{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,19]],"date-time":"2025-11-19T17:17:57Z","timestamp":1763572677593,"version":"build-2065373602"},"reference-count":48,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2022,12,13]],"date-time":"2022-12-13T00:00:00Z","timestamp":1670889600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Polish NCN","award":["UMO-2011\/01\/B\/NZ5\/04224"],"award-info":[{"award-number":["UMO-2011\/01\/B\/NZ5\/04224"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Abnormalities in hematological parameters of peripheral blood have been noted in patients with endogenous Cushing\u2019s Syndrome (CS) in the corticotropin (ACTH)-dependent and ACTH-independent forms. Nevertheless, the exact mechanism of glucocorticoids (GCs) action on human hematopoiesis is still not entirely clear. The aim of the study was to determine whether endogenous excessive production of GCs could affect apoptosis of CD34+ cells enriched in hematopoietic stem and progenitor cells (HSPCs) collected from the peripheral blood of newly diagnosed CS patients. Flow cytometry, Annexin-V enzyme-linked immunosorbent assay, TUNEL assay, real-time quantitative PCR, and microarray RNA\/miRNA techniques were used to characterize CS patients\u2019 HSPCs. We found that the glucocorticoid receptor (GR) protein expression levels in CS were higher than in healthy controls. A complex analysis of apoptotic status of CS patients\u2019 HSPC cells showed that GCs significantly augmented apoptosis in peripheral blood-derived CD34+ cells and results obtained using different methods to detect early and late apoptosis in analyzed cell population were consistent. CS was also associated with significant upregulation in several members of the BCL-2 superfamily and other genes associated with apoptosis control. Furthermore, global gene expression analysis revealed significantly higher expression of genes associated with programmed cell death control in HSPCs from CS patients. These findings suggest that human endogenous GCs have a direct pro-apoptotic activity in hematopoietic CD34+ cells derived from CS subjects before treatment.<\/jats:p>","DOI":"10.3390\/ijms232415794","type":"journal-article","created":{"date-parts":[[2022,12,13]],"date-time":"2022-12-13T02:59:01Z","timestamp":1670900341000},"page":"15794","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Apoptosis Evaluation in Circulating CD34+-Enriched Hematopoietic Stem and Progenitor Cells in Patients with Abnormally Increased Production of Endogenous Glucocorticoids in Course of Cushing\u2019s Syndrome"],"prefix":"10.3390","volume":"23","author":[{"given":"Mi\u0142osz P.","family":"Kawa","sequence":"first","affiliation":[{"name":"Department of General Pathology, Pomeranian Medical University in Szczecin, 72 Powstancow Wlkp. Street, 70-111 Szczecin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8155-7470","authenticated-orcid":false,"given":"Anna","family":"Sobu\u015b","sequence":"additional","affiliation":[{"name":"Department of General Pathology, Pomeranian Medical University in Szczecin, 72 Powstancow Wlkp. Street, 70-111 Szczecin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ewa","family":"Pius-Sadowska","sequence":"additional","affiliation":[{"name":"Department of General Pathology, Pomeranian Medical University in Szczecin, 72 Powstancow Wlkp. Street, 70-111 Szczecin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4008-7473","authenticated-orcid":false,"given":"Karolina","family":"\u0141uczkowska","sequence":"additional","affiliation":[{"name":"Department of General Pathology, Pomeranian Medical University in Szczecin, 72 Powstancow Wlkp. Street, 70-111 Szczecin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8884-837X","authenticated-orcid":false,"given":"Dorota","family":"Rogi\u0144ska","sequence":"additional","affiliation":[{"name":"Department of General Pathology, Pomeranian Medical University in Szczecin, 72 Powstancow Wlkp. Street, 70-111 Szczecin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Szymon","family":"Wn\u0119k","sequence":"additional","affiliation":[{"name":"Department of General Pathology, Pomeranian Medical University in Szczecin, 72 Powstancow Wlkp. Street, 70-111 Szczecin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7052-9741","authenticated-orcid":false,"given":"Edyta","family":"Paczkowska","sequence":"additional","affiliation":[{"name":"Department of General Pathology, Pomeranian Medical University in Szczecin, 72 Powstancow Wlkp. Street, 70-111 Szczecin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mieczys\u0142aw","family":"Walczak","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Endocrinology, Diabetology, Metabolic Diseases and Cardiology of the Developmental Age, Pomeranian Medical University in Szczecin, 1 Unii Lubelskiej Street, 71-252 Szczecin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anhelli","family":"Syrenicz","sequence":"additional","affiliation":[{"name":"Department of Endocrinology, Metabolic Diseases and Internal Diseases, Pomeranian Medical University in Szczecin, 1 Unii Lubelskiej Street, 71-252 Szczecin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bogus\u0142aw","family":"Machali\u0144ski","sequence":"additional","affiliation":[{"name":"Department of General Pathology, Pomeranian Medical University in Szczecin, 72 Powstancow Wlkp. Street, 70-111 Szczecin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1007\/s11102-013-0522-0","article-title":"Elevated white blood cell counts in Cushing\u2019s disease: Association with hypercortisolism","volume":"17","author":"Robenshtok","year":"2014","journal-title":"Pituitary"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2307","DOI":"10.1161\/01.CIR.98.21.2307","article-title":"Glucocorticoid-induced granulocytosis: Contribution of marrow release and demargination of intravascular granulocytes","volume":"98","author":"Nakagawa","year":"1998","journal-title":"Circulation"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3181","DOI":"10.1182\/blood.V86.8.3181.3181","article-title":"Glucocorticoids inhibit apoptosis of human neutrophils","volume":"86","author":"Liles","year":"1995","journal-title":"Blood"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1016\/j.autrev.2010.11.009","article-title":"The kaleidoscope of glucorticoid effects on immune system","volume":"10","author":"Zen","year":"2011","journal-title":"Autoimmun. 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