSMC) appears to enhance atherosclerosis in a mouse model. In angiotensin II (AngII) plus BAPN model of mouse AAA, silencing of VSMC AngII type 1 rec receptors attenuated AAA development and rupture which was associated with less immune cell infiltration to the aortas. However, the role of VSMC GLUT1 in AAA remains elusive. Here we tested our hypothesis that VSMC GLUT1 deletion attenuates AAA development and rupture in mice. Rat aortic VSMC were obtained by the explant method. Media glucose consumption of cultured VSMCs was evaluated in the presence or absence of angiotensin II (AngII, 100 nM) stimulation and GLUT1 inhibitor BAY-876. GLUT1 floxed mice were crossed with tamoxifen-inducible SMMHC-Cre mice. At 6 weeks of age, mice were injected with 2 mg/kg tamoxifen for 5 days. Significant vascular GLUT1 silencing was confimed with immunofluorescent chemistry. As a control, tamoxifen-treated SMMHC-Cre male mice were utilized. At 8 weeks, AngII (1 μg/kg/min) was delivered for 4 weeks. In addition, mice were received 1 mg/mL BAPN during the first 2 weeks of AngII infusion. After 4weeks, all animals were euthanized and maximal external diameter of the abdominal aorta was measured. In cultured VSMC, glucose consumption increased by AngII was attenuated by pretreatment of 100 nM BAY-876. AngII plus BAPN significantly developed AAA in control mice. However, silencing of VSMC GLUT1 did not affect AAA development by AngII plus BAPN. The survival rate due to aortic rupture in VSMC GLUT1 silenced mice and control mice treated with BAPN and AngII were 60.0% and 57.1%, respectively. AngII induced hypertension in control mice and VSMC GLUT1 silenced mice. Although GLUT1 may be a primary VSMC GLUT to enhance glucose utilization in response to AngII stimulation, VSMC GLUT1 silencing did not alter AngII-dependent AAA development or rupture in a mouse model. Since pharmacological inhibition of GLUT

attenuated AAA and associated macrophage infiltration, it is possible that GLUT1 or other GLUT expressed in macrophages may contribute to development of AAA. In conclusion, VSMC GLUT1 silencing did not alter AngII-dependent AAA development or rupture in a mouse model.
Abstract P228: Kdm6a Regulates Extracellular Matrix Genes During Arterial Stiffening and High Blood Pressure in Female Mice
Authors: Sakamuri Anil , Barris Candee , Kamau Anne , Sengottaian Divya , Belin De Chantemele Eric , Ogola Benard
Published: October 2024

Arterial stiffness measured by pulse wave velocity (PWV) increases steeply during menopause due to estradiol decline. Longitudinal studies show higher PWV in postmenopausal women compared to age-matched men. Previously, we have demonstrated increased PWV in ovariectomized (OVX) and middle-aged female mice. However, the impact of sex chromosomes on PWV is unknown in female mice. We hypothesize that a decline in estradiol unleashes the female sex chromosomes effect that promotes arterial stiffening. This study used four core genotype mice (n=6-10/group) of XX or XY sex chromosomes left intact or OVX for 8 weeks. Arterial stiffening was assessed with pulse wave Doppler and 24-hour blood pressure by radiotelemetry. Carotid artery passive myography and vascular reactivity of 2
nd
order mesenteric artery were assessed by pressure myography. To induce hypertension, XX and XY mice were treated with deoxycorticosterone acetate (DOCA; 50 mg/pellet) for 21 days and 1% saline. In vitro, isolated aortic vascular smooth muscle cells (VSMC) were stretched on collagen matrix or treated with Kdm6a inhibitor GSKJ-1. PWV was significantly increased in OVX than in gonadal intact XX and XY mice (P<0.001). However, an interaction effect was indicated with higher PWV in XX than XY OVX mice (4.9±0.1 vs. 4.2±0.2 m/s; P<0.001), indicating a sex chromosome effect. Carotid artery stress-strain curves showed a leftward shift of XX than XY mice, suggesting increased structural stiffness. Masson’s trichrome staining of the aorta showed increased collagen in the adventitia of XX than XY (30% vs. 26%; P=0.005) mice. Higher systolic blood pressure (SBP) was indicated in XX than in XY mice (Baseline; 123±3 vs. 116±4 mmHg; P=0.02). Aortic RNAseq showed increased X-linked genes, including Kdm6a, in XX compared to XY mice. Stretched aortic VSMC showed increased Col1a1 and decreased Itga4 and Lama2 genes reversed by GSKJ-1. DOCA-induced hypertension blunted the PWV difference between XX and XY mice; however, SBP was higher in XX than in XY mice (Week 1: 135±13 vs.127±3 mmHg; P=0.03), but not weeks 2 and 3. We observed impaired acetylcholine-mediated relaxation in hypertensive XX than XY mice (47% vs. 24%; P=0.03). Our data suggested that OVX mice with XX sex chromosomes promote arterial stiffening and high blood pressure, and Kdm6a regulates extracellular matrix genes in XX female VSMC. DOCA-induced hypertension promotes endothelial dysfunction and blunts PWV in ovariectomized XX and XY mice.
Abstract P229: Neurovascular and Cognitive Dysfunction in Hypertension at the Single-Cell Level
Authors: Pacholko Anthony , Schaeffer Samantha , Santisteban Monica , Wang Gang , Racchumi Gianfranco , Faraco Giuseppe , Garcia-Bonilla Lidia , Anrather Josef , Iadecola Costantino
Published: October 2024

Hypertension (HTN) is the leading cause of vascular cognitive impairment, which has been attributed to altered microvascular regulation, neural network dysfunction and white matter (WM) damage. However, the neurovascular cells and cellular networks through which microvascular alterations lead to neuronal dysfunction and WM damage require elucidation. We employed unbiased single-cell RNA sequencing of the neocortex in 10-week-old C57BL/6 male mice treated with AngII (600ng/kg/min s.c.) or vehicle, to map out the transcriptomic changes induced by HTN. Transcriptomes (3 mice/group) were obtained for 39,451 cells before (day 3 after AngII) and after development of neurovascular and cognitive dysfunction (day 42). Profound transcriptional alterations were already seen at day 3 in venular endothelial cells (EC), GABAergic interneurons expressing neuronal nitric oxide synthase (nNOS), and oligodendrocyte precursors (OPC) (EdgeR; FDR< 0.05, logFC> 2). Gene ontology (GO) analysis (p <0.05) revealed terms consistent with senescence of venular EC, validated by immunolabeling with senescence markers (β-galactosidase, p27, p21, p16). GO also flagged terms reflecting suppression of neuronal NO synthesis and synaptic dysfunction in interneurons, which were confirmed by demonstrating reduced NMDA-evoked NO release in brain slices (20±0.03% Veh vs. 0±0.03% AngII;p<0.0001;n=60 neurons from 5 slices/group) and loss of the vesicular GABA transporter, VGAT. In addition, GO revealed early impairments in OPC differentiation, which could underlie later WM disruption. Consistent with this prediction, transcripts suggestive of impaired myelination were overrepresented at 42 days in mature oligodendrocytes (e.g.,
Hes5
), and these WM alterations were confirmed in vivo by an increase in internodal distance (Nav1.6/CASPR) in axons (0.09±0.014 µm
3
Veh vs. 0.16±0.01 µm
3
AngII;n=4/group;p<0.05). Ligand-receptor interaction analysis (CellChat) indicated molecular reprogramming of oligodendrocytes by senescent EC via suppression of signaling promoting myelination (CD39-AdeRA1) and upregulation of signaling arresting OPC development (Endothelin-ER2). These data reveal a previously unappreciated susceptibility of venular EC, OPC and nNOS interneurons to the deleterious effects of AngII HTN, and point to a novel pathogenic interaction between senescent EC and OPC, as well as to early nNOS interneuron dysfunction, the disease relevance and therapeutic potential of which requires validation.
Abstract P230: Elevated Blood Pressure Promotes Perivascular Adipocyte Progenitor Proliferation and Alters Their Spatial Distribution
Authors: Rendon Javier , Rondini Elizabeth , Garver Hannah , Lauver Adam , Fink Gregory , Krieger-Burke Teresa , Granneman Jammes , Watts Stephanie , Contreras Andres
Published: October 2024

Perivascular adipose tissue (PVAT), the outermost layer of blood vessels, regulates vascular function. Hypertension (HTN) induces vascular remodeling, a process characterized by fibrosis and cell proliferation. However, this process is poorly understood in PVAT. Adipocytes in PVAT are maintained by continuous adipogenesis of progenitor cells (APC) expressing PDGFRα. Single-cell studies in non-PVAT depots identified 3 APC subtypes that may develop into adipocytes vs fibroblasts and are differentially distributed in AT (ie, parenchyma, fascia). Changes in APC distribution may contribute to fibrosis and consequently vascular remodeling. Currently, the contribution of APC subtypes to HTN-induced PVAT fibrosis is unknown. This study determined HTN effect on APC abundance and phenotype in thoracic PVAT. We hypothesize that elevated blood pressure increases the abundance of profibrotic APC in PVAT. Male mice (PDGFRa-CreER
T2
-tdTomato n=12) were used to trace APC (PDGFRα
+
). HTN was induced using 1) aorta coarctation using a ligature (Coarct), creating a hypertensive (Upstream: Up) and hypotensive (Downstream: Down) gradient. Sham animals underwent surgery without ligature. 2) Angiotensin-II (or Saline) infusion. Blood pressure was measured by telemetry under anesthesia or tail cuff. Blood flow velocity was measured with Doppler ultrasound (mm/sec; reported as ratio between Up:Down). APC proliferation was determined by PDGFRα
+
/BrdU
+
colocalization (reported as % of total APC), and APC subtypes’ spatial distribution was evaluated with SABER-FISH.
Cxcl14
defined: APC1,
Pi16
: APC2, and
Gdf10
: APC3 (reported as % of total nuclei). Coarctation created a ~19mmHg mean arterial pressure gradient across the ligature. Coarct had a faster blood flow ratio (2.05±0.18 vs sham 1.04±0.03). APC3 abundance increased in the parenchyma of coarcted Up (2.7%±1.1) compared to Down (0%); whereas sham animals had no APC3 in the parenchyma of both sites. Similarly, coarct adventitia had more APC3 Up (18.84%± 6.8) than Down (4.5%±2.2). Ang-II increased mean arterial pressure by ~56mmHg compared to saline. APC proliferated in Ang II-treated animals (72%±2.8%) compared to control (45%±8.9%). Our results demonstrate that HTN increased APC abundance, specifically APC3. In conclusion, we identify APC3 as a possible driver of PVAT remodeling during HTN (Figure). Future studies will evaluate how APC subtypes contribute to fibrosis in vitro.
Abstract P232: Estradiol Supplementation Prevents Ovariectomy-Induced Arterial Stiffness and Vascular Fibrosis
Authors: Turner Casey , de Oliveira Karla , Breton Jade , Martin Gregory , Dupont Jennifer
Published: October 2024

Background:
Arterial stiffness is an independent predictor of cardiovascular events and mortality. Clinical data shows arterial stiffness increases rapidly after menopause, but mechanisms that mediate this change are not well known. The effect of ovariectomy (ovx) in rodent models is inconsistent in the literature. We hypothesize that the loss of estradiol (E2) contributes to ovx-induced arterial stiffening in a mouse model via fibrotic mechanisms, and that restoration of E2 will prevent these changes.

Methods:
Eight-week-old female C57/Bl6J mice received bilateral ovx or sham operation. Subcutaneous placebo or E2 (0.36 mg/day) pellets were implanted 2 weeks later to achieve three groups (sham+placebo, ovx+placebo, and ovx+E2). Aortic pulse wave velocity (PWV) was measured as an index of arterial stiffness. Blood pressure (BP) was measured by radiotelemetry. Aortic fibrosis was assessed by histological quantification of Masson’s trichrome staining and expressed as fold change in %fibrosis from sham. Pro-fibrotic genes were assessed in aortic tissue via quantitative PCR. Data presented are mean ± SEM.

Results:
Successful ovx was confirmed by reduction in uterine weight in the ovx group compared with other groups (
p
<0.0001). At 6-weeks-post-surgery (N=16-18/group), PWV was increased in ovx (5.2±0.2 mm/ms) versus sham (3.4±0.1 mm/ms,
p
<0.0001) and ovx+E2 (3.5±0.1 mm/ms,
p
<0.0001) groups. In biweekly measures (N=4-5/group), PWV was greater in the ovx group at 4- and 6-weeks-post-surgery compared with sham (
p
<0.0001) and ovx+E2 (
p
<0.0001) groups, while PWV in the ovx+E2 group did not differ from sham at any time (
p
≥0.6). At 6-weeks post-surgery (N=5/group), there were no observed differences between groups for dark or light cycle systolic or diastolic BP, mean arterial pressure, or pulse pressure. Aortic fibrosis was increased in the ovx group (1.6-fold) compared with sham (
p
=0.03) and ovx+E2 (0.9-fold,
p
=0.02) groups (N=10-12/group). Aortic fibrosis correlated with PWV at 6-weeks-post-surgery (R
2
=0.25,
p
<0.01). Aortic mRNA expression of collagen1a1 was increased in ovx versus ovx+E2 mice (
p
=0.04, N=4-6/group).

Conclusion:
In conclusion, the restoration of E2 prevents ovx-driven arterial stiffening in female mice via a reduction in fibrotic remodeling of the aorta. The results also support that ovx is an appropriate model to study the mechanisms of menopause-induced arterial stiffening and that anti-fibrotic therapeutics may be beneficial in aging women.
Abstract P300: Modulation of Osmolyte Levels by ACE Inhibitors and SGLT-2 Inhibitors in Normotensive Rats
Authors: Rybka Mateusz , Mazurek Lukasz , Szudzik Mateusz , Samborowska Emilia , Ufnal Marcin , Konop Marek
Published: October 2024

Hypertension is associated with disturbances in the water-electrolyte balance, leading to changes in osmotic pressure. Osmolytes, such as taurine among other compounds, play a critical role in maintaining cellular integrity under osmotic stress.

This study evaluated the effects of ACE inhibitors (ACE-I) and SGLT-2 inhibitors (SGLT2i) on osmolyte homeostasis. Twelve-week-old male Sprague-Dawley (SD, n = 38) rats were divided into five groups: control (water), low and high doses of canagliflozin, and low and high doses of captopril. Metabolic studies were conducted in metabolic cages. Using liquid chromatography coupled with mass spectrometry (LC-MS), we measured osmolyte concentrations in blood, urine, kidney, liver and heart tissues.

The obtained results demonstrated significant increases in liver taurine concentrations (13.34 ± 1.39 vs. 10.49 ± 1.45, p=0.029) and urinary taurine excretion (25.08 ± 2.81 vs. 16.54 ± 3.77, p=0.001) in the high-dose canagliflozin group compared to controls. Both treatments enhanced the total urinary excretion of L-alanine, β-alanine, choline, glycine, taurine, and carnitine. Additionally, canagliflozin increased the excretion of betaine and glycerophosphorylcholine (GPC). A reduction in serum β-alanine was noted in the captopril-treated rats.

These findings highlight the novel effects of SGLT-2 inhibitors and ACE-I on osmolyte levels in normotensive rats, suggesting potential mechanisms contributing to their cardioprotective effects. Further research is required to elucidate the clinical significance of osmolyte changes in enhancing the protective effects of these drug classes.
Abstract P302: Impact of Publication of Hypertension Guidelines on Inappropriate Use of Intravenous (IV) Antihypertensive Medications for Hypertensive Urgencies (HU)
Authors: Peixoto Aldo , Ghazi Lama , SIMONOV MICHAEL , Yamamoto Yu , Wilson Francis
Published: October 2024

Background:
Hypertensive urgencies are common in hospitalized patients. In late 2017 and 2018, the publication of major U.S and European hypertension guidelines included specific recommendations against the use of IV antihypertensive medications for the management of severe acute BP elevations in the absence of acute target organ damage (HU).

Aim:
To study the impact of the publication of guidelines on local utilization of IV medications for HU.

Methods and Results:
We analyzed 224,265 non-ICU, non-obstetric, non-pediatric hospital admissions to 5 U.S. hospitals (total 2,681 beds) in a medium-sized U.S. healthcare system between January 2016 and March 2020. These included one large academic medical center with two campuses (Hosp A), one medium sized teaching community hospital (Hosp B), one small teaching community hospital (Hosp C), and two small non-teaching hospitals (Hosp D and E). Among all admitted patients, 22,929 (10.2%) developed HU (SBP>180 or DBP >110 mmHg sustained for at least 1 hour without evidence of acute target organ damage). A total of 2,914 HU patients (12.7%) received at least 1 intravenous medication within 6 hours of acute BP elevation, most commonly hydralazine (N 1473, 50.5%], labetalol (N 749, 25.7%) or metoprolol (N 632, 21.7%). The fraction of patients receiving IV antihypertensives decreased during the five years of the study across the five hospitals, but the absolute change was small (2016=13.7%, 2017=12.8%, 2018=12.5%, 2019=11.4%, 2020=11.7%, P value for overall change = 0.003). The Figure displays temporal trends for each hospital. There was significant heterogeneity across hospitals (P<0.0001). Trends were not related to teaching (vs. nonteaching) status of each hospital.

Conclusion:
In a medium sized U.S. hospital system, the publication of guidelines recommending against IV antihypertensive use in HU was temporally associated with very modest reductions in IV medication use in HU. Process improvement initiatives are needed to optimize guideline-congruent practice.
Abstract P305: Resolution of Resistant Hypertension with Unilateral Adrenalectomy in Bilateral Primary Aldosteronism: A Case Report from Jordan
Authors: Al Kayed Hadeel , qussous jwana , Hajjaj Nada , Alwahedy Saba , Maaita Mahmoud , Alsuradi Awni , Tabbalat Ramzi
Published: October 2024

Introduction:
Primary aldosteronism (PA) is a common cause of resistant hypertension. The standard of care for bilateral hyperaldosteronism is medical management, while surgery is often preferred for unilateral cases.

Case Presentation:
We report the case of a 53-year-old female with an 18-year history of hypertension. She presented with uncontrolled blood pressure despite adherence to seven antihypertensive medications.

Blood tests, including creatinine, urea, and electrolytes, were within normal limits, as was the renal ultrasound. Serum aldosterone was 136.3 pg/mL, and plasma renin activity was 0.23 ng/mL/hr, with a ratio of 593. Her antihypertensive medications were changed to hydralazine, moxonidine, and verapamil.

A Seated Saline Suppression Test performed a month later, with 2 liters of saline infused over 4 hours, showed that serum aldosterone levels failed to suppress, confirming the diagnosis of PA.

Three-Phase adrenal CT revealed a well-defined adenoma in the left adrenal gland measuring 4.5 x 3.4 cm, and a small adenoma in the right adrenal gland measuring 1.2 x 1 cm. Adrenal Vein Sampling indicated excess aldosterone secretion from both adrenal glands, confirming bilaterality.Therefore, high-dose spironolactone was added to her antihypertensive regimen; however, her BP remained elevated after 2 months of therapy.

The option of surgically removing the adrenal gland with the larger adenoma in hopes of reducing the total burden of aldosterone was discussed with the patient as the only remaining option of improving her BP control. The patient agreed and a laparoscopic left adrenalectomy was performed. Histopathological examination ruled out malignancy.

Immediate preoperative BP was 151/90 mmHg. The surgery was uneventful. All antihypertensive medications were withheld postoperatively. On the third postoperative day her BP was consistently below 130 mmHg systolic and below 85 mmHg diastolic. Her BP remained well controlled with gradual resumption of bisoprolol 5 mg/day and spironolactone 50 mg/day

Conclusion:
Despite the bilaterality of primary aldosteronism in this case, the patient's hypertension did not respond to medical treatment, necessitating surgical intervention. The unilateral adrenalectomy of the larger adenoma proved sufficient to significantly improve her persistently elevated blood pressure. This case underscores the potential efficacy of targeted surgical treatment in managing resistant hypertension associated with bilateral adrenal disease.
Abstract P307: A Review Of The Burden, Management And Outcomes Of Patients With Aldosterone Dysregulation
Authors: Luan Shan , Agiro Abiy , Daniel Ian , McKendrick Jan , Davis Harrison , Huang Joanna , linganathan Karthik
Published: October 2024

Introduction:
Aldosterone dysregulation (excess aldosterone production) is a complex physiological driver of uncontrolled hypertension as well as cardiovascular (CV) and kidney outcomes. Understanding the clinical and economic burden of excess aldosterone in routine practice is critical to optimizing treatment and improving long-term outcomes. This review examines the burden of excess aldosterone production as a driver of disease and its impact on clinical and economic outcomes.

Hypothesis:
Excess aldosterone production has a broad impact on long-term outcomes, and left untreated, contributes to worsened patient outcomes.

Methods:
A targeted literature review was conducted to identify articles on the burden, management and outcomes of patients with excess aldosterone production (2013 – 2024) with 27 manuscripts included.

Results:
More studies document the impact of excess aldosterone on organ function in the cardiovascular system and kidney (10, 8 manuscripts) compared to liver (1 manuscript). Excess aldosterone levels and aldosterone-induced oxidative stress and inflammation are associated with increased risk of CV events including myocardial infarction, heart failure, stroke, and increased morbidity and mortality. For example, Framingham Offspring Study showed increased CV disease risk with each rise in serum aldosterone levels (HR, 1.11; 95% CI, 1.02–1.21). Kidney, hepatic and metabolic manifestations are also well-documented, with implications for progression to chronic kidney disease, liver disease and metabolic syndrome. Another study reported an 11% increased risk of CKD progression associated with each doubling of serum aldosterone. Current pharmacological interventions for hypertension, including mineralocorticoid receptor antagonists, were not reported to restore normal aldosterone levels. No literature was found on the economic burden of excess aldosterone or the association between excess aldosterone production and clinical outcomes independent of blood pressure control.

Conclusions:
Currently available literature emphasizes the broad impact of excess aldosterone production on CV, kidney, hepatic, and metabolic outcomes. The impact on outcomes indicates that excess aldosterone production is underappreciated, with prognostic implications beyond primary aldosteronism. Current hypertension treatments do not address excess aldosterone production. These findings underscore the unmet need for novel management strategies that target excess aldosterone production.
Abstract P313: Diastolic to Systolic Arterial stiffening has Greater Prognostic Power than Diastolic Stiffness in 20-60 Years Old Patients Undergoing 24-Hour Ambulatory Blood Pressure Monitoring
Authors: Gavish Benjamin , Bursztyn Michael , Ruilope Luis , Williams Bryan , DeLa Sierra Alejandro
Published: October 2024

The 24-hour ambulatory pulse pressure (PP), can be expressed as a sum of two ‘PP components’: ‘elastic’ PP (elPP) and ‘stiffening’ PP (stPP) associated, respectively, with arterial stiffness at the diastolic pressure (somewhat equivalent to pulse wave velocity being measured in the diastole), and the relative stiffness change from the diastolic- to the systolic pressure. Here we assess and compare the hazard ratio (HR) of elPP and stPP for outcomes in a large cohort of young and middle-age patients.

We analyzed adequate quality 24h ambulatory blood pressure monitoring records from the Spanish Ambulatory Blood Pressure Registry. For standard deviation(SD), K=SD(SBP)/SD(DBP), elPP=PPxln(K)/(K-1) and stPP=PP-elPP. HRs were determined for age 20-40, 41-50, and 51-60 years, using Cox regression models applied to standardized elPP & stPP (combined), adjusted for age, sex, BMI, smoking status, diabetes, dyslipidemia, mean DBP, pulse rate, SD(SBP), antihypertensive treatment, and previous cardiovascular disease (CVD). The endpoints were total mortality, and death from CVD, or coronary heart disease (CHD).

Of the 24,260 patients mean age (SD) 48.0(9.0) years, 34% male] 845 deaths occurred during period of 9.6(2.2) years. The Table shows that mean elPP slightly decreased from younger to older age, while stPP strongly increased. At age 41-60 years the adjusted HRs of stPP appeared significant and greater than those of elPP for almost all cases. The most contributing adjustors in the Cox regression were smoking, for all age classes, and sex for age over 40 years.

Thus, the stiffening component may have greater predictive power than the elastic component for total mortality, and CVD&CHD death, in middle-age patients.
Abstract P315: Using Home Blood Pressure Monitors in the Office Setting as an Alternative to Automated Office Blood Pressure: A Quality Improvement Project
Authors: Gallagher Benjamin , Huot Stephen
Published: October 2024

Background:
Accurate blood pressure (BP) measurement is key to the diagnosis and effective treatment of hypertension (HTN). The U.S. HTN guideline recommends having patients rest for 5 minutes before measuring BP and obtaining at least two measurements 1-2 minutes apart. This is difficult to achieve in a busy primary care practice, and while automated office BP (AOBP) has been advanced as a solution, the required device may be cost-prohibitive. As home BP monitors are much less expensive, a protocol employing them in the office setting could prove to be an attractive alternative to AOBP.

Objective:
To promote the recording of multiple BP readings in primary care visits by using home BP monitors for measurement.

Methods:
Our primary care practice hosts 110 internal medicine residents and completes more than 35,000 patient visits annually. Prior to the intervention, medical assistants (MAs) obtained vital signs (VS) at every visit using a VS cart. In the first version of the new protocol, MAs measured all VS except BP using the VS cart, then measured BP once using an Omron 5 series BP monitor and recorded that reading in the chart. MAs left the cuff on the patient’s arm and instructed him/her to rest quietly for 5 minutes before taking a second BP reading him/herself while waiting for the provider after the MA exited the room. The patient took a third reading 1-2 minutes after the second. The provider recorded the second and third readings in the chart.

Results:
We implemented the new protocol in November 2023. In February 2024, we met with MAs to solicit feedback. They found the protocol easy to follow overall but had two concerns: (1) It was cumbersome to use two different devices to obtain a full set of VS; and (2) since many patients had normal BP, it did not seem worthwhile to obtain multiple BP measurements for all patients. After this meeting, we began measuring heart rate and oxygen saturation with a portable pulse oximeter instead of the VS cart, which is now only used in special circumstances. We also modified the protocol so that patients with systolic BP < 120 mm Hg and diastolic BP < 80 mm Hg are not required to obtain any repeat BP measurements.

Conclusions:
A new BP measurement protocol using home BP monitors to obtain multiple readings at each visit was feasible in our large academic primary care practice. We plan to investigate its effect on frequency of repeat BP measurement and likelihood of intensifying therapy for uncontrolled HTN.
Abstract P318: English and Spanish-Speaking Patient Preferences on Home Blood Pressure Monitors: A Qualitative Study in an Urban Safety Net Setting
Authors: Kwok Vivian , Shih Jonathan , Luna Isabel , Kim Hyunjin , Garcia Faviola , Gutierrez Christian , Miles Mahal , Lyles Courtney , Khoong Elaine
Published: October 2024

Background:
Self-measured blood pressure (SMBP) monitoring improves hypertension (HTN) control. Adoption of SMBP monitoring is lower among low-income or minoritized patients, who have higher rates of uncontrolled HTN. Little is known about these patients’ preferences for or challenges with SMBP monitoring.

Objective:
We explored patient perspectives on SMBP monitoring among patients receiving care at an urban safety-net system.

Methods:
We interviewed English and Spanish-speaking patients with HTN about preferences for different BP monitoring devices and their accompanying mobile apps, past experiences sharing BP data with their clinical care team, and knowledge about home BP monitoring. Interviews were conducted in the participants’ preferred language. We also asked participants to demonstrate a home BP measurement to evaluate their measurement technique. We presented two BP monitoring devices: a Bluetooth-enabled device and a cellular-enabled device. Rapid qualitative data analysis was used to analyze data.

Results:
Of the 15 participants (8 English-speaking; 7 Spanish-speaking), eight identified as Latine, four as Black or African American, one as American Indian/Native American, one as Asian/Pacific Islander, and one as multi-ethnic. Highest educational attainment varied: five less than high school, five high school/GED, and five college. Most required assistance with Bluetooth pairing to share data and navigating app features. All participants valued tracking their BP and were motivated to engage in SMBP monitoring but desired more training. Nearly all participants demonstrated inadequate BP knowledge and incorrect BP measurement techniques based on AHA/AMA recommendations. Few reported receiving training on SMBP monitoring techniques. Spanish speakers reported that using apps was challenging due to language discordance and wanted translated apps and resources. Costs related to device features were a key factor in preferences.

Conclusions:
Patient-reported barriers to SMBP adoption include cost, insufficient training, digital exclusion, and language discordance. Addressing these challenges may increase adoption of SMBP monitoring in safety-net populations. Clinicians should consider patients’ preferences when recommending SMBP monitoring. Cellular SMBP devices that automatically transmit BP readings may mitigate challenges with digital literacy and promote data sharing with clinical care teams, though future studies are needed to evaluate usability.