N. V. Yunusova, D. A. Svarovsky, E. S. Kolegova, O. V. Cheremisina, D. N. Kostromitsky, I. V. Kondakova, E. A. Sidenko, A. Yu. Dobrodeev, A. E. Grigor’eva
{"title":"Concentration and Composition of Circulating Adipocyte-Derived Extracellular Vesicles in Patients with Colonic Polyps and Colorectal Cancer","authors":"N. V. Yunusova, D. A. Svarovsky, E. S. Kolegova, O. V. Cheremisina, D. N. Kostromitsky, I. V. Kondakova, E. A. Sidenko, A. Yu. Dobrodeev, A. E. Grigor’eva","doi":"10.1134/s0022093024040069","DOIUrl":null,"url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>Extracellular vesicles (EVs) are a heterogeneous population\nof membrane-bound nanoparticles (< 1 µm in size) secreted by\nvarious cell types. Most of EVs circulating in human blood are derived\nfrom platelets, leukocytes, erythrocytes, and endotheliocytes. The\ncomposition of circulating adipocyte-derived EVs under various pathological\nconditions has been virtually unknown. Small EVs were isolated by\nultrafiltration and double ultracentrifugation from blood plasma\nof patients with colorectal cancer (CRC) and colonic polyps with\nobesity or metabolic syndrome. The composition of adipocyte-derived EVs\nwas analyzed by immunoprecipitation combined with Western blotting\nand flow cytometry. EVs fractions (FABP4- and CD11b-immunoprecipitated\nEVs, as well as EVs contained in the supernatant after removal of\nCD11b-positive EVs) contained a complex of adipocyte markers (FABP4,\nPPAR-γ, perilipin 1). In CRC patients without obesity, monocyte/macrophage-derived\nEVs precipitated on CD11b-coated particles were characterized by\na combined overexpression of FABP4 and perilipin 1, while such an\noverexpression was not typical for CRC patients with metabolic syndrome\nor obesity. The fraction of true adipocyte-derived EVs (supernatant\nafter removal of CD11b-positive EVs) was characterized by the presence\nof a complex of adipocyte markers with a predominant expression\nof FABP4 in all patients both with metabolic syndrome/metabolically\nhealthy obesity and without metabolic disorders. To correctly characterize\ncirculating EVs in patients without obesity, it is necessary first\nto remove the fraction of CD11b-positive monocyte/macrophage-derived\nEVs from EV preparations by immunoprecipitation or similar methods,\nand then, after removal/sorption of precipitated EVs, to analyze\nthe composition of adipocyte-derived EVs in the supernatant using\na set of above-mentioned adipocyte markers. Moreover, in patients\nwith metabolic disorders, given the insignificant FABP4 expression\nin CD11b-immunoprecipitated EVs, the pre-depletion of EV preparations\ndoes not appear to be that necessary.</p>","PeriodicalId":0,"journal":{"name":"","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1134/s0022093024040069","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
Extracellular vesicles (EVs) are a heterogeneous population
of membrane-bound nanoparticles (< 1 µm in size) secreted by
various cell types. Most of EVs circulating in human blood are derived
from platelets, leukocytes, erythrocytes, and endotheliocytes. The
composition of circulating adipocyte-derived EVs under various pathological
conditions has been virtually unknown. Small EVs were isolated by
ultrafiltration and double ultracentrifugation from blood plasma
of patients with colorectal cancer (CRC) and colonic polyps with
obesity or metabolic syndrome. The composition of adipocyte-derived EVs
was analyzed by immunoprecipitation combined with Western blotting
and flow cytometry. EVs fractions (FABP4- and CD11b-immunoprecipitated
EVs, as well as EVs contained in the supernatant after removal of
CD11b-positive EVs) contained a complex of adipocyte markers (FABP4,
PPAR-γ, perilipin 1). In CRC patients without obesity, monocyte/macrophage-derived
EVs precipitated on CD11b-coated particles were characterized by
a combined overexpression of FABP4 and perilipin 1, while such an
overexpression was not typical for CRC patients with metabolic syndrome
or obesity. The fraction of true adipocyte-derived EVs (supernatant
after removal of CD11b-positive EVs) was characterized by the presence
of a complex of adipocyte markers with a predominant expression
of FABP4 in all patients both with metabolic syndrome/metabolically
healthy obesity and without metabolic disorders. To correctly characterize
circulating EVs in patients without obesity, it is necessary first
to remove the fraction of CD11b-positive monocyte/macrophage-derived
EVs from EV preparations by immunoprecipitation or similar methods,
and then, after removal/sorption of precipitated EVs, to analyze
the composition of adipocyte-derived EVs in the supernatant using
a set of above-mentioned adipocyte markers. Moreover, in patients
with metabolic disorders, given the insignificant FABP4 expression
in CD11b-immunoprecipitated EVs, the pre-depletion of EV preparations
does not appear to be that necessary.
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