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Arrangement of myofibroblastic and smooth muscle-like cells in superficial peritoneal endometriosis and a possible role of transforming growth factor beta 1 (TGFβ1) in myofibroblastic metaplasia

  • Mohamed Gamal Ibrahim
  • Martin Sillem
  • Johanna Plendl
  • Eliane T. Taube
  • Andreas Schüring
  • Martin Götte
  • Vito Chiantera
  • Jalid Sehouli
  • Sylvia Mechsner
General Gynecology
  • 18 Downloads

Abstract

Purpose

Superficial peritoneal endometriotic (pEM) lesions are composed of endometrial glands and stroma, in addition to a third component—myofibroblasts and smooth muscles (SM)-like cells. The latter develops secondary to a metaplasia. In this study, we characterised the third component cells in pEM according to differentiation markers in different micro-compartments. Furthermore, a possible effect of TGFβ1 on myofibroblastic metaplasia in endometriotic epithelial cells was studied.

Methods

Seventy-six premenopausal patients were included. Peritoneal biopsies were excised from EM patients (n = 23), unaffected peritoneum (peritoneum from EM patients but without EM components, n = 5/23) and non-EM patients (n = 10). All peritoneal biopsies were immunolabeled for ASMA, calponin, collagen I, desmin, TGFß receptor 1 (R1), R2 and R3 in addition to ultrastructure examination by transmission electron microscopy (TEM) (n = 1). TGFß1 level was measured in peritoneal fluid (PF) (EM, n = 19 and non-EM, n = 13) collected during laparoscopy. Furthermore, TGFß1 effect on myofibroblastic metaplasia was studied in vitro.

Results

At the centre of pEM lesions, calponin immunolabeling outweighs the collagen I while in the periphery the reverse occurs. SM-like cells expressing desmin predominate at the periphery, while ASMA immunolabeling was detectable in all micro-compartments. Both indicate an abundance of myofibroblasts at the centre of pEM lesions and SM-like cells in the periphery. Although activated TGFß1 in PF did not differ between EM and non-EM, it inhibited the cell proliferation of the endometriotic epithelial cells and induced an upregulation in ASMA and collagen IA2 expression as well.

Conclusion

The abundance of the myofibroblasts and SM-like cells points to a myofibroblastic metaplasia in pEM. Both cells are differentially arranged in the different micro-compartments of pEM lesions, with increasing cell maturity towards the periphery of the lesion. Furthermore, TGFß1 may play a role in the myofibroblastic metaplasia of the endometriotic epithelial cells. These findings provide a better insight in the micro-milieu in EM lesions, where most of the disease dynamics occur.

Keywords

Peritoneal endometriosis Myofibroblastic metaplasia TGFβ1 Smooth muscle-like cells 

Notes

Acknowledgements

We would like to thank the Ernst Schering Foundation, Germany, and the Humboldt University in Berlin, Germany, for the doctoral scholarships and the FAZIT foundation, Germany, for the travel grant awarded to the first author. We would like to thank Professor Anna Starzinski-Powitz for supplying the cell line.

Author contribution

MG Ibrahim participated in the study design, execution (collected the samples, carrying out the experiments), analysis, manuscript drafting and critical discussion. VC helped in sample collection. ETT, MS, AS and JS did manuscript editing and critical discussion. ETT was the expert of the histopathological staining. JP was the expert for TEM, manuscript editing and critical discussion. MG carried out the real-time PCR and manuscript editing. SM helped with the study design, supervision, manuscript editing and critical discussion.

Compliance with ethical standards

Conflict of interest

The first author was granted scholarships from the Ernst Schering Foundation, the Humboldt University in Berlin and FAZIT foundation in the course of his doctoral work.

Informed consent

All patients included in this study were operated on via laparoscopy at Charité University of Medicine and gave written informed consent. The study was approved by the local research and ethics committee at the Charité University of Medicine, Berlin-Germany (EA4/071/07).

Supplementary material

404_2018_4995_MOESM1_ESM.docx (2.9 mb)
Supplementary material 1 (docx 2981 kb)
404_2018_4995_MOESM2_ESM.docx (12 kb)
Supplementary material 2 (docx 12 kb)

References

  1. 1.
    Dunselman GA, Vermeulen N, Becker C, Calhaz-Jorge C, D’Hooghe T, De Bie B, Heikinheimo O, Horne AW, Kiesel L, Nap A, Prentice A, Saridogan E, Soriano D, Nelen W (2014) ESHRE guideline: management of women with endometriosis. Hum Reprod 29(3):400–412.  https://doi.org/10.1093/humrep/det457 CrossRefPubMedGoogle Scholar
  2. 2.
    Mehasseb MK, Bell SC, Pringle JH, Habiba MA (2010) Uterine adenomyosis is associated with ultrastructural features of altered contractility in the inner myometrium. Fertil Steril 93(7):2130–2136.  https://doi.org/10.1016/j.fertnstert.2009.01.097 CrossRefPubMedGoogle Scholar
  3. 3.
    Kennedy S, Bergqvist A, Chapron C, D’Hooghe T, Dunselman G, Greb R, Hummelshoj L, Prentice A, Saridogan E (2005) ESHRE guideline for the diagnosis and treatment of endometriosis. Hum Reprod 20(10):2698–2704CrossRefPubMedGoogle Scholar
  4. 4.
    Anaf V, Simon P, Fayt I, Noel J (2000) Smooth muscles are frequent components of endometriotic lesions. Hum Reprod 15(4):767–771CrossRefPubMedGoogle Scholar
  5. 5.
    Mechsner S, Bartley J, Loddenkemper C, Salomon DS, Starzinski-Powitz A, Ebert AD (2005) Oxytocin receptor expression in smooth muscle cells of peritoneal endometriotic lesions and ovarian endometriotic cysts. Fertil Steril 83(Suppl 1):1220–1231CrossRefPubMedGoogle Scholar
  6. 6.
    Itoga T, Matsumoto T, Takeuchi H, Yamasaki S, Sasahara N, Hoshi T, Kinoshita K (2003) Fibrosis and smooth muscle metaplasia in rectovaginal endometriosis. Pathol Int 53(6):371–375CrossRefPubMedGoogle Scholar
  7. 7.
    van Kaam KJ, Schouten JP, Nap AW, Dunselman GA, Groothuis PG (2008) Fibromuscular differentiation in deeply infiltrating endometriosis is a reaction of resident fibroblasts to the presence of ectopic endometrium. Hum Reprod 23(12):2692–2700CrossRefPubMedGoogle Scholar
  8. 8.
    Sopha SC, Rosado FG, Smith JJ, Merchant NB, Shi C (2015) Hepatic uterus-like mass misdiagnosed as hepatic abscess. Int J Surg Pathol 23(2):134–139.  https://doi.org/10.1177/1066896914534465 CrossRefPubMedGoogle Scholar
  9. 9.
    Flieder DB, Moran CA, Travis WD, Koss MN, Mark EJ (1998) Pleuro-pulmonary endometriosis and pulmonary ectopic deciduosis: a clinicopathologic and immunohistochemical study of 10 cases with emphasis on diagnostic pitfalls. Hum Pathol 29(12):1495–1503CrossRefPubMedGoogle Scholar
  10. 10.
    Ibrahim MG, Delarue E, Abesadze E, Haas M, Sehouli J, Chiantera V, Mechsner S (2016) Abdominal wall endometriosis: myofibroblasts as a possible evidence of metaplasia: a case report. Gynecol Obstet Investig.  https://doi.org/10.1159/000452101 CrossRefGoogle Scholar
  11. 11.
    Barcena de Arellano ML, Gericke J, Reichelt U, Okuducu AF, Ebert AD, Chiantera V, Schneider A, Mechsner S (2011) Immunohistochemical characterization of endometriosis-associated smooth muscle cells in human peritoneal endometriotic lesions. Hum Reprod 26(10):2721–2730.  https://doi.org/10.1093/humrep/der253 CrossRefPubMedGoogle Scholar
  12. 12.
    Liu Y, Dong Z, Liu H, Zhu J, Liu F, Chen G (2015) Transition of mesothelial cell to fibroblast in peritoneal dialysis: EMT, stem cell or bystander? Perit Dial Int 35(1):14–25.  https://doi.org/10.3747/pdi.2014.00188 CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    De Vriese AS, Tilton RG, Mortier S, Lameire NH (2006) Myofibroblast transdifferentiation of mesothelial cells is mediated by RAGE and contributes to peritoneal fibrosis in uraemia. Nephrol Dial Transplant 21(9):2549–2555.  https://doi.org/10.1093/ndt/gfl271 CrossRefPubMedGoogle Scholar
  14. 14.
    Bartley J, Julicher A, Hotz B, Mechsner S, Hotz H (2014) Epithelial to mesenchymal transition (EMT) seems to be regulated differently in endometriosis and the endometrium. Arch Gynecol Obstet 289(4):871–881.  https://doi.org/10.1007/s00404-013-3040-4 CrossRefPubMedGoogle Scholar
  15. 15.
    Matsuzaki S, Darcha C (2012) Epithelial to mesenchymal transition-like and mesenchymal to epithelial transition-like processes might be involved in the pathogenesis of pelvic endometriosis. Hum Reprod 27(3):712–721.  https://doi.org/10.1093/humrep/der442 CrossRefPubMedGoogle Scholar
  16. 16.
    Young VJ, Brown JK, Saunders PT, Duncan WC, Horne AW (2014) The peritoneum is both a source and target of TGF-beta in women with endometriosis. PLoS ONE 9(9):e106773.  https://doi.org/10.1371/journal.pone.0106773 CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Wynn TA (2008) Cellular and molecular mechanisms of fibrosis. J Pathol 214(2):199–210CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Gazvani R, Templeton A (2002) Peritoneal environment, cytokines and angiogenesis in the pathophysiology of endometriosis. Reproduction 123(2):217–226CrossRefGoogle Scholar
  19. 19.
    Ibrahim MG, Sillem M, Plendl J, Chiantera V, Sehouli J, Mechsner S (2017) Myofibroblasts are evidence of chronic tissue microtrauma at the endometrial-myometrial junctional zone in uteri with adenomyosis. Reprod Sci.  https://doi.org/10.1177/1933719116687855 CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Daimon E, Shibukawa Y, Wada Y (2013) Calponin 3 regulates stress fiber formation in dermal fibroblasts during wound healing. Arch Dermatol Res 305(7):571–584.  https://doi.org/10.1007/s00403-013-1343-8 CrossRefPubMedGoogle Scholar
  21. 21.
    Small JV, Gimona M (1998) The cytoskeleton of the vertebrate smooth muscle cell. Acta Physiol Scand 164(4):341–348.  https://doi.org/10.1046/j.1365-201X.1998.00441.x CrossRefPubMedGoogle Scholar
  22. 22.
    Miano JM, Olson EN (1996) Expression of the smooth muscle cell calponin gene marks the early cardiac and smooth muscle cell lineages during mouse embryogenesis. J Biol Chem 271(12):7095–7103CrossRefPubMedGoogle Scholar
  23. 23.
    Daimon E, Shibukawa Y, Wada Y (2013) Calponin 3 regulates stress fiber formation in dermal fibroblasts during wound healing. Arch Dermatol Res 305(7):571–584CrossRefPubMedGoogle Scholar
  24. 24.
    Rahmioglu N, Fassbender A, Vitonis AF, Tworoger SS, Hummelshoj L, D’Hooghe TM, Adamson GD, Giudice LC, Becker CM, Zondervan KT, Missmer SA (2014) World Endometriosis Research Foundation Endometriosis Phenome and Biobanking Harmonization Project: III. Fluid biospecimen collection, processing, and storage in endometriosis research. Fertil Steril 102(5):1233–1243.  https://doi.org/10.1016/j.fertnstert.2014.07.1208 CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Fassbender A, Rahmioglu N, Vitonis AF, Vigano P, Giudice LC, D’Hooghe TM, Hummelshoj L, Adamson GD, Becker CM, Missmer SA, Zondervan KT (2014) World Endometriosis Research Foundation Endometriosis Phenome and Biobanking Harmonisation Project: IV. Tissue collection, processing, and storage in endometriosis research. Fertil Steril 102(5):1244–1253.  https://doi.org/10.1016/j.fertnstert.2014.07.1209 CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Mechsner S, Grum B, Gericke C, Loddenkemper C, Dudenhausen JW, Ebert AD (2010) Possible roles of oxytocin receptor and vasopressin-1alpha receptor in the pathomechanism of dysperistalsis and dysmenorrhea in patients with adenomyosis uteri. Fertil Steril 94(7):2541–2546.  https://doi.org/10.1016/j.fertnstert.2010.03.015 CrossRefPubMedGoogle Scholar
  27. 27.
    Richardson KC, Jarett L, Finke EH (1960) Embedding in epoxy resins for ultrathin sectioning in electron microscopy. Stain Technol 35:313–323CrossRefPubMedGoogle Scholar
  28. 28.
    Zeitvogel A, Baumann R, Starzinski-Powitz A (2001) Identification of an invasive, N-cadherin-expressing epithelial cell type in endometriosis using a new cell culture model. Am J Pathol 159(5):1839–1852.  https://doi.org/10.1016/S0002-9440(10)63030-1 CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Veres-Szekely A, Pap D, Sziksz E, Javorszky E, Rokonay R, Lippai R, Tory K, Fekete A, Tulassay T, Szabo AJ, Vannay A (2017) Selective measurement of alpha smooth muscle actin: why beta-actin cannot be used as a housekeeping gene when tissue fibrosis occurs. BMC Mol Biol 18(1):12.  https://doi.org/10.1186/s12867-017-0089-9 CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Koks CA, Groothuis PG, Dunselman GA, de Goeij AF, Evers JL (2000) Adhesion of menstrual endometrium to extracellular matrix: the possible role of integrin alpha(6)beta(1) and laminin interaction. Mol Hum Reprod 6(2):170–177CrossRefPubMedGoogle Scholar
  31. 31.
    Merrill JA (1966) Endometrial induction of endometriosis across millipore filters. Am J Obstet Gynecol 94(6):780–790PubMedGoogle Scholar
  32. 32.
    Nakayama K, Masuzawa H, Li SF, Yoshikawa F, Toki T, Nikaido T, Silverberg SG, Fujii S (1994) Immunohistochemical analysis of the peritoneum adjacent to endometriotic lesions using antibodies for Ber-EP4 antigen, estrogen receptors, and progesterone receptors: implication of peritoneal metaplasia in the pathogenesis of endometriosis. Int J Gynecol Pathol 13(4):348–358CrossRefPubMedGoogle Scholar
  33. 33.
    Mai KT, Yazdi HM, Perkins DG, Parks W (1997) Pathogenetic role of the stromal cells in endometriosis and adenomyosis. Histopathology 30(5):430–442CrossRefPubMedGoogle Scholar
  34. 34.
    Clement PB, Scully RE (1992) Endometrial stromal sarcomas of the uterus with extensive endometrioid glandular differentiation: a report of three cases that caused problems in differential diagnosis. Int J Gynecol Pathol 11(3):163–173CrossRefPubMedGoogle Scholar
  35. 35.
    Lessey BA, Higdon HL, Miller SE, Price TA (2012) Intraoperative detection of subtle endometriosis: a novel paradigm for detection and treatment of pelvic pain associated with the loss of peritoneal integrity. J Visual Exp.  https://doi.org/10.3791/4313 CrossRefGoogle Scholar
  36. 36.
    Witz CA, Dechaud H, Montoya-Rodriguez IA, Thomas MR, Nair AS, Centonze VE, Schenken RS (2002) An in vitro model to study the pathogenesis of the early endometriosis lesion. Ann NY Acad Sci 955:296–307 (discussion 340-292, 396-406) CrossRefPubMedGoogle Scholar
  37. 37.
    Witz CA, Thomas MR, Montoya-Rodriguez IA, Nair AS, Centonze VE, Schenken RS (2001) Short-term culture of peritoneum explants confirms attachment of endometrium to intact peritoneal mesothelium. Fertil Steril 75(2):385–390CrossRefPubMedGoogle Scholar
  38. 38.
    Oosterlynck DJ, Meuleman C, Waer M, Koninckx PR (1994) Transforming growth factor-beta activity is increased in peritoneal fluid from women with endometriosis. Obstet Gynecol 83(2):287–292PubMedGoogle Scholar
  39. 39.
    Liu Y, Hu J, Shen W, Wang J, Chen C, Han J, Zai D, Cai Z, Yu C (2011) Peritoneal fluid of patients with endometriosis promotes proliferation of endometrial stromal cells and induces COX-2 expression. Fertil Steril 95(5):1836–1838.  https://doi.org/10.1016/j.fertnstert.2010.11.039 CrossRefGoogle Scholar
  40. 40.
    Callegari EA, Ferguson-Gottschall S, Gibori G (2005) PGF2alpha induced differential expression of genes involved in turnover of extracellular matrix in rat decidual cells. Reprod Biol Endocrinol 3:3.  https://doi.org/10.1186/1477-7827-3-3 CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Komiyama S, Aoki D, Komiyama M, Nozawa S (2007) Local activation of TGF-beta1 at endometriosis sites. J Reprod Med 52(4):306–312PubMedGoogle Scholar
  42. 42.
    Slater M, Quagliotto G, Cooper M, Murphy CR (2005) Endometriotic cells exhibit metaplastic change and oxidative DNA damage as well as decreased function, compared to normal endometrium. J Mol Histol 36(4):257–263.  https://doi.org/10.1007/s10735-005-3802-9 CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Mohamed Gamal Ibrahim
    • 1
    • 5
  • Martin Sillem
    • 2
  • Johanna Plendl
    • 3
  • Eliane T. Taube
    • 4
  • Andreas Schüring
    • 5
  • Martin Götte
    • 5
  • Vito Chiantera
    • 1
  • Jalid Sehouli
    • 1
  • Sylvia Mechsner
    • 1
  1. 1.Clinic for GynaecologyCharité University of MedicineBerlinGermany
  2. 2.Universitäts-Frauenklinik Homburg/Saar und Praxisklinik am RosengartenMannheimGermany
  3. 3.Department of Veterinary Medicine, Institute of Veterinary AnatomyFree University of BerlinBerlinGermany
  4. 4.Institute for PathologyCharité University of MedicineBerlinGermany
  5. 5.Department of Gynecology and Obstetrics, UKM Fertility CenterUniversity Hospital of MuensterMünsterGermany

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