{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T20:23:28Z","timestamp":1787603008928,"version":"build-2736575974"},"reference-count":34,"publisher":"Oxford University Press (OUP)","issue":"11","license":[{"start":{"date-parts":[[2022,9,27]],"date-time":"2022-09-27T00:00:00Z","timestamp":1664236800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"name":"Australian Government Research Training Program"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2022,10,31]]},"abstract":"Abstract<\/jats:title>\n \n STUDY QUESTION<\/jats:title>\n Is circulating cell-free DNA (cirDNA) from the endometrium elevated during menstruation and in endometriosis?<\/jats:p>\n <\/jats:sec>\n \n SUMMARY ANSWER<\/jats:title>\n Endometrial cirDNA does not increase during menstruation and is not elevated in endometriosis.<\/jats:p>\n <\/jats:sec>\n \n WHAT IS KNOWN ALREADY<\/jats:title>\n Changes in cirDNA associated with common benign conditions are a potential source of false positives in cancer diagnostic applications, but also present an opportunity for biomarker development for diseases such as endometriosis. Elevated cirDNA has been reported in endometriosis patients compared to healthy community controls, but no difference in total or endometrial cirDNA has been found between patients with endometriosis and patients with other gynaecological conditions. Likewise, menstruation is a potential driver of changes in cirDNA levels and tissue profile, but total and endothelial cirDNA do not increase during menstruation.<\/jats:p>\n <\/jats:sec>\n \n STUDY DESIGN, SIZE, DURATION<\/jats:title>\n For endometriosis comparisons, 59 participants with surgically confirmed endometriosis and 27 laparoscopic patients without endometriosis (hospital controls) were prospectively recruited, while 25 healthy community participants (healthy controls) were recruited in a university setting. Total and endometrial cirDNA and cirDNA fragmentation were measured across the three groups. For menstrual comparisons, 36 matched non-menstruating and menstruating samples were collected from healthy women recruited within a university setting, and the endometrial cirDNA was compared between the two groups.<\/jats:p>\n <\/jats:sec>\n \n PARTICIPANTS\/MATERIALS, SETTING, METHODS<\/jats:title>\n cirDNA was extracted from venous blood plasma then quantitated by quantitative PCR of ALU repetitive element (115\u2009bp) and TP53 gene sequence (105\u2009bp) for total concentration. cirDNA derived from the endometrium was quantitated by methylation-specific droplet digital PCR of a FAM101A region (69\u2009bp) after bisulfite conversion of the DNA. A cirDNA size fragmentation ratio was obtained by quantifying a long segment of ALU repetitive element (247\u2009bp) and expressing the amount relative to the 115\u2009bp ALU target.<\/jats:p>\n <\/jats:sec>\n \n MAIN RESULTS AND THE ROLE OF CHANCE<\/jats:title>\n No differences in cirDNA level were found in any comparison populations in this study. Mean total cirDNA was unchanged between healthy controls (ALU-115\u20133.31\u2009ng\/ml; TP53\u20132.73\u2009ng\/ml), hospital controls (ALU-115\u20133.47\u2009ng\/ml; TP53\u20132.83\u2009ng\/ml) and endometriosis patients (ALU-115\u20133.35\u2009ng\/ml; TP53\u20132.66\u2009ng\/ml). Likewise, endometrial cirDNA was unchanged between healthy controls (18.3 copies\/ml), hospital controls (20.6 copies\/ml) and endometriosis patients (22 copies\/ml). Endometrial cirDNA did not change during menstruation (non-menstruating: 38 copies\/ml; menstruating: 33 copies\/ml). Irrespective of endometriosis diagnosis, blood from patients undergoing laparoscopy (hospital controls: 0.77; endometriosis patients: 0.79), had a significantly higher cirDNA size ratio than community-recruited healthy controls (0.64), indicating increased abundance of long cirDNA fragments.<\/jats:p>\n <\/jats:sec>\n \n LIMITATIONS, REASONS FOR CAUTION<\/jats:title>\n It was not possible to completely match the age, BMI and parity between the three cohorts investigated, however of these, only age has been shown to influence circulating DNA levels and not within the age range of our cohort. Blood from community-recruited healthy women and women undergoing laparoscopy was collected via antecubital vein venepuncture (processed within 3\u2009h) and with either peripheral cannula or venepuncture (processed within 6\u2009h), respectively, which could potentially impact the size distribution of circulating DNA fragments. For the collection of non-menstruating phase blood samples, we did not differentiate between follicular phase, ovulation and luteal phase. Thus, only the mensturating samples were collected at a consistent phase, and any fluctuations in cirDNA that occur at the other phases may have obscured small changes during menstruation.<\/jats:p>\n <\/jats:sec>\n \n WIDER IMPLICATIONS OF THE FINDINGS<\/jats:title>\n There is no evidence that cirDNA has potential as a diagnostic biomarker for endometriosis. Endometriosis, representing a common benign gynaecological condition, and menstruation, representing a normal physiological occurrence in women, should not affect methylation-based diagnostics in other disease areas, including oncology.<\/jats:p>\n <\/jats:sec>\n \n STUDY FUNDING\/COMPETING INTEREST(S)<\/jats:title>\n N.L.Y.: Australian Government Research Training Program (RTP) Stipend through The University of New South Wales, Translational Cancer Research Network PhD Scholarship Top-Up Award via the Cancer Institute NSW, Beth Yarrow Memorial Award in Medical Science, UNSW Completion Scholarship; C.E.H.: Gynaecological Oncology Fund of the Royal Hospital for Women; K.W.: Ovarian Cancer Research Foundation and CAMILLA AND MARC. C.E.F.: UNSW Women\u2019s Wellbeing Academy and the Australian Human Rights Institute. We declare the following competing interest: K.W. holds stock in Guardant Health, Exact Sciences and Epigenomics AG. No other authors have competing interests.<\/jats:p>\n <\/jats:sec>\n \n TRIAL REGISTRATION NUMBER<\/jats:title>\n N\/A.<\/jats:p>\n <\/jats:sec>","DOI":"10.1093\/humrep\/deac198","type":"journal-article","created":{"date-parts":[[2022,9,27]],"date-time":"2022-09-27T19:39:24Z","timestamp":1664307564000},"page":"2560-2569","source":"Crossref","is-referenced-by-count":7,"title":["Circulating cell-free endometrial DNA level is unaltered during menstruation and in endometriosis"],"prefix":"10.1093","volume":"37","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9515-3020","authenticated-orcid":false,"given":"N L","family":"Yuwono","sequence":"first","affiliation":[{"name":"Department of Obstetrics and Gynaecology, Gynaecological Cancer Research Group, Adult Cancer Program, School of Women\u2019s and Children\u2019s Health, Faculty of Medicine and Health, University of New South Wales , Sydney, NSW, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A","family":"Alonso","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynaecology, Gynaecological Research and Clinical Evaluation (GRACE) Unit, Royal Hospital for Women , Sydney, NSW, Australia"},{"name":"Department of Obstetrics and Gynaecology, School of Women\u2019s and Children\u2019s Health, Faculty of Medicine and Health, University of New South Wales , Sydney, NSW, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J","family":"Abbott","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynaecology, Gynaecological Research and Clinical Evaluation (GRACE) Unit, Royal Hospital for Women , Sydney, NSW, Australia"},{"name":"Department of Obstetrics and Gynaecology, School of Women\u2019s and Children\u2019s Health, Faculty of Medicine and Health, University of New South Wales , Sydney, NSW, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S","family":"Houshdaran","sequence":"additional","affiliation":[{"name":"Department of Obstetrics, Gynecology and Reproductive Sciences, Center for Reproductive Sciences, University of California, San Francisco , San Francisco, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C E","family":"Henry","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynaecology, Gynaecological Cancer Research Group, Adult Cancer Program, School of Women\u2019s and Children\u2019s Health, Faculty of Medicine and Health, University of New South Wales , Sydney, NSW, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R","family":"Rodgers","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynaecology, School of Women\u2019s and Children\u2019s Health, Faculty of Medicine and Health, University of New South Wales , Sydney, NSW, Australia"},{"name":"Department of Gynaecology and Reproductive Medicine, Royal Hospital for Women , Sydney, NSW, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C E","family":"Ford","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynaecology, Gynaecological Cancer Research Group, Adult Cancer Program, School of Women\u2019s and Children\u2019s Health, Faculty of Medicine and Health, University of New South Wales , Sydney, NSW, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9129-9081","authenticated-orcid":false,"given":"K","family":"Warton","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynaecology, Gynaecological Cancer Research Group, Adult Cancer Program, School of Women\u2019s and Children\u2019s Health, Faculty of Medicine and Health, University of New South Wales , Sydney, NSW, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2022,9,27]]},"reference":[{"key":"2022110206062825700_deac198-B1","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1016\/j.jmig.2019.12.003","article-title":"An abundance of studies but dearth of evidence regarding endometriosis and adenomyosis","volume":"27","author":"Abbott","year":"2020","journal-title":"J Minim Invasive Gynecol"},{"key":"2022110206062825700_deac198-B2","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1016\/j.rbmo.2021.11.006","article-title":"Comparison of total and endometrial circulating cell-free DNA in women with and without endometriosis","volume":"44","author":"Alonso","year":"2022","journal-title":"Reprod Biomed Online"},{"key":"2022110206062825700_deac198-B3","doi-asserted-by":"crossref","first-page":"16253","DOI":"10.1038\/s41598-020-73389-2","article-title":"Endometriosis and chronic pelvic pain have similar impact on women, but time to diagnosis is decreasing: an Australian survey","volume":"10","author":"Armour","year":"2020","journal-title":"Sci Rep"},{"key":"2022110206062825700_deac198-B4","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1148\/rg.2018170093","article-title":"Atypical sites of deeply infiltrative endometriosis: clinical characteristics and imaging findings","volume":"38","author":"Chami\u00e9","year":"2018","journal-title":"Radiographics"},{"key":"2022110206062825700_deac198-B5","doi-asserted-by":"crossref","first-page":"2313","DOI":"10.1111\/jan.14078","article-title":"Drawing blood from peripheral intravenous cannula compared with venepuncture: a systematic review and meta-analysis","volume":"75","author":"Coventry","year":"2019","journal-title":"J Adv Nurs"},{"key":"2022110206062825700_deac198-B6","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1016\/j.cca.2013.05.022","article-title":"Circulating cell free DNA: preanalytical considerations","volume":"424","author":"El Messaoudi","year":"2013","journal-title":"Clin Chim Acta"},{"key":"2022110206062825700_deac198-B7","doi-asserted-by":"crossref","first-page":"2389","DOI":"10.1056\/NEJMcp1000274","article-title":"Clinical practice. Endometriosis","volume":"362","author":"Giudice","year":"2010","journal-title":"N Engl J Med"},{"key":"2022110206062825700_deac198-B8","doi-asserted-by":"crossref","first-page":"e462","DOI":"10.1097\/TXD.0000000000000902","article-title":"The measurement of donor-specific cell-free DNA identifies recipients with biopsy-proven acute rejection requiring treatment after liver transplantation","volume":"5","author":"Goh","year":"2019","journal-title":"Transplant Direct"},{"key":"2022110206062825700_deac198-B9","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1016\/j.jpba.2013.06.010","article-title":"Assessment of diurnal changes and confounding factors that affect circulating cell death biomarker levels: a short communication","volume":"84","author":"Greystoke","year":"2013","journal-title":"J Pharm Biomed Anal"},{"key":"2022110206062825700_deac198-B10","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1097\/GRF.0b013e3181db7ce8","article-title":"Invasive and noninvasive methods for the diagnosis of endometriosis","volume":"53","author":"Hsu","year":"2010","journal-title":"Clin Obstet Gynecol"},{"key":"2022110206062825700_deac198-B11","doi-asserted-by":"crossref","first-page":"236","DOI":"10.1038\/s41398-018-0264-x","article-title":"Cell-free DNA release under psychosocial and physical stress conditions","volume":"8","author":"Hummel","year":"2018","journal-title":"Transl Psychiatry"},{"key":"2022110206062825700_deac198-B12","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1136\/ijgc-2020-001656","article-title":"Clear cell carcinoma of the ovary: a clinical and molecular perspective","volume":"31","author":"Iida","year":"2021","journal-title":"Int J Gynecol Cancer"},{"key":"2022110206062825700_deac198-B13","doi-asserted-by":"crossref","first-page":"1878","DOI":"10.1038\/bjc.2014.29","article-title":"Risk and prognosis of ovarian cancer in women with endometriosis: a meta-analysis","volume":"110","author":"Kim","year":"2014","journal-title":"Br J Cancer"},{"key":"2022110206062825700_deac198-B14","doi-asserted-by":"crossref","first-page":"1167","DOI":"10.1016\/j.annonc.2021.05.806","article-title":"Clinical validation of a targeted methylation-based multi-cancer early detection test using an independent validation set","volume":"32","author":"Klein","year":"2021","journal-title":"Ann Oncol"},{"key":"2022110206062825700_deac198-B15","doi-asserted-by":"crossref","first-page":"1614","DOI":"10.1373\/clinchem.2017.272401","article-title":"DNA of erythroid origin is present in human plasma and informs the types of anemia","volume":"63","author":"Lam","year":"2017","journal-title":"Clin Chem"},{"key":"2022110206062825700_deac198-B16","doi-asserted-by":"crossref","first-page":"E1826","DOI":"10.1073\/pnas.1519286113","article-title":"Identification of tissue-specific cell death using methylation patterns of circulating DNA","volume":"113","author":"Lehmann-Werman","year":"2016","journal-title":"Proc Natl Acad Sci USA"},{"key":"2022110206062825700_deac198-B17","doi-asserted-by":"crossref","first-page":"745","DOI":"10.1016\/j.annonc.2020.02.011","article-title":"Sensitive and specific multi-cancer detection and localization using methylation signatures in cell-free DNA","volume":"6","author":"Liu","year":"2020","journal-title":"Ann Oncol"},{"key":"2022110206062825700_deac198-B18","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1186\/s12872-020-01587-x","article-title":"Cardiac-specific methylation patterns of circulating DNA for identification of cardiomyocyte death","volume":"20","author":"Liu","year":"2020","journal-title":"BMC Cardiovasc Disord"},{"key":"2022110206062825700_deac198-B19","first-page":"7689","article-title":"Association between ovarian cancer and advanced endometriosis","volume":"5","author":"Matalliotakis","year":"2018","journal-title":"Oncol Lett"},{"key":"2022110206062825700_deac198-B20","doi-asserted-by":"crossref","first-page":"1087","DOI":"10.1007\/s00421-020-04593-1","article-title":"Changes in plasma hydroxyproline and plasma cell-free DNA concentrations after higher- versus lower-intensity eccentric cycling","volume":"121","author":"Mavropalias","year":"2021","journal-title":"Eur J Appl Physiol"},{"key":"2022110206062825700_deac198-B21","doi-asserted-by":"crossref","first-page":"307","DOI":"10.5500\/wjt.v10.i11.307","article-title":"Donor-specific cell-free DNA as a biomarker in liver transplantation: a review","volume":"10","author":"McClure","year":"2020","journal-title":"World J Transplant"},{"key":"2022110206062825700_deac198-B22","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1111\/1471-0528.16447","article-title":"Prevalence and incidence of endometriosis in Australian women: a data linkage cohort study","volume":"128","author":"Rowlands","year":"2021","journal-title":"BJOG"},{"key":"2022110206062825700_deac198-B23","doi-asserted-by":"crossref","first-page":"1863","DOI":"10.1093\/humrep\/deq141","article-title":"Non-invasive diagnosis of endometriosis: the goal or own goal?","volume":"25","author":"Somigliana","year":"2010","journal-title":"Hum Reprod"},{"key":"2022110206062825700_deac198-B24","doi-asserted-by":"crossref","first-page":"e69805","DOI":"10.1371\/journal.pone.0069805","article-title":"Complete genes may pass from food to human blood","volume":"8","author":"Spis\u00e1k","year":"2013","journal-title":"PLoS One"},{"key":"2022110206062825700_deac198-B25","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1002\/ijgo.12521","article-title":"An evidence-based approach to assessing surgical versus clinical diagnosis of symptomatic endometriosis","volume":"142","author":"Taylor","year":"2018","journal-title":"Int J Gynecol Obstet"},{"key":"2022110206062825700_deac198-B26","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.cca.2009.08.011","article-title":"Characterisation of circulating DNA by parallel tagged sequencing on the 454 platform","volume":"409","author":"van der Vaart","year":"2009","journal-title":"Clin Chim Acta"},{"key":"2022110206062825700_deac198-B27","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1007\/978-1-4939-8973-7_14","article-title":"Circulating cell-free DNA in physical activities","volume":"1909","author":"Vittori","year":"2019","journal-title":"Methods Mol Biol"},{"key":"2022110206062825700_deac198-B28","doi-asserted-by":"crossref","first-page":"476","DOI":"10.1186\/1471-2164-15-476","article-title":"Methylation-capture and next-generation sequencing of free circulating DNA from human plasma","volume":"15","author":"Warton","year":"2014","journal-title":"BMC Genomics"},{"key":"2022110206062825700_deac198-B29","doi-asserted-by":"crossref","first-page":"931","DOI":"10.1016\/j.cjca.2019.10.016","article-title":"Clinical significance of circulating cardiomyocyte-specific cell-free DNA in patients with heart failure: a proof-of-concept study","volume":"36","author":"Yokokawa","year":"2020","journal-title":"Can J Cardiol"},{"key":"2022110206062825700_deac198-B30","doi-asserted-by":"crossref","first-page":"e0250561","DOI":"10.1371\/journal.pone.0250561","article-title":"Total and endothelial cell-derived cell-free DNA in blood plasma does not change during menstruation","volume":"16","author":"Yuwono","year":"2021","journal-title":"PLoS One"},{"key":"2022110206062825700_deac198-B31","doi-asserted-by":"crossref","first-page":"e69679","DOI":"10.7554\/eLife.69679","article-title":"The influence of biological and lifestyle factors on circulating cell-free DNA in blood plasma","volume":"10","author":"Yuwono","year":"2021","journal-title":"eLife"},{"key":"2022110206062825700_deac198-B32","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1016\/S1472-6483(10)60100-9","article-title":"Circulating cell-free DNA as a potential biomarker for minimal and mild endometriosis","volume":"18","author":"Zachariah","year":"2009","journal-title":"Reprod Biomed Online"},{"key":"2022110206062825700_deac198-B33","doi-asserted-by":"crossref","first-page":"1443","DOI":"10.1038\/s41467-018-03961-y","article-title":"Non-invasive detection of human cardiomyocyte death using methylation patterns of circulating DNA","volume":"9","author":"Zemmour","year":"2018","journal-title":"Nat Commun"},{"key":"2022110206062825700_deac198-B34","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1038\/s41467-020-20162-8","article-title":"Pan-cancer circulating tumor DNA detection in over 10,000 Chinese patients","volume":"12","author":"Zhang","year":"2021","journal-title":"Nat Commun"}],"container-title":["Human Reproduction"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/humrep\/article-pdf\/37\/11\/2560\/46694782\/deac198.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/humrep\/article-pdf\/37\/11\/2560\/46694782\/deac198.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,11,2]],"date-time":"2022-11-02T06:07:34Z","timestamp":1667369254000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/humrep\/article\/37\/11\/2560\/6724348"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,9,27]]},"references-count":34,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2022,9,27]]},"published-print":{"date-parts":[[2022,10,31]]}},"URL":"https:\/\/doi.org\/10.1093\/humrep\/deac198","relation":{},"ISSN":["0268-1161","1460-2350"],"issn-type":[{"value":"0268-1161","type":"print"},{"value":"1460-2350","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2022,11,1]]},"published":{"date-parts":[[2022,9,27]]}}}