Abstract

Background: Senescence is a terminal differentiation state that can be induced in cells in response to different stresses, including exposure to glucocorticoids. Accumulation of senescent cells in tissues induce functional impairment of different organs and age-related pathologies, due to the secretion of factors that create a pro-inflammatory microenvironment and induce further senescence in neighbour cells.Aim: We hypothesize that persistence of senescent cells in glucocorticoid target tissues can mediate pathological changes long time after cure of Cushing syndrome (CS). Quantifying the presence of these cells could have prognostic value and allow the stratification of patients into groups that could receive senotherapies (drugs specific to reduce senescent cell accumulation) as adjuvant treatment. Methods: Fractionation and unbiased mass spectrometry screen to detect proteins related to senescence in glucocorticoid target tissues (muscle, brown and white adipose tissue, liver, adrenals, hypothalamus and pituitary) and plasma in 12 weeks old male C5BL/6J mice with active CS induced by chronic oral cortiscosterone administration (5 weeks) and mice with cured CS (10 weeks after treatment) as compared to CTR mice. Proteins have been extracted by digestion of the tissue samples and tryptic peptides measured by OPA assay prior to be injected on column to a Waters nanoacquity nanoUPLC coupled to a Waters Synapt G2Si. Data has been acquired using Ion mobility enabled data independent acquisition (HDMSE). Results: Raw data extraction and analysis using Progenesis QI for Proteomics identified a tissue-specific profile of proteins linked to senescence and/or ageing persistent after remission of CS in target mice tissue, which can be used for diagnostic and therapeutic purposes.