Ovary Syndrome for Efficient Diagnosis and Targeted Therapy

Study Overview
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Brief Summary
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OBJECTIVES: Analysis of DNA methyl transferases (DNMT1, DNMT3A and DNMT3B) and Histone deacetylases (HDAC 1,2,3 and SIRTs) polymorphisms (Somatic and germ line variations). Analysis of differential mRNA and protein expression of epigenetic markers in ovarian tissues obtained from PCOS patients. miRNA regulated epigenetic mechanisms in PCOS Epigenetic regulation of endocrine genes in PCOS DESIGN : A Case Control study.Sample size:200

Detailed Description
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We will take both blood and ovarian tissue samples used to diagnosis. We will take approximately 3 ml of blood which will be used from the blood sample collected for routine preoperative tests (surgical profile) before their planned surgery or for disease evaluation. Small tissue samples (from ovary) will be taken from the tissue which is surgically excised as a part of treatment.

Official Title
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Understanding the Epigenetic Mechanisms in the Pathophysiology of Polycystic Ovary Syndrome for Efficient Diagnosis and Targeted Therapy

Conditions
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Polycystic Ovary Syndrome

Intervention / Treatment
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* Procedure: laparoscopy / laparotomy
* Other: NO INTERVENTION


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: All the subjects would be of Indian origin (controls  100, cases  100). They would be in their reproductive age (18-45 yrs). Rotterdam consensus would be used to diagnose PCOS and cases will be selected based on it. Criteria for the diagnosis of PCOS would include oligo-ovulation cycles longer than 35 days or less than 26 days, elevated free testosterone levels (0.5 ng/dl; the cutof level for free testosterone level was the mean±2 SD according to normal levels in controls), oligomenorrhea or amenorrhea. A Ferriman-Gallwey (FG) score of≥7 would be taken as indicator for the presence of hirsutism. In accordance with the above criteria, polycystic ovary morphology would be determined by transvaginal ultrasonography, which defines PCOS as the presence of 12 or more small (2-9 mm) follicles in each ovary. Control subjects would have no signs of menstrual dysfunction and their androgen levels should be within the normal range, with normal glucose tolerance, and no family history of hirsutism, type 2 diabetes mellitus, and infertility. Exclusion Criteria: Women with other causes of hyperandrogenism such as hyperprolactinemia, androgen-secreting tumors, Cushing syndrome and nonclassic congenital hyperplasia would be excluded from this study.

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 45 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: This is a Case control study with two parallel groups
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: 100 women undergoing surgery (laparoscopy / laparotomy) with presumed diagnosis of PCOS (<br>We will take both blood and ovarian tissue samples used to diagnosis. We will take approximately 3 ml of blood which will be used from the blood sample collected for routine preoperative tests (surgical profile) before their planned surgery or for disease evaluation. Small tissue samples (from ovary) will be taken from the tissue which is surgically excised as a part of treatment. | Procedure: laparoscopy / laparotomy<br>* We will take both blood and ovarian tissue samples used to diagnosis. We will take approximately 3 ml of blood which will be used from the blood sample collected for routine preoperative tests (surgical profile) before their planned surgery or for disease evaluation. Small tissue samples (from ovary) will be taken from the tissue which is surgically excised as a part of treatment.<br>Other: NO INTERVENTION<br>* .We will take both blood and ovarian tissue samples used to diagnosis. 2.We will take approximately 3 ml of blood which will be used from the blood sample collected for routine preoperative tests (surgical profile) before their planned surgery or for disease evaluation. 3. Small tissue samples (from ovary) will be taken from the tissue which is surgically excised as a part of treatment.<br>|
| No Intervention: 100 women undergoing surgery for gynaecological disorder<br>We will take both blood and ovarian tissue samples used to diagnosis. We will take approximately 3 ml of blood which will be used from the blood sample collected for routine preoperative tests (surgical profile) before their planned surgery or for disease evaluation. Small tissue samples (from ovary) will be taken from the tissue which is surgically excised as a part of treatment. | |

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Evaluate the Analysis of DNA methyl transferases | DNA will be extracted as per the protocol followed routinely in our lab from blood and ovarian tissues. | 3 YEARS | 
| Evaluate the Analysis of differential mRNA and protein expression of epigenetic markers in ovarian tissues | Total RNA from ovarian tissue will be extracted using TRIzol reagent according to the manufacturer's instructions (Invitrogen Life Technologies) and reverse transcribed in the presence of random hexamers. Quantitative real-time PCR reactions will be carried out in an ABI 7000 Thermal Cycler (Applied Biosystems) to analyse the expression of epigenetic markers including the androgen regulated miRNAs and their downstream targets. | 3 YEARS | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| understand miRNA regulated epigenetic mechanisms | The present study would investigate the expression pattern of DNMTs, HDACs, miRNAs and their downstream targets in Ovarian tissues (samples of normal adult ovaries and polycystic ovaries) by western blot and immunohistochemical analysis (IHC). | 3 YEARS |