Marker, kit and application for determining initial dose of FSH for polycystic ovary syndrome

Abstract

The present invention discloses a marker for determining the starting dose of FSH for polycystic ovary syndrome and its application. The marker includes BMI, bFSH, bLH, AMH and AFC. The marker of the present invention will help clinicians select the most appropriate starting dose of FSH to obtain the best number of retrieved oocytes while avoiding the risk of ovarian hyperstimulation syndrome.

Description

[0001] This application claims the priority of Chinese Patent Application No. 2023104280405 filed on April 20, 2023, and the entire text of this patent application is hereby incorporated by reference into this application.

[0002] This application also claims the priority of Chinese Patent Application No. 2023105443024 filed on May 15, 2023, and the entire text of this patent application is hereby incorporated by reference into this application. Technical Field

[0003] The present invention belongs to the field of biomedical technologies, and particularly relates to a biomarker, a kit and their applications for determining the starting dose of FSH for polycystic ovary syndrome. Background Art

[0004] Polycystic ovary syndrome (PCOS) is the most common reproductive endocrine and metabolic disorder in women. Approximately 15 - 20% of infertile women suffer from polycystic ovary syndrome, which is the main cause of anovulatory infertility in 80% of patients.

[0005] If PCOS patients still cannot conceive after receiving clomiphene citrate / letrozole for ovulation induction, or in the case of combined tubal and / or male factor infertility, in vitro fertilization (IVF) / intracytoplasmic sperm injection (ICSI)-embryo transfer (ET) is an effective treatment method to meet the fertility requirements of PCOS infertile patients. Among them, the gonadotropin-releasing hormone antagonist (GnRH-ant) protocol is the first choice for PCOS women because of its short ovarian stimulation time, reduced treatment dose, and low incidence of ovarian hyperstimulation syndrome (OHSS). However, a lower starting dose of follicle-stimulating hormone (FSH) will lead to poor follicle growth, fewer retrieved oocytes, and an increased cycle cancellation rate, while a higher FSH starting dose greatly increases the risk of ovarian hyperstimulation. At the same time, high levels of estrogen may also affect the endometrial receptivity, thereby affecting the clinical pregnancy rate. Therefore, an appropriate FSH starting dose is considered to significantly improve the clinical outcomes of IVF / ICSI-ET in PCOS patients.

[0006] Based on existing domestic and international evidence, it is considered that 15 retrieved oocytes is the optimal number. In the past, clinicians usually determined the FSH starting dose based on age, body weight, body mass index, ovarian response in previous IVF cycles, and ovarian reserve indicators including basal FSH level (bFSH), AMH, and AFC. In the past, a few researchers proposed to calculate the most suitable FSH starting dose for the ultra-long protocol in IVF cycles according to the patient's age, bFSH, AFC, or AMH level to help infertile patients obtain the best response. So far, no one has studied the optimal FSH starting dose for the antagonist protocol in the PCOS population. Therefore, this study has important clinical significance and practical value. Summary of the Invention

[0007] To make up for the deficiencies of the prior art, the purpose of the present invention is to provide a biomarker for determining the starting dose of FSH in polycystic ovary syndrome and its application. To achieve the purpose of the present invention, the following technical solutions are proposed:

[0008] In the first aspect of the present invention, a kit for determining the starting dose of FSH in polycystic ovary syndrome is provided. The kit includes reagents for quantitatively detecting BMI, bFSH, bLH, AMH, and AFC in patients with polycystic ovary syndrome. The starting dose of FSH refers to the starting dose of FSH in in vitro fertilization (IVF) / intracytoplasmic sperm injection (ICSI)-embryo transfer (ET) treatment.

[0009] In one embodiment, the age of the patients with polycystic ovary syndrome is 20 - 40 years old.

[0010] In a preferred embodiment, the calculation method of the starting dose of FSH is FSH = (0.491×BMI + 1.312×bFSH - 0.291×bLH - 0.455×AMH - 0.075×AFC - 8.576) / 0.042, where the unit of FSH is IU / day, the unit of BMI is kg / m 2 , the unit of bFSH is IU / L, the unit of bLH is IU / L, the unit of AMH is ng / mL, and the unit of AFC is number.

[0011] In the second aspect of the present invention, a method for determining the starting dose of FSH in polycystic ovary syndrome is provided. The method includes quantitatively detecting BMI, bFSH, bLH, AMH, and AFC of the patient.

[0012] In a preferred embodiment, the starting dose of FSH is FSH = (0.491×BMI + 1.312×bFSH - 0.291×bLH - 0.455×AMH - 0.075×AFC - 8.576) / 0.042, where the unit of FSH is IU / day, the unit of BMI is kg / m 2 , the unit of bFSH is IU / L, the unit of bLH is IU / L, the unit of AMH is ng / mL, and the unit of AFC is number.

[0013] The beneficial effect of the present invention is to determine the most suitable starting dose of FSH to obtain the best number of retrieved eggs (15) to ensure a good clinical outcome, and at the same time avoid the occurrence of ovarian hyperstimulation syndrome. Brief Description of the Drawings

[0014] Figure 1It is a Nomogram for calculating the starting dose of FSH. In this example, for a PCOS patient with a body mass index (BMI) of 28 kg / m 2 , bFSH = 6 IU / L, bLH = 8 IU / L, AMH = 8 ng / mL, and AFC = 24, the predicted starting dose of FSH is 125 IU / day.

[0015] Figure 2 The difference between the predicted starting dose of FSH and the actual starting dose of FSH calculated by the training set of the Nomogram in this study was used to test the effectiveness of this Nomogram. Among PCOS patients with 15 retrieved oocytes, the predicted starting dose of FSH we calculated was almost the same as the actual dose; when the number of retrieved oocytes was less than 15, the actual dose was significantly lower than the predicted dose; similarly, when the number of retrieved oocytes was more than 15, the actual dose was significantly higher than the predicted dose.

[0016] Figure 3 The difference between the predicted starting dose of FSH and the actual starting dose of FSH calculated by the test set of the Nomogram in this study was used to test the effectiveness of this Nomogram. Among PCOS patients with 15 retrieved oocytes, the predicted starting dose of FSH we calculated was almost the same as the actual dose; when the number of retrieved oocytes was less than 15, the actual dose was significantly lower than the predicted dose; similarly, when the number of retrieved oocytes was more than 15, the actual dose was significantly higher than the predicted dose. Detailed implementation manners

[0017] To further understand the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0018] Unless otherwise specified, the reagents involved in the embodiments of the present invention are all commercially available products and can be obtained through commercial channels.

[0019] Screening of Predictive Indicators for the Optimal Initial FSH Dose in Example 1

[0020] Based on the clinical relevant indicators of 1898 PCOS women (aged 20 - 40 years) who received the GnRH-ant protocol in the Reproductive Center of Peking University Third Hospital during their first IVF / ICSI cycles, a set of predictive indicators suitable for calculating the optimal starting dose of FSH was screened.

[0021] In this study, 1898 PCOS patients from 2017 to 2021 were strictly included according to the inclusion and exclusion criteria (all patients had signed informed consent forms).

[0022] Inclusion criteria: (1) Reproductive-aged women aged 20 to 40 years; (2) Women diagnosed with polycystic ovary syndrome according to the Rotterdam diagnostic criteria revised in 2003, meeting at least 2 of the 3 criteria - oligo-ovulation and / or anovulation, clinical and / or biochemical indices of hyperandrogenism, and polycystic ovarian changes (≥12 follicles in unilateral / bilateral ovaries), and excluding other etiologies (congenital adrenal hyperplasia, androgen-secreting tumors, Cushing's syndrome, etc.); (3) Women undergoing their first IVF / ICSI cycle; (4) Women undergoing ovulation induction with the GnRH-ant protocol; and (5) Patients with complete clinical and IVF cycle information.

[0023] Exclusion criteria: (1) Chromosomal abnormalities in either spouse; (2) History of ovarian diseases or surgeries, such as ovarian endometriotic cysts, ovarian tumors, ovarian drilling, or history of ovarian cyst or tumor resection; (3) In vitro maturation cycles.

[0024] GnRH-ant protocol: Starting from the 2nd - 3rd day of the menstrual cycle, gonadotropin (Gn, FSH in this study) is injected daily. Transvaginal ultrasound examination is performed starting from the 5th day of Gn injection to monitor follicular development for adjusting the drug dosage according to the ovarian response. There are two options for the administration time of GnRH-ant: (1) Fixed administration protocol, adding the antagonist on the 5th - 7th day after Gn administration; (2) Flexible administration protocol, adding the antagonist when the dominant follicle diameter reaches 12 mm until the HCG day. When at least 3 follicles have a diameter > 17 mm, human chorionic gonadotropin (hCG) is injected, with a dose of 5000 - 10000 IU or 250 μg recombinant hCG. Patients at high risk of OHSS are triggered with 0.2 mg GnRH-a or in combination with a low dose of hCG to reduce the risk of OHSS. Egg retrieval is performed 36 - 38 hours later.

[0025] The following factors that may be related to the number of retrieved oocytes were analyzed by univariate analysis, including age, infertility type, infertility duration, parity, BMI, basal FSH (bFSH) level, basal LH (bLH) level, AMH level, and AFC. It was found that age, parity, BMI, bFSH, bLH, AMH, and AFC were significantly correlated with the number of retrieved oocytes, and the results are shown in Table 1.

[0026] Table 1: Results of univariate and multivariate analyses of factors related to the number of retrieved oocytes

[0027]

[0028] Furthermore, the indicators with significant differences in the univariate analysis were further analyzed by multivariate analysis, and it was found that only BMI and ovarian reserve markers (including bFSH, bLH, AMH, and AFC) were significantly correlated with the number of retrieved oocytes. Accordingly, this study successfully screened out the predictive indicators (BMI, bFSH, bLH, AMH, AFC) applicable to calculating the optimal starting dose of FSH, and further proposed a predictive model for the optimal starting dose of FSH for PCOS patients undergoing IVF / ICSI with the GnRH-ant protocol:

[0029] Optimal starting dose of FSH = (0.491 × BMI + 1.312 × bFSH - 0.291 × bLH - 0.455 × AMH - 0.075 × AFC - 8.576) / 0.042.

[0030] This model helps guide clinicians to select the most appropriate starting dose of FSH in this part of the population in the future. Taking PCOS patients with body mass index BMI = 28 kg / m 2 , bFSH = 6 IU / L, bLH = 8 IU / L, AMH = 8 ng / mL, and AFC = 24 as an example, the predicted optimal starting dose of FSH is 125 IU / day ( Figure 1 , Table 2).

[0031] Table 2 Nomogram for calculating the starting dose of FSH.

[0032]

[0033] Note: β is the slope, representing the change value of the dependent variable (number of retrieved oocytes) when the independent variable changes by one unit. If the P value of the slope is less than 0.05, it proves that the slope is statistically significant.

[0034] Utility Evaluation of the Prediction Model (Training Set) in Example 2

[0035] To test the utility of the prediction model in this study, we applied this model to a cohort of 1898 PCOS patients. The lowest initial FSH dose for PCOS women undergoing ovulation induction was set at 112.5 IU / day based on clinical experience to prevent follicle non-growth and cancellation of the ovulation induction cycle.

[0036] Among PCOS patients with the number of retrieved oocytes equal to 15, the predicted starting dose of FSH using this model was almost the same as the actual dose used ( Figure 2 ). When the number of retrieved oocytes was less than 15, the actual dose used clinically was significantly lower than the predicted dose. Similarly, when the number of retrieved oocytes exceeded 15, the actual dose used was significantly higher than the predicted dose. These results further verified the reliability of the prediction model in this study.

[0037] Clinical Validation of the Prediction Model (Test Set) in Example 3

[0038] An independent cohort consisting of 593 patients treated from 2020 to 2021 was assigned to the test set to validate the above model. These 593 patients also met the inclusion and exclusion criteria described in Example 1 above. The relevant information of the 1898 training set described in Example 1 and the test set described in Example 2 is shown in Table 3 below.

[0039] Table 3: Patient and IVF cycle characteristics of the training set and the test set

[0040]

[0041]

[0042] For the test set, the minimum initial FSH dose for ovulation induction in PCOS women was set at 112.5 IU / day according to clinical experience to prevent follicle non-growth and cancellation of the ovulation induction cycle. As can be seen from Figure 3 Among PCOS patients with 15 retrieved oocytes, the predicted starting FSH dosage using this model was almost the same as the actual dosage. When the number of retrieved oocytes was less than 15, the actual clinical dosage was significantly lower than the predicted dosage. Similarly, when the number of retrieved oocytes exceeded 15, the actual dosage was significantly higher than the predicted dosage. These results further verified the reliability of the prediction model in this study.

[0043] The preferred embodiments of the present invention have been described above, but they are not intended to limit the present invention. Those skilled in the art can make improvements and changes to the embodiments disclosed herein without departing from the scope and spirit of the present invention.

Claims

1. A method for determining the starting dose of FSH for polycystic ovary syndrome, characterized in that, The method comprises quantitatively detecting the patient's BMI, bFSH, bLH, AMH and AFC, wherein the patient is a PCOS patient receiving IVF or ICSI of a GnRH-ant regimen, and the starting dose of FSH is FSH=(0.491×BMI+1.312×bFSH-0.291×bLH-0.455×AMH-0.075×AFC-8.576) / 0.042, wherein the unit of FSH is IU / day, and the unit of BMI is kg / m 2 , the unit of bFSH is IU / L, the unit of bLH is IU / L, the unit of AMH is ng / mL, and the unit of AFC is ng / mL.

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