bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 035: Novel Metabolites Associated With Blood Pressure During Dietary Protein And Carbohydrate Interventions
Authors: Changwei Li , Bundy Joshua D , Tian Ling , Zhang Ruiyuan , Chen Jing , Kelly Tanika N , He Jiang
Published: September 2022

Introduction:
Dietary protein and carbohydrate intake have been associated with blood pressure (BP). However, molecular mechanisms for these associations are not well studied.

Hypothesis:
We hypothesize that novel metabolites – intermediate products of metabolic reactions of diet or genetic factors, may play a role in BP regulation during dietary protein and carbohydrate interventions.

Methods:
In the Protein and Blood Pressure (ProBP) trial, 80 participants were randomly assigned to take 40 grams/day of soy protein, milk protein, or carbohydrate supplementation each for 8 weeks in a cross-over design. We analyzed the associations of serum metabolites with BP at baseline and each dietary intervention, respectively. Age, sex, race, education, smoking, drinking, and body mass index were adjusted in all analyses. ProBP findings were evaluated for replication among 1206 participants of the Bogalusa Heart Study.

Results:
After Bonferroni correction for 77 independent metabolite clusters, we identified 17 metabolites (1 at baseline, 8 at carbohydrate diet, 4 at milk protein diet, and 5 at soy protein diet) positively associated with BP (P<6.49E-4). Furthermore, 12 of these metabolite-BP associations identified at dietary interventions were replicated in the BHS (P<0.05,
Table
). Of the 12 metabolites,
glycerol
and
hexadecadienoate (16:2n6)
were associated with BP in previous studies, and all other metabolite-BP associations were novel.

Conclusions:
Several novel metabolites may play an important role in BP regulation during dietary protein and carbohydrate intake.
Abstract 037: Serum Chemerin As A Novel Biomarker For Preeclampsia Complications: Effects Of Statins On Its Release From The Placenta
Authors: Tan Lunbo , Broekhuizen Michelle , Neuman Rugina I. , Schoenmakers Sam , Lu Xifeng , Danser A.H. Jan , Verdonk Koen
Published: September 2022

Background:
The adipokine chemerin is increased in preeclampsia (PE), possibly because it is released from the placenta. We aimed to evaluate the potential of chemerin as novel biomarker to predict PE or PE-related complications.

Methods:
Making use of a secondary analysis of a prospective cohort study that evaluated the usefulness of the sFlt-1 (soluble Fms-like tyrosine kinase-1)/PlGF (placental growth factor), we measured chemerin in 371 women with suspected or confirmed PE. In addition, we obtained placentas from 25 women (15 healthy controls, 10 PE) to determine whether statins are capable of reducing chemerin release from isolated perfused placentas and placental explants. For this purpose, chemerin, sFlt-1 and PlGF were measured in the perfusate before and after the addition of 1000 ng/mL pravastatin or 6000 ng/mL fluvastatin to the maternal side, and in explant cultured medium before and after exposure to pravastatin (10, 100 or 1000 μmol/L) or fluvastatin (10 or 100 μmol/L) for 72 hours at 37°C with 5% O
2
and 5% CO
2
.

Results:
At time of biomarker measurement, women had a median gestational age of 35.6 weeks (range 31.7-38.0). Serum chemerin levels were elevated (P<0.001) in women with confirmed PE (n=85) and gestational hypertension (n=68) in comparison to those without hypertensive disease of pregnancy (n=218; 243±13 and 205±19 vs. 185±6 ng/mL). Serum chemerin correlated positively with the sFlt-1/PlGF ratio (r=0.27, P<0.001), the protein/creatinine ratio (r=0.27, P<0.001), and mean arterial blood pressure (r=0.20, P<0.001), and negatively with birth weight (r=-0.24, P<0.001). Levels of chemerin were significantly higher in maternal perfusate of preeclamptic placentas in comparison to healthy placentas. Pravastatin, but not fluvastatin, suppressed chemerin and sFlt-1 release from healthy placentas, and upregulated PlGF release. Studies in placental explants confirmed this observation.

Conclusions:
Chemerin is increased in both serum and placental perfusate of women with PE, and should therefore be investigated as a potential biomarker for PE or PE-related complications. Moreover, pravastatin lowered placental chemerin release in parallel with sFlt-1, suggesting that the 2 might be causally linked.
Abstract 046: Induction Of Endothelial Sirt3 After Onset Of Hypertension Rescues Endothelial Function
Authors: Dikalova Anna , Tkachuk Liliya , Ao Mingfang , Fehrenbach Daniel , Arbiser Jack , Dikalov Sergey
Published: September 2022

Endothelial dysfunction plays a key role in the pathogenesis of hypertension and represents a major risk factor for cardiovascular disease. Despite treatment with multiple drugs, only 1 in 4 patients has blood pressure under control. We discovered a new synergistic control point with the potential to address this problem. We have shown that Sirt3 level is reduced in essential hypertension and endothelial Sirt3 overexpression attenuates endothelial dysfunction and hypertension. We proposed that induction endothelial Sirt3 after onset of hypertension rescues endothelial function and reduces hypertension. To test this hypothesis, we used genetic- and pharmacological Sirt3 induction. To genetically induce the expression of endothelial Sirt3 we used Sirt3
flox/flox
mice crossed with VeCad-Cre mice treated with low dose of 4-hydroxytamoxifen (4HOT, i.p. 0.3 mg/20 g daily, 5 days) which does not have off-target cardiovascular effects. Mice underwent telemetry placement and ten days later received 4-week osmotic pumps containing vehicle (saline) or angiotensin II (0.7 mg/kg/day). 14-days later half of mice received 4HOT to induce endothelial Sirt3 overexpression. Treatment of mice with 4HOT after onset of hypertension slightly reduced blood pressure but most importantly completely rescued endothelial-dependent relaxation, normalized mitochondrial O
2
.
- and restored endothelial nitric oxide. Second, we tested if pharmacological induction of Sirt3 by hexafluoro (i.p. 8 mg/kg, 4 days) after onset of angiotensin II-induced hypertension improves endothelial function and reduces hypertension. It was found that hexafluoro substantially reduced systolic blood pressure, significantly diminished vascular mitochondrial O
2
.
- and improved endothelial nitric oxide. To test the role of endothelial Sirt3 in pharmacological effect of hexafluoro we used endothelial specific Sirt3 knockout mice and angiotensin II model of hypertension. Interestingly, hexafluoro was not effective in endothelial Sirt3 deficient mice supporting critical role of endothelial Sirt3. These in vivo studies demonstrate therapeutic potential of genetic and pharmacological endothelial Sirt3 induction after onset of hypertension for treatment of endothelial dysfunction.
Abstract 051: Exploiting Angiotensin Sensitive Vagal Afferents To Understand And Alleviate Hypertension: A Closed Loop System For Blood Pressure Control
Authors: Baumer Harrison Caitlin , Penaloza Aponte Jesus , Elsaafien Khalid , Johnson Dominique N , de Araujo Alan , Smith Justin A , Scott Karen , de Lartigue Guillaume , Krause Eric G , De Kloet Annette D
Published: September 2022

Cardiovascular homeostasis is maintained by neural signals arising from arterial baroreceptors (BRs) that apprise the brain of vascular stretch. Here, we test the overall hypothesis that angiotensin-II (AngII) type 1a receptor expressing neurons within the nodose ganglia (NDG
AT1aR
) serve as BR that undergo plasticity during chronic elevations in BP. We found that NDG
AT1aR
receive input from the aortic arch, project to the caudal nucleus of the solitary tract (cNTS), and synthesize Piezo-1/-2. AT1aR-GCaMP6s mice were used to determine whether NDG
AT1aR
couple their firing to elevated BP. Two-photon intravital imaging revealed that phenylephrine (PE; i.v.) increased GCaMP6s fluorescence within NDG
AT1aR
(ΔF/F
max
= 0.65±0.05*, Time
ΔF/Fmax
= 92s) and this response was delayed in mice given DOCA-salt (Time
ΔF/Fmax
= 124s) or chronic AngII (Time
ΔF/Fmax
= 155s). To determine if optical perturbation of NDG
AT1aR
affects the baroreflex, expression of excitatory channelrhodpsin2 (ChR2) or inhibitory halorhodopsin (Halo) was directed to AT1aR cells. Optical excitation of NDG
AT1aR
at their axon terminals in the cNTS elicited frequency-dependent decreases in BP (ΔSBP @ 0.5, 1, 2, 3 Hz = -6.74±3.42
#
, -9.75±2.33
#
, -23.24±4.39
#
, -25.27±4.38
#
) and HR (ΔHR @ 0.5, 1, 2, 3 Hz = -7.81±3.14
#
, -14.86±3.22
#
, -23.39±5.02
#
, -23.66±4.13
#
). Conversely, optical inhibition of NDG
AT1aR
at axon terminals in the cNTS blunted pressor responses to PE (ΔHR
PE(Min)
= -36.15±5.31, ΔHR
PE+Light(Min)
= -23.11±3.33
+
). ChR2-expressing mice were then subjected to DOCA-salt. Optical excitation elicited hypotensive (ΔSBP
Min
= -21.11±4.05
+
) and bradycardic (ΔHR
Min
= -235.6±46.69
+
) responses; however, the duration of these effects was altered. We developed a closed loop system to target NDG
AT1aR
at axon terminals in the cNTS of ChR2-expressing mice to determine if these neurons can be exploited to alleviate hypertension. Mice were subjected to acute and chronic stimuli to elevate BP and stimulated by the system until BP returned below threshold. Collectively, these studies reveal the structure and function of NDG
AT1aR
and suggest that such neurons may be exploited to discern and relieve hypertension. (
p
< 0.05, * time by treatment,
#
time by frequency, or
+
time by light interaction)
Abstract 052: Central Gain For Sympathetic Outflow Is Increased In Postural Tachycardia Syndrome
Authors: Urechie Vasile , Rigo Stefano , Shibao Cyndya A , Okamoto Luis E , Biaggioni Italo , Gamboa Alfredo , Wahba Amr , Elkholey Khaled , Giesecke Morgane , Muldowney James , Mohr Abigail , Diedrich Andre
Published: September 2022

White noise perturbations using neck suction (NS) can be applied to identify open loop transfer function in a closed loop baroreflex system. It allows an estimation of central arc transfer function (H1) which represents how afferent baroreceptor information is translated into sympathetic outflow and peripheral arc transfer function (H2) which describes how muscle sympathetic nerve activity (MSNA) affects blood pressure (BP). We hypothesize that patients with postural tachycardia syndrome (POTS) have elevated central gain compared to healthy controls (CONT). We recorded raw MSNA and continuous finger BP during randomized NS (0 to -50 mmHg, 0.03-0.3 Hz) and controlled breathing at 0.25 Hz for 6 minutes in 10 female POTS and 9 female CONT. Both groups were matched for age (28.4±2.1 vs. 31.9±3.0 p=0.80) and BMI (23.0±0.7 vs 24.5±1.0, p=0.25). We derived the continuous spike rate using wavelet-based denoising. We estimated median central gain in H1 and peripheral gain in H2 for low (LF: 0.04-0.15 Hz) and high (HF: 0.15-0.4 Hz) frequency ranges (Figure). We found that POTS have significantly higher central gain in LF range (2.44±0.38 vs 1.53±0.24 spikes/s/mmHg, p=0.028, MN±SE) but not in HF range (9.02±3.02 vs. 5.06±0.52 spikes/s/mmHg, p=0.604). Median peripheral gains were comparable in LF (0.68±0.15 vs 0.58±0.09 mmHg/spikes/s, p>0.99) and HF ranges (0.18±0.02 vs 0.21±0.03 mmHg/spikes/s, p=0.497). Our results indicate that POTS have higher recruitment of neural sympathetic spike activity during BP oscillations in the LF range. This could explain why higher sympathetic activation/modulation and greater BP fluctuations in the LF range are found during upright in POTS.
Abstract 053: Increased Arterial Pressure Precedes Renal Dysfunction And Elevated Renal Sympathetic And Afferent Nerve Activity In Deoxycorticosterone-Salt Rat
Authors: Gauthier Madeline M , Parvin Irin , Dennis Melissa R , Leung Christian H , Banek Christopher T
Published: September 2022

Hypertension (HTN) is a primary risk factor for cardiovascular disease - the leading cause of death worldwide. HTN is multifaceted but a key factor is aberrant autonomic signaling, including renal sympathetic (RSNA) and afferent nerve activity (ARNA). ARNA is elevated in several models of HTN and renal afferent nerve ablation can mitigate the HTN. It is unknown if ARNA rises before changes in blood pressure (BP) or renal function in HTN. This study aimed to determine when renal nerve activity changes
vis a vis
BP and renal function in a model of salt sensitive HTN. We hypothesized increased ARNA and RSNA would coincide with a rise in BP and decline in renal function. To test this hypothesis, 24 uninephrectomized male Sprague Dawley rats (300g) received a deoxycorticosterone acetate (DOCA; 100mg, SC) implant and 0.9% NaCl drinking water. Renal function (serum creatinine; Cr) and systolic BP (SBP; tail cuff) were measured weekly. Animals were randomly allocated to acute renal nerve recording on protocol day (D) 0, 7, 14, or 21. Multiunit RSNA and ARNA were recorded serially, as ARNA was isolated after sectioning proximal side. RSNA is expressed as bursts/heartbeat, ARNA and expressed as %ARNA
max
(response to intrapelvic capsaicin). Data were analyzed by one-way ANOVA with Bonferroni post hoc. Data presented as mean±SEM; *p<.05 vs D0. SBP (mmHg) progressively increased on D7 (*152 ± 3), D14 (*171 ± 3), and D21 (*197 ± 7) vs D0 (138 ± 3). ARNA (%) increased at D14 (*23.0 ± 4.7) and D21 (*34.2 ± 4.4) vs D0 (10.1 ± 0.8), but no changes were detected at D7 (13.4 ± 2.1). No changes in RSNA from D0 were detected. Cr (mg/dL) increased at D14 (*1.2 ± 0.1) and D21 (*1.2 ± 0.1) vs D0 (0.9 ± 0.0), indicating a decline in kidney function. These data do not support the hypothesis that ARNA and RSNA increase in parallel to increasing BP. Rather, BP appears to initially increase independent of detectable changes in RSNA, ARNA or renal dysfunction. Interestingly, renal dysfunction and elevated ARNA were detected on D14, which coincides with our previous report that renal inflammation is first detected at D14. These data do not support a causal role of ARNA in this model. Rather, ARNA may subsequently drive disease progression. Further studies on the cause of increased ARNA and inflammation interaction are underway.
Abstract 057: Macula Densa Stimulation Acutely Increases Afferent Renal Nerve Activity And Blood Pressure
Authors: Gyarmati Georgina , Stocker Sean D , Deepak Sachin , Izuhara Audrey , Peti-Peterdi Janos
Published: September 2022

Macula densa (MD) cells in the kidney are known to regulate renal hemodynamics and the renin-angiotensin system, and indirectly systemic blood pressure (BP). However, neuron-like differentiation and activity as non-traditional MD functions are emerging. This study addressed the hypothesis that MD cells function similarly to peripheral ganglia, form neuronal networks that are part of the peripheral (renal) nervous system and synapse with renal afferent/efferent nerves as a new pathway for the direct control of sympathetic tone and BP. Single cell MD Ca
2+
signaling was analyzed in vivo using intravital multiphoton microscopy (MPM) of MD-G6 mice that selectively express the ratiometric Ca
2+
reporter GCaMP6f/tdTomato in MD cells. Afferent renal nerve activity (ARNA) was isolated and measured via recording electrodes. Systemic BP was monitored via the cannulated carotid artery. Optogenetic MD stimulation was induced by blue light (470nm) exposure of the whole left kidney of MD-Ai27 mice that selectively express the depolarizing channelrhodopsin ChR2 in MD cells. Pacemaker-like regular Ca
2+
oscillations (4-fold elevations in baseline Ca
2+
and frequency of 0.03 Hz) and their cell-to-cell propagation via long, axon-like processes were exclusively confined to the MD area. Bulk and scRNA seq and MD transcriptome analysis identified the high expression of genes related to synaptic transmission that was validated by immunohistochemistry. Acute intra-renal infusion of arginine-vasopressin (AVP) produced 3-fold increase in MD Ca
2+
firing frequency (via the MD-specific AVPR1a) and a dose-dependent increase in ARNA and BP (saline: -1±1; 0.01ug/mL: 2±1; 0.1ug/mL: 7±2; 0.25ug/mL: 7±3; 0.5ug/mL: 10±3; 1.0ug/mL: 23±13). IV administration of AVP (0.25ug/mL) did not significantly alter ARNA (108±3%, n=3) but produced a significant increase in arterial blood pressure (40±6mmHg, n=3). Blue light exposure of MD-Ai27 mouse kidneys acutely and reversibly increased systemic BP by 22±5mmHg (P<0.05, n=5). In summary, MD cells send interoceptive signals to the brain via renal sensory afferent nerves to directly control BP. Targeting of novel MD mechanisms may result in exciting new therapeutic opportunities for renal and cardiovascular diseases and hypertension.
Abstract 059: Continuous Positive Airway Pressure In The Treatment Of Nocturnal Supine Hypertension And Orthostatic Hypotension In Autonomic Failure
Authors: Okamoto Luis E , Celedonio Jorge E , Smith Emily C , Paranjape Sachin , Black Bonnie , Wahba Amr T , Park Jin-Woo , Shibao Cyndya A , Diedrich Andre , Biaggioni Italo
Published: September 2022

Supine hypertension affects most patients with autonomic failure (AF), complicates the treatment of orthostatic hypotension (OH) and induces nocturnal pressure diuresis, resulting in nocturnal volume depletion and worsening of daytime OH. However, it is often untreated for fear of worsening OH. We previously showed that continuous positive airway pressure (CPAP 4, 8, 12 and 16 cm H
2
O, each for 3 min) had an acute, dose-dependent, blood-pressure (BP)-lowering effect in these patients (maximal systolic BP [SBP] drop 22±4 mmHg with CPAP 16) driven by reductions in stroke volume (-16+3%) and cardiac output (-14±3%), suggesting a Valsalva-like effect. In this study, we hypothesized that overnight CPAP therapy can improve nocturnal supine hypertension, nocturia and daytime OH. Eleven AF patients with supine hypertension (age 74±2 years, 8 men, supine SBP 179±7 mmHg) had placebo or CPAP (8-12 cm H
2
O) applied for 8 hours (10pm-6am) in 2 separate nights. Supine BP was measured every 2 hr from 8pm-8am. Morning orthostatic tolerance was assessed at 8am. SBP significantly decreased during overnight CPAP therapy compared to placebo (Figure; P=0.044 by mixed-effects model) with a maximal reduction of 25±5 mmHg at 4 hours of CPAP. This BP effect was associated with lower nighttime diuresis (609±84 versus placebo 1004±160 mL; P=0.004) and improved morning orthostatic tolerance (AUC
upright
SBP
642±121 versus placebo 410±109 mmHg*min; P=0.014). In conclusion, CPAP is potentially a novel non-pharmacologic approach to treat the supine hypertension of AF that can improve nocturia and daytime OH.
Abstract 060: Angiotensin Type-2 Receptors In The Median Preoptic Nucleus Demarcate A Novel Depressor Neural Circuit In Mice
Authors: Elsaafien Khalid , Eikenberry Sophia A , Harden Scott W , Sheng Wanhui , Scott Karen A , Frazier Charles J , Sumners Colin , Krause Eric G , De Kloet Annette D
Published: September 2022

Blood pressure (BP) is under constant control and regulation by an extensive neural network that spans a number of brain regions. This includes the median preoptic nucleus (MnPO) which directly projects to the pre-autonomic neurons of the hypothalamus to increase BP. While angiotensin type-1 receptors (AT
1
R) are expressed in the MnPO and project to the hypothalamus to increase BP, the role of angiotensin type-2 receptors (AT
2
R) is not clear. Here, we combine the use of AT
2
R-Cre mice, neuroanatomical techniques,
in vivo
optogenetics, pharmacology, and cardiovascular physiology to test the hypothesis that AT
2
R neurons of MnPO (MnPO
AT2R
) elicit cardioprotective effects. We found an abundance of AT
2
R-containing neurons in the MnPO that expressed the mRNA(s) for AT
2
R with high fidelity (~77%; 241 of 313 neurons; n=7). Using RNAscope
in situ
hybridization, we found that ~ 42% (153 of 364 neurons; n=8) of MnPO
AT2R
expressed genetic markers for GABA, an inhibitory neurotransmitter. Next, we delivered a Cre-inducible AAV to direct expression of channelrhodopsin-2 (ChR2) and/or enhanced yellow fluorescent protein (eYFP) within MnPO
AT2R
of AT
2
R-Cre mice. Using
in vitro
electrophysiology, we confirmed functional expression of ChR2 within MnPO
AT2R
, with pulses of light reliably producing action potentials (n= 12 neurons/3 animals). Subsequently, we found that activation of MnPO
AT2R
elicited inhibitory post-synaptic currents in neighboring AT
2
R-negative neurons. The
in vivo
optogenetic excitation of MnPO
AT2R
elicited depressor responses in both adult male (ΔSBP: +2.8 ± 1.3 vs. -4.1 ± 1.9 mmHg, p<0.05 eYFP vs. ChR2, n=15) and female mice (ΔSBP: +0.8 ± 2.2 vs.-5.3 ± 1.9 mmHg, p<0.05 eYFP vs. ChR2, n=5). These responses were abolished in the presence of the autonomic ganglion blocker, hexamethonium. Finally, to assess whether the depressor effects were AT
2
R-mediated, we administered the selective AT
2
R-agonist, compound 21, into the MnPO and observed depressor responses (ΔSBP: +0.5 ± 0.3 vs.-4.9 ± 1.0 mmHg, p<0.05 Saline vs. C21, n=7). In conclusion, our results demonstrate that MnPO
AT2R
are depressor inhibitory neurons that can be targeted with compound 21 to lower blood pressure.
Abstract 061: Renal A20 Protects Against The Cisplatin-induced Kidney Injury
Authors: Lu Xiaohan , Chen Yanting , Yang Ting , Rianto Fitra , Hammer Gianna , Crowley Steven D
Published: September 2022

We recently reported that the ubiquitin-editing protein A20 in myeloid cells can limit the severity of hypertension. However, whether A20 in the kidney regulates renal injury without impacting systemic immune responses remains unclear. To address this question, we bred

A20
flox/flox

mice with the Pax8-rtTA and Tet-On lines, generating inducible renal epithelial cell A20 knockout mice (A20 iKKO). Mice with all 3 transgenes were used as the A20 iKKO group, whereas mice lacking the Pax8-rtTA or Tet-On transgene were wild-type (WT) controls. Before the experiments, mice were given 2mg/ml of doxycycline in water containing 5% (5 of 100) sucrose for 2 weeks so that A20 in renal tubular cells was ablated. 3 days after a single dose of cisplatin (20mg/kg), A20 iKKO mice exhibited more severe kidney injury compared to WTs (blinded injury score, 3.2±0.41 vs. 2.1±0.35 au, p=0.039). A20 iKKOs also had upregulated renal mRNA levels for NGAL (6.0±1.91 vs. 1.0±0.18 au, p=0.018) and KIM-1(3.3±1.2 vs. 1.0±0.29 au, p=0.07). Injured kidneys from the A20 iKKOs contained greater absolute numbers of T cells (123.6±8.5 vs. 53.8±4.7, x10
4
cells per gram kidney, p<0.001), macrophages (159.7±22.5 vs. 52.7±9.9, x10
4
cells per gram kidney, p<0.001), and dendritic cells (66.7±10.6 vs. 32.6±6.1, x10
4
cells per gram kidney, p=0.015) than WTs by flow cytometric analysis. In turn, mRNA levels for TNF-α (9.1±2.62 vs. 1.0±0.18 au; p=0.006) and IL-1β (4.1±1.06 vs. 1.0±0.13 au; p=0.009) were increased in the injured A20 iKKO injured kidneys vs WTs. Thus, A20 in the kidney epithelium protects against cisplatin-induced kidney injury by constraining renal immune cell accumulation and inflammatory cytokine release.
Abstract 062: Bone Marrow-derived Mesenchymal Stromal Cells Engineered To Release Anti-fibrotic Cargo, Evoke Renoprotective Effects Against Hypertensive Chronic Kidney Disease
Authors: Li Yifang , Broughton Brad R , Ferens Dorota M , Widdop Robert E , Ricardo Sharon , Samuel Chrishan
Published: September 2022

Human bone marrow-derived mesenchymal stromal cells (BM-MSCs) have emerged as a promising treatment option for chronic kidney disease (CKD), based on their anti-hypertensive, immunomodulatory and tissue-reparative effects. However, the build-up of fibrosis (scar tissue) in CKD significantly impairs the viability and efficacy of implanted BM-MSCs. To address this, we previously combined the separate administration of BM-MSCs with the anti-fibrotic drug, serelaxin (RLX), which enhanced the renoprotective effects of BM-MSCs in normotensive and hypertensive models of CKD. To advance the clinical application of this combined therapy, we have now engineered BM-MSCs to express RLX and GFP (BM-MSCs-RLX+GFP) or GFP alone (BM-MSCs-GFP) using lentiviral transduction, to deliver the combined treatments via the same route simultaneously. The therapeutic effects of BM-MSCs-RLX+GFP were then investigated in a high salt (HS; 2% NaCI)-induced model of hypertension. Flow cytometry revealed a transduction efficiency of 76% and 93% at a multiplicity of infection of 1 or 2, for BM-MSCs-RLX+GFP and BM-MSCs-GFP, respectively. When cultured for 3- and 7-days
in vitro
, 1x10
6
BM-MSCs-RLX+GFP produced 29±3 and 31±3ng/ml of RLX, respectively. Subgroups of 10-12 week-old male Balb/C mice were subjected to HS loading for 8 weeks. During the final 2 weeks (weeks 7-8), sub-groups of mice were treated with i) RLX (0.5mg/kg/day; s.c) and BM-MSCs (RLX+BM-MSCs); ii) BM-MSCs-GFP; iii) BM-MSCs-RLX+GFP (all 1x10
6
cells/mouse; i.v); or iv) the ACE inhibitor, perindopril (2mg/kg/day, p.o). HS loading increased systolic blood pressure (SBP) to 140-145mmHg but reduced creatine clearance (23mL/24h), compared to the 115-120mmHg and 50mL/24h, respectively, in mice given normal drinking water (both p<0.05). All treatments blunted SBP to 123-130mmHg (no different to baseline levels), whilst creatinine clearance was partially restored by all cell-based treatments (to 34-40mL/24h), but not perindopril. The treatment-induced effects on renal inflammation and fibrosis are currently being evaluated and will be presented. These findings suggested that the genetic engineering of BM-MSCs that released anti-fibrotic cargo (RLX) may be a novel treatment option for hypertensive CKD.
Abstract 069: Methylation Of The Androgen Receptor In White Adipose Tissue Contributes To Female Obesity And Hypertension In BPH/5, A Model Of Superimposed Preeclampsia
Authors: Sones Jennifer L , Flanagan Juliet , Beckers Kalie , Liu ChinCHi
Published: September 2022

Preeclampsia (PE) is a hypertensive disorder of pregnancy that occurs in 10% of women with obesity. The effects of PE on offspring are chronic with increased incidence of cardiometabolic disease into adulthood. We hypothesize that a maternal obesogenic environment plays a role in pregnancy outcomes and offspring cardiometabolic disease in a sex-dependent manner through epigenetic modifications on the X chromosome. To test our hypothesis, we used the obese BPH/5 female mouse that spontaneously exhibits late-gestational hypertension, fetal growth restriction, and excessive gestational weight gain. BPH/5 offspring have sexually dimorphic cardiometabolic phenotypes with females being hyperphagic, obese, and hypertensive, while males are only hypertensive. BPH/5 dams were pair-fed (PF) to C57 control dams beginning at conception. Male and female BPH/5 offspring were fed an ad libitum (lib) diet until analyses. Whole genome bisulfite sequencing was performed to reveal differentially methylated regions between BPH/5 male and female genomic DNA. BPH/5 PF dams had attenuation of late gestational hypertension. BPH/5 offspring had improved fetal growth and significantly reduced adult female BPH/5 obesity, 14.7% BW and visceral white adipose tissue (WAT) from 1366 to 229.7 mg (n=7-32, p<0.05). Despite detectable levels of serum testosterone (2.04±0.6ng/dL, n=16), serum estrogen in BPH/5 adult females during proestrus is significantly lower than controls (3.3±1 v. 7.3±0.5 pg/mL, n=6, p<0.05). This is associated with hypermethylation of the androgen receptor on the X chromosome in adult BPH/5 females. Because androgen response elements are in the promoter region of the thermogenic marker, uncoupled protein 1 (Ucp1), an interaction has been proposed between sex steroid hormones and WAT energy homeostasis. Ucp1 mRNA expression is significantly increased in PF adult BPH/5 female mice compared to ad lib (n=3, p<0.05). Reduction in the maternal obesogenic environment may play a role in BPH/5 sex-dependent offspring differences via epigenetic modifications of the X chromosome. Future studies are needed to understand the transgenerational methylation of cardiometabolic genes in PE offspring to prevent the lifecycle of disease associated with obesity.
Abstract 079: Long-term Circadian Disruption Via Food Availability Leads To Kidney Damage
Authors: Benjamin Jazmine I , Pollock David M
Published: September 2022

Circadian disruption (CD) is a risk factor in cardiovascular disease although we know little about its effects on the kidney. Previous work from our group showed that short-term CD in mice by restricting food availability to the daytime is sufficient to invert diurnal blood pressure rhythms yet kidney excretory function remained aligned with the light-dark cycle. Thus, we hypothesize that long-term CD will result in kidney damage. After putting C57BL/6 mice on a regular 12:12 light:dark cycle were placed on an inactive time-restricted feeding (iTRF, 12h food access during the day/lights on) or
ad libitum
feeding protocol for 8 weeks. During the final week, male and female iTRF mice ate significantly less during the 24-hour day compared to
ad lib
feeding (male iTRF 6.1±1.4g/day vs.
ad lib
9.1±0.9g/day, p<.0001, n=9 for both groups; female iTRF 6.4 ±0.7 g/day, vs.
ad lib
8.3±1.5g/day, p<0.01, n=9 and 8, respectively). Both sexes had an inversion in their diurnal water intake rhythms but maintained normal diurnal urinary water excretion. Aldosterone (aldo) and endothelin-1 (ET-1) contribute to circadian timing of sodium transport systems in the collecting duct. While male iTRF mice maintained their diurnal rhythm of aldosterone excretion (2.0±1.4ng/hr in light, p=0.06; 7.8±3.1ng/hr in dark, p=0.37, n=6), female iTRF mice lost their diurnal rhythm (4.4±2.7ng/hr in light, p=0.85; 4.4±3.1ng/hr in dark, p=0.11, n=6). Male mice with iTRF had significantly lower ET-1 excretion, but maintained the diurnal variation. In contrast, female iTRF mice demonstrated an inverted ET-1 excretion pattern (18.5±6.3ng/hr in light vs. 7.3±2.9ng/hr in dark, p=0.03, n=3) compared to
ad lib
fed mice (18.9±2.1ng/hr in light vs. 33.7±5.6ng/hr in dark, p=0.01, n=3). Picrosirus red staining of kidneys revealed a striking increase in glomerular and interstitial fibrosis in iTRF male mice as compared to their
ad lib
counterparts. In contrast, iTRF female mice appeared to be completely protected from increased fibrosis. These results support our hypothesis that long-term CD due to timing of food intake exacerbates kidney damage. However, females were protected against renal fibrosis, perhaps due in part to an ability to better modulate kidney rhythms in response to food timing.
Abstract 086: Impact Of Ambulatory Blood Pressure Monitoring On Hypertension Diagnosis In Primary Care
Authors: Flatow James , Sanchez Jose , Singer Jessica , Moise Nathalie , Shimbo Daichi , Kronish Ian M
Published: September 2022

Introduction:
Hypertension (HTN) screening guidelines recommend using ambulatory blood pressure monitoring (ABPM) to exclude white coat HTN prior to newly diagnosing HTN. Yet, little is known about the impact of ABPM testing on HTN diagnosis decisions in primary care. Unfamiliarity with how to interpret ABPM results could lead to overtreatment or diagnostic inertia, the failure to diagnose a disease despite clinical evidence.

Methods:
We examined medical records of primary care patients with elevated office BP (≥140/90 mmHg) who were referred to an ABPM testing service. Patients were eligible for analysis if they were ≥18 years old, had elevated office BP without a HTN diagnosis and not prescribed BP medications, were referred by a primary care clinician from clinics affiliated with Columbia University Medical Center, and completed ABPM testing between 2016-2019. Using mean awake BP of 135/85 mmHg as the threshold for elevated ABPM, we compared the BP testing outcome (white coat HTN, in which only office BP was elevated, or sustained HTN, in which both office and ambulatory BP were elevated) to the physician’s action at the subsequent scheduled primary care visit (diagnosed or did not diagnose HTN per manual review of electronic medical records).

Results:
Overall, 111 patients with newly elevated office BP completed ABPM testing during the analysis period. Patients were referred by 60 physicians (50% trainees) from 6 clinics. Patients had a mean (SD) age of 53 (15) years; 73% were women. Fifty-nine patients (53%) had white coat HTN, and 52 (88%) were not diagnosed with HTN at their next primary care visit. The remaining 52 patients (47%) had sustained HTN, and 44 (85%) were diagnosed with HTN at their next visit. Medication was started for 31 of these 44 (70%). Overall, physicians diagnosed HTN concordantly with ABPM results for 96/111 patients (86%).

Conclusions:
More than half of primary care patients with elevated office BP referred for ABPM had white coat HTN. There was high concordance between ABPM results and physician HTN diagnoses, suggesting that ABPM reduces but does not eliminate overdiagnosis and diagnostic inertia in HTN. More research is needed to understand the reasons for discordance and how to optimize the implementation of ABPM testing into primary care.
Abstract 092: Activation Of Neuronal At
1
R Exacerbates Hypertension-associated Mci Through Vascular And Neural Mechanisms
Authors: Wu Hanxue , Zhuang Jinzhu , Wang Jiawei , Yuan Shenglan , Sun Hui , Xu Jiaxi
Published: September 2022

Hypertension (HTN) has now been associated with cognitive impairment and considered as one of the risk factors for mild cognitive impairment (MCI). To investigate whether there is neuronal mechanism, HTN was induced in C57BL/6j male mice (~22 weeks) by DOCA-salt treatment (DS, 1.4mg/g+1%NaCl, 28 days), a neurogenic model. Firstly, BP was monitored by tail-cuff (sham vs. DS: SBP= 116.1±7.6
vs.
145.0±8.2 mmHg). Then, behavioral study was performed to assess the cognitive function (CFN). In whole, compared to sham, the DS mice performed worse in all 4 tasks, including object location recognition (before
vs.
after: NLR%= 29.34±13.73
vs.
-20.81±10.70), novel object recognition (NOR%= 48.60±22.62
vs.
9.09±25.50), Barnes maze (latency T= 26.55±16.52
vs.
116.80±57.67s), and nesting (score= 4.00±0.92
vs.
2.43±1.55). Later, CBF was measured by laser doppler. The rise of CBF upon whisker-stimulation (WS) was lower in the DS mice (27.83±13.86
vs.
51.12±18.22 PU), while their basal CBF was not affected, suggesting that DS-induced MCI could be related to impairment of neuro-vascular coupling (NVC). Previously, we have showed the overactivation of brain renin-angiotensin system (RAS) in the onset of DS HTN. To identify the possible contribution from brain RAS, neuronal AT
1a
R were deleted (AT1N). In AT1N mice, DS-related impairments in CFN and NVC were attenuated, as evidenced by higher nesting scores (AT1N+DS
vs.
NT+DS: 3.75±1.89
vs.
2.43±1.55 ), better performance in Barnes maze (61.28±54.47
vs.
116.80±57.67 s) and grater augment in CBF upon WS (41.49±10.65
vs.
27.83±13.86 PU), compared to NT. Through LTP recording in CA1 neurons using both field and whole-cell modes, we identified that activation of neuronal AT
1
R could contribute directly to the development of MCI, thereby neuronal AT
1
R was selectively knocked down in the hippocampus of NT mice (AT1hN). After DS treatment, AT1hN showed no difference in SBP compared with the sham NT (149.0±14.7
vs.
150.4±6.9 mmHg), while their CFN showed improvement in both NOR and Barnes maze (NOR%= 11.70±25.78
vs.
-10.15±42.60; latency T= 71.72±47.60
vs.
113.70±65.64 s). In summary, our data suggest that neuron-expressing AT
1
R could participate in the onset of hypertension-associated MCI via both vascular and neural mechanisms.
Abstract 093: Association Of Preterm Birth And Low Birth Weight With The Circulatory And Urinary Renin-angiotensin System In Youth With Newly Diagnosed Primary Hypertension
Authors: Vincent Carol , Chen Ashton , South Andrew M
Published: September 2022

Background:
While the RAS is altered in youth born preterm and may play a role in development of HTN, this has not been investigated in youth with primary HTN. We hypothesized that lower gestational age (GA) and lower birth weight (BW) would be associated with higher Ang II, lower Ang-(1-7), and higher Ang II/Ang-(1-7) in plasma and urine.

Methods:
This is a secondary analysis of baseline data from a pilot prospective cohort study of 30 youth aged 5-17 years with newly diagnosed primary HTN per national guidelines. Exclusion criteria were heart or chronic kidney disease and diabetes mellitus. Exposures were GA, BW, and BW z-score on the continuous scale and as binary variables, preterm birth (GA <37 weeks) and low BW (<2500 g). RAS outcomes were Ang II and Ang-(1-7) measured in plasma and urine per well-validated RIAs; urinary values standardized to urine creatinine (Cr). Plasma renin activity and aldosterone were measured per well-validated methods. We estimated associations with unadjusted generalized linear models.

Results:
Median GA was 39 weeks [IQR 36, 40] with 28% (8/30) born preterm. Mean BW was 3.12 kg (
SD
0.65) with low BW in 17% (5/30). Preterm birth was associated with lower urine Ang-(1-7)/Cr and higher aldosterone/renin ratio and urine Ang II/Ang-(1-7) (Figure). Low BW was associated with higher plasma Ang-(1-7) and urine Ang II/Cr.

Conclusion:
Early-life risk factors were associated with intrarenal RAS alterations favoring Ang II activation and Ang-(1-7) suppression in youth with primary HTN. Our findings provide novel insight into the mechanistic role that the RAS may play in HTN development in youth born preterm or with low BW that differs from youth born at term who have HTN.
Abstract 100: Effects Of Upregulation Of Cyp4a In Dahl Salt-sensitive Rats And Ko Of Cyp4a2 And 4a3 In Ss.5
Bn
Rats On The Development Of Hypertension And Renal Injury
Authors: Strong Luke B , Murphy Sydney , Border Jane , Wang Shaoxun , Fan Letao , Fang Xing , Fan Fan , Roman Richard J
Published: September 2022

Mutations in CYP4F2 (rs2108622) and CYP4A11 (rs1126742) that inhibit the production of 20-HETE have been linked to hypertension in human genetic studies. We confirmed that these same variants are associated with hypertension and cognitive dysfunction in 4,286 elderly subjects in the
Atherosclerosis Risk in Communities Neurocognitive Study
. These studies establish that mutations in CYP enzymes that reduce 20-HETE promote hypertension, but the mechanisms remain controversial since 20-HETE has both pro- and antihypertensive actions. To address this question, we identified a homologous genetic deficiency in the formation of 20-HETE in Dahl S (SS) rats and created SS.5
BN
consomic and CYP4A transgenic SS rats to restore CYP4A expression and 20-HETE production, and CYP4A2 and CYP4A3 KO rats on the rescued SS.5
BN
background. Mean arterial pressure (MAP), renal and cerebral blood flow, proteinuria, and renal injury were compared in SS versus SS.5
BN
and CYP4A transgenic SS rats and in SS.5
BN
versus CYP4A2 and CYP4A3 KO rats. MAP increased from 117±2 to 158±5 mmHg (n=34) in SS rats fed a high salt (HS) diet for 3 weeks. Proteinuria rose from 50±4 to 403±30 mg/day. MAP increased less, from 108
+
5 to 133
+
5 mmHg (n=7) in CYP4A1 transgenic SS rats and from 109
+
2 to 132
+
2 mmHg (n=23) in SS.5
BN
rats. Proteinuria (141±15 and 169±11 mg/day), glomerular injury, and renal fibrosis were all significantly reduced in the CYP4A transgenic and SS.5
BN
rats compared to SS rats. KO of CYP4A2 had no effect on MAP (134
+
2 mmHg, n=23) or proteinuria (248
+
23 mg/day) relative to SS.5
BN
rats fed an HS diet for 3 weeks. In contrast, MAP and proteinuria were elevated to 148
+
3 mmHg and 350
+
23 mg/day in CYP4A3 KO rats (n=23). The myogenic response of renal and cerebral arteries and autoregulation of RBF and CBF were impaired in SS and CYP4A3 KO rats but were intact in CYP4A transgenic SS rats, SS.5
BN
and CYP4A2 KO rats. These findings indicate that a deficiency in the formation of 20-HETE that impairs the myogenic response of renal and cerebral arteries, autoregulation of RBF and CBF, and increases glomerular capillary pressure, promotes the development of hypertension, proteinuria, and renal injury in genetically susceptible individuals and SS rats.
Abstract 102: Mitoquinone Reduces Preeclamptic Phenotypes Associated With Syncytiotrophoblast-Specific Gq Signaling
Authors: Opichka Megan A , Livergood M C , Gibson-Corley Katherine N , Fogo Agnes B , Kwitek Anne E , Sigmund Curt D , McIntosh Jennifer J , Grobe Justin L
Published: September 2022

Syncytiotrophoblast (STB) stress is theorized to drive the pathogenesis of preeclampsia (PE), and Gq-activating hormones implicated in PE have been associated with placenta oxidative damage, mitochondrial dysfunction, and maternal symptomology. As the causes and consequences of STB stress remain largely unknown, we used an in vivo model to determine whether excess STB-specific Gq signaling is sufficient to cause phenotypes of PE and if a mitochondrial-targeted antioxidant (mitoquinone) mitigates these effects. Activation of Gq signaling in STB cells was achieved by crossing dams harboring a cre-dependent Gq-coupled DREADD (hM3Dq) with
Gcm1
-Cre
+/-
sires and injecting clozapine N-oxide (CNO) or saline mid-gestation (GD12.5-14.5) before tissue collection at GD14.5. Gq activation resulted in maternal proteinuria (n=9-11 CNO 43±4 vs saline 28±3mg/day; p=0.01), mild segmental congestion of glomerular capillaries, and elevated circulating IP-10 (p=0.03), MCP-5 (p=0.04), and IL-12p40 (p=0.01). hM3Dq stimulation reduced labyrinth vascularization (n=10 Cre
+
19±1%, Cre
-
24±1%, saline 23±1%; p<0.01 Cre
+
vs each), spiral artery diameter (n=7-11 Cre
+
110±11μm, Cre
-
135±9, saline 149±12 μm; p=0.03), placenta mass (n=10-13 Cre
+
0.12±0.01, Cre
-
0.14±0.01, saline 0.13±0.01g; p<0.01), and fetal mass (n=10-13 Cre
+
0.23±0.03, Cre
-
0.30±0.02, saline 0.29±0.02g; p<0.01). Preliminary data suggest a placental oxidative defense response to Gq induction based on increased protein levels of SOD2 and catalase, and decreased malondyaldehyde. Mitoquinone (5mg/kg) corrected maternal proteinuria (n=8 22±4mg/day, p<0.01), decreased circulating IL-12p40 (p=0.03), and protected both labyrinth vascularization (n=8 23±1%, p=0.05) and spiral artery diameter (n=8 164±18μm, p=0.02). These data indicate that elevated Gq signaling in the STB layer is sufficient to cause pathological features of PE, via a mechanism involving mitochondrial stress.
Abstract 106: In Utero Morphine Exposure Dysregulates The Endogenous Opioid System (EOS) And Increases Risk Factors For Cardiovascular Disease In The Adulthood Offspring
Authors: Ahmed Nermin , KASSIS ALANA , MALONE JENA , ZAMZAMI ESRAA , Gordon Scott M , Dalmasso Carolina , Loria Analia S
Published: September 2022

Chronic opioid exposure and opioid use disorder are associated with nearly a 2-fold increased risk of CVD and 16% increase in coronary artery disease. Opioid abuse during pregnancy increases 5 fold the incidence of neonatal opioid withdrawal syndrome (NOWS) and is associated with low birth weight; however, the effect of OUD among pregnant women on their offspring health is understudied. Previously, we have shown that both male and female offspring from dams exposed to mu-opioid receptor agonist morphine (MOR) during the whole pregnancy are smaller and display high blood pressure. This study aimed to investigate the potential underlying mechanisms associated with increased blood pressure in MOR- exposed offspring. Sprague Dawley dams were treated with escalating doses of MOR (5-20 mg/kg/day, s.c) or saline (VEH) from gestational day 1 to 18, and pups were weaned on standard diet. Lipid profile was assessed as a risk factor for coronary artery disease using FPLC. MOR exposed offspring showed increased LDL (F-MOR vs. VEH ~2 fold; M-MOR vs. VEH ~1.5 fold mg/dL; n=6-8, p<0.05). Leptin levels, normalized to body fat, were not different between groups (F-MOR 219.8±35 vs F-VEH 181.4±15.8 pg/g fat; M-MOR 207.2±26.7 vs M-VEH 200.3±23.3 pg/g fat, n=7-8). Although proteinuria was not different in MOR exposed rats vs. VEH (F-MOR 4.6±0.8 vs F-VEH 5.5±1mg/ml; M-MOR 29.1±1.4 vs M-VEH 37.5±2.95 mg/mL; n=7-8), creatinine clearance was reduced in MOR-exposed males but not females (M-MOR 0.51±0.1 vs M-VEH 0.87±0.3 mL/min, n=7-8). MOR-exposed offspring showed a greater reduction in the pressor response to mecamylamine (F-MOR -46.25±6 vs F-VEH -18.3±1.3, M-MOR -50.5±0.28 vs M-VEH -25±5.1 delta mmHg, p<0.05). Proenkephalin, an endogenous opioid peptide that has been shown to inhibit the beta adrenergic receptor signaling, showed a ~3-fold reduction in heart and a ~7.5-fold reduction in the kidney cortex and outer medulla mRNA expression vs. VEH (-2ddCt, n=6-8, p<0.05). Maternal MOR exposure increases the sympathetic tone, which could be associated with increased sympathetic outflow to the kidney and the heart. Reduced proenkephalin could be involved in potential crosstalk between adrenergic and opioid receptor signaling exacerbating the actions of catecholamines.
Abstract 107: Adoptive Transfer Reveals That T Cell-Derived Reactive Oxygen Species Contributes To Pregnancy-Induced Renal Damage In The Dahl SS Rat
Authors: Dasinger John Henry , Abais-Battad Justine M , Walton Samuel , Burns Emily , Shaw Mary , Mattson David L
Published: September 2022

Previous work from our lab demonstrated that Dahl SS rats develop a pregnancy-specific increase in blood pressure and proteinuria, also known as maternal syndrome (MS). Both T cells and reactive oxygen species (ROS) have been implicated in MS, and genetic deletion of either resulted in protection from MS in the Dahl SS rat. Therefore, we tested the hypothesis that ROS produced from T cells contributes to the development of MS in the Dahl SS. An adoptive transfer of splenocytes was utilized to reconstitute T cells from wild type Dahl SS (SS→CD247) or SS
p67phox-/-
(p67phox→CD247), which lack the ability to generate NOX2-derived ROS, into SS
CD247-/-
rats by intraperitoneal injection at postnatal day 5 to investigate the oxidative capacity of T cells. Injected SS
CD247-/-
rats were mated at 9-11 weeks of age with naïve littermate males, and virgin controls were run in parallel. Baseline protein excretion rates were comparable between mated groups (72±12 vs 66±9 mg/day,
p>
0.05, n=10/group, SS→CD247 vs p67phox→CD247, respectively), yet the SS→CD247 develop an increase in proteinuria throughout pregnancy that is significantly elevated relative to p67phox→CD247 rats (217±65 vs 114±14 mg/day,
p<
0.05). There was a trending decrease in creatinine clearance in the SS→CD247 vs p67phox→CD247 at the end of the study (0.37±0.03 vs 0.46±0.06,
p
=0.1), but MAP did not differ between groups. There was however, an increase in uterine artery resistance index as assessed by Doppler ultrasound at GD20 in the SS→CD247 relative p67phox→CD247 (0.72±0.03 vs 0.61±0.02,
p
<0.01) indicating an impairment in blood flow leading to the placenta. While, there was no difference observed in the number of infiltrating leukocytes into the kidneys between SS→CD247 vs p67phox→CD247, a significant correlation was observed between the number of infiltrating CD4+ T cells and the late pregnancy proteinuria levels (R
2
=0.7973) in the SS→CD247 mated group while no correlation was observed in the p67phox→CD247 (R
2
=0.0472). Taken together, adoptive transfer of T cells lacking NOX2 into SS
CD247-/-
rats fails to induce pregnancy associated renal damage and uterine artery blood flow impairment indicating a potential role for ROS in MS.