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EP-4740024-A1 - ENDOMETRIOSIS MARKERS AND METHODS OF DIAGNOSING ENDOMETRIOSIS

EP4740024A1EP 4740024 A1EP4740024 A1EP 4740024A1EP-4740024-A1

Abstract

The present disclosure relates to methods and kits for detecting levels of androgens, androgen precursors and androgen metabolites. In particular, the present disclosure relates to methods and kits for diagnosing and/or prognosing endometriosis based on the levels of these androgens, androgen precursors and androgen metabolites.

Inventors

  • GIBSON, DOUGLAS
  • SAUNDERS, Philippa Tansy Kemp
  • HORNE, ANDREW

Assignees

  • The University Court Of The University of Edinburgh

Dates

Publication Date
20260513
Application Date
20240704

Claims (20)

  1. 1. A method of diagnosing and/or prognosing endometriosis, the method comprising: probing a test sample for the presence of one or more steroid(s) to determine a level of the one or more steroid(s) in the test sample; wherein the one or more steroid(s) is selected from 11 -ketotestosterone (11 KT), 11 -ketoandrostenedione (11 KA4), 11 -hydroxyandrostenedione (11OHA4), 11- hydroxytestosterone (11OHT), 11 -hydroxydihydrotestosterone (11OHDHT), 11- ketodihydrotestosterone (11 KDHT), 11 -hydroxyandrosterone (HOHAndro), 17- hydroxypregnanolone (17HP), and combinations thereof; wherein the level of the one or more steroid(s) in the test sample is indicative of the likelihood and/or severity of endometriosis.
  2. 2. The method according to claim 1 , wherein the one or more steroids is selected from 11 -ketotestosterone (11 KT), 11 -ketoandrostenedione (11 KA4), 11- hydroxyandrostenedione (11OHA4), 11 -hydroxytestosterone (11OHT), and combinations thereof.
  3. 3. The method according to claim 2, wherein the method comprises additionally probing the test sample for the presence of one or more additional steroid(s) selected from: 17-hydroxypregnenolone (17OHP5), 17-hydroxyprogesterone (17OHP4), dehydroepiandrosterone (DHEA), androstenedione (A4), testosterone (T), 11- deoxycortisol (S), 11 -deoxycorticosterone (DOC), androstanedione (5-adione), androsterone (An), etiocholanolone (Et), 11 -hydroxyandrosterone (HOHAndro), 5- pregnenediol (5-PD), 5-pregnenetriol (5-PT), pregnanediol (PD), pregnanetriol (PT), pregnanetriolone (PTONE), 17-OH-pregnanolone (17HP), 3a5a-17-OH-pregnanolone (3a5a17HP), tetrahydro-11 -deoxycortisol (THS), and combinations thereof.
  4. 4. The method according to any one of the preceding claims, wherein the method comprises probing a test sample for the presence of 11 -ketotestosterone (11 KT) to determine a level of 11 KT in the test sample.
  5. 5. The method according to claim 4, wherein the method comprises probing a test sample for the presence of 11 -ketotestosterone (11 KT) to determine a level of 11- ketotestosterone (11 KT) in the test sample, and additionally probing the test sample for the presence of one or more additional steroid(s) selected from: 17-hydroxypregnenolone (17OHP5), 11 -hydroxyandrostenedione (11OHA4), 11- ketoandrostenedione (11 KA4), 17-hydroxyprogesterone (17OHP4), dehydroepiandrosterone (DHEA), androstenedione (A4), testosterone (T), 11- hydroxytestosterone (11OHT), 11 -hydroxydihydrotestosterone (11OHDHT), 11- ketodihydrotestosterone (11 KDHT), 11 -deoxycortisol (S), 11 -deoxycorticosterone (DOC), androstanedione (5-adione), androsterone (An), etiocholanolone (Et), 11- hydroxyandrosterone (HOHAndro), 5-pregnenediol (5-PD), 5-pregnenetriol (5-PT), pregnanediol (PD), pregnanetriol (PT), pregnanetriolone (PTONE), 17-OH- pregnanolone (VHP), 3a5a-17-OH-pregnanolone (3a5a17HP), tetrahydro-11- deoxycortisol (THS), and combinations thereof.
  6. 6. A method according to any one of the preceding claims, wherein the method comprises probing a test sample for the presence of 11 -ketotestosterone (11 KT) to determine a level of 11 -ketotestosterone (11 KT) in the test sample, and additionally probing the test sample for the presence of one or more additional steroid(s) selected from: 11 -ketoandrostenedione (11 KA4), 11 -hydroxyandrostenedione (11OHA4), 11- hydroxytestosterone (11OHT), and combinations thereof.
  7. 7. A method according to any one of claims 1 to 6, wherein the method comprises probing the test sample for the presence of at least the following steroids: (i) 11 KT and 11 KA4; (ii) 11OHT and 11OHA4; (iii) 11OHA4 and A4; (iv) HOHT and T; (v) 11 KA4 and 11OHA4; or (vi) 11 KT and 11OHT.
  8. 8. A method according to any one of the preceding claims, wherein the method further comprises comparing the level of the one or more steroid(s) detected for the test sample with a reference or baseline level of the same or corresponding one or more steroid(s) in a healthy subject.
  9. 9. A method according to claim 8, wherein when the level of the one or more steroid(s) in the test sample is higher than the reference or baseline level, the test sample is determined to have been derived from a subject suffering from endometriosis, optionally wherein when the level of 11 KT and/or 11OHT is higher than the reference or baseline level, the test sample is determined to have been derived from a subject suffering from endometriosis.
  10. 10. The method according to claim 8, wherein when the level of the one or more steroid(s) in the test sample is lower than the reference or baseline level, the test sample is determined to have been derived from a subject suffering from endometriosis; optionally wherein when the level of 11OHA4 and/or 11 KA4 is lower than the reference or baseline level, the test sample is determined to have been derived from a subject suffering from endometriosis.
  11. 11. A method according to any one of the preceding claims, wherein the method comprises detecting at least two steroids in a test sample, and wherein the method further comprises: (i) probing the test sample to determine the relative levels of a steroid pair selected from the following: (a) 11 KT and 11 KA4; (b) 11OHT and 11OHA4; (c) androstenedione (A4) and 11OHA4; (d) 11OHT and T; (e) 11 KA4 and 11OHA4; and (f) 11 KT and 11OHT.
  12. 12. The method according to claim 11 , wherein the method further comprises comparing the relative levels of the steroid pair in the test sample with a baseline or reference relative level of the same or corresponding steroid pair.
  13. 13. The method according to claim 12, wherein when the relative levels of the steroid pair in the test sample is different or altered to the baseline or reference relative level, the test sample is determined to have been derived from a subject suffering from endometriosis.
  14. 14. A method according to any one of claims 11 to 13, further comprising determining a ratio of: (a) 11 KT to 11 KA4; (b) 11OHT to 11OHA4; (c) 11OHA4 to androstenedione (A4); (d) 11OHT to T; (e) 11 KA4 to 11OHA4; and (f) 11 KT to 11OHT; in the test sample.
  15. 15. A method according to claim 14, further comprising: comparing the ratio of 11 KT to 11 KA4 with a baseline or reference ratio of 11 KT to 11 KA4, optionally wherein when the ratio of 11 KT to 11 KA4 is increased in the test sample relative to the baseline or reference ratio, the sample is determined to have been derived from a subject suffering from endometriosis.
  16. 16. A method according to claim 14, further comprising: comparing the ratio of 11OHT to 11OHA4 in the test sample with a baseline or reference ratio of 11OHT to 11OHA4, optionally wherein when the ratio of 11OHT to 11OHA4 is increased in the test sample relative to the baseline or reference ratio, the test sample is determined to have been derived from a subject suffering from endometriosis.
  17. 17. A method according to claim 14, further comprising: (i) comparing the ratio of 11OHA4 to androstenedione in the test sample with a baseline or reference ratio of 11OHA4 to androstenedione, optionally wherein when the ratio of 11OHA4 to androstenedione is decreased in the test sample relative to the baseline or reference ratio, the test sample is determined to have been derived from a subject suffering from endometriosis; (ii) comparing the ratio of 11OHT to testosterone in the test sample with a baseline or reference ratio of 11OHT to testosterone, optionally wherein when the ratio of 11OHT to testosterone is decreased in the test sample relative to the baseline or reference ratio, the test sample is determined to have been derived from a subject suffering from endometriosis; (iii) comparing the ratio of 11 KA4 to 11OHA4 in the test sample with a baseline or reference ratio of 11 KA4 to 11OHA4, optionally wherein when the ratio of 11 KA4 to 11OHA4 is increased in the test sample relative to the baseline or reference ratio, the test sample is determined to have been derived from a subject suffering from endometriosis; or (iv) comparing the ratio of 11 KT to 11OHT in the test sample with a baseline or reference ratio of 11 KT to 110HT, optionally wherein when the ratio of 11 KT to 110HT is increased in the test sample relative to the baseline or reference ratio, the test sample is determined to have been derived from a subject suffering from endometriosis.
  18. 18. A method according to any one of the preceding claims, wherein the test sample is a biological fluid selected from blood (e.g. serum or plasma), urine, saliva and peritoneal fluid.
  19. 19. A method according to claim 1 , wherein the test sample is serum.
  20. 20. A method of diagnosing and/or prognosing endometriosis comprising detecting one or more steroid(s) in a urine sample, the method comprising: probing the urine sample for the presence of the one or more steroid(s) to determine a level of the one or more steroid(s) in the urine sample; wherein the one or more steroid(s) is selected from androsterone (An), etiocholanolone (Et), 11 -hydroxyandrosterone (HOHAndro), 17-hydroxypregnanolone (17HP), and combinations thereof; wherein the level of the one or more steroid(s) in the test sample is indicative of the likelihood or severity of endometriosis; optionally wherein the method comprises probing the urine sample for the presence of at least 11 -hydroxyandrosterone (HOHAndro).

Description

Endometriosis markers and methods of diagnosing endometriosis TECHNICAL FIELD The present disclosure relates to methods and kits for detecting levels of androgens, androgen precursors and androgen metabolites. The present disclosure further relates to methods and kits for diagnosing and/or prognosing endometriosis based on the levels of these androgens, androgen precursors and androgen metabolites. BACKGROUND Endometriosis is a chronic condition affecting the reproductive system of females. It is characterized by the presence of endometrial-like tissue outside the uterus. Endometriosis is associated with various symptoms including dysmenorrhoea, dyspareunia, pelvic pain and infertility. Endometriosis is understood to be an estrogen-dependent disease in which endometriotic tissue proliferates in response to estrogen. Consequently, many of the clinical treatments of endometriosis rely on hormone-based therapies which look to reduce levels of estrogen. Huhtinen et al describe a study in which local estradiol (E2) and estrone (E1) concentrations in the endometrium and different types of endometriotic lesions were measured in endometriosis patients, and also mRNA expression of the estrogen- metabolizing enzymes. The study indicated that endometrial or endometriotic E2 concentrations are actively regulated by local estrogen metabolism in the tissue (“Endometrial and Endometriotic Concentrations of Estrone and Estradiol are determined by local metabolism rather than circulating levels”, J. Clin. Endocrinol. Metab. November 2012, 97(11), pages 4228-4235). Huhtinen et al further describe a study which sought to evaluate whether the tissue steroid hormone concentrations in endometriosis differ from the endometrium or serum (“Intra-tissue steroid profiling indicates differential progesterone and testosterone metabolism in the endometrium and endometriosis lesions”, J. Clin. Endocrinol. Metab. November 2014, 99(11), pages E2188 to E2197). In the described study, steroid analysis of serum and tissue specimens of women with endometriosis and healthy controls were measured. The authors concluded that endometriosis lesions present with progestin and androgen metabolism which are different from that of the endometrium, and the lesions are characterized by high levels of testosterone (T) in the tissue and a loss of cyclical changes in the concentration of progesterone (P4) in the tissue. Currently, the only reliable method to diagnose endometriosis is by performing a laparoscopy. This is an expensive and invasive procedure and so endometriosis has significant diagnostic delays for patients, typically an average of eight years from onset of symptoms. Some attempts to use various biomarkers for diagnosing endometriosis have been described in US11001892B1, US7879562B2, US20150132295A1 and US8541173B2. However, it has proved difficult to identify biomarkers that can reliably identify the presence of the disease and/or distinguish patients from healthy controls. Therefore, there remains a need for quick and/or minimally invasive methods of diagnosing and/or prognosing endometriosis. SUMMARY The present disclosure is based on the finding that certain steroids and their metabolites can provide useful biomarkers of endometriosis. In particular, the present inventors have identified that certain androgens, androgen precursors and androgen metabolites may be elevated in females suffering or at risk of developing endometriosis. Indeed, the present inventors have observed that the hormone metabolism signature (particularly the androgen metabolism signature) is different in patients suffering from endometriosis compared to healthy controls. Accordingly, the present disclosure provides methods and kits for detecting levels of certain androgens, androgen precursors and androgen metabolites, and also methods and kits for diagnosing and/or prognosing endometriosis based on these insights. Described herein is a method for detecting one or more steroid(s) in a test sample, the method comprising: probing a test sample for the presence of the one or more steroid(s) to determine a level of the one or more steroid(s) in the test sample; wherein the one or more steroid(s) is selected from an androgen, an androgen precursor, an androgen metabolite, and combinations thereof. In some examples, the method may comprise a step of comparing the level of the one or more steroid(s) detected for the test sample with a reference or baseline level of the same one or more steroid(s) in a healthy subject (e.g. one that is not suffering from endometriosis). This reference or baseline level may be the level of the one or more steroid(s) detected in a sample obtained from the healthy subject. Where the level of the one or more steroid(s) is different for the test sample to the reference or baseline level, the test sample is determined to have been derived from a subject suffering from endometriosis. In particular, in many cases, where the level of the one or m