bluuebunny/crossref_metadata_2025 huggingface dataset:
PS-BPC06-5: EXOGENOUS THYROXINE INCREASES GLUT4 TRANSLOCATION IN MEMBRANE AND IMPROVES CARDIAC METABOLISM INDEPENDENT OF REDUCED SBP IN INSULIN RESISTANT OLETF RATS
Authors: Mendez Dora Alicia , Organis Jose Guadalupe Sonanez , Anaya Guillermo Vasquez , Nakano Daisuke , Nishiyama Akira , Ortiz Rudy Martin
Published: January 2023

Objective:

During insulin resistance the heart undergoes a metabolic shift where fatty acids (FA) account for about 99% of the ATP production, indicative of impaired glucose metabolism. This results in increased reactive oxygen species and lipotoxicity leading to impaired mitochondrial function. Thyroid disorders such as hyperthyroidism and hypothyroidism both have the potential to lead to hypertension. Thyroid hormones (TH) may improve glucose intolerance by increasing glucose reabsorption and metabolism and therefore improve cardiac function and lower the risk of developing hypertension. However, incongruent data exists showing that TH promote oxidative stress and cardiac hypertrophy. Thus, the beneficial effects of TH in the heart during insulin resistance remains controversial. The glucose transport protein, GLUT4, is a downstream target of the TH response element indicating a possible mechanism by which TH regulate glucose metabolism.

Design:

Insulin resistant, Otsuka Long Evans Tokushima Fatty (OLETF) rats were used to assess the effects of exogenous thyroxine (T4) on glucose metabolism in cardiac tissue. Rats were assigned to four groups: 1) lean, Long Evans Tokushima Otsuka (LETO; n = 6), 2) LETO + T4 (8 μg/100 g BM/d × 5 wks; n = 7), 3) untreated OLETF (n = 6), and 4) OLETF + T4 (n = 7). T4 was administered via osmotic minipumps implanted subcutaneously. Systolic blood pressure (SBP) was measured weekly in conscious rats by tail-cuff plethysmography. Animals were provided water and food ad libitum.

Results:

SBP increased by 15% in OLETF compared to lean LETO (138 ± 2 vs. 117 ± 1 mm Hg), but T4 did not alter SBP is OLETF strain. T4 increased GLUT4 gene expression by 85% and GLUT4 protein translocation to the membrane by 328% in OLETF. Additionally, T4 increased phosphofructokinase-1, the rate limiting step in glycolysis, by 98% and decreased CD36 gene expression for FA transport by 20% in OLETF. We expect our recent metabolomics and lipidomics data to help better explain these molecular changes.

Conclusions:

Five-week dose of exogenous T4 did not reduce SBP is OLETF. The results suggest that increased T4 has the potential to increase glucose reabsorption and metabolism in the heart of insulin resistant rats and restore impaired substrate metabolism, but these improvements in substrate metabolism do not translate to sustained reduction in arterial pressure.
PS-BPC06-9: THE EFFECT OF COMMON SURGICAL MASKS ON UNATTENDED BLOOD PRESSURE CHANGES IN DRUG TREATED HYPERTENSIVE PATIENTS
Authors: Polyzos Dimitris , Drogkaris Sotiris , Tatakis Fotis , Manta Eleni , Fragkoulis Christos , Siafi Eirini , Filippou Christina , Kariori Maria , Chalmoukou Konstantina , Konstantinidis Dimitris , Thomopoulos Costas , Tsioufis Konstantinos
Published: January 2023

Objective:

While there are several studies that have focused on the role of face masks in preventing airborne transmission of SARS-CoV-2, few data are available on their effects on physiological measures, and no study has examined their effects on blood pressure (BP). The purpose of our study was to investigate the effect of surgical masks on BP in drug-treated hypertensive patients who had a routine follow-up visit to a university hospital outpatient hypertension clinic.

Design and method:

The study included already treated hypertensive patients aged > 18 years, while the exclusion criteria were atrial fibrillation or any other arrhythmia affecting the BP measurement, an arm circumference > 42 cm, mental disorders, Parkinson disease, pregnancy, intolerance to the BP measurement method, or unwillingness to participate. A new surgical mask was provided to all participants to replace the face mask that was already in use. After the routine mask-on office BP measurement, patients were left alone and randomized to automated office BP measurement, with measurements taken after first wearing a mask for 10 min, then without wearing the mask for 10 min, and vice versa.

Results:

A total number of 265 patients were included in the study. Among the participants, 115 were women (43.4%), the mean age was 62 ± 12 years, and the mean office BP was 134 ± 15 / 81 ± 12 mmHg. There was no significant difference between mask-on unattended systolic BP (133 ± 15 mmHg) and mask-off unattended systolic BP (132 ± 15 mmHg) (P = 0.13) or between mask-on unattended diastolic BP (77 ± 13 mmHg) and mask-off unattended diastolic BP (76 ± 13 mmHg) (P = 0.32). Moreover, there was no difference in the heart rate (mask-on first, 69 ± 11 bpm; mask-off first, 69 ± 11 bpm, P = 0.7).

Conclusions:

Common surgical masks do not affect systolic/diastolic BP levels during unattended BP measurements in treated hypertensive patients.
PS-BPC07-1: PREEXISTING DIURETIC THERAPY DOES NOT ATTENUATE THE ANTIHYPERTENSIVE EFFECT OF EMPAGLIFLOZIN IN TYPE 2 DIABETIC PATIENTS WITH UNCONTROLLED HYPERTENSION
Authors: Kuzior Agnieszka , Hernandez-Lazaro Alba , Leon-Durango Ricardo de , Rios-Gomez Carlos , Santana-Ojeda Borja , Perez-Rivero Jennifer Maria , Perdomo-Herrera Esperanza , Tocino-Hernandez Alba Lucia , Gonzalez-Diaz Paula , Arnas-Leon Claudia , Acosta-Calero Carmen , Martinez-Martin Francisco Javier
Published: January 2023

Objective:

The antihypertensive effect of SGLT2 inhibitors has been mostly overlooked, but in patients with type 2 diabetes mellitus (T2DM) and uncontrolled hypertension empagliflozin elicits a powerful systolic blood pressure (SBP) lowering effect, often attributed to diuretic/natriuretic effect and/or weight loss (WL), but other mechanisms are possible. We sought to retrospectively study the impact of the initiation of empagliflozin therapy on SBP in patients living with T2DM and uncontrolled hypertension in a real-world setting, and to assess its relationship with previous diuretic and other classes of antihypertensive therapy, and with weight loss during treatment.

Design and Methods:

Retrospective analysis of the SBP, weight and antihypertensive medication data in patients with T2DM who initiated empagliflozin therapy while having sitting office SBP > 140 mmHg. Mediation analyses with WL as mediator were performed post-hoc to quantify the relative contribution of WL to SBP reduction. Consent was obtained from all included subjects. Patients who changed their antihypertensive medication simultaneously with empagliflozin initiation were excluded.

Results:

Data were obtained from 198 patients (59.6% female, age 63.2 ± 8.9 years) from the visit in which empagliflozin (largely 10 mg/day) was started and the following one (3–4 months afterwards). Baseline BMI was 31.7 ± 8.5 Kg/m2. Baseline SBP was 166 ± 17 mmHg. 112 patients had ACEI/ARB, 76 diuretic, 69 calcium antagonist, 32 alpha-blocker and 29 beta-blocker therapy. The SBP reduction was 7.2 ± 5.7 mmHg (p = 0.012, paired t-test). The Figure shows comparisons by number and classes of antihypertensive drugs, with no significant differences for SBP reduction. The WL was 3.2 ± 1.7 kg, and it correlated significantly with the SBP reduction (Pearson's r: 0.22, p = 0.0012). The mediation analyses showed that a SBP reduction of 1.5 ± 0.9 mmHg was mediated by WL while 5.7 ± 4.9 mmHg was WL-independent.

Conclusions:

Empagliflozin initiation elicited a clinically relevant SBP decrease in patients with T2DM and uncontrolled hypertension. The number and classes of previous antihypertensive effects did not modify the SBP reduction. In particular, previous diuretic treatment does not attenuate it, suggesting that the diuretic effect is not the main driver of SBP reduction with empagliflozin. Also, the contribution of WL seems modest, implying alternative mechanisms for SBP reductions with empagliflozin; improvement of endothelial function and/or arterial stiffness and many others have been suggested.
PS-BPC10-2: A NOVEL PREDICTION MODEL OF ACUTE KIDNEY INJURY BASED ON COMBINED BLOOD VARIABLES IN STEMI
Authors: Goriki Yuhei , Tanaka Atsushi , Node Koichi
Published: January 2023

Background:

Development of acute kidney injury (AKI) is associated with poor prognosis in patients with ST-segment elevation myocardial infarction (STEMI).

Objective:

We sought to investigate whether a combination of pre-procedural blood tests could predict the incidence of AKI in patients with STEMI.

Methods:

A total of 908 consecutive Japanese patients with STEMI who underwent primary percutaneous coronary intervention within 48 hours of symptom onset were recruited and divided into derivation (n = 617) and validation (n = 291) cohorts. A risk-score model was created based on a combination of parameters assessed on routine blood tests on admission.

Results:

In the derivation cohort, multivariate analysis showed that the following 4 variables were significantly associated with AKI: blood sugar ≧ 200 mg/dL (odds ratio [OR] 2.07), high-sensitive troponin I > 1.6 ng/mL (upper limit of normal × 50) (OR 2.43), albumin ≧ 3.5 mg/dL (OR 2.85) and estimated glomerular filtration rate < 45 mL/min/1.73 m2 (OR 2.64) (Table 1). Zero to four points were given according to the number of those factors. Incremental risk scores were significantly associated with a higher incidence of AKI in both cohorts (p < 0.001) (Figure 1). Receiver operating characteristic curve analysis of risk models showed adequate discrimination between patients with and without AKI (derivation cohort: area under the curve [AUC] 0.754, 95% confidence interval [CI] 0.733 - 0.846; validation cohort: AUC 0.754, 95% CI 0.644 - 0.839) (Figure 2).

Conclusions:

Our novel laboratory-based model might be useful for early prediction of the post-procedural risk of AKI in patients with STEMI.
PS-BPC11-5: INCIDENCE OF CLINICAL HYPERTENSION IN YOUNG TRANSGENDER PEOPLE AFTER A FIVE-YEAR FOLLOW-UP: ASSOCIATION WITH GENDER-AFFIRMING HORMONAL THERAPY
Authors: Hernandez-Lazaro Alba , de Leon-Durango Ricardo , Rios-Gomez Carlos , Santana-Ojeda Borja , Perez-Rivero Jennifer Maria , Fernandez-Trujillo-Comenge Paula Maria , Santana-Suarez Ana Delia , Gonzalez-Diaz Paula , Arnas-Leon Claudia , Acosta-Calero Carmen , Perdomo-Herrera Esperanza , Kuzior Agnieszka , Sanchez-Bacaicoa Maria del Sol , Perez-Garcia Maria del Pino , Martinez-Martin Francisco Javier
Published: January 2023

Introduction and Objective:

There is a paucity of data on the long-term cardiovascular side effects of hormonal therapy in transgender people. The most overlooked issue is the cardiovascular risk associated with the use of androgen blockers. While some short-term data on blood pressure effects of gender-affirming hormonal therapy are available, the risk of hypertension development is unknown. We sought to assess this risk in our transgender patients.

Design and Methods:

Retrospective analysis of the clinical records of consecutive transgender patients who began hormonal therapy in our Outpatient Gender Identity Clinic with < 30 years of age and have a follow-up of at least 5 years. Patients who had office sitting BP > 140/90 mmHg at least twice, had an ABPM or systematic HBPM register with > 135/85 mmHg or were taking antihypertensive medication were diagnosed with hypertension. Mediation analyses with weight increase as mediator were performed post-hoc to quantify its relative contribution to the increase in blood pressure. Patients who were already living with hypertension at the onset of hormonal therapy, withdrew from it or switched androgen blockers were excluded. All subjects gave informed consent.

Results:

129 transgender women treated with estradiol and 153 transgender men treated with testosterone were included. Their onset and 5-year follow-up age, blood pressure and body weight are shown in the Table (women are stratified by type of androgen blocker) along with the incidence of hypertension. The increase in body weight was strongly correlated with the SBP increase (Spearman's r: 0.361, p < 0.001). Mediation analyses showed that 31.2% - 42.3% (95% CI) of the SBP increase was mediated by body weight increase but the rest was body weight-independent.

Conclusions:

The annual incidence of hypertension in young transgender men (1.17%) seems comparable to that of the general population, but in young transgender women the choice of androgen blocker has a remarkable effect on the incidence of hypertension, with apparent protective effect of spironolactone but a striking risk increase with cyproterone acetate (close to 5% yearly). Taken together with the known risks of this drug (meningioma, depression, low libido, liver toxicity, thrombosis…), the use of cyproterone acetate for this indication should be reconsidered. Body weight increase is undoubtedly related to the increase of BP transgender people on gender-affirming hormone therapy but is probably not the main factor.
PS-BPP01-3: DIFFERENTIAL ASSOCIATION OF CIRCULATING PROPROTEIN CONVERTASE SUBTILISIN/KEXIN TYPE 9 WITH THE SUBCLINICAL CEREBRAL SMALL VESSEL DISEASE AND INTRACRANIAL ARTERY STENOSIS
Authors: Kunimura Ayako , Yano Yuichiro , Torii Sayuki , Kondo Keiko , Hisamatsu Takashi , Kadota Aya , Fujiyoshi Akira , Okamura Tomonori , Watanabe Yoshiyuki , Shiino Akihiko , Nozaki Kazuhiko , Ueshima Hirotsugu , Miura Katsuyuki
Published: January 2023

Background and aims:

Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a new target for reducing low density lipoprotein cholesterol (LDL C) and incident cardiovascular disease, including stroke. However, how PCSK9 work on developing stroke is not fully elucidated. In this study we assessed the association of circulating PCSK9 levels with subclinical cerebrovascular disease, including cerebral small vessel diseases (CSVD) and intracranial artery stenosis (ICAS), in a general population.

Methods:

In community dwelling Japanese men (n = 526) aged 46–82 years, without a history of lipid lowering medications or cardiovascular disease, we assessed the associations of serum PCSK9 levels with CSVD and ICAS using magnetic resonance imaging. CSVD included lacunar infarction, deep and subcortical white matter hyperintensity, periventricular hyperintensity, cerebral microbleeds. Multivariable Poisson regression with robust error variance was performed to estimate the relative risk (RR) and 95% confidence interval (CI) of the serum PCSK9 levels for the presence of CSVD and ICAS. Additionally, ordinal logistic regression was performed to estimate odds ratio (OR) and 95% CI for ICAS, with graded detectable stenosis, mild (1%-50%) and severe (> 50%).

Results:

The average (standard deviation: SD) age at baseline and median (interquartile range) PCSK9 were 68 (9) years and 240 (205 - 291) ng/mL, respectively. After multivariable adjustment including traditional cardiovascular risk factors (e.g., LDL C), a 1- SD increase of PCSK9 levels was associated with ICAS (adjusted RR 1.18, 95% CI 1.02 - 1.37), and the ordinal logistic regression model also showed a similar association (adjusted OR 1.31, 95% CI 1.04 - 1.64). However, there was no significant association between serum PCSK9 levels and CSVD.

Conclusions:

Higher circulating PCSK9 level was associated with the higher prevalence of ICAS but not with CSVD, and the association was independent of traditional cardiovascular risk factors including LDL-C. The results of this study imply the direct and independent association of PCSK9 with cerebral atherosclerotic plaque formation.
PS-BPR01-10: PHEOCHROMOCYTOMA MASQUERADING AS A LONG-TERM SEVERE ANXIETY DISORDER WITH PANIC ATTACKS REFRACTORY TO PSYCHIATRIC TREATMENT
Authors: Santana-Ojeda Borja , Hernandez-Lazaro Alba , de Leon-Durango Ricardo , Rios-Gomez Carlos , Perez-Rivero Jennifer Maria , Kuzior Agnieszka , Gonzalez-Diaz Paula , Arnas-Leon Claudia , Acosta-Calero Carmen , Martinez-Martin Francisco Javier
Published: January 2023

Introduction and Objective:

Pheochromocytoma is often diagnosed in the workup of a patient with severe, refractory hypertension, but nowadays many silent pheochromocytomas are diagnosed in the workup of an incidental adrenal mass. Anxiety is also a common symptom in patients with pheochromocytoma and occasionally may be the only clinical manifestation. Hereby we describe the case of a patient with a severe refractory anxiety disorder as a bizarre presentation of pheochromocytoma.

Design and Methods:

Review of the patient's records and of the relevant literature.

Results (Case Description):

A 67-year old woman was referred to our Endocrinology Clinic for the workup of an incidental left adrenal mass found in an abdominal CT ordered in the nephrolithiasis workup after a renal colic. The patient had no previous relevant history of somatic disease. She had been suffering from anxiety disorder since she was 50, beginning with climacteric symptoms (hot flashes, irritability and mood swings) followed by severe anxiety, frequent panic attacks, insomnia and inability to concentrate. Hormonal replacement therapy had been ineffective. In the following 17 years she had consulted many public and private psychiatrists and had been treated with combinations of anxiolytic and antidepressant drugs including lormetazepam, clonazepam, bromazepam, vortioxetine, quetiapine, venlafaxine, agomelatine, risperidone, mirtazapine, but the patient's anxiety symptoms kept deteriorating. BP up to 180/110 mmHg and tachycardia up to 130 bpm had been recorded during anxiety crises but ABPM ruled out sustained hypertension or tachycardia. After the finding of the adrenal lesion the diagnostic process was straightforward. It was a solid, heterogeneous mass of about 6 cm diameter, with necrotic areas and mean densitometry of 86 UH. The lab workup showed normal creatinine and potassium, suppressed cortisol after 1 mg dexamethasone, normal ACTH, aldosterone, renin and aldosterone/renin ratio, but plasma metanephrines were above 5x the upper normal limit (634/2423 pg/mL). A MIBG scan confirmed intense uptake in the left adrenal area without abnormal extra-adrenal deposits. After standard preparation, the tumor was laparoscopically removed without complications. 1 month after the surgery plasma metanephrines were normal and the anxiety symptoms had vanished almost totally, but the patient kept taking a low dose of risperidone (1.5 mg/day) as a maintenance therapy; BP and HR remained normal

Conclusions:

Pheochromocytoma/paraganglioma should be included in the workup of patients with severe anxiety disorder, especially when refractory to treatment.
PS-B03-3: THE CD153-CPG VACCINE IS A NOVEL SENOTHERAPEUTIC OPTION TO REMOVE SENESCENCE-ASSOCIATED T CELLS
Authors: Yoshida Shota , Nakagami Hironori , Rakugi Hiromi
Published: January 2023

Cellular senescence in primary culture cells has allowed us to understand basic individual tissue aging and cancer inhibition from the view of cellular aging. Senescent cells accumulate gradually and systemically with age, and secrete various inflammatory cytokines or chemokines, which is different from the cell fate in apoptosis. These phenomena describe the so-called senescence-associated secretory phenotype, and chronic inflammation and carcinogenesis are induced in the surrounding tissues through senescence-associated secretory phenotype factors. The senescence-associated T cells do not proliferate in response to T-cell receptor stimulation, and produce abundant inflammatory cytokines including osteopontin. Senescence-associated T cells are also increased in adipose tissue under a high-fat diet, which might be related to the progress of obesity or diabetes.

We evaluated the long-lasting effect of using CD153 vaccination to remove senescent T cells from high-fat diet (HFD)-induced obese C57BL/6J mice. We administered a CD153 peptide-KLH (keyhole limpet hemocyanin) vaccine with CpG oligodeoxynucleotide (ODN) 1585 (CD153-CpG) and confirmed an increase in anti-CD153 antibody levels that was sustained for several months. After being fed a HFD for 10–11 weeks, adipose senescent T cell accumulation was significantly reduced in the VAT of CD153-CpG-vaccinated mice, accompanied by glucose tolerance and insulin resistance. A histological analysis also indicated that the accumulation of macrophages and senescence-associated T cells in adipose tissues successfully reduced in the CD153-CpG-vaccinated mice.

The CD153-CpG vaccine is an optional tool for senolytic therapy. We further modify the vaccine to remove the target cells now.
PS-B05-11: IMPAIRED CELL INTERACTIONS AT THE PRE-ECLAMPTIC MATERNAL-FOETAL INTERFACE
Authors: Nonn Olivia , Debnath Olivia , Valdes Daniela , Sallinger Katja , Secener Ali Kerim , Haider Sandra , Tiesmeyer Sebastian , Coscia Fabian , Heliebi Amin El , Tilaganathan Basky , Mueller Dominik N , Huppertz Berthold , Gauster Martin , Ishaque Naveed , Herse Florian
Published: January 2023

Hypertensive disorders in pregnancy, of which the multisystem syndrome pre-eclampsia is most severe, leading to preterm delivery, maternal mortality and life-long complications1. Pre-eclampsia lacks early screening tools2,3 and causal therapies4,5. To elucidate early disease dynamics, we present the first spatio-temporal study comparing single-nuclei transcriptomes of human preterm pre-eclamptic placentae and healthy controls, contextualising this in a comprehensive study including early and late gestational placentae and implementing spatial multi-omics methods.

Here we show trophoblast pre-fusion cells and their developmental trajectory running towards a novel trophoblast cell-state we identify as juvenile syncytiotrophoblast. We found that disturbed trophoblast development, reconstructed by spatial multi-omics and including early dysregulation in differentiation-driving transcription factor p300, likely initiates pre-eclampsia. We suggest pre-eclampsia to be a disease of dysregulated syncytiotrophoblast differentiation: Disease-specific premature differentiation of the maternal-foetal barrier leads to a senescent syncytiotrophoblastic interaction through maternal circulation with maternal vessels. This initiates a disrupted decidual maternal response via LEP, GDF15 and INHBA.

Our results highlight a disturbed cellular communication from foetal to maternal side during the development of pre-eclampsia starting with a disturbed trophoblast maturation. We provide new targets for potential early disease prevention in order to protect mother and child from increased gestational mortality and morbidity but also from life-long increased cardiovascular disease risk.
PS-B06-8: EXPLORING THE ROLE OF NECROPTOSIS IN HYPERTENSION
Authors: ODonnell Joanne A , Dinakis Evany , Jones Dakota Rhys , Muralitharan Rikeish M , Marques Francine Z
Published: January 2023

Objective:

Altered immune cell activation plays a key role in promoting hypertension. A major question remains as to the immune system becomes activated during hypertension. A possibility is necroptosis, which a newly described form of cell death alerts the immune system to dying cells, causing inflammation. Using mice genetically unable to undergo necroptosis, we aimed to determine whether necroptosis contributes to inflammation and fibrosis or alters blood pressure during hypertension. To determine whether factors present during hypertension impact necroptosis, we examined whether Angiotensin II sensitizes human and murine cells to necroptosis.

Design and Methods:

Monocytes were sorted from peripheral blood mononuclear cells from 3 normotensive and 3 untreated hypertensive participants diagnosed with 24-h blood pressure (BP) monitoring. Necroptosis was induced in human monocytes using hTNFα (10ug/ml), SMAC mimetic (500 nM), zVAD (20uM), and in primary neonatal murine cardiomyocytes and fibroblasts from 3 litters of C57BL/6J mice using mTNFα (10ug/ml), SMAC mimetic (100 nM), zVAD (10uM), in the presence or absence of angiotensin II (Ang-II, 5 ug/ml). Viability of human cells was measured using propidium iodide by flow cytometry, and in murine cells using CellTitre-Glo Luminescent Cell Viability Assay. Hypertension was induced in male Mlkl-/- mice, which cannot undergo necroptosis, and wild-type (WT) littermate controls by implanting minipumps containing Ang-II (0.5 mg/kg/day for 28 days; n = 6/genotype). Blood pressure (BP) was measured weekly by tail-cuff.

Results:

Monocytes from hypertensive participants were not more susceptible to necroptosis than normotensive participants (% viability ± SD; 48% ± 0.11 vs 51% ± 0.04, P = 0.79). This was not altered by Ang-II treatment in normotensive (51% ± 0.04 vs 56% ± 0.11 with Ang-II, P = 0.98) or hypertensive participants (48% ± 0.11 vs 56% ± 0.18 with Ang-II, P = 0.92). Moreover, Ang-II did not alter the viability of primary murine cardiomyocytes in vitro (42% ± 23 vs 40.1 ± 17.8 with Ang-II, P = 0.79). Primary mouse cardiac fibroblasts were unable to undergo necroptosis. In vivo, there was no difference in BP of Ang-II-treated Mlkl-/- and WT control mice at 4 weeks (P = 0.87).

Conclusions:

Our findings showed that, compared to normotensive participants, monocytes from hypertensive participants were not more susceptible to necroptosis. Furthermore, Ang-II did not directly act on the necroptosis pathway in both mouse cardiac fibroblasts or human monocytes. Changes in BP were not observed between WT and MLKL knockout mice upon Ang-II treatment, suggesting necroptosis does not impact BP regulation. Further analyses are being performed to determine whether inflammation or tissue fibrosis are altered in the absence of necroptosis.
PS-B06-9: SINGLE-CELL TRANSCRIPTOMIC PROFILING OF AORTIC B CELLS DURING EXPERIMENTAL HYPERTENSION
Authors: Galvao Hericka Bruna Figueiredo , Jelinic Maria , Dona Malathi S I , Farrugia Gabriella E , Hughes Tayla G , Diep Henry , Pinto Alexander , Drummond Grant R , Vinh Antony
Published: January 2023

Objective:

Inflammation and immunity are known to strongly influence aortic fibrosis leading to stiffening and remodelling. We have shown that genetic and pharmacological ablation of B cells affords protection from the development of aortic stiffening and remodelling associated with hypertension. Whether this protective phenotype is due to systemic effects such as reduced antibody secretion, or local B cell effector functions within the aorta remains unclear. Using single-cell transcriptomics, this study aimed to characterise the B cell populations present in the mouse aorta and identify phenotypes/effector functions which may drive inflammation and aortic remodelling.

Design and Methods:

Hypertension was induced in 12-week-old C57BL6/J mice by infusion of angiotensin II (0.7 mg/kg/day; s.c.) for 28-days via osmotic minipumps. Normotensive mice received vehicle infusion (0.5% NaCl, 0.1% acetic acid). Weekly systolic blood pressure (SBP) measurements were acquired by tail-cuff plethysmography. High-resolution ultrasound imaging was used for pulse wave velocity (PWV) measurements on day 28. Thoracic and abdominal aortas were then isolated and digested for single-cell transcriptomic analyses. Aortic suspensions were stained with Calcein Blue and Vybrant Ruby to sort for metabolically active, nucleated cells. RNA libraries were generated using the 10x Chromium platform.

Results and Conclusions:

Angiotensin II caused significant increases in SBPs (∼180 mmHg) compared to vehicle treated mice (∼120 mmHg). Hypertensive (HT) mice had increased aortic PWV (∼4 m/s) compared to normotensive (NT) mice (∼1 m/s). Single-cell transcriptomic analyses revealed a near 2-fold increase in B cells present in the aorta of hypertensive mice. Leiden-based community detection identified 7 clusters within the aortic B cell population, including pro-B cells, pre-BCR-independent B cells, transitional B cell populations, B1 and B2 B cells, as well as antibody secreting cells. A greater number of B2 B cells was observed in the hypertensive aorta (58% HT vs 36% NT), without an observable increase in B1 B cell numbers. Gene ontology (GO) analysis of aortic B2 B cells from angiotensin II-treated mice revealed an upregulation in biological processes associated with B cell proliferation (GO:0042100), B cell activation (GO:0042113) and leukocyte differentiation (GO:0002521) compared to vehicle controls. Interestingly, the majority of antibody secreting cells in the aorta appear to be derived from B1 B cells in both vehicle (∼82%) and angiotensin II-treated mice (∼96%). In summary, hypertension is associated with a divergent aortic B cell transcriptome reflective of an augmented immune response, with observable shifts in composition which may be associated with inflammation and aortic remodelling.
PS-B08-4: CHRONIC INTERMITTENT HYPOXIA INDUCES RENAL FIBROSIS BY MR ACTIVATION
Authors: Zhang Cui juan , Yang Fan , Chang Yi , Xiong Yun zhao , Shimosawa Tatsuo , Xu Qing you
Published: January 2023

Purpose:

Obstructive sleep apnea is mainly characterized by chronic intermittent hypoxia (CIH), which is associated with chronic kidney disease. Macrophage is known to play an important role in renal fibrosis. However, little is known of the potential role of macrophage-to-myofibroblast transition (MMT) in hypoxic conditions. The aim of this study was to understand how infiltrating macrophages may contribute to renal fibrosis in hypoxic conditions.

Methods:

In this study, 30 adult male sprague dawley (SD) rats were randomly divided into normoxic control(NC), chronic intermittent hypoxia(CIH), and eplerenone(EPL) groups-each containing ten rats. The CIH and EPL groups were exposed to hypoxia in a commercial hypoxic chamber for 5 weeks. Rats in the EPL group were administered EPL orally via diet for 5 weeks. All rats were then sacrificed and serum, 24-hour urine and tissue specimens were collected for renal function, histopathology, western blot, immunohistochemistry. The RAW264.7 macrophages were subjected to 24 hours of hypoxia and 12 hours of reoxygenation. Cells were collected for immunofluorescence staining.

Results:

In addition to blood pressure, renal aldosterone level was also increased in CIH rats. Massons trichrome and Sirius red staining showed significantly higher collagen deposition and fibrosis in CIH rats. A markedly increased co-expression of α-SMA and CD68 in vitro and in vivo after CIH exposure. MR nuclear translocation was seen in hypoxic macrophages. Western blot showed that the expression of SGK-1 and p-SGK-1 was upregulated. These effects were reversed by MR blocker (EPL).

Conclusions:

CIH-induced macrophage infiltration, an important source of myofibroblasts, might participate in the development of renal interstitial fibrosis. These effects were reversed by EPL. Therefore, we speculate MR activation may be related to CIH-induced renal fibrosis.
PS-B08-6: ROLE OF THE HMGB-1/RAGE AXIS AND MINERALOCORTICOID RECEPTOR IN THE PATHOGENESIS OF ACUTE KIDNEY INJURY
Authors: Otsuka Tomoyuki , Ueda Seiji , Nagasawa Hajime , Okuma Teruyuki , Nakata Masami , Sato Koji , Kobayashi Takashi , Murakoshi Maki , Gohda Tomohito , Matsui Takanori , Yamagishi Shoichi , Suzuki Yusuke
Published: January 2023

Background:

Endothelial dysfunction plays a central role in the pathogenesis of cardio-renal syndrome. High mobility group box-1 (HMGB-1) is a protein with various roles in different cellular compartments, and indirectly regulates the activity of transcription and DNA repair in the nucleus. On the other hand, during tissue damage, it is released into the extracellular environment as damage-associated molecular patterns (DAMPs). HMGB-1 is elevated in the pathogenesis of acute kidney injury (AKI) and be involved in endothelial dysfunction through binding to toll like receptor (TLR) and receptor for advanced glycation end products (RAGE). In addition, we recently demonstrated that RAGE-mediated Rac1 activated mineralocorticoid receptor (MR) and subsequently resulted in podocytes damage. In the present study, we hypothesized that crosstalk between HMGB-1/ RAGE and Rac1-MR pathways could contribute to endothelial dysfunction in AKI.

Methods:

In the present study, we performed a renal ischemia-reperfusion model (I/R group) on C57BL/6J mice to evaluate the expression of Rac1-MR pathway and organ damage. In addition, the I/R group was pretreated with MR blocker (esaxerenone) and evaluated in the same way. In vitro study, we investigated whether HMGB-1 could activate Rac1-MR axis and thus induce endothelial injury in cultured human umbilical vein endothelial cells (HUVECs) by assessing expression levels of genes for MCP-1 and NF-κB with or without administration of RAGE aptamer or MR blocker.

Results:

In the I/R group, MR activation assessed by translocation into the nucleus and enhanced RAGE expression were observed along with HMGB-1 and RAGE elevation, and pretreatment with MR blocker suppressed the elevation of serum creatinine levels as well as tubular necrosis. Further, the loss of peritubular capillaries observed in I/R kidney was markedly suppressed by MR blocker. In vitro study, HMGB-1 supplementation significantly increased RAGE expression, GTP-bound Rac1 and enhanced MR translocation into the nucleus in HUVECs as well. We also found that HMGB-1 upregulated MCP-1 and NF-κB, all of which were significantly blocked by pretreatment of RAGE aptamer and MR blocker.

Conclusion:

These results suggest that there may be a close relationship between HMGB-1/ RAGE axis and Rac1/ MR activation, thus contributing endothelial injury. Using RAGE aptamer or MR blocker could be novel therapeutic strategies against endothelial dysfunction in patients with AKI.
PS-B08-8: THERAPEUTIC STRATEGIES TARGETING RAGE IMPROVES RENAL OUTCOME IN LUPUS NEPHRITIS
Authors: Taguchi Kensei , Ito Sakuya , Yokota Yunosuke , Kodama Goh , Higashimoto Yuichiro , Fukami Kei
Published: January 2023

Introduction:

Lupus nephritis (LN) is present in 60% of patients with systemic lupus erythematosus (SLE). Despite of current development of immunosuppressant agents, LN still impairs the survival and quality of life in SLE patients. Receptor for advanced glycation endproducts (RAGE) is a multi-ligand receptor that belongs to the immunoglobulin superfamily, which is strongly associated with innate immune system. In the present study, we examined whether RAGE is involved in the development of LN. Further, we explored the therapeutic impact of DNA-aptamer directed against RAGE on lupus-related kidney injury.

Methods:

[Protocol 1] RAGE expression in kidneys and urinary RAGE excretion (uRAGE) were determined at 8, 12, 16, 18, 20-week-old by real-time PCR and ELISA, respectively in MRL/lpr, SLE-prone mice. [Protocol 2] LN was induced by peritoneally injecting pristane in wild type and RAGE globally knockout mice. [Protocol 3] MRL/lpr mice were subcutaneously administrated with DNA-aptamer raised against RAGE (RAGE-apt) or Control-aptamer (Ctrl-apt) for 10weeks.

Results:

[Protocol 1] uRAGE levels, but not urinary protein excretion, were significantly increased in 8-week-old MRL/lpr mice when compared to those in control MRL/MPJ mice, suggesting that uRAGE could be an early marker of LN.. Immunofluorescence staining demonstrated that RAGE was upregulated in distal tubules but not in glomerulus in 8-week-old mice. Glomerular RAGE expression was increased from 12-week-old with mesangial matrix expansion and formation of cellular crescents. RAGE expression was also found in endothelium and infiltrated macrophages in glomerulus while RAGE was merged with cathepsine D, a lysosomal aspartyl protease, and Rab7, a marker of late endocytosis in tubular epithelial cells. [Protocol 2] Pristane-induced increase in systolic blood pressure was reduced with improvement of renal dysfunction in RAGE knockouts. RAGE knockouts also showed less fibrosis with decrease in macrophage infiltration when compared to wild type mice in pristane-induced lupus nephritis. [Protocol 3] Administration of RAGE-apt ameliorated mesangial expansion, crescent formation, and macrophage infiltration into glomerulus in MRL/lpr mice. RAGE-apt, but not Ctrl-apt, reduced the level of pro-inflammatory cytokines gene expression including IL-6, TNF-α, and MCP-1 in renal cortex of MRL/lpr mice.

Conclusion:

RAGE could be involved in the pathogenesis of LN, and RAGE-DNA aptamer might be one of the promising therapeutic strategies for preventing the development of LN.
PS-B08-13: PLASMIN INHIBITION ALLEVIATES HYPERTENSION AND PODOCYTE INJURY IN DAHL SALT-SENSITIVE RATS
Authors: Deng Qinyuan , Kakizoe Yutaka , Iwata Yasunobu , Nakagawa Terumasa , Miyasato Yoshikazu , Nakagawa Miyuki , Nishiguchi Kayo , Nagayoshi Yu , Adachi Masataka , Narita Yuki , Izumi Yuichiro , Kuwabara Takashige , Tsuda Yuko , Mukoyama Masashi
Published: January 2023

Objective:

The epithelial sodium channel (ENaC) in the kidney plays pivotal roles in blood pressure regulation, and the γ subunit is activated by extracellular serine proteases. In proteinuric renal diseases, plasmin filtered through injured glomeruli proteolytically activates γENaC. In addition, filtered plasmin directly causes podocyte injury. We previously reported that Dahl salt-sensitive (DS) rats fed a high-salt (HS) diet developed severe hypertension and proteinuria together with γENaC activation, and that a synthetic serine protease inhibitor, camostat mesilate, mitigated these changes. However, the role of plasmin in DS rats remains unclear. In this study, we evaluated the relationship between plasmin and hypertension as well as glomerular injury and the effects of plasmin inhibitors in DS rats.

Design and method:

Five-week-old male DS rats were divided into normal-salt (NS) diet, HS diet, and HS + plasmin inhibitors [tranexamic acid (TA, 2 mg/mL TA in drinking water) and synthetic plasmin inhibitor YO-2 (4 mg/kg/day, intraperitoneal injection)] groups. After systolic blood pressure measurement and 24-h urine collection over time for 5 weeks, the rats were sacrificed for biochemical examination.

Results:

The HS group displayed severe hypertension and proteinuria together with activation of plasmin in urine and γENaC in the kidney, which were not attenuated by TA. On the other hand, YO-2 mitigated both hypertension and proteinuria [SBP (mmHg): NS, 130.8 ± 7.5; HS, 209.0 ± 14.2; HS+YO-2, 183.8 ± 8.4; Urinary protein (mg/day): NS, 15.4 ± 3.2; HS 259.0 ± 129.3; HS+YO-2, 94.5 ± 31.5]. YO-2 inhibited the attachment of plasmin(ogen) to podocytes and alleviated podocyte injury by reducing glomerular apoptotic cells. Furthermore, YO-2 suppressed the upregulation of protease-activated receptor-1 and phosphorylated ERK1/2 as well as mRNA expression of inflammatory and pro-fibrotic cytokines in the kidney.

Conclusion:

These results indicate that plasmin plays important roles in the development of salt-sensitive hypertension and glomerular injuries in a rat model of hypertension, and suggest that plasmin inhibition could be a potential therapeutic strategy against salt-sensitive hypertension.
PS-B09-1: ENOS/NO SIGNALING REGULATES RENAL INTERSTITIAL FIBROSIS BY INCREASING CYTOPLASMIC BETA-CATENIN DEGRADATION
Authors: Takasu Masanobu , Kadoya Hiroyuki , Nagasu Hajime , Kishi Seiji , Sasaki Tamaki , Kashihara Naoki
Published: January 2023

Renal interstitial fibrosis is a final common pathway in progressive renal disease. Recently, activation of the wnt/beta-catenin signaling pathway has been demonstrated to play an important role in the development of renal fibrosis. It is also well known that endothelial dysfunction is associated with chronic kidney disease. However, the role of endothelial dysfunction in the progression of renal fibrosis remains largely elusive. We hypothesized that protein kinase G (PKG) activation by nitric oxide (NO) could phosphorylate beta-catenin and downregulate beta-catenin signaling in the process of renal fibrosis. We showed that beta-catenin levels in the cytoplasm were decreased by activation of the NO-PKG pathway. Renal fibrotic changes were not observed in wild type (WT)-lithium (Li; glycogen synthase kinase-3beta inhibitor) mice. However, renal interstitial fibrosis developed in endothelial nitric oxide synthase knockout (eNOSKO)-Li mice compared with WT-Li mice. beta-catenin protein expression was remarkably increased in eNOSKO-Li mice. These changes were suppressed in eNOSKO-Li/Bay (soluble guanylate cyclase [sGC] stimulator) mice. Moreover, increased interstitial renal fibrosis was observed in eNOSKO mice subjected to unilateral ureteral obstruction (eNOSKO-UUO) compared with WT-UUO mice. beta-catenin protein expression was significantly increased in eNOSKO-UUO mice. These phenotypic changes in eNOSKO-UUO mice were suppressed by Bay treatment. Activation of eNOS signaling could be protective against renal interstitial fibrosis. These findings indicate the potential therapeutic utility of eNOS-NO-PKG stimulation for the amelioration of renal fibrosis.
PS-B09-3: EXAMINATION OF ALBUMINURIA REDUCTION EFFECT THROUGH IMPROVEMENT OF ENOS UNCOUPLING BY SGLT2 IN TYPE 2 DIABETIC MICE
Authors: Iwakura Tsukasa , Kondo Megumi , Kidokoro Kengo , Kashihara Naoki
Published: January 2023

Background and Aims:

One of the main mechanisms of the renoprotective effects of SCLT2is in DKD is considered the ability of these inhibitors to improve glomerular hyperfiltration. We previously demonstrated that the adenosine/adenosine A1 receptor pathway played a pivotal role in the tubuloglomerular feedback(TGF) system in a type 1 diabetic model, Akita mice (Circulation, 2019). We also reported that the uncoupling of endothelial nitric oxide (NO) synthase (eNOS) via oxidation of tetrahydrobiopterin (BH4), a cofactor required for NO production, played a major role in generation of oxidative stress (AJPRP, 2005; JASN, 2013). In the present study, we explored the renal protective effects of SGLT2 inhibition, with a focus on glomerular oxidative stress and glomerular hemodynamics.

Methods:

This study used type 2 diabetic db/db mice and db/m+ mice as a control (male, 8wk old). In the first experiment, we measured the glomerular filtration rate of single nephrons (SNGFRs) using in vivo imaging technique in 12 wk-old db/db and db/m+ mice to confirm glomerular hyperfiltration. Next, we evaluated the SNGFRs change before and after the administration of a single dose of canagliflozin (CANA) (10 mg/kg). The SNGFRs, glomerular permeability of macromolecules, glomerular reactive oxygen species (ROS) and NO production, and tetrahydrobiopterin (BH4) level in serum and kidney were evaluated after the CANA treatment for 8 wk. Finally, human glomerular endothelial cells (hGECs) were exposed to normal glucose (5 mmol/L), high glucose (30 mmol/L of D-glucose), or a hyperosmotic control (5 mmol/L of D-glucose plus 25 mmol/L of L-glucose) in the presence or absence of CANA (10umol/L).

Results:

The CANA treatment ameliorated glomerular hyperfiltration in the db/db mice. In the db/db mice, glomerulus ROS production increased, and NO production decreased as compared with the levels in the control mice. CANA improved the imbalance between ROS and NO production. The serum and kidney concentrations of BH4 declined in the non-treated db/db mice, whereas the CANA treatment preserved the BH4 level. Leakage of 70-kD FITC-labeled albumin into the urinary space was observed in the db/db mice. The CANA treatment reduced the amount of FITC-labeled albumin in the urinary space of the db/db mice. The CANA treatment also alleviated vascular endothelial damage in glomeruli. BH4 levels decreased in the hGECs exposed to high glucose. CANA did not improved BH4 level in the hGECs exposed to high glucose.

Conclusions:

SGLT2i improved glomerular ROS/NO imbalance by suppressing eNOS uncoupling of glomerular endothelial cells in type 2 diabetic mice.
PS-B09-7: NON-DIPPER BLOOD PRESSURE CAUSED BY INTERMITTENT HYPOXIA AND ITS POSSIBLE MECHANISM IN THE KIDNEY
Authors: Ueda Kohei , Yeerbolati Alimila , Lijuan Liang , Ogura Sayoko , Takase Mai , Goda Nobuhito , Shimosawa Tatsuo
Published: January 2023

Background:

Non-dipper, less than 10% decrease of blood pressure in nighttime than daytime, is one of the strongest risk factor of cardiovascular diseases among hypertensive disorders. High prevalence of non-dipper among patients of sleep-apnea syndrome suggests intermittent hypoxia in nighttime as the mechanism of non-dipper, although it is still to be revealed.

Purpose:

To identify the mechanism of non-dipper caused by intermittent hypoxia in nighttime.

Method:

C57Bl/6-background male mice were kept under 8AM-8PM of light and 8PM-8AM dark cycle in closed air-conditioned system which could decrease the O2 level from 21% room air to 5% every 3 minutes. Mice were divided into two groups, normoxia (NX) or intermittent hypoxia (IH), and were kept for two weeks under room air thoroughly or under the IH only during 8 AM to 5 PM respectively. The mice were euthanized at 12PM and kidneys were obtained. Blood pressure (BP) of left carotid artery was recorded by radio-telemetry during the latter one week. To evaluate diurnal change in BP, BPs of 8AM-5PM (inactive) and 5PM-8AM (active) were compared in each of NX and IH mice. Kidney lysate was fractionated by ultracentrifuge to obtain membrane fraction of protein.

Result:

BP of NX mice in the active phase was significantly higher than the inactive phase, although such diurnal change was not observed in IH mice, demonstrating a development of non-dipper pattern by IH. In the kidney lysates, membrane protein expression levels of phosphorylated-Na+-Cl- cotransporter (NCC) and Na + -K+-Cl- cotransporter 2 (NKCC2) were significantly increased in IH mice compared to NX mice, but those of full-length and cleaved form of ENaCα were not changed. Transcriptional expression levels of Sgk1, Gilz, Per1/2 and Cry1/2 were significantly increased and those of Bmal1 and Clock were decreased in IH mice compared to those of NX mice.

Conclusion:

These data suggest that defect in molecular clock of the kidney and sodium reabsorption augmented by NCC and/or NKCC2 in the kidney contributes to the development of non-dipper.
PS-C18-2: BODY MASS INDEX TRAJECTORY AND TECHNIQUE FAILURE OR DEATH IN PATIENTS INITIATING PERITONEAL DIALYSIS
Authors: Kojima Daiki , Uchiyama Kiyotaka , Hama Eriko Yoshida , Nagasaka Tomoki , Kusahana Ei , Nakayama Takashin , Morimoto Kohkichi , Kanda Takeshi , Washida Naoki , Itoh Hiroshi
Published: January 2023

Objective:

A decreased body mass index (BMI) over time, as a nutritional status marker, is associated with the poor prognosis of patients undergoing hemodialysis; however, this association has not been studied in patients with peritoneal dialysis (PD). Thus, this study aimed to evaluate the association between BMI trajectory and survival of patients initiating PD.

Design and Method:

BMI trajectory was defined as percent changes in BMI before PD catheter insertion until six months after insertion. The association of BMI trajectory with technique failure or all-cause death at six months after PD catheter insertion until October 2021 was analyzed.

Results:

This retrospective cohort study included 122 patients (aged 61.1 ± 12.1 years; 90 males) who underwent PD catheter insertion between January 2008 and March 2020. The median follow-up period was 43.1 (interquartile range, 21.2 - 78.8) months. The mean 6-month percent change in BMI was - 1.98% ± 6.91%, and patients were categorized into tertiles based on BMI trajectory. The adjusted Cox regression analysis for age, sex, Charlson comorbidity index, estimated glomerular filtration rate, and geriatric nutritional risk index revealed a significantly higher rate of technical failure or all-cause mortality (hazard ratio, 2.48; 95% confidence interval: 1.41 - 4.37) in patients with the lowest tertile (T1, BMI trajectory: < - 4.13%) compared to patients with middle tertile, the reference group (T2, BMI trajectory: - 4.13 - 0.67%). The risk was not significantly higher in patients with the highest tertile (T3, BMI trajectory: > 0.67%) than T2 group (hazard ratio: 1.18; 95% confidence interval: 0.66 - 2.11).

Conclusions:

A decreased BMI (not increased) over time is independently associated with technique failure or all-cause death among patients initiating PD, highlighting the importance of BMI trajectory as a novel prognostic marker. Moreover, further studies are warranted to investigate the interventions, including BMI optimization, as a novel therapeutic target to improve technique and patient survival.