Cathepsin Protease Inhibition Reduces Endometriosis Lesion Establishment

Endometriosis is a gynecologic disease characterized by the ectopic presence of endometrial tissue on organs within the peritoneal cavity, causing debilitating abdominal pain and infertility. Current treatments alleviate moderate pain symptoms associated with the disorder but exhibit limited ability to prevent new or recurring lesion establishment and growth. Retrograde menstruation has been implicated for introducing endometrial tissue into the peritoneal cavity, but molecular mechanisms underlying attachment and invasion are not fully understood. We hypothesize that cysteine cathepsins, a group of powerful extracellular matrix proteases, facilitate endometrial tissue invasion and endometriosis lesion establishment in the peritoneal wall and inhibiting this activity would decrease endometriosis lesion implantation. To test this, we used an immunocompetent endometriosis mouse model and found that endometriotic lesions exhibited a greater than 5-fold increase in active cathepsins compared to tissue from peritoneal wall or eutopic endometrium, with cathepsins L and K specifically implicated. Human endometriosis lesions also exhibited greater cathepsin activity than adjacent peritoneum tissue, supporting the mouse results. Finally, we tested the hypothesis that inhibiting cathepsin activity could block endometriosis lesion attachment and implantation in vivo. Intraperitoneal injection of the broad cysteine cathepsin inhibitor, E-64, significantly reduced the number of attached endometriosis lesions in our murine model compared to vehicle-treated controls demonstrating that cathepsin proteases contribute to endometriosis lesion establishment, and their inhibition may provide a novel, nonhormonal therapy for endometriosis. Similar content being viewed by others

Eskenazi B, Warner ML. Epidemiology of endometriosis. Obstet Gynecol Clin North Am. 1997;24(2):235–258. Giudice LC. Clinical practice. Endometriosis. N Engl J Med. 2010;362(25):2389–2398. Giudice LC, Kao LC. Endometriosis. Lancet. 2004;364:1789–1799. Soares SR, Martinez-Varea A, Hidalgo-Mora JJ, Pellicer A. Pharmacologic therapies in endometriosis: A systematic review. Fertil Steril. 2012;98(3):529–555. Sampson JA. Metastatic or embolic endometriosis, due to the menstrual dissemination of endometrial tissue into the venous circulation. Am J Pathol. 1927;3(2):93–110. 143. Halme J, Hammond MG, Hulka JF, Raj SG, Talbert LM. Retrograde menstruation in healthy women and in patients with endometriosis. Obstet Gynecol. 1984;64(2):151–154. Candiani GB, Fedele L, Vercellini P, Bianchi S, Di Nola G. Repetitive conservative surgery for recurrence of endometriosis. Obstet Gynecol. 1991;77(3):421–424. Namnoum AB, Hickman TN, Goodman SB, Gehlbach DL, Rock JA. Incidence of symptom recurrence after hysterectomy for endometriosis. Fertil Steril. 1995;64:898–902. Witz CA, Montoya-Rodriguez IA, Cho S, Centonze VE, Bonewald LF, Schenken RS. Composition of the extracellular matrix of the peritoneum. J Soc Gynecol Investig. 2001;8(5):299–304. Sillem M, Hahn U, Coddington CC III, Gordon K, Runnebaum B, Hodgen GD. Ectopic growth of endometrium depends on its structural integrity and proteolytic activity in the cynomolgus monkey (Macaca fascicularis) model of endometriosis. Fertil Steril. 1996;66(3):468–473. Bergqvist A, Ferno M, Mattson S. A comparison of cathepsin D levels in endometriotic tissue and in uterine endometrium. Fertil Steril. 1996;65(6):1130–1134. Suzumori N, Ozaki Y, Ogasawara M, Suzumori K. Increased concentrations of cathepsin D in peritoneal fluid from women with endometriosis. Mol Hum Reprod. 2001;7(5):459–462. Chung HW, Lee JY, Moon HS, et al. Matrix metalloproteinase-2, membranous type 1 matrix metalloproteinase, and tissue inhibitor of metalloproteinase-2 expression in ectopic and eutopic endometrium. Fertil Steril. 2002;78(4):787–795. Bruner KL, Matrisian LM, Rodgers WH, Gorstein F, Osteen KG. Suppression of matrix metalloproteinases inhibits establishment of ectopic lesions by human endometrium in nude mice. J Clin Invest. 1997;99(12):2851–2857. Yasuda Y, Li Z, Greenbaum D, Bogyo M, Weber E, Bromme D. Cathepsin V, a novel and potent elastolytic activity expressed in activated macrophages. J Biol Chem. 2004;279(35):36761–36770. Garnero P, Borel O, Byrjalsen I, et al. The collagenolytic activity of cathepsin K is unique among mammalian proteinases. J Biol Chem. 1998;273(48):32347–32352. Chen B, Platt MO. Multiplex zymography captures stage-specific activity profiles of cathepsins K, L, and S in human breast, lung, and cervical cancer. J Transl Med. 2011;9:109. Liu J, Sukhova GK, Yang JT, et al. Cathepsin L expression and regulation in human abdominal aortic aneurysm, atherosclerosis, and vascular cells. Atherosclerosis. 2006;184(2):302–311. Cheng XW, Shi G-P, Kuzuya M, Sasaki T, Okumura K, Murohara T. Role for cysteine protease cathepsins in heart disease: Focus on biology and mechanisms with clinical implication. Circulation. 2012;125:1551–1562. Mohamed MM, Sloane BF. Cysteine cathepsins: Multifunctional enzymes in cancer. Nat Rev Cancer. 2006;6:764–775. Vasiljeva O, Reinheckel T, Peters C, Turk D, Turk V, Turk B. Emerging roles of cysteine cathepsins in disease and their potential as drug targets. Curr Pharm Des. 2007;13:387–403. Costa AG, Cusano NE, Silva BC, Cremers S, Bilezikian JP. Cathepsin K: Its skeletal actions and role as a therapeutic target in osteoporosis. Nat Rev Rheumatol. 2011;7:447–456. Desmarais S, Masse F, Percival MD. Pharmacological inhibitors to identify roles of cathepsin K in cell-based studies: A comparison of available tools. Biol Chem. 2009;390:941–948. Bromme D, Lecaille F. Cathepsin K inhibitors for osteoporosis and potential off-target effects. Expert Opin Investig Drugs. 2009;18:585–600. Wieser F, Wu J, Shen Z, Taylor RN, Sidell N. Retinoic acid suppresses growth of lesions, inhibits peritoneal cytokine secretion, and promotes macrophage differentiation in an immunocompetent mouse model of endometriosis. Fertil Steril. 2012;97(6): 1430–1437. Hirata T, Osuga Y, Yoshino O, et al. Development of an experimental model of endometriosis using mice that ubiquitously express green fluorescent protein. Hum Reprod. 2005;20(8): 2092–2096. Pierzchalski K, Taylor RN, Nezhat C, et al. Retinoic acid biosynthesis is impaired in human and murine endometriosis. Biol Reprod. 2014;91(4):84. Revised american society for reproductive medicine classification of endometriosis: 1996. Fertil Steril. 1997;67(5):817–821. Wilder CL, Park KY, Keegan PM, Platt MO. Manipulating substrate and pH in zymography protocols selectively distinguishes cathepsins K, L, S, and V activity in cells and tissues. Arch Biochem Biophys. 2011;516(1):52–57. Li WA, Barry ZT, Cohen JD, et al. Detection of femtomole quantities of mature cathepsin K with zymography. Anal Biochem. 2010;401(1):91–98. Park KY, Li WA, Platt MO. Patient specific proteolytic activity of monocyte-derived macrophages and osteoclasts predicted with temporal kinase activation states during differentiation. Integr Biol (Camb). 2012;4(12): 1459–1469. Bromme D, Li Z, Barnes M, Mehler E. Human cathepsin V functional expression, tissue distribution, electrostatic surface potential, enzymatic characterization, and chromosomal localization. Biochemistry. 1999;38(8):2377–2385. Keegan PM, Surapaneni S, Platt MO. Sickle cell disease activates peripheral blood mononuclear cells to induce cathepsins K and V activity in endothelial cells. Anemia. 2012;2012: 201781. Keegan PM, Wilder CL, Platt MO. Tumor necrosis factor alpha stimulates cathepsin K and V activity via juxtacrine monocyteendothelial cell signaling and JNK activation. Mol Cell Biochem. 2012;367(1–2):65–72. Nair AS, Nair HB, Lucidi RS, et al. Modeling the early endometriotic lesion: Mesothelium-endometrial cell co-culture increases endometrial invasion and alters mesothelial and endometrial gene transcription. Fertil Steril. 2008;90(4 suppl): 1487–1495. Lebovic DI, Mueller MD, Taylor RN. Immunobiology of endometriosis. Fertil Steril. 2001;75(1):1–10. Yin M, Soikkeli J, Jahkola T, Virolainen S, Saksela O, Holtta E. TGF-beta signaling, activated stromal fibroblasts, and cysteine cathepsins B and L drive the invasive growth of human melanoma cells. Am J Pathol. 2012;181(6):2202–2216. Wijkmans J, Gossen J. Inhibitors of cathepsin K: a patent review (2004 - 2010). Expert Opin Ther Pat. 2011;21(10):1611–1629. Lee-Dutra A, Wiener DK, Sun S. Cathepsin S inhibitors: 2004–2010. Expert Opin Ther Pat. 2011;21(3):311–337. Author information Authors and Affiliations Corresponding author Rights and permissions About this article Cite this article Porter, K.M., Wieser, F.A., Wilder, C.L. et al. Cathepsin Protease Inhibition Reduces Endometriosis Lesion Establishment. Reprod. Sci. 23, 623–629 (2016). https://doi.org/10.1177/1933719115611752 Published: Issue date: DOI: https://doi.org/10.1177/1933719115611752