Advertisement

Comparison of total and endometrial circulating cell-free DNA in women with and without endometriosis

  • Author Footnotes
    # Contributed equally to this study.
    Anais Alonso
    Footnotes
    # Contributed equally to this study.
    Affiliations
    Gynaecological Research and Clinical Evaluation (GRACE) Unit, Royal Hospital for Women, Sydney NSW, Australia

    School of Women's and Children's Health, Faculty of Medicine and Health, University of New South Wales, Sydney NSW, Australia
    Search for articles by this author
  • Author Footnotes
    # Contributed equally to this study.
    Nicole Laurencia Yuwono
    Footnotes
    # Contributed equally to this study.
    Affiliations
    Gynaecological Cancer Research Group, Adult Cancer Program, School of Women's and Children's Health, Faculty of Medicine and Health, University of New South Wales, Sydney NSW, Australia
    Search for articles by this author
  • Sahar Houshdaran
    Affiliations
    Center for Reproductive Sciences, Department of Obstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco, USA
    Search for articles by this author
  • Jason Abbott
    Affiliations
    Gynaecological Research and Clinical Evaluation (GRACE) Unit, Royal Hospital for Women, Sydney NSW, Australia

    School of Women's and Children's Health, Faculty of Medicine and Health, University of New South Wales, Sydney NSW, Australia
    Search for articles by this author
  • Rachael Rodgers
    Affiliations
    Center for Reproductive Sciences, Department of Obstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco, USA

    Department of Gynaecology and Reproductive Medicine, Royal Hospital for Women, Sydney NSW, Australia
    Search for articles by this author
  • Caroline Elizabeth Ford
    Affiliations
    Gynaecological Cancer Research Group, Adult Cancer Program, School of Women's and Children's Health, Faculty of Medicine and Health, University of New South Wales, Sydney NSW, Australia
    Search for articles by this author
  • Kristina Warton
    Correspondence
    Corresponding author.
    Affiliations
    Gynaecological Cancer Research Group, Adult Cancer Program, School of Women's and Children's Health, Faculty of Medicine and Health, University of New South Wales, Sydney NSW, Australia
    Search for articles by this author
  • Author Footnotes
    # Contributed equally to this study.
Published:November 12, 2021DOI:https://doi.org/10.1016/j.rbmo.2021.11.006

      Abstract

      Research question

      Do women with laparoscopically confirmed endometriosis have higher plasma concentrations of circulating cell-free DNA (cirDNA) than those without endometriosis?

      Design

      Prospective study of women aged 18–45 years undergoing benign gynaecological laparoscopy at two tertiary hospitals. Venous blood was collected immediately before surgery, and women were allocated to the endometriosis or control groups based on surgical findings. Total plasma cirDNA and cirDNA integrity were measured by quantitative polymerase chain reaction (qPCR) targeting short (115 bases) and long (247 bases) ALU segments. Endometrial-derived cirDNA was measured by qPCR of bisulfite-treated cirDNA using primers selective for a FAM101A sequence uniquely unmethylated in endometrial tissue. Five cirDNA parameters were compared between the control and endometriosis cohorts: total cirDNA concentration, long-stranded cirDNA concentration, integrity ratio, endometrial cirDNA concentration and endometrial cirDNA proportion.

      Results

      Twenty-eight endometriosis and 15 control samples were included. Women with and without endometriosis had cirDNA concentrations of 2.24 ± 0.89 ng/ml and 2.56 ± 0.92 ng/ml, respectively. Analysis by phenotype of endometriosis revealed a significantly higher endometrial cirDNA concentration in women with superficial disease (n = 10) compared with deep endometriosis (n = 18) (mean difference 0.14 ng/ml; 95% CI 0.15 to 0.26; P = 0.025), but not with controls.

      Conclusions

      No significant differences were found in any of the cirDNA parameters between women with and without endometriosis. The low statistical power and heterogenous pelvic pathology in the control group render it difficult to determine whether the negative results reflect a true lack of increase in cirDNA in endometriosis.

      KEYWORDS

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic and Personal

      Subscribe:

      Subscribe to Reproductive BioMedicine Online
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Abbott J.A.
        • Hawe J.
        • Clayton R.D.
        • Garry R.
        The Effects And Effectiveness Of Laparoscopic Excision Of Endometriosis: A Prospective Study With 2-5 Year Follow-Up.
        Hum. Reprod. 2003; 18: 1922-1927
        • Abdelal I.T.
        • Zakaria M.A.
        • Sharaf D.M.
        • Elakad G.M.
        Levels Of Plasma Cell-Free Dna And Its Correlation With Disease Activity In Rheumatoid Arthritis And Systemic Lupus Erythematosus Patients.
        The Egyptian Rheumatologist. 2016; 38: 295-300
        • Alghofaili L.
        • Almubarak H.
        • Gassem K.
        • Islam S.S.
        • Coskun S.
        • Kaya N.
        • Karakas B.
        Cell-Free Dna Levels Of Twins And Sibling Pairs Indicate Individuality And Possible Use As A Personalized Biomarker.
        Plos One. 2019; 14E0223470
        • Anaf V.
        • Chapron C.
        • El Nakadi I.
        • De Moor V.
        • Simonart T.
        • Noel J.C.
        Pain, Mast Cells, And Nerves In Peritoneal, Ovarian, And Deep Infiltrating Endometriosis.
        Fertil. Steril. 2006; 86: 1336-1343
        • Andersen R.F.
        • Spindler K.-L.G.
        • Brandslund I.
        • Jakobsen A.
        • Pallisgaard N.
        Improved Sensitivity Of Circulating Tumor Dna Measurement Using Short Pcr Amplicons.
        Clinica Chimica Acta. 2015; 439: 97-101
        • Arko-Boham B.
        • Aryee N.A.
        • Blay R.M.
        • Owusu E.D.A.
        • Tagoe E.A.
        • Doris Shackie E.S.
        • Debrah A.B.
        • Adu-Aryee N.A.
        Circulating Cell-Free Dna Integrity As A Diagnostic And Prognostic Marker For Breast And Prostate Cancers.
        Cancer Genet. 2019; 235-236: 65-71
        • Baker S.G.
        • Kramer B.S.
        • Mcintosh M.
        • Patterson B.H.
        • Shyr Y.
        • Skates S.
        Evaluating Markers For The Early Detection Of Cancer: Overview Of Study Designs And Methods.
        Clin. Trials. 2006; 3: 43-56
        • Catarino R.
        • Ferreira M.M.
        • Rodrigues H.
        • Coelho A.
        • Nogal A.
        • Sousa A.
        • Medeiros R.
        Quantification Of Free Circulating Tumor Dna As A Diagnostic Marker For Breast Cancer.
        Dna Cell Biol. 2008; 27: 415-421
        • Dmowski W.P.
        • Ding J.
        • Shen J.
        • Rana N.
        • Fernandez B.B.
        • Braun D.P.
        Apoptosis In Endometrial Glandular And Stromal Cells In Women With And Without Endometriosis.
        Hum. Reprod. 2001; 16: 1802-1808
        • Duffy J.M.
        • Arambage K.
        • Correa F.J.
        • Olive D.
        • Farquhar C.
        • Garry R.
        • Barlow D.H.
        • Jacobson T.Z.
        Laparoscopic Surgery For Endometriosis.
        Cochrane Database Syst. Rev. 2014; Cd011031
        • Frishman G.N.
        • Salak J.R.
        Conservative Surgical Management Of Endometriosis In Women With Pelvic Pain.
        J. Minim. Invasive Gynecol. 2006; 13: 546-558
        • Hashimoto T.
        • Yoshida K.
        • Hashimoto N.
        • Nakai A.
        • Kaneshiro K.
        • Suzuki K.
        • Kawasaki Y.
        • Shibanuma N.
        • Hashiramoto A.
        Circulating Cell Free Dna: A Marker To Predict The Therapeutic Response For Biological Dmards In Rheumatoid Arthritis.
        Int. J. Rheum. Dis. 2017; 20: 722-730
        • Holoch K.J.
        • Lessey B.A.
        Endometriosis And Infertility.
        Clin. Obstet. Gynecol. 2010; 53: 429-438
        • Houshdaran S.
        • Zelenko Z.
        • Irwin J.C.
        • Giudice L.C.
        Human Endometrial Dna Methylome Is Cycle-Dependent And Is Associated With Gene Expression Regulation.
        Molecular Endocrinology. 2014; 28: 1118-1135
        • Hussein N.A.
        • Mohamed S.N.
        • Ahmed M.A.
        Plasma Alu-247, Alu-115, And Cfdna Integrity As Diagnostic And Prognostic Biomarkers For Breast Cancer.
        Appl. Biochem. Biotechnol. 2019; 187: 1028-1045
        • Kamat A.A.
        • Sood A.K.
        • Dang D.
        • Gershenson D.M.
        • Simpson J.L.
        • Bischoff F.Z.
        Quantification Of Total Plasma Cell-Free Dna In Ovarian Cancer Using Real-Time Pcr.
        Ann. N. Y. Acad. Sci. 2006; 1075: 230-234
        • Kim J.H.
        • Han E.
        Endometriosis And Female Pelvic Pain.
        Semin. Reprod. Med. 2018; 36: 143-151
        • Liu Q.
        • Ma J.
        • Deng H.
        • Huang S.J.
        • Rao J.
        • Xu W.B.
        • Huang J.S.
        • Sun S.Q.
        • Zhang L.
        Cardiac-Specific Methylation Patterns Of Circulating Dna For Identification Of Cardiomyocyte Death.
        Bmc Cardiovasc. Disord. 2020; 20: 310
        • Liu X.
        • Ren J.
        • Luo N.
        • Guo H.
        • Zheng Y.
        • Li J.
        • Tang F.
        • Wen L.
        • Peng J.
        Comprehensive Dna Methylation Analysis Of Tissue Of Origin Of Plasma Cell-Free Dna By Methylated Cpg Tandem Amplification And Sequencing (Mcta-Seq).
        Clin. Epigenetics. 2019; 11: 93
        • Lo Y.M.
        • Zhang J.
        • Leung T.N.
        • Lau T.K.
        • Chang A.M.
        • Hjelm N.M.
        Rapid Clearance Of Fetal Dna From Maternal Plasma.
        Am. J. Hum. Genet. 1999; 64: 218-224
        • Meresman G.F.
        • Vighi S.
        • Buquet R.A.
        • Contreras-Ortiz O.
        • Tesone M.
        • Rumi L.S.
        Apoptosis And Expression Of Bcl-2 And Bax In Eutopic Endometrium From Women With Endometriosis.
        Fertil. Steril. 2000; 74: 760-766
        • Moss J.
        • Magenheim J.
        • Neiman D.
        • Zemmour H.
        • Loyfer N.
        • Korach A.
        • Samet Y.
        • Maoz M.
        • Druid H.
        • Arner P.
        • Fu K.Y.
        • Kiss E.
        • Spalding K.L.
        • Landesberg G.
        • Zick A.
        • Grinshpun A.
        • Shapiro A.M.J.
        • Grompe M.
        • Wittenberg A.D.
        • Glaser B.
        • Shemer R.
        • Kaplan T.
        • Dor Y.
        Comprehensive Human Cell-Type Methylation Atlas Reveals Origins Of Circulating Cell-Free Dna In Health And Disease.
        Nat. Commun. 2018; 9: 5068
        • Nnoaham K.E.
        • Hummelshoj L.
        • Webster P.
        • D'hooghe T.
        • De Cicco Nardone F.
        • De Cicco Nardone C.
        • Jenkinson C.
        • Kennedy S.H.
        • Zondervan K.T.
        • World Endometriosis Research Foundation Global Study Of Women's Health, C
        Impact Of Endometriosis On Quality Of Life And Work Productivity: A Multicenter Study Across Ten Countries.
        Fertil. Steril. 2011; 96 (E8): 366-373
        • Pölcher M.
        • Ellinger J.
        • Willems S.
        • El-Maarri O.
        • Höller T.
        • Amann C.
        • Wolfgarten M.
        • Rudlowski C.
        • Kuhn W.
        • Braun M.
        Impact Of The Menstrual Cycle On Circulating Cell-Free Dna.
        Anticancer Res. 2010; 30: 2235-2240
        • Ransohoff D.F.
        • Feinstein A.R.
        Problems Of Spectrum And Bias In Evaluating The Efficacy Of Diagnostic Tests.
        N. Engl. J. Med. 1978; 299: 926-930
        • Rowlands I.J.
        • Abbott J.A.
        • Montgomery G.W.
        • Hockey R.
        • Rogers P.
        • Mishra G.D.
        Prevalence And Incidence Of Endometriosis In Australian Women: A Data Linkage Cohort Study.
        Bjog. 2020; (N/A)
        • Rykova E.
        • Sizikov A.
        • Roggenbuck D.
        • Antonenko O.
        • Bryzgalov L.
        • Morozkin E.
        • Skvortsova K.
        • Vlassov V.
        • Laktionov P.
        • Kozlov V.
        Circulating Dna In Rheumatoid Arthritis: Pathological Changes And Association With Clinically Used Serological Markers.
        Arthritis Res. Ther. 2017; 19: 85
        • Szymanowski K.
        Apoptosis Pattern In Human Endometrium In Women With Pelvic Endometriosis.
        Eur. J. Obstet. Gynecol. Reprod. Biol. 2007; 132: 107-110
        • Tran L.V.
        • Tokushige N.
        • Berbic M.
        • Markham R.
        • Fraser I.S.
        Macrophages And Nerve Fibres In Peritoneal Endometriosis.
        Hum. Reprod. 2009; 24: 835-841
        • Tug S.
        • Helmig S.
        • Menke J.
        • Zahn D.
        • Kubiak T.
        • Schwarting A.
        • Simon P.
        Correlation Between Cell Free Dna Levels And Medical Evaluation Of Disease Progression In Systemic Lupus Erythematosus Patients.
        Cell Immunol. 2014; 292: 32-39
        • Umetani N.
        • Kim J.
        • Hiramatsu S.
        • Reber H.A.
        • Hines O.J.
        • Bilchik A.J.
        • Hoon D.S.B.
        Increased Integrity Of Free Circulating Dna In Sera Of Patients With Colorectal Or Periampullary Cancer: Direct Quantitative Pcr For Alu Repeats.
        Clinical Chemistry. 2006; 52: 1062-1069
        • Xu Y.
        • Song Y.
        • Chang J.
        • Zhou X.
        • Qi Q.
        • Tian X.
        • Li M.
        • Zeng X.
        • Xu M.
        • Zhang W.
        • Cram D.S.
        • Liu J.
        High Levels Of Circulating Cell-Free Dna Are A Biomarker Of Active Sle.
        Eur. J. Clin. Invest. 2018; 48: E13015
        • Yokokawa T.
        • Misaka T.
        • Kimishima Y.
        • Shimizu T.
        • Kaneshiro T.
        • Takeishi Y.
        Clinical Significance Of Circulating Cardiomyocyte-Specific Cell-Free Dna In Patients With Heart Failure: A Proof-Of-Concept Study.
        Canadian Journal Of Cardiology. 2020; 36: 931-935
        • Zachariah R.
        • Schmid S.
        • Radpour R.
        • Buerki N.
        • Fan A.X.-C.
        • Hahn S.
        • Holzgreve W.
        • Zhong X.Y.
        Circulating Cell-Free Dna As A Potential Biomarker For Minimal And Mild Endometriosis.
        Reproductive Biomedicine Online. 2009; 18: 407-411
        • Zanetti-Dallenbach R.A.
        • Schmid S.
        • Wight E.
        • Holzgreve W.
        • Ladewing A.
        • Hahn S.
        • Zhong X.Y.
        Levels Of Circulating Cell-Free Serum Dna In Benign And Malignant Breast Lesions.
        Int. J. Biol. Markers. 2007; 22: 95-99
        • Zemmour H.
        • Planer D.
        • Magenheim J.
        • Moss J.
        • Neiman D.
        • Gilon D.
        • Korach A.
        • Glaser B.
        • Shemer R.
        • Landesberg G.
        • Dor Y.
        Non-Invasive Detection Of Human Cardiomyocyte Death Using Methylation Patterns Of Circulating Dna.
        Nat. Commun. 2018; 9: 1443
        • Zhang Q.
        • Hu G.
        • Yang Q.
        • Dong R.
        • Xie X.
        • Ma D.
        • Shen K.
        • Kong B.
        A Multiplex Methylation-Specific Pcr Assay For The Detection Of Early-Stage Ovarian Cancer Using Cell-Free Serum Dna.
        Gynecol. Oncol. 2013; 130: 132-139

      Biography

      Anais Alonso is a fifth-year medical student at the University of New South Wales, Australia, having recently completed her BSc (Med) Hons degree.
      Key message
      Endometriosis affects the lives of many pre-menopausal women; however, biomarkers to detect the disease non-invasively are lacking. Circulating cell-free DNA in plasma offers a potential substrate for a diagnostic test; however, no significant increase in total or endometrial-derived cell-free DNA in women with endometriosis was observed.