bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P148: Transcriptomic and Proteomic Analysis of CD4
+
T Cells to Identify Sex Differences in Angiotensin II Signaling Pathways
Authors: Uhlorn Joshua A , Husband Nathaniel A , Romero-Aleshire Melissa J , Moffett Caitlin K , Kelly Amy C , Langlais Paul R , Nikolich-Zugich Janko , Brooks Heddwen L
Published: September 2019

T cells are involved in hypertension pathogenesis in both males and postmenopausal females while premenopausal females are resistant to T cell-mediated Ang II-induced hypertension. The goals of this study were (1) to identify T cell specific proteomic pathways associated with postmenopausal susceptibility to hypertension (2) to identify T cell specific transcriptomic changes associated with premenopausal protection from hypertension. Proteomic analysis was performed on splenic CD4
+
T cells isolated from premenopausal and postmenopausal females (VCD, 160 mg/kg/day i.p. 20d) following Ang II infusion (800 ng/kg/min 14d). 384 proteins from CD4
+
T cells were identified as differentially expressed following Ang II infusion in premenopausal females. 285 proteins from CD4
+
T cells were identified as differentially expressed between premenopausal and postmenopausal females following Ang II infusion. Gene ontology (GO) analysis of pre vs. postmenopausal proteins identified enriched pathways associated with RNA binding, chaperone activity and cellular stress responses. Transcriptomic changes were analyzed, via RNAseq, on isolated splenic CD4
+
T cells from premenopausal females, with and without Ang II infusion. Thirty-four genes were identified as differentially expressed in CD4
+
T cells following Ang II infusion. GO analysis of Ang II upregulated genes revealed an enrichment of five distinct molecular functions, including antioxidant activity. In a subsequent study to validate the RNAseq, we confirmed that Ang II increased CD4
+
T cell mRNA expression of calprotectin (S100a8/S100a9), a calcium and zinc binding protein complex that contributes to antioxidant defense (S100a8: Con 1.0 ± 0.4 vs Ang II 5.0 ± 0.8*; S100a9: Con 1.0 ± 0.4 vs Ang II 6.0 ± 0.8*, *P<0.05 vs Con). Furthermore, we determined that Ang II did not increase calprotectin expression in mice lacking estrogen receptor α (ERKO) (S100a8: ERKO 0.1 ± 0.4 vs ERKO/Ang II 0.3 ± 0.7; S100a9: ERKO 0.2 ± 0.5 vs ERKO/Ang II 0.3 ± 0.6). The current studies demonstrate a role for estrogen in Ang II-induced T cell gene expression and signal transduction, and begin to elucidate the molecular mechanisms of female protection from T cell-mediated hypertension.
Abstract P150: Genes of a Brown Norway Chromosome 2 Fragment Introgressed Into Hypertensive Dahl Salt-Sensitive Background Exert Pro-Inflammatory Effects When Stimulated by a High-Salt Diet
Authors: Berillo Olga , Ouerd Sofiane , Huo Ku-Geng , Rehman Asia , Richer Chantal , Sinnett Daniel , Kwitek Anne E , Paradis Pierre , Schiffrin Ernesto L
Published: September 2019

Background:
Chromosome 2 (Chr2) introgression from normotensive Brown Norway (BN) rats into hypertensive Dahl salt-sensitive (SS) background (consomic S
B
2) reduced blood pressure (BP) and vascular inflammation under normal-salt diet (NSD). We hypothesized that BN Chr2 contains anti-inflammatory genes that could reduce BP elevation and vascular inflammation in rats fed NSD and high-salt diet (HSD).

Method:
Four- to 6-week old male SS and congenic rats containing the BN Chr2 distal portion (S
B
2a) and middle segment (S
B
2b) were fed NSD or HSD (4% NaCl) for 8 weeks. We determined systolic BP (SBP) by telemetry, reactive oxygen species (ROS) generation using dihydroethidium staining, and vascular cell adhesion molecule 1 (VCAM-1), monocyte chemoattractant protein-1 (MCP-1) expression and monocyte/macrophage (MoMϕ) infiltration by immunofluorescence in aorta or perivascular fat (PVAT). RNA was extracted from aorta and used for small and total RNA sequencing and data were analyzed using a systems biology approach. Differentially expressed genes (DEGs) were identified with fold change >1.3 and fold discovery rate <0.05 and some of them were validated with RT-qPCR.

Results:
SS SBP was 145±2 mm Hg under NSD and 168±1 mm Hg under HSD, which was lower in S
B
2a and S
B
2b SBP (125±3 and 127±6,
P
<0.05) under NSD but similar under HSD. Examination of ROS generation, VCAM-1 and MCP-1 expression and MoMϕ infiltration revealed that S
B
2a present less and more inflammation under NSD and HSD, respectively, compared to SS. DEGs were identified in S
B
2a vs SS uniquely under NSD (15↑ and 8↓) and HSD (318↑ and 221↓) and under both diet (3↑ and 3↓), and in S
B
2b vs SS uniquely under NSD (43↑ and 65↓) and HSD (4↑ and 7 ↓) and under both diet (3↑ and 4 ↓). DEGs encoded within BN Chr2a were uniquely identified under NSD (1↑ and 1↓) and HSD (6↑ and 1↓), and in both diets (1↑ and 1↓). Gene enrichment analysis revealed that under NSD, 2 BN Chr2a DEGs are involved in regulation of BP, and under HSD, 7 BN Chr2a DEGs in cell proliferation, cell differentiation, signal transduction and the immune system. RT-qPCR validated DEGs encoded within BN Chr2a under NSD (2) and HSD (4).

Conclusions:
DEGs encoded within BN Chr2a fragment are associated with anti- and pro-inflammatory effects under NSD and HSD, respectively.
Abstract P152: Forkhead Box Subclass O Transcription Factors Differentially Regulate Soluble Guanylyl Cyclase Expression in Smooth Muscle
Authors: Galley Joseph , Sanker Subramaniam , Miller Megan , Straub Adam
Published: September 2019

Nitric oxide (NO) stimulates soluble guanylyl cyclase (sGC) activity leading to elevated cyclic guanosine 3′, 5′-monophosphate (cGMP) and vascular smooth muscle relaxation. Downregulation of sGC expression attenuates vascular dilation and contributes to the pathogenesis of cardiovascular disease, however, it is not well understood how sGC transcription is regulated. We recently published evidence that inhibition of the Forkhead box subclass O (FoxO) transcription factors resulted in reduced sGC mRNA, protein expression, and cGMP production in cultured smooth muscle. Furthermore, we showed that FoxO inhibitor treatment in
ex-vivo
aortas decreased sGC β protein expression and impaired NO-dependent vasorelaxation. We now seek to determine which FoxO transcription factor, FoxO1, FoxO3a and/or FoxO4 in vascular smooth muscle cells regulate(s) sGC transcription. In this study, cultured rat aortic smooth muscle cells (RASMC) were treated with adenoviral shRNA constructs targeting FoxO1, FoxO3a, or FoxO4. Western blot analyses measured protein expression and phosphorylation, while RT-qPCR was quantified mRNA expression. Each FoxO shRNA construct produced a variable effects on smooth muscle. FoxO1 shRNA treatment significantly increased both sGC α and β mRNA expression by 3-fold (n=3) and showed a 22% increase in sGC β protein expression (n=3), while FoxO3a shRNA increased both sGC α and β mRNA expression by 2-fold (n=3) and increased sGC β protein expression by 48% (n=3). FoxO4 shRNA treatment, however, decreased sGC α and β mRNA by 50% and 59%, respectively, and also decreased sGC β protein expression by 49% (n=3). FoxO4 shRNA treatment also phosphorylated 38% less vasodilator stimulated protein (pVASP) at its cGMP-dependent phosphorylation site on serine 239 after stimulation with 500 nM treatment with the nitric oxide donor molecule DEA-NONOate (n=4) and 30% less after 1 μM stimulation with DEA-NONOate (n=4) than controls. Taken together, these data suggest a complex FoxO-dependent mechanism wherein FoxO1/3a act antagonistically to FoxO4 regulate sGC gene expression. Future studies will include luciferase promoter-reporter assays and chromatin immunoprecipitation assays to identify integral FoxO binding sites on the sGC promoter.
Abstract P166: Paracrine Endothelial Microrna214 Exosomes Modulate Smooth Muscle Cell Function in Pulmonary Arterial Hypertension
Authors: Sahoo Sanghamitra , Pagano Patrick
Published: September 2019

Background:
Communication among various cell types is known to play a pivotal role in PAH. Endothelial cell (EC)-derived exosomes have emerged as important mediators of intercellular communication and are known to influence the function of recipient cells. A role for miR-214 in EC-derived exosomes in PAH, however, is entirely unknown. In this study, we hypothesize that hypoxia-induced EC injury releases exosomes enriched in miR-214 that mediates phenotype switching of smooth muscle cells (SMCs) resulting in increased SMC proliferation in PAH.

Methods and Results:
We subjected human pulmonary artery endothelial cells (hPAECs) to hypoxia vs. normoxia (norm) to recapitulate the PAH phenotype
in vitro
. Quantitative RT-PCR of hPAECs showed that hypoxia upregulated miR-214 by 1.51±0.24-fold (p<0.05 vs. norm). Moreover, exosomes derived from hypoxic hPAECs were found to be enriched in miR-214 (1.73±0.16-fold vs. norm, p<0.05). TEM and Nanotracking Analysis System determined the size of EC-derived exosomes to be within a characteristic 30-100nm. Furthermore, Z-stack confocal imaging demonstrated that FITC-PHK67-labeled EC-derived exosomes were taken up by hPASMCs. Western blot of hPASMCs treated with hypoxia-induced EC-derived exosomes showed upregulation of phospho-Akt (1.65±0.29-fold vs. norm,
p
<0.05), phospho-ERK1/2 (1.68±0.27-fold vs. norm,
p
<0.05), and Bcl2 (1.63±0.19-fold vs. norm, NS). Similar treatment also decreased SMC-specific genes, such as MYH11, calponin and smoothelin, indicating that EC-derived hypoxic exosomes can induce switching of PASMCs to a synthetic, pro-proliferative phenotype. Moreover, scratch assay demonstrated that application of EC-derived exosomes laden with exogenous miR-214 increased SMC migration (1.28±0.08-fold vs. control,
p
=0.05).

Conclusions & Discussion:
We found that miR-214 expression was induced in hypoxia both in ECs and in exosomes released from them. Furthermore, EC-derived hypoxic or miR-214-laden exosomes were able to transduce changes in SMC, which result in a proliferative and synthetic phenotype. These findings demonstrate that EC-derived miR-214 can direct a proliferative fate for smooth muscle cells, and may contribute to SMC-associated pathophysiological changes in PAH.
Abstract P185: The Effect of Anti-Interleukin-17A Biologic Therapies on Cardiovascular Outcomes in Patients With Psoriasis
Authors: Smart Charles D , Shuey Megan , Elijovich Fernando , Cox Nancy , Madhur Meena S
Published: September 2019

We and others have demonstrated that interleukin-17A (IL-17A) plays a critical role in the pathogenesis of cardiovascular disease and hypertension. Patients with psoriasis, an IL-17A mediated autoimmune disease, are at elevated risk for cardiovascular events. New biologic therapies for psoriasis have been developed that target IL-17A (secukinumab) or its receptor (brodalumab), but their impact on incident hypertension and its associated complications is unknown and remain unexplored outside of short-term safety in clinical trials. Our goal is to perform a retrospective case-control study utilizing the Synthetic Derivative (Vanderbilt’s de-identified version of the electronic health record) to determine whether incident hypertension or other major adverse cardiovascular events is reduced in patients with psoriasis receiving IL-17A targeted therapies versus other therapeutics. Patients are defined as cases by 2 or more mentions of a psoriasis ICD9 or ICD10 code and at least one documented prescription for secukinumab or brodalumab. Controls are defined by the presence of 2 or more psoriasis ICD codes, no documented exposure to the drugs of interest, and a mention of “psoriasis” in their Problem List. Using these definitions, we identified 571 cases and 4484 controls in the Synthetic Derivative. Of the 571 potential cases identified, 65 were confirmed by manual review for an initial validation study. Of the manually confirmed cases, 22 (34%) were male and 43 (66%) were female. Manually reviewed cases had a mean record length of 5.0 years with an average of 29.9 discrete ICD mentions. Twenty-three cases (35%) had an ICD code for hypertension, which predated the first documented psoriasis code in eight patients. Thus, we are able to use the Synthetic Derivative to identify patients with psoriasis receiving biologic therapies and assess temporal relationships relevant to hypertension. We plan to expand this study to the larger cohort identified and analyze rates of hypertension and a composite outcome of hypertension complications, including stroke, myocardial infarction, heart failure, and chronic kidney disease, in cases and controls matched for age, gender, race, and BMI.
Abstract P193: A Novel Mechanism for the Pathogenesis of Preeclampsia: Cell-Free Mitochondrial DNA Mediated Toll Like Receptor 9 Activation
Authors: Rappelt Matthew , Beyer Andreas , Gutterman David , Norwood Toro Laura , Derayunan Alexa , McIntosh Jennifer
Published: September 2019

Background:
Preeclampsia (PreE) is a significant global health burden which impacts 10 million pregnancies annually resulting in over a half million lost lives. Mitochondrial damage has previously been demonstrated to be present in PreE. A novel relationship may exist between preeclampsia and mitochondrial damage via the release of cell-free mitochondrial DNA (cf-mtDNA). Previous work in models of systemic hypertension shows cf-mtDNA mediated activation of TLR9 to be a key part of disease development. We aim to evaluate the level of TLR9 activation and ROS production in patients with and without PreE.

Methods:
To investigate whether PreE plasma contains increased levels of cell free DNA that can activate TLR9, HEK-Blue
TM
-hTLR9 reporter cell line (Invivogen) was treated with 3% plasma from either control or PreE patients. The TLR9 activator ODN2006 served as positive control (Fig.1, A&B). To investigate whether PreE plasma can stimulate production of ROS, HUVECs were treated in similar fashion and ROS was evaluated using Mitosox
TM
(Fig. 1, C&D).

Results:
Cells treated with PreE plasma demonstrated a 1.7-fold increase in TLR9 activation vs control (p=0.005, Fig. 1A&B). In addition, Mitosox
TM
signal indicated a 1.8-fold increase in ROS in PreE vs. control (p=0.066, Fig. 1C&D).

Conclusion:
We conclude that plasma from PreE patients contains higher levels of cf-mtDNA that stimulates inflammation response via TLR9 that promotes mitochondrial ROS production. This mechanism may present a novel target for treatment of Pre-E.
Abstract P198: High Glucose Degrades (PRO) Renin Receptor by Autophagy and Ubiquitin Proteasome System in Inner Medullary Collecting Duct Cells
Authors: Luo Tao , Siragy Helmy
Published: September 2019

Previously we reported high glucose (HG) regulation of PRR expression in the kidney. However, in contrast to the observed increase in PRR expression in mesangial cells and podocytes, its expression decreased in distal tubules in response to HG. We hypothesize that HG degrades PRR through autophagy and ubiquitin proteasome system in inner medullary collecting duct cells. Mouse inner medullary collecting duct cells (mIMCDs) were cultured for 3 days in normal glucose (NG, 5 mmol/L) or HG (25 mmol/L). The mRNA and protein expressions of PRR were evaluated in response to HG alone and in presence of the proteasome inhibitor, MG-132 (10 mM) during the HG exposure. In addition, we monitored protein expressions of PRR and the autophagy activity markers, LC3-II and p62, in response to the autophagy inhibitor, bafilomycin A1 (160 nM). HG significantly increased PRR mRNA level after HG treatment (28% vs NG, P<0.05). In contrast, PRR protein expression was significantly decreased (31% vs NG, P<0.05). Pretreatment of MG-132 for 4 hours increased PRR expression after 3 days HG exposure (66% vs HG, P<0.05). Similarly, HG decreased LC3-II and p62 at 3 days of HG exposure (66% and 50% respectively, P<0.05). Bafilomycin A1 pretreatment for 2 hours attenuated the HG induced reduction in PRR protein and increased the expression of LC3-II and p62 (250% and 71% respectively, P<0.05). We conclude that HG degrades mIMCDs PRR protein via increased autophagy and ubiquitin-proteasome activity.
Abstract P2003: Loss of Trpa1 in Mice Leads to Age-Related Dilated Cardiomyopathy
Authors: Ma Shuangtao , Qian Chunqi , Wang Donna H
Published: September 2019

Activation of the transient receptor potential ankyrin 1 (TRPA1) channel, a mechanical and oxidative stress sensor, has been shown to slow down aging process and promote longevity in invertebrate animal models like
Caenorhabditis elegans
. Although physiologic sensory functions of TRPA1 are conservative across species, the role of TRPA1 in regulation of organ development and function in mammals in the aging process is unknown. The present study tests the hypothesis that TRPA1 ablation leads to cardiac dilation and cardiomyopathy in aging mice. Echocardiography was performed in male 12-week-old (young) and 52-week-old (older) Trpa1 gene knockout (
Trpa1
-/-
) mice and wild-type (WT) littermates on a mixed C57BL/6 x 129 genetic background. Echocardiogram data showed that, compared to young WT littermates, young
Trpa1
-/-
mice had similar cardiac dimensions and left ventricular function, as demonstrated by left ventricular end-diastolic internal dimension (LVIDd, WT 4.29±0.62 vs.
Trpa1
-/-
4.50±0.41 mm, P>0.05), left ventricular ejection fraction (LVEF, a parameter of systolic function, WT 66.73±9.50 vs.
Trpa1
-/-
63.39±12.76 %, P>0.05), and E/A ratio (a parameter of diastolic function, WT 1.23±0.03 vs.
Trpa1
-/-
1.39±0.14, P>0.05). In contrast, older
Trpa1
-/-
mice developed dilated cardiomyopathy with dilated left ventricle (LVIDd: WT 4.60±0.16 vs.
Trpa1
-/-
5.00±0.37 mm, P<0.05) and impaired left ventricular systolic function (LVEF: WT 58.04±9.27 vs.
Trpa1
-/-
41.79±7.19 %, P<0.05) when compared to older WT mice. In addition, older
Trpa1
-/-
mice had impaired left ventricular myocardial performance index (LV MPI, a parameter of ventricular systolic and diastolic function, WT 0.62±0.15 vs.
Trpa1
-/-
0.89±0.18, P<0.05) albeit left ventricular diastolic function was not different between old
Trpa1
-/-
and WT mice (E/A ratio: WT 1.53±0.22 vs.
Trpa1
-/-
1.99±0.92, P>0.05). These results show that genetic knockout of TRPA1 leads to age-related left ventricular dilation and systolic dysfunction. These findings suggest that TRPA1 activation protects against age-related dilated cardiomyopathy.
Abstract P2008: The Efficacy of Spironolactone in the Management of Uncontrolled Hypertension in Patients With Left Ventricular Hypertrophy and Diastolic Dysfunction in a Community Practice
Authors: Saxena Naveen R , Srivastava Vinita , Saxena Anju
Published: September 2019

Uncontrolled hypertension with left ventricular hypertrophy and diastolic dysfunction causes major cardiovascular mortality and morbidity. There are few studies in medical literature examining the use of spironolactone in this high-risk population. The aim of this study is to evaluate the efficacy of spironolactone in this high-risk population in a community practice. The current study is a retrospective analysis of chart review of 92 patients with uncontrolled hypertension with left ventricular hypertrophy and diastolic dysfunction. All patients underwent 2-D echocardiography. Left ventricular hypertrophy and diastolic dysfunction were diagnosed using standard criteria from American Society of Echocardiography. Within the study population, 65% were already taking 3 antihypertensive drugs before spironolactone was added to the regimen. Blood pressure was recorded in the office after patient rested for 5 minutes in a quiet room. The patient follow-ups occurred every 12 to 16 weeks. Spironolactone was added as an add-on therapy with a dosage ranging from 25 to 50 mg daily. The goal blood pressure was defined as systolic pressure less than 135 mm Hg and diastolic pressure less than 85 mm Hg. Total patients with uncontrolled hypertension were 92 and consisted of 52 females and 40 males. The racial distribution of the population included 50 African Americans, 39 Caucasians, and 3 Asians. Patients with diabetes were 53% of the study population. A total of 62 patients out of the 92 (67%) patients reached the goal blood pressure. There were 32 out of 50 (64%) African Americans who reached the goal and 30 out of 39 (77%) Caucasians reached the goal. Hyperkalemia was seen in 7.6% of the population and gynecomastia was not seen.Given the data, we conclude that spironolactone therapy was effective in the management of uncontrolled hypertension with left ventricular hypertrophy and diastolic dysfunction in both Caucasian and African American populations in this study. Using a hypothesis test to compare the difference in proportion of two samples, we show statistical significance at a greater than 99% confidence level that the proportion of Caucasian patients who reached the goal was greater than the proportion of African American patients who reached the goal.
Abstract P2009: Dysbiotic 1-Carbon Metabolism in Hypertensive Cardiac Remodeling
Authors: Tyagi Suresh C. , George Akash K , Singh Mahavir , Pushpakumar Sathnur B , Homme Rubens P , Metreveli Naira
Published: September 2019

Unless there is a genetic defect, most diseases start in our gut. Balance in gut’s microbiome; symbiosis vs dysbiosis need to be maintained during health, however; in diseases such metabolic syndrome, bowl diseases, and hypertension, dysbiosis prevails, but mechanisms are unclear. Our research focuses on homocysteine (Hcy) that occupies a center-stage (Figure). For example, dysbiosis methylate promoters in turn inhibiting genes. Interestingly; product of 1-carbon metabolism is Hcy, unequivocally. Recently, studies on host resistance to antibiotics have shown to be related with inverton-promoter inhibition, presumably because of methylation, thus causing modification promoters by bacterial antigens. These changes force host to lose its drug sensitivity. In this regard, we focus on the role of high methionine diet (HMD, rich in red meat) and measure the effects of a probiotic on cardiac muscle. We employed wild type and hypertensive cystathionine beta synthase (CBS) heterozygote KO mice with and without HMD and with and without a probiotic (PB, Lactobacillus) treatment in drinking water for 16 weeks. Results suggest that matrix metalloproteinase-2 activity was robust in CBS and HMD which was attenuated by PB intervention. Cardiomyocyte contractility and ECHO data suggest mitigation of cardiac dysfunction in CBS and HMD mice with PB. Also, gut microbiota reconstitution resulted in decreased interstitial fibrosis in probiotic mice hearts. Expression of key regenerative factors: DAAM1, DAAM2, REDD1 and BHMT were also improved by PB. In conclusion, our data suggest that probiotic mitigates dysbiotic 1-carbon metabolism induced hypertensive cardiac remodeling.
Abstract P2010: Silencing Epidermal Growth Factor Receptor Reduces P44/42 Mitogen-Activated Protein Kinase Signaling in the Hypothalamic Paraventricular Nucleus and Ameliorates Sympathetic Excitation in Rats With Heart Failure After Myocardial Infarction
Authors: Yu Yang , Wei Shun-Guang , Weiss Robert M , Felder Robert B
Published: September 2019

The p44/42 mitogen-activated protein kinase (MAPK) signaling pathway is activated in the brain in myocardial infarction (MI)-induced heart failure (HF). Activation of brain p44/42 MAPK signaling can induce endoplasmic reticulum (ER) stress, increase proinflammatory cytokine expression and upregulate renin-angiotensin system (RAS) activity in the brain, contributing to sympathetic excitation. Multiple excitatory agonists (e.g., angiotensin II, aldosterone and pro-inflammatory cytokines) activate brain MAPK signaling in HF, but the mechanisms are poorly understood. We tested the involvement of the epidermal growth factor receptor (EGFR), which has recently been shown to mediate MAPK signaling in peripheral cardiovascular tissues. Male rats received bilateral microinjections of a lentiviral vector encoding a small interfering RNA (siRNA) for EGFR, or a scrambled siRNA, into the hypothalamic paraventricular nucleus (PVN), a critical cardiovascular and autonomic center. One week later, some rats were euthanized to confirm the effectiveness of EGFR knockdown in PVN. mRNA for EGFR was significantly (*P<0.05) lower in rats that received the EGFR siRNA (0.54±0.17* vs 1.10±0.20, fold change). Other rats underwent coronary artery ligation (CL) to induce HF. Four weeks later, HF rats treated with siRNA for EGFR, compared with HF rats treated with scrambled siRNA, had lower protein levels of phosphorylated p44/42 MAPK (0.20±0.08* vs 0.34±0.06), lower mRNA levels of the ER stress markers BIP (0.77±0.10* vs 1.00±0.12) and ATF4 (0.71±0.14* vs 1.03±0.25), the inflammatory mediators TNF-α (0.73±0.11* vs 1.01±0.15), IL-1β (0.65±0.14* vs 1.02±0.21) and COX-2 (0.71±0.16* vs 1.00±0.14), and the RAS components ACE (0.68±0.26* vs 1.06±0.37) and AT
1a
R (0.70±0.11* vs 1.03±0.28) in the PVN, lower urinary norepinephrine levels (28±9* vs 39±7 ng/ml), and improved peripheral manifestations of HF as indicated by reduced right ventricular/body weight and lung/body weight and left ventricular end-diastolic pressure. These results suggest that EGFR in the PVN activates p44/42 MAPK, as well as RAS, neuroinflammation and ER stress, in MI-induced HF. Brain EGFR may be a novel target for therapeutic intervention in the MI-induced HF syndrome.
Abstract P2012: Combining the Anti-Fibrotic Actions of Serelaxin With the Anti-Oxidant, N-Acetylcysteine, Abrogates Cardiac Fibrosis in an Isoprenaline-Induced Murine Model of Cardiomyopathy
Authors: Wang Chao , Kemp-Harper Barbara K , Samuel Chrishan S
Published: September 2019

Oxidative stress is a significant contributor to cardiac fibrosis, particularly in the setting of hypertension. While current frontline medications (such as ACE inhibitors) and anti-oxidants (such as N-Acetylcysteine; NAC) can attenuate the impact of elevated blood pressure (BP) on cardiac fibrosis, it is unclear to what extent established fibrosis can be reduced in the absence of BP regulation. The drug-version of the ovarian and cardiovascular hormone, relaxin, serelaxin (RLX) mediates rapid-occurring anti-fibrotic effects by inhibiting the effects of TGF-β1 on myofibroblast-induced collagen production, independently of BP regulation. In this study, we compared and combined the effects of RLX to the anti-oxidant, NAC, in a normotensive model of heart disease. 14-16 week old adult 129sv mice were subjected to isoprenaline (ISO)-induced cardiomyopathy, which along with saline-treated controls demonstrated equivalent systolic BP (SBP) measurements (122±3 vs 124±4 mmHg, respectively; as determined by tail cuff plethysmography). Sub-groups of ISO-injured mice were treated from days 7-14 post-injury with RLX (0.5mg/kg/day), NAC (25mg/kg/day; s.c) or both combined via osmotic mini-pumps. At day 14 post-injury, the left ventricle (LV) of ISO-injured mice presented with increased macrophage infiltration, phosphorylated IkB levels (a marker of NF-kB activity), oxidative stress, NOX2, NOX4 and TGF-β1 expression, myofibroblast differentiation and interstitial fibrosis (all p<0.05 vs saline-treated controls). RLX decreased cardiac fibrosis (by ~40%; p<0.05 vs ISO alone) independently of effects on macrophage infiltration, NOX2/NOX4 expression or SBP. NAC alone reduced interstitial fibrosis (by ~58%; p<0.01 vs ISO alone; no different to ISO+RLX treatment), while when added in combination, RLX and NAC completely abrogated cardiac fibrosis, returning levels to those measured in saline-treated controls. These finding suggest that NAC can reduce fibrosis without necessarily impacting on the contribution of BP to disease progression, and that the combined effects of RLX and an anti-oxidant may offer superior therapeutic efficacy in the treatment of cardiac fibrosis.
Abstract P2018: PRR Mediates P53 Activation in Aristolochic Acid Induced Renal Proximal Tubule Cell Injury
Authors: Culver Silas A , McDevitt Rebecca K , Siragy Helmy M
Published: September 2019

Aristolochic acid (AA) induces renal proximal tubular injury leading to inflammation, atrophy and fibrosis. These effects of AA were reported to be mediated by p53 induced cell cycle arrest and apoptosis. Recently, our laboratory demonstrated that the (pro)renin receptor (PRR) promotes podocyte inflammation and apoptosis. We hypothesized that PRR mediates AA induced proximal tubular injury via upregulation of p53 activation. Mouse immortalized proximal tubule cells (PTCs) were grown to 70% confluence and transfected with either scramble (Scr) or PRR siRNA followed by treatment with either AA 10ug/mL or culture media alone (control) for 48 hours prior to harvesting for RNA and protein extraction (n=3 for all treatment groups). Compared to control, AA treatment alone significantly increased PRR mRNA expression by 202% (p<0.05). Compared to Scr treatment, PRR siRNA reduced PRR mRNA expression by 49% (p<0.05). Compared to AA treatment, AA+PRR siRNA ameliorated the observed increase in PRR mRNA. Total p53 mRNA expression was not significantly changed by treatment with PRR siRNA or AA. However, compared to the control group, p53 phosphorylation (activity) increased by 167% in the AA treated group (p<0.05). PRR shRNA attenuated (40% reduction) the observed increase in p53 phosphorylation that was induced by AA treatment (p<0.05). We conclude that PRR mediates increased activity of p53 in response to AA in proximal tubule cells.
Abstract P2022: The Jak3 Pathway is Involved in the Benefit of the Mineralocorticoid Receptor Antagonist Finerenone in Acute and Chronic Kidney Injury Following Ischemia-Reperfusion
Authors: Jaisser Frederic , Estela Gabriel R , Kolkhof Peter , Barrera-Chimal Jonatan
Published: September 2019

Introduction:
Mineralocorticoid receptor antagonists (MRA) prevent ischemic acute kidney injury (AKI) and its transition to chronic kidney disease (CKD). The specific contribution of interleukin-4 (IL-4) signaling pathway to this effect is unknown.

Objective:
To gain insight into the role of IL-4 signaling and specifically the Jak3 pathway in the MRA protective effect of Finerenone.

Methods:
Male C57/B6 mice (48) were divided in: sham, renal ischemia for 22.5 min (IR), IR plus treatment with the non-steroidal MRA finerenone (10 mg/kg) and IR-finerenone plus Tofacitinib (15mg/kg), a JAK3 inhibitor at -48, -24 and -1 h before IR. The mice were followed-up for 24 hours or 4 weeks to evaluate the AKI to CKD transition.

Results:
At 24h post-injury, finerenone showed benefits on various parameters while concomitant Jak3 inhibition blunted these effects: plasma creatinine (uM/l, 40.8±6.1 IR, 13.2±1.3 Fine, 31.9±5.4 Jak3i, mean+/-SEM) and urea (mM/l; 39.5±6.1 IR, 10.7±0.5 Fine, 25.4±4.1 Fine+Jak3i, mean+/-SEM) and protects from acute tubular necrosis (rel. values: 1.34±0.19 IR, 0.47±0.14 Fine, 1.18±0.17 Fine+Jak3i, mean+/-SEM) We previously showed that finerenone has an important role in changing the macrophage phenotype from M1 (pro-inflammatory) to M2 (anti-inflammatory) profile. Finerenone treatment decreased expression of M1 markers, an effect blunted by Jak3 inhibition. One month after injury, the AKI to CKD transition was evidenced by a 40% increase in plasma creatinine, in interstitial fibrosis and increased mRNA levels of various profibrotic markers. Finerenone treatment protected against these alterations while the JAK3 inhibitor reversed the Finerenone protective effects on interstitial fibrosis (rel. values: 1.95±0.15 IR, 0.33±0.08 Fine, 0.91±0.14 Fine+Jak3i, mean+/-SEM) and on profibrotic markers (rel. values, mean±SEM): fibronectin (17.18±3.93 IR, 3.40±0.77 Fine, 12.62±4.32 Fine+Jak3i), col1a (8.76±2.07 IR, 4.33±0.08 Fine, 19.24±1.55 Fine+Jak3i), α-sma (5.42±1.27 IR, 1.77±0.15 Fine, 6.88±1.61 Fine+Jak3i)

Conclusion:
MR antagonism with finerenone counteracts the acute and chronic deleterious effects after renal ischemic injury. The IL-4 receptor-JAK3 signaling pathway is involved in the benefit of the MRA finerenone.
Abstract P2030: DOCA-Salt Diminishes Brain RAS Activity In Parallel With Plasma And Renal RAS Activity - No Evidence For Selective Brain RAS Activation
Authors: Uijl Estrellita , Ren Liwei , Mirabito Colafella Katrina M , Poglitsch Marko , Domenig Oliver , Zlatev Ivan , Kim Jae B , Huang Stephen , Melton Lauren , Foster Don , Danser A.H. Jan
Published: September 2019

Deoxycorticosterone acetate (DOCA)-salt is suggested to increase blood pressure via selective upregulation of the brain renin-angiotensin system (RAS), while suppressing the circulating RAS. Yet, we have observed parallel downregulation of plasma and brain renin in mice. Here, we quantified brainstem angiotensinogen (AGT) and angiotensin (Ang) levels in rats exposed to DOCA (200mg; 60-day release) and 0.9% NaCl as drinking water for 7 weeks. To determine the contribution of AGT (liver vs. kidney/brain), Ang II type 1 receptors (AT1R) and blood pressure to tissue Ang II content, rats received vehicle, liver-targeted AGT siRNA (10 mg/kg fortnightly; s.c.), valsartan (31 mg/kg/day; s.c.) or spironolactone (80 mg/kg/day; s.c.) during the final 3 weeks, the latter fully normalizing blood pressure (n=3-9 per group). Plasma renin and AGT were determined by enzyme-kinetic assay, tissue AGT by western blotting, and Ang I and II by LC-MS/MS. Plasma renin, AGT, Ang I and II in rats not exposed to DOCA-salt were 24±8ng Ang I/mL per h, 731±51nM, 123±38pM and 109±43pM. AGT was present in liver, kidney and brainstem. Kidney Ang I and II were 540±135and 510±97fmol/g, while brainstem Ang I and II were undetectable in all and 50% of the rats, respectively (<6 and <12±4 fmol/g). DOCA-salt lowered plasma and renal Ang I and II by ≈90%, while brainstem Ang II became undetectable in 70% of the rats. AGT siRNA reduced plasma AGT by 97±1%, with similar reductions in the liver and kidney, while brain AGT was unaltered. Neither valsartan nor spironolactone affected AGT at any site. Spironolactone increased plasma renin, and hence plasma, renal and brain Ang I and II also rose modestly. Both siRNA and valsartan tended to further lower renal Ang levels, while neither drug affected plasma renin or Ang I and II in plasma and brain. In conclusion, DOCA-salt suppresses plasma, renal and brain Ang II in parallel. Renal Ang II is determined by hepatic AGT and relies on AT1R-mediated internalization. Although brain AGT is liver-independent, it is unaltered by DOCA-salt and does not permit selective brain RAS upregulation. The RAS upregulation induced by spironolactone, combined with its blood pressure-normalizing effect, argues against the (brain) RAS as a major blood pressure determinant in this model.
Abstract P2054: Experiencing the Primary Composite Cardiovascular Outcome is More Strongly Associated With Serious Adverse Events Than Intensive Treatment in the Systolic Blood Pressure Intervention Trial
Authors: Botchway Albert , Buhnerkempe Michael G , Prakash Vivek , Al-Akchar Mohammad , Adekola Oritsegbubemi , Flack John M
Published: September 2019

Background:
SPRINT was the first large-scale trial to investigate the benefits of intensive versus standard blood pressure treatment (goal of <120 vs <140 mmHg, respectively). Participants who were treated intensively experienced fewer cardiovascular events. About 38% (3529/9361) of participants had a serious adverse event (SAE), leading to debate about the role of intensive treatment in causing of SAEs. However, no study has examined cardiovascular events as an independent predictor of SAEs, accounting for other variables.

Methods:
SAEs were predicted using multivariable cox models. The variables were CVD composite (the original SPRINT primary outcome, included in this analyses to capture the effect of having a study event), treatment (intensive or standard), albumin (normal, micro, macro), BMI (<30 vs ≥30), estimated glomerular filtration rate (eGFR, <60 vs ≥60), statin, and aspirin use. Analyses were stratified by tertiles of baseline 10-year ASCVD risk score in those without and with baseline CVD. Thirty-eight participants were excluded due to incomplete data resulting in a sample of 9323 (mean age 68 years, 35% female).

Results:
In all groups, including the in-study CVD composite primary endpoint improved the harrell’s C-index of model fit. In the multivariable models, CVD composite was a predictor of SAEs in ASCVD tertiles (T1, T2, T3) and the clinical CVD group (Hazard Ratio [HR] 5.01 CI 3.86-6.51; HR 5.10 CI 4.23-6.28; HR 4.40 CI 3.79-5.10; and HR 3.67 CI 3.18-4.23, respectively). Kidney function as measured by eGFR <60 predicted SAEs in T1, T2, T3 and clinical CVD groups (HR 1.52 CI 1.26-1.84; HR 1.25 CI 1.06-1.47; HR 1.24 CI 1.10-1.40; and HR 1.20 CI 1.03-1.38, respectively). BMI ≥ 30 emerged as a predictor in T1, T2, and T3 but not in the clinical CVD group. Albuminuria, statin and aspirin use were not consistent predictors across groups. Intensive treatment predicted SAEs in T2 (HR 1.20 CI 1.04-1.38), but not in the T1, T3 and clinical CVD groups. No interaction was detected between ASCVD tertile and randomized treatment group assignment.

Conclusions:
These analyses demonstrated that experiencing the primary composite CVD endpoint was the strongest predictor of SAEs in all groups and that intensive treatment was a modest, infrequent predictor of SAEs.
Abstract P2055: Incremental Impact of Stressful Situations in Stage 1 vs. Stage 2 Hypertensives: Insights From a Large Digital BP Management Service and Registry
Authors: Peretto Giovanni , Bordignon Tommaso , Del Mauro Gaia , Fabi Martina , Colombo Eros , Cianflone Domenico
Published: September 2019

Stressful conditions induce an increase in BP levels. It is often difficult to systematically record BP levels under those conditions. The current availability of digital data collection platforms may provide new insights on the changes of BP level under multiple conditions on a large scale. This study leverages the 298,621 BP values of 8992 subjects (2702 females, 48.8 ±13.4 y) consecutively accrued on a digital platform. This is a software-as-a-service (SAAS) platform for BP management leveraging innovative proprietary AI expert systems algorithms providing to patients progressive interpretation of the evolution of their BP and delivering highly personalized lifestyle interventions. Users access the service via App (iOS and Android). This service fosters the Class IA indication for HBPM of the 2017 AHA Hypertension Guidelines. Upon BP data entry (through connected devices, the smartphone health app, or manual entry after extensive validation) users are required to indicate any additional condition at the time of BP measurement including work, after exercising, during intense stress, during headache. Overall, in all subjects, we observed a significantly higher systolic +10.3±0.4mmHg and diastolic +5.1±0.3mmHg BP vs rest (Mean±SEM;95% C.I.) [9.4-11.25, p< 0,0001 and 5.1-11.25, p< 0,001; respectively]. Out of 8992 subjects, 1986 subscribed the lifestyle intervention program after self-declaring their usual BP range. Thus, according to the AHA Hypertension Guidelines, 325 subjects declared normal BP (Group N) values, while 1641 were normotensives (Group H). We analyzed the BP changes during stress in both normotensive and hypertensives, and we observed significant increase of both systolic (N: +6.7±0.7; [2.99-10.4] p=0.0004; H: +10.2±1.4 [7.49-12.9] p<0.0001) and diastolic BPs (N: +2.2±0.3; [2.99-10.4] p=0.0004; H: +6.0±0.3 [7.49-12.9] p<0.0001). These data support the notion of a more intense response to stress in hypertensives. These insights are useful for an appropriate and personalized treatment strategy in hypertensives. A digital SAAS platform is a strong adjunctive tool for a better management of hypertension.
Abstract P2066: The Difference of With or Without Blood Pressure Adjustment for Pulse Wave Velocity as a Risk Factor for Cardiovascular Disease
Authors: Nakano Hiroki , Tomiyama Hirofumi , Yamashina Akira , Study Group FMD-J
Published: September 2019

Background:
Arterial stiffness, assessed by pulse wave velocity (PWV), is an independent predictor of future cardiovascular (CV) events. While blood pressure (BP) functionally increases the value of PWV, the significance of this increase as a risk for CV disease (CVD) has not been fully clarified. The stiffness-index beta has been used as a less BP-influenced marker to assess arterial stiffness. Then, the present study examined whether BP-adjusted value of the PWV improve or attenuate the predictive value for the development of the hypertension, retinopathy, and renal dysfunction, or of future CV events as compared to brachial-ankle PWV (baPWV).

Methods and Results:
Study 1: In the 7-year prospective observation conducted in 3274 Japanese men without hypertension at the baseline for the development of hypertension and also in 3490 Japanese men for the development of organ damage, baPWV, BP, estimated glomerular filtration rate (eGFR), and fundus findings were measured at baseline and end of study period. During the study period, 474 subjects developed to hypertension, 29 subjects had renal dysfunction, and 91 subjects had the development of retinopathy. The area under the curve (AUC), in the receiver operator characteristic curve analysis, of baPWV was higher than that of BP-adjusted baPWV for the development of hypertension (0.73 vs. 0.59; p<0.01), for eGFR <60 mL/min/1.73m
2
and for the development of retinopathy. The baPWV rather than BP-adjusted baPWV showed a significant odds ratio for the development of hypertension (1.84 per standard deviation increase, p<0.01) and that of retinopathy (1.25 per standard deviation increase, p<0.01). Study 2: During the 3-year observation period in 511 patients with coronary artery disease, 72 CV events were confirmed. While the AUCs of both markers for the development of CV events were similar, the Cox proportional hazards model analysis revealed that baPWV rather than BP-adjusted baPWV showed a significant hazard ratio (1.38, P<0.01) for the development of CV events.

Conclusions:
The BP adjustment, based on the concept of stiffness-index beta, may attenuate the significance of baPWV as a risk marker for the development of CVD, especially in its development related pathophysiological abnormalities in male subjects.
Abstract P2067: Trends in Ambulatory and Home Blood Pressure Monitoring Use for Confirmation of Hypertension or Resistant Hypertension Among the Commercially-Insured in the U.S., 2008-2017
Authors: Smith Steven M , Desai Raj , Dietrich Eric , Park Haesuk
Published: September 2019

Objectives:
Out-of-office BP measurement, including ambulatory and home BP monitoring (ABPM, HBPM, respectively) are considered the gold standard for hypertension and treatment-resistant hypertension (TRH) confirmation, but their real-world use is not well-studied. We examined recent trends in submitted claims for ABPM and HBPM among commercially-insured U.S. adults with suspected hypertension or TRH.

Methods:
Using Truven commercial claims data (2008–2017), we identified adults (age ≥18 years) with hypertension, and ≥1 antihypertensive medication fill. We evaluated CPT codes for claims submitted for ABPM and HBPM from 6 months before to 1 month after initial antihypertensive drug use (incident treated hypertension [ITH] cohort) or the first occurrence of overlapping use of ≥4 antihypertensive drugs (incident TRH cohort). We excluded persons without continuous enrollment during the 7-month observation period.

Results:
Overall, 3,378,645 patients with ITH and 335,200 with incident TRH met inclusion criteria. Of those with ITH, 13,063 (3.9 per 1,000 persons) had ≥1 ABPM or HBPM claim between 6 months prior to and 1 month after initiating treatment. The annual proportion of patients with ≥1 claim ranged from 1.6 to 3.0 per 1,000 for ABPM and 1.3 to 2.1 per 1,000 for HBPM. Among those with incident TRH, 1126 (3.4 per 1,000) had ≥1 ABPM or HBPM claim between 6 months prior to and 1 month after initiating a fourth agent. The annual proportion with ≥1 claim ranged from 1.2 to 2.0 per 1,000 persons for ABPM and 1.1 to 2.9 per 1,000 for HBPM. From 2008 to 2017, use of ABPM modestly increased in the ITH cohort and decreased in the TRH cohort; no time trends were observed with HBPM. Sensitivity analyses suggested that ABPM was used most often for diagnosis confirmation (especially in the ITH cohort), followed by treatment monitoring, whereas HBPM was distributed more evenly.

Conclusion:
Our findings suggest ABPM is used rarely for guiding treatment initiation decisions among those with incident hypertension or apparent TRH. Claim submissions for HBPM were similarly low,

but a significant proportion of HBPM may occur without attempting reimbursement. Future research is needed to understand the factors responsible for low utilization, billing and coverage of these services.
Abstract P2070: Renal, Cardiovascular, And Mortality Impacts Of Refractory Hypertension In Patients With Chronic Kidney Disease: Insights From The Chronic Renal Insufficiency Cohort (CRIC).
Authors: Prakash Vivek , Buhnerkempe Michael G , Botchway Albert , Adekola Bemi , Al-Akchar Mohammad , Flack John M
Published: September 2019

Background:
Refractory hypertension can be defined as uncontrolled blood pressure (BP at or above 140/90 mmHg) when taking five or more classes of antihypertensive medication, including a diuretic for patients with chronic kidney disease. The prognosis for patients with refractory hypertension is not understood.

Methods:
Refractory hypertension was assessed in the Chronic Renal Insufficiency Cohort (CRIC). A total of 3314 treated hypertensive patients were included, and patients were assigned a hypertensive phenotype (i.e., refractory hypertension, apparent treatment resistant hypertension [aTRH], hypertension) based on their baseline visit. Cox proportional hazards models were used to analyze differences in outcome risk across hypertensive phenotypes after controlling for age, race, diabetes, smoking, BMI, urine protein, eGFR, systolic blood pressure, education, and study site.

Results:
Refractory hypertension was found in 4.5% (149/3314) of treated hypertensive patients. When compared with those with aTRH, patients with refractory hypertension were significantly more likely to experience renal events - composite of end-stage renal disease and/or 50% drop in eGFR (hazard ratio [95% confidence interval], 1.64 [1.29, 2.09]); and cardiovascular events - composite of myocardial infarction, stroke, peripheral arterial disease, and congestive heart failure (1.34 [1.04, 1.73]); but showed no significant difference in all-cause mortality risk (1.06 [0.80, 1.41]).

Conclusions:
Refractory hypertension increases the risk for renal and cardiovascular disease over that previously observed for aTRH in patients with chronic kidney disease.