Abstract 4: Molecular Profile Of Peripheral Blood Mononuclear Cells In Hypertensive Adolescents With Target Organ Damage

Hypertension ◽  
2020 ◽  
Vol 76 (Suppl_1) ◽  
Author(s):  
Fnu MOHAMMED ARIF ◽  
Elaine M Urbina ◽  
Sakthivel Sadayappan ◽  
Richard Becker
Keyword(s):  
Growth Factor ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Target Organ Damage ◽  
Organ Damage ◽  
Left Ventricular ◽  
Target Organ ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells

Introduction: Primary hypertension (PH) is a multifactorial disease influenced by genetic, epigenetic, and environmental factors. Despite the occurrence of PH-associated cardiovascular events in youth, the molecular mechanism(s) of target organ damage (TOD) are unknown. Objectives: (1) To identify an epigenetic signature and gene expression profiles in adolescents with low blood pressure (BP) and normal left ventricular mass (LVM) compared to those with high BP and high LVM; and (2) to determine novel gene targets and associated signaling pathways for future investigation and intervention. Methods and Results: A total of 397 participants (mean age 15.6 ±1.7 years, 59% male, 63% Caucasian) were enrolled across the distribution of BP. The average daytime ambulatory systolic BP recorded in healthy and hypertensive participants was 112 ±9.71 and 133 ±7.2 mmHg ( p<0.05 ) respectively. Clinical measures revealed higher body mass index (26.8 ±7.02 vs 29.6 ±7.88 Kg/m 2 ; p<0.05 ), and abnormal circulatory HDL (47.4 ±12.1 vs 43.4 ±11.7 mg/dL; p<0.05 ), glucose (87.8 ±7.98 vs 90.8 ±8.17 mg/dL; p<0.05 ), insulin (17.8 ±14.3 vs 23.7 ±19 μIU/dL; p<0.05 ), creatinine (0.718 ±0.13 vs 0.727 ±0.17 mg/dL; p<0.05 ), uric acid (5.4 ±1.63 vs 6.04 ±1.52 mg/dL; p<0.05 ), CRP (1.35 ±1.8 vs 1.92 ±2.14 mg/dL; p<0.05 ), and left ventricular hypertrophy (LVM/ht 2.7 ; 31.4 ±6.74 vs 33.5 ±7.15 g/m 2.7 ; p<0.05 ), and arterial stiffness (Pulse wave velocity; 4.83 ±0.69 vs 5.35 ±0.92 m/sec; p<0.05 ). Using peripheral blood mononuclear cells, mRNA-Seq, miRNA-Seq, and whole-genome DNA methylation analysis revealed master genes, and regulatory pathways related to BP regulation, tissue fibrosis and cardiovascular remodeling. Our study reveals a novel PH-associated TOD mechanism, showing angiogenesis inhibition mediated by VASH1 (Vasohibin-1) upregulation and downtrends in VASH2 (Vasohibin-2), VEGFC (Vascular endothelial growth factor C), HIF1α (Hypoxia-inducible factor 1-alpha), and IGF1 (Insulin Like Growth Factor 1). Moreover, VASH1 targeting miRNA hsa-miR-30e-5p is inversely regulated. Conclusion: Angiogenesis inhibition in the presence of common demographic and clinical intermediate-phenotypes may contribute to the development of TOD in hypertensive youth.

Abstract MP44: A Novel And 2 Known Differentially Expressed MicroRNAs Were Identified In Peripheral Blood Mononuclear Cells Of Patients With Hypertension Associated With Metabolic Syndrome

Hypertension ◽  
2021 ◽  
Vol 78 (Suppl_1) ◽  
Author(s):  
Olga Berillo ◽  
Kugeng Huo ◽  
Julio C Fraulob-Aquino ◽  
Chantal Richer ◽  
Na Li ◽  
...  
Keyword(s):  
Metabolic Syndrome ◽  
Peripheral Blood Mononuclear Cells ◽  
Immune Cells ◽  
Mononuclear Cells ◽  
Target Organ Damage ◽  
Organ Damage ◽  
Target Organ ◽  
Rna Seq ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells

Background: Hypertension (HTN) is associated with subclinical target organ damage including cardiac, vascular and kidney injury. The immune system plays a role in hypertension and target organ damage. Activation of T cells has been reported among peripheral blood mononuclear cells (PBMCs) of patients with HTN. MicroRNAs (miRNAs) are crucial post-transcriptional regulators of immune cells. Whether miRNAs play a role in the activation of immune cells in hypertension complicated by target organ damage in humans remains unknown. We aimed to address this question by identifying differentially expressed (DE) miRNAs and their mRNA targets in PBMCs of patients with hypertension complicated or not with metabolic syndrome (MetS) or chronic kidney disease (CKD). Methods: Normotensive subjects and patients with hypertension (HTN) associated or not with at least 2 other features of MetS or CKD were studied (n=15-16). PBMCs were isolated from blood, RNA extracted for small and total RNA sequencing (RNA-seq) using Illumina HiSeq-2500 and data were analyzed using a systems biology approach. MiRDeep2 was used for novel miRNAs prediction, miRNA annotation and counting. TargetScan 7.07 was used to predict DE miRNA targets with weighted context score percentile >50%. DE genes miRNAs and mRNAs were identified with fold change (FC) >1.5 and P <0.005. DE miRNAs with FC>2 and mean read count number (MRCM) >500, and with predicted targets with MRCM>300 were validated by reverse transcription-quantitative PCR (RT-qPCR). Results: DE miRNAs, mRNAs and non-coding RNAs were identified in HTN (22, 19 and 0), MetS (57, 401 and 11) and CKD (6, 26 and 2) compared to NTN. TargetScan predicted that 7 miRNAs target 3 mRNAs in NTN, 57 miRNAs target 55 mRNAs in MetS and 3 miRNAs target 2 mRNAs in CKD. DE miR-409-5p (FC: 0.54±0.10 vs 1.00±0.09, P <0.05), miR-411-5p (FC: 0.40±0.06, vs 1.00±0.11, P <0.001) and the novel miR-pl-86 (FC: 1.96±0.17 vs 1.00±0.15, P <0.05) in MetS vs NTN were validated by RT-qPCR. RNA-seq data were correlated with RT-qPCR for miR-409-5p (R 2 =0.40, P <2.4E-07, n=55), miR-411-5p (R 2 =0.55, P <1.1E-10, n=55), miR-pl-86 (R 2 =0.37, P <5.5E-07, n=56). Conclusion: This study showed that DE miR-409-5p, miR-411-5p and miR-pl-86 may play a role in HTN associated with MetS.

scholarly journals A Growth Factor-Free Co-Culture System of Osteoblasts and Peripheral Blood Mononuclear Cells for the Evaluation of the Osteogenesis Potential of Melt-Electrowritten Polycaprolactone Scaffolds

2019 ◽  
Vol 20 (5) ◽  
pp. 1068 ◽  
Author(s):  
Andreas Hammerl ◽  
Carlos Diaz Cano ◽  
Elena De-Juan-Pardo ◽  
Martijn van Griensven ◽  
Patrina Poh
Keyword(s):  
Growth Factor ◽  
Bone Regeneration ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Culture System ◽  
Mononuclear Cells ◽  
Regeneration Potential ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells

Scaffolds made of biodegradable biomaterials are widely used to guide bone regeneration. Commonly, in vitro assessment of scaffolds’ osteogenesis potential has been performed predominantly in monoculture settings. Hence, this study evaluated the potential of an unstimulated, growth factor-free co-culture system comprised of osteoblasts (OB) and peripheral blood mononuclear cells (PBMC) over monoculture of OB as an in vitro platform for screening of bone regeneration potential of scaffolds. Particularly, this study focuses on the osteogenic differentiation and mineralized matrix formation aspects of cells. The study was performed using scaffolds fabricated by means of a melt electrowriting (MEW) technique made of medical-grade polycaprolactone (PCL), with or without a surface coating of calcium phosphate (CaP). Qualitative results, i.e., cell morphology by fluorescence imaging and matrix mineralization by von Kossa staining, indicated the differences in cell behaviours in response to scaffolds’ biomaterial. However, no obvious differences were noted between OB and OB+PBMC groups. Hence, quantitative investigation, i.e., alkaline phosphatase (ALP), tartrate-resistant acid phosphatase (TRAP) activities, and gene expression were quantitatively evaluated by reverse transcription-polymerase chain reaction (RT-qPCR), were evaluated only of PCL/CaP scaffolds cultured with OB+PBMC, while PCL/CaP scaffolds cultured with OB or PBMC acted as a control. Although this study showed no differences in terms of osteogenic differentiation and ECM mineralization, preliminary qualitative results indicate an obvious difference in the cell/non-mineralized ECM density between scaffolds cultured with OB or OB+PBMC that could be worth further investigation. Collectively, the unstimulated, growth factor-free co-culture (OB+PBMC) system presented in this study could be beneficial for the pre-screening of scaffolds’ in vitro bone regeneration potential prior to validation in vivo.

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