Proteomics & Glyco-Proteomic Analysis of Follicular Fluid

The recruitment status of this study is unknown because the information has not been verified recently.
Verified December 2011 by University of Cincinnati.
Recruitment status was  Not yet recruiting
Sponsor:
Collaborator:
Merck Sharp & Dohme Corp.
Information provided by (Responsible Party):
Steven Lindheim, University of Cincinnati
ClinicalTrials.gov Identifier:
NCT01487486
First received: November 16, 2011
Last updated: December 5, 2011
Last verified: December 2011
  Purpose

To the best of the investigators knowledge, exhaustive characterization of the low and high abundant proteins and glyco-proteins of the Follicular Fluid (FF) has not yet been achieved. Such an analysis may provide critical molecular data on the role of the FF in oocyte maturation and may identify specific changes in the FF proteome of patients with gynecologic problems, such as Polycystic Ovary Syndrome (PCOS).

Specific Aims

  1. To perform a comprehensive analysis of normal human FF using sensitive mass spectrometry in combination with conventional approaches for proteomic evaluation and using HPLC and Western blot for glyco-proteomic analysis.
  2. Characterize differential proteomic and glyco-proteomic patterns of the FF in normal women compared to lean and obese women with PCOS.
  3. To supplement the differential proteomic and glyco-proteomic analysis with steroid hormone analysis in all FF samples.

Condition Intervention
Polycystic Ovary Syndrome
Normal Volunteers
Drug: IVF Antagonist Protocol

Study Type: Observational
Study Design: Observational Model: Case Control
Time Perspective: Prospective
Official Title: Proteomics & Glyco-Proteomic Analysis of Follicular Fluid Derived From Health Patient/Donors and Polycystic Ovary Syndrome Patients

Resource links provided by NLM:


Further study details as provided by University of Cincinnati:

Primary Outcome Measures:
  • Proteomic analysis [ Time Frame: Participants will be followed for one IVF cycle including pregnancy outcomes, on average this will be 6-8 weeks. ] [ Designated as safety issue: No ]
    For proteomic analysis the follicular fluid samples will be either directly analyzed by MS or will be processed to deplete albumin which is likely to be present in very high abundance in the FF.


Secondary Outcome Measures:
  • Hormone analysis [ Time Frame: Participants will be followed for one IVF cycle including pregnancy outcomes, on average this will be 6-8 weeks. ] [ Designated as safety issue: No ]
    An aliquot of FF from each patient will be analyzed for the following steroid hormones: progesterone, 17-alpha-hydroxyprogesterone, androstenedione, testosterone, estradiol, and dihydrotestosterone.


Biospecimen Retention:   Samples Without DNA

Follicular Fluid


Estimated Enrollment: 30
Study Start Date: December 2011
Estimated Study Completion Date: January 2014
Estimated Primary Completion Date: December 2013 (Final data collection date for primary outcome measure)
Groups/Cohorts Assigned Interventions
Normal patients
Women with infertility diagnosis of male factor only or women who are oocyte donors
Drug: IVF Antagonist Protocol
  1. Ovulation Induction: Achieved with recombinant FSH (Follistim®) with or without HMG (Menopur®) at total doses of 75-450 IU/day subcutaneous (SC) for 9-14 days.
  2. Ovulation Suppression: GnRH Antagonist (Ganirelix® - 250microgram 0.5ml) will be initiated following ovulation induction when lead follicle >14mm diameter on ultrasound and continued through the day of hCG (Novirel® or Ovidrel ®) injection
  3. hCG Injection: Once patient has met criteria for oocyte retrieval, she will inject either Novarel® (5,000-10,000 units Intramuscular) or Ovidrel® (250microgram - 500microgram SC) 35 hours prior to oocyte retrieval.
Polycystic Ovary Syndrome, High BMI
Women with Polycystic Ovary Syndrome with a BMI between 30-35
Drug: IVF Antagonist Protocol
  1. Ovulation Induction: Achieved with recombinant FSH (Follistim®) with or without HMG (Menopur®) at total doses of 75-450 IU/day subcutaneous (SC) for 9-14 days.
  2. Ovulation Suppression: GnRH Antagonist (Ganirelix® - 250microgram 0.5ml) will be initiated following ovulation induction when lead follicle >14mm diameter on ultrasound and continued through the day of hCG (Novirel® or Ovidrel ®) injection
  3. hCG Injection: Once patient has met criteria for oocyte retrieval, she will inject either Novarel® (5,000-10,000 units Intramuscular) or Ovidrel® (250microgram - 500microgram SC) 35 hours prior to oocyte retrieval.
Polycystic Ovary Syndrome, Low BMI
Women with Polycustic Ovary Syndrom with a BMI between 20 & 25
Drug: IVF Antagonist Protocol
  1. Ovulation Induction: Achieved with recombinant FSH (Follistim®) with or without HMG (Menopur®) at total doses of 75-450 IU/day subcutaneous (SC) for 9-14 days.
  2. Ovulation Suppression: GnRH Antagonist (Ganirelix® - 250microgram 0.5ml) will be initiated following ovulation induction when lead follicle >14mm diameter on ultrasound and continued through the day of hCG (Novirel® or Ovidrel ®) injection
  3. hCG Injection: Once patient has met criteria for oocyte retrieval, she will inject either Novarel® (5,000-10,000 units Intramuscular) or Ovidrel® (250microgram - 500microgram SC) 35 hours prior to oocyte retrieval.

Detailed Description:

In this study, we plan to utilize ultrasensitive mass spectrometry (MS) and other conventional proteomic approaches to identify the low and high abundant proteins present in human FF. Additionally, we plan to use high-performance liquid chromatography (HPLC) and Western blot techniques to evaluate the Neu5Gc and glycan array based ELISA techniques to detect anti-Neu5Gc antibody profile in human FF. This analysis will be performed on FF samples obtained from normal women undergoing In-Vitro Fertilization and Embryo Transfers (IVF-ET) for a male factor alone and oocyte donors from our 3rd Party Reproduction Program and from lean and obese women with PCOS. This study will provide information on protein, glycoprotein, and steroid hormone expression during normal folliculogenesis and during the pathologic condition of PCOS, which should also provide basic scientific information on normal and abnormal oocyte development.

Human FF bathes the developing oocyte. Previous studies indicate that the FF contains cytokines, steroidal and protein hormones, and growth factors. The presence of proteins with such significant biological properties implies a paracrine and autocrine role for the FF in promoting normal oocyte development. Furthermore, the presence of any antigenic sialic acid Neu5Gc and the presence of antibodies targeting these antigenic glycoconjugates (glycolipid and glycoproteins decorated with sialic acid) may interfere with oocyte development, hormonal expression, fertilization, and possibly implantation. Here we hypothesize that an exhaustive proteomic and glyco-proteomic characterization of human FF is essential for a thorough understanding of its biological significance. We also hypothesize that PCOS may have differential expression of the FF protein and glyco-protein milieu, and that the expression may differ further between lean and obese women with PCOS. PCOS represents a heterogeneous disorder. The severity of hyperandrogensim, metabolic and menstrual disturbance, and obesity is variable with up to 40% not clinically expressing signs of classic hyperandrogenism. On the other hand, these atypical, often lean, PCOS women can have impaired glucose tolerance and diabetes. Reports suggest that these lean PCOS women have altered serum IGFBP-1, a characteristic endocrine feature of patients with obese PCOS, and related to hyperinsulinemia and/or obesity. The lean phenotype of PCOS and its significance is unclear but may represent a cryptic or unexpressed form of PCOS or may be a prelude to individuals who will later manifest clinical signs of obese/overweight PCOS. Changes in expression may be expected because of the different amounts of steroidal hormones and inflammatory markers in the FF derived from women with PCOS.

  Eligibility

Ages Eligible for Study:   18 Years and older
Genders Eligible for Study:   Female
Accepts Healthy Volunteers:   No
Sampling Method:   Non-Probability Sample
Study Population

Women undergoing In Vitro Fertilization who have:

  1. male factor only infertility diagnosis or are oocyte donors
  2. PCOS
Criteria

Inclusion criteria: Inclusion Criteria All

  1. Female patients undergoing controlled ovarian hyperstimulation (COH), transvaginal oocyte aspiration (TVA), and Saline Infused Sonography (SIS) with UL collection
  2. Age <35 y/o at time of in vitro fertilization (IVF) cycle
  3. Normal ovarian function defined Day 3 Follicular Stimulating Hormone (FSH) <8 pg/ml or Anti-Mullerian Hormone (≥ 1.0 ng/ml)

Inclusion Criteria Controls:

  1. Female patients undergoing COH and TVA donating her oocytes
  2. Female patients undergoing COH and TVA for male factor infertility only (i.e. no female causes of infertility)
  3. Normal menstrual cycles

Inclusion Criteria Lean PCOS:

  1. Diagnosis of PCOS by Rotterdam Criteria
  2. BMI ≤ 25 kg/m2 Inclusion Criteria Classic PCOS

1. Diagnosis of PCOS by Rotterdam Criteria 2. BMI > 30 kg/m2

Exclusion criteria:

  1. Age ≥ 35 y/o
  2. Female partners with infertility associated diagnosis (i.e. tubal factor, cervical factor, endometriosis)
  3. Unexplained infertility
  Contacts and Locations
Choosing to participate in a study is an important personal decision. Talk with your doctor and family members or friends about deciding to join a study. To learn more about this study, you or your doctor may contact the study research staff using the Contacts provided below. For general information, see Learn About Clinical Studies.

Please refer to this study by its ClinicalTrials.gov identifier: NCT01487486

Contacts
Contact: Steven Lindheim, MD, MMM 513-585-0063 steven.lindheim@uc.edu
Contact: Julie Sroga, MD 513-585-2355 julie.sroga@uc.edu

Locations
United States, Ohio
Center for Reproductive Health Not yet recruiting
Cincinnati, Ohio, United States, 45219
Contact: Steven Lindheim, MD, MMM    513-585-0063    steven.lindheim@uc.edu   
Contact: Julie Sroga, MD    513-585-2355    julie.sroga@uc.edu   
Principal Investigator: Steven Lindheim, MD,MMM         
Sponsors and Collaborators
University of Cincinnati
Merck Sharp & Dohme Corp.
Investigators
Principal Investigator: Steven Lindheim, MD, MMM University of Cincinnati
  More Information

Publications:

Responsible Party: Steven Lindheim, Professor, University of Cincinnati
ClinicalTrials.gov Identifier: NCT01487486     History of Changes
Other Study ID Numbers: Proteomics of FF in PCOS
Study First Received: November 16, 2011
Last Updated: December 5, 2011
Health Authority: United States: Institutional Review Board

Keywords provided by University of Cincinnati:
Polycystic Ovary Syndrome
PCOS
Follicular Fluid
Proteomics
Glyco-proteomics

Additional relevant MeSH terms:
Polycystic Ovary Syndrome
Syndrome
Adnexal Diseases
Cysts
Disease
Endocrine System Diseases
Genital Diseases, Female
Gonadal Disorders
Neoplasms
Ovarian Cysts
Ovarian Diseases
Pathologic Processes

ClinicalTrials.gov processed this record on October 23, 2014