1. Flow Cytometric Quantification of Peripheral Blood Cell β-Adrenergic Receptor Density and Urinary Endothelial Cell-Derived Microparticles in Pulmonary Arterial Hypertension
- Author
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Kewal Asosingh, Kimberly Queisser, Hoi I. Cheong, Patrick Barrett, Corrine Hite, Nicholas Wanner, Serpil C. Erzurum, Sathyamangla V. Naga Prasad, Margaret M. Park, and Jonathan A. Rose
- Subjects
0301 basic medicine ,Male ,Pathology ,Physiology ,Urine ,Biochemistry ,Epithelium ,chemistry.chemical_compound ,Immunophenotyping ,Spectrum Analysis Techniques ,Cell-Derived Microparticles ,Animal Cells ,Leukocytes ,Medicine and Health Sciences ,Pulmonary Arteries ,Multidisciplinary ,medicine.diagnostic_test ,Heart ,Arteries ,Middle Aged ,Flow Cytometry ,3. Good health ,Body Fluids ,Spectrophotometry ,Medicine ,Female ,Cytophotometry ,Anatomy ,Cellular Types ,Research Article ,Adult ,Cell Binding ,medicine.medical_specialty ,Cell Physiology ,Cardiac Ventricles ,Urinary system ,Hypertension, Pulmonary ,Science ,Biology ,Research and Analysis Methods ,Flow cytometry ,03 medical and health sciences ,Right ventricular hypertrophy ,medicine.artery ,Receptors, Adrenergic, beta ,medicine ,Humans ,Peripheral blood cell ,Alprenolol ,Blood Cells ,Biology and Life Sciences ,Endothelial Cells ,Epithelial Cells ,Cell Biology ,medicine.disease ,Pulmonary hypertension ,030104 developmental biology ,Biological Tissue ,chemistry ,Pulmonary artery ,Cardiovascular Anatomy ,Blood Vessels ,Biomarkers - Abstract
Pulmonary arterial hypertension (PAH) is a heterogeneous disease characterized by severe angiogenic remodeling of the pulmonary artery wall and right ventricular hypertrophy. Thus, there is an increasing need for novel biomarkers to dissect disease heterogeneity, and predict treatment response. Although β-adrenergic receptor (βAR) dysfunction is well documented in left heart disease while endothelial cell-derived microparticles (Ec-MPs) are established biomarkers of angiogenic remodeling, methods for easy large clinical cohort analysis of these biomarkers are currently absent. Here we describe flow cytometric methods for quantification of βAR density on circulating white blood cells (WBC) and Ec-MPs in urine samples that can be used as potential biomarkers of right heart failure in PAH. Biotinylated β-blocker alprenolol was synthesized and validated as a βAR specific probe that was combined with immunophenotyping to quantify βAR density in circulating WBC subsets. Ec-MPs obtained from urine samples were stained for annexin-V and CD144, and analyzed by a micro flow cytometer. Flow cytometric detection of alprenolol showed that βAR density was decreased in most WBC subsets in PAH samples compared to healthy controls. Ec-MPs in urine was increased in PAH compared to controls. Furthermore, there was a direct correlation between Ec-MPs and Tricuspid annular plane systolic excursion (TAPSE) in PAH patients. Therefore, flow cytometric quantification of peripheral blood cell βAR density and urinary Ec-MPs may be useful as potential biomarkers of right ventricular function in PAH.
- Published
- 2016