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We aimed to study the associations of 5 adipocytokines, two endothelial damage markers, and hs-CRP with the MetS components to distinguish the most significant cytokines likely related to distinct metabolic profiles. Methods: Cross-sectional study with 202 Chilean subjects (18–65 years old), categorized by MetS, and No-MetS according to Harmonizing ATP III. Adipocytokines profiling included adiponectin, leptin, hs-CRP, CTRP-1, PAI-1, FABP4, and metalloproteinase (MMP)-9 and MMP-2 activity. Results: Subjects with MetS showed higher levels of the most proinflammatories molecules but significantly lower adiponectin than subjects with No-MetS. Among the studied adipocytokines, PAI-1 and adiponectin showed the strongest associations with most of MetS components. PAI-1 was associate with MetS OR 1.107 [1.065–1.151], p< 0.0001, and adiponectin inversely associated with MetS OR 0.710 [0.610 -0.825], p< 0.0001). Following adjustment by sex, age, BMI, and 24 h sodium urinary excretion in a multivariate analysis, the association of PAI-1 OR 1.090 [1.044–1.137], p< 0.0001) and adiponectin OR 0.634 [0.519 - 0.775], p < 0.0001) with MetS remained significant. Multivariate analyses support a model where PAI-1associate to waist_hip, SBP, DBP, and glucose (all p< 0.0001) and adiponectin associate to TG (p=0.03) and HDL-cholesterol (p=0.0001). Conclusion: PAI-1 and Adiponectin rendered the most robust associations with MetS components in a general population, indicating that unfavourable adipose tissue performance is a key contributor to these metabolic anomalies. Further prospective analyses should allow establishing whether these adipocytokines can anticipate the progress of MetS and cardiovascular risk. Conflict of interest: The authors declared no conflict of interest. Funding: This work was supported by Chilean grants CONICYT Fondo Nacional de Desarrollo Científico y Tecnológico, (FONDECYT) 1160695(CEF) and 1190419(RB); FONDECYT Post-doctorado 3200646(ATC); Millenium Institute of Immunology and Immunotherapy - ICM (P09/16-F)(AK-CEF).

In obesity, the increase in the number and / or size of adipocytes leads to chronic low-grade systemic inflammation that conditions the development and evolution of a series of pathologies such as coronary heart disease or cancer. The change from hyperplasia to hypertrophy has been characterized by the expression of early and late adipogenic biomarkers, but little is known about the communications between cells. Exosomes are small extracellular vesicles (30-100 nm in diameter) released by exocytosis in almost all cell types. Due to its wide distribution and absorption by different cell types, exosomes have been considered an attractive tool for diagnosis, therapy and response evaluation. Objective: To characterize the exosomes released by human fat cells during differentiation and the effect of hyperplasia on the function of these. Material and methods: Human cells (SW-872) were cultured for 24h in DMEM-F12- with FBS serum (free of steroids and exosomes), differentiated with adipogenic cocktail for 7 days, and hypertrophy for further 24h G1 agonist treatment. Exosomes were isolated from conditioned medium of SW872 cells by ultracentrifugation and characterized by immunoblotting against exosomal markers, (Nanoparticle Tracking Analysis (NTA) and transmission electron microscopy (TEM). Cancer initiating cells (CICs) were isolated from HeyA8 ovarian cancer cells using culture selecting conditions. CICs were 24h treated with exosomes (1x1011 particles/mL) and then seeded over matrigel to carry out 3Dmigration assays. Results: SW872 cells showed the morphological characteristics described for this cell line and MR expression was observed. Successful isolation of SW872-derived exosomes was confirmed by assessing the particle size distribution by NTA, the morphology by TEM and the presence of exosome markers (Alix, HSP70, TSG101, and CD36) by immunoblotting. Preadipocyte differentiation showed a significant decrease in the exosome concentration (pre,1.3x1011 particles/ml vs adipo, 1.5x1010 particles/ml p

Mineralocorticoid arterial hypertension has risen as one of the most prevalent causes of secondary hypertension, where 11β-hydroxysteroid dehydrogenase type-2 partial deficiency (also known as non-classical AME (NC-AME)), could reach a prevalence of 7% in Chilean subjects. The phenotype of NC-AME is associated to high serum cortisol (F) to cortisone (E) ratio &low E, high potassium excretion and low-renin activity (Tapia-Castillo et al, JCEM 2018). However, local metabolic changes in NC-AME affecting protein, RNA and miRNA expression have not been studied. Nowadays, exosomes technology allows to identify those biomolecules in specific biofluids. Aim: To identify the transcriptomic (miRNA) and proteomic profile in urinary exosomes of subjects with non-classical AME and healthy subjects. Subjects and methods: A cross-sectional study was carried out in 24 subjects (10-65 years). The subjects were classified as NC-AME (F/E ratio> percentile 75th, and E < percentile 25th) and healthy controls. The levels of F, E, aldosterone and plasma renin activity were quantified. The exosomes were obtained from morning urine samples by differential ultracentrifugation and were characterized with the NS300 nanoparticle analyzer, electron microscopy and western-blot for CD63 and TSG101. The total exosomal RNA was isolated with Trizol reagent. The Illumina TruSeq Small RNA kit was used and sequenced by Illumina NextSeq-500. Additionally, an exosomal proteomic profile was obtained by LC-MS/MS 5600 Triple TOF (ABSciex, Framingham, USA). Bioinformatic analyzes were performed with miRdeep2, PANTHER and STRING. Results: In urinary exosomes, we found355 from 2822 predicted miRNAs , of which 170 miRNA were found to be upregulated and 185 miRNA were dowregulated in subjects with NC-AME vs controls. We also found miR-204-5p (change times = 0.115; p = 0.001) and have miR-192-5p (change times = 0.246; p = 0.03), present a significantly lower expression in NC-AME subjects vs controls, both confirmed by Taqman PCR. Genetic ontology analyses indicate that both miRNAs would have a role in steroid biosynthesis (hsa00100). We identified around 350 exosomal proteins, of which 79 proteins were commonly expressed in both groups and 23 proteins were exclusively expressed in subjects with NC-AME, highlighting the 14-3-3 (YWHAE, YWHAZ), RHOA and CDC42 protein. Conclusion: This is the first study on urinary exosomes showing the differential expression of miRNA (miR-204 and miR-192) and proteins (14-3-3, RHOA and CDC42) which highlight its potential role as biomarkers and regulators of the high mineralocorticoid activity in NC-AME subjects. Acknowledgements: This study was supported by grants CONICYT-FONDECYT 1150437, 1160695, 1160836, CONICYT-FONDEQUIP EQM150023, IMII P09/16-F, & CETREN-UC. Unless otherwise noted, all abstracts presented at ENDO are embargoed until the date and time of presentation. For oral presentations, the abstracts are embargoed until the session begins. Abstracts presented at a news conference are embargoed until the date and time of the news conference. The Endocrine Society reserves the right to lift the embargo on specific abstracts that are selected for promotion prior to or during ENDO.