Exosomes derived from endometriotic stromal cells

Authors: Djana Harp Adel Driss Sharifeh Mehrabi Indrajit Chowdhury Wei Xu Dong Liu Minerva GarciaBarrio Robert N Taylor Bert Gold Samantha Jefferson Neil Sidell Winston Thompson

Publish Date: 2016/02/03

Volume: 365, Issue: 1, Pages: 187-196

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Abstract

Our objective has been to establish a proangiogenic role for exosomes in endometriosis and to determine whether a differential expression profile of cellular and exosomal microRNAs miRNAs exists in endometriosis We performed an in vitro study of human primary endometrial stromal cells ESCs and human umbilical vein endothelial cells HUVECs We isolated and characterized exosomes from ESCs from five endometriosis patients and five phasematched controls Exosomes were characterized by transmission electron microscopy and NanoSight technology MiRNA was assessed by deep sequencing and reverse transcription with quantitative polymerase chain reaction Exosome uptake studies were achieved by means of confocal microscopy The proangiogenic experiments were executed by treating HUVECs with ESCderived exosomes We observed differential profiles of exosomal miRNA expression between exosomes derived from endometriosis lesion cells and diseased eutopic stromal cells compared with exosomes derived from control ESCs We also demonstrated autocrine cellular uptake of exosomes and paracrine functional angiogenic effects of exosomes on HUVECs The results of this study support the hypothesis that exosomes derived from ESCs play autocrine/paracrine roles in the development of endometriosis potentially modulating angiogenesis The broader clinical implications are that Sampson’s theory of retrograde menstruation possibly encompasses the finding that exosomes work as intercellular communication modulators in endometriosisThis work was supported by NIH grants 5U01HD066439 and 1R01HD057235 This publication was also supported by the Research Centers in Minority Institutions Grant Number 8G12MD007602 from the National Institute of Minority Health and Health Disparities NIMHD and 8U54MD007588Endometriosis is a benign gynecological disorder characterized by the presence of endometrial glandular tissue and stroma outside of their normal uterine location Up to 10  of reproductiveage women in the United States are estimated to have endometriosis which is associated with debilitating pain menstrual irregularities and infertility in as many 30  of the affected women Bulun 2009 Verkauf 1987 The precise etiology of endometriosis remains to be elucidated Commonly accepted theories of the causes of endometriosis include retrograde menstruation coelomic metaplasia and lymphatic and vascular dissemination Macer and Taylor 2012 Patients with endometriosis often have delayed 6–11 years diagnosis because symptoms are variable and often mimic other gynecological diseases Fassbender et al 2015 Leone Roberti Maggiore et al 2016 Nogales et al 1993 A definitive diagnosis of endometriosis requires surgery Gordts et al 2015 The development of new ways of diagnosis by using exosomal biomarkers might enable earlier diagnosis of the disease Fassbender et al 2015Angiogenesis the formation of new blood vessels has increasingly been recognized as a key pathological factor in endometriosis Human endometrial tissue is known to be angiogenic throughout the menstrual cycle Maas et al 2001 Sampson’s theory of retrograde menstruation posits that endometrial tissue travels retrogradely through the Fallopian tubes and is deposited into the peritoneal cavity where it establishes a blood supply proliferates and forms an implant that is clinically recognized as a lesion Sampson 1927 Women with endometriosis have been found to have increased retrograde menstruation Halme et al 1984 Successful implantation of endometriotic lesions depends on neovascularization fibrosis adhesion formation apoptosis evasion and neuronal infiltration Rocha et al 2013 The exact mechanisms underlying the sprouting and sustainment of endometriosis neovascularization need further clarification Recent studies however demonstrate that inhibitors of angiogenesis block the progression of endometriosis Edwards et al 2013 Nap et al 2004 Wang et al 2014Exosomes have been found to play physiological roles as mediators of intercellular cell signaling between neighboring cells and even amongst distant tissues and they may act independently but synergistically with soluble growth factors and hormones Colombo et al 2014 Exosomes are vesicles ranging from 30 to 150 nm in diameter are derived from the fusion of multivesicular bodies with the plasma membrane and are secreted by a variety of living cells they are composed of a lipid bilayer membrane and contain functionally active proteins mRNA and microRNA miRNA that can be released rendering them important mediators of intercellular communication Colombo et al 2014 The recent discovery of these extracellular organelles containing genetic cargo with a unique signature raises the hope that exosomes will serve as diagnostic markers or therapeutic vehicles Krause et al 2015 MomenHeravi et al 2015 Tang and Wong 2015Among all molecules the miRNAs that are contained within exosomes Valadi et al 2007 could affect gene expression and function in target cells including vascular endothelial cells Several miRNAs such as miR21 and miR126 have been shown previously to play critical roles in angiogenesis Wang and Olson 2009 We have chosen to screen these two miRNAs because in previous literature they have been widely used by the scientific community as proangiogenic indicators in other systems Bao et al 2012 GuduricFuchs et al 2012 Xu et al 2015

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