Methylation of the PPARg Promoter Region in Pregnancy

NCT02395835 | Completed

• 1 other identifier: 05-06-2009
• Study Type: observational
• Target: 41
• Locations: 0 countries
• Sites: N/A

Brief Summary

The main objective of this study was to assess whether clinical, anthropometric, and biochemical variables of the mother were associated with changes in the methylation of the PPARg promoter region (-351 to -260). Methodology: This was a matched cohort study with two groups: a) normal weight (NW) pregnant women (n = 21) and their offspring, and b) overweight (OW) pregnant women (n = 20) and their offspring. DNA was extracted from leukocytes (4000-10,000 cells) in the MagnaPure (Roche) using the MagNAPure LC DNA Isolation Kit 1 (Roche, Germany). The treatment of DNA (2 µg) was performed with sodium bisulfite (EZ DNA Methylation-Direct Kit, ZymoResearch). Real-time polymerase chain reaction (qPCR) was performed in a LightCycler 2.0 (Roche) using the SYBR® Advantage® qPCR Premix Kit (Clontech).

Conditions

Body Weight

Keywords

body mass index; methylation; PPAR gamma; pregnancy

Outcome Measures

Primary Outcomes (1)

• Evidence of the effect of Body Mass Index (BMI) on the methylation status of the PPAR gamma promoter region (-351 to -260).

  The treatment of DNA (2 µg) was performed with sodium bisulfite (EZ DNA Methylation-Direct Kit, ZymoResearch). For the control group, we used purified human methylated and unmethylated DNA (Zymo Research) with specific oligonucleotides. Lymphocyte DNA from healthy donors was used as negative control, and methylated DNA "in vitro" with Sss I enzyme (New England Biolabs) was used as positive control for methylation. The methylated (M3) and unmethylated (U3) primers used were those proposed by Pancione et al.

  Pregnant women were followed until delivery.

Study Arms (2)

Normal weight (NW) pregnant women

Pregnant women with Body Mass Index (BMI) \> = 30. Body weight and height were measured in an overnight fasting status using an adult scale (Seca, Hamburg, Germany). Prepregnancy BMI was calculated as weight in kg divided by height in meters squared based on the prenatal chart or on the self-reported weight of women with no prenatal chart. Dietetic treatment was calculated according to height, weeks of gestation, and weight, considering an energy intake of 30 kcal/kg of ideal weight and a macronutrient distribution of: 55-65% carbohydrates, 10-20% fat, and the remainder as proteins.

Overweight (OW) pregnant women

Pregnant women with Body Mass Index (BMI) \< 30. Body weight and height were measured in an overnight fasting status using an adult scale (Seca, Hamburg, Germany). Prepregnancy BMI was calculated as weight in kg divided by height in meters squared based on the prenatal chart or on the self-reported weight of women with no prenatal chart. Dietetic treatment was calculated according to height, weeks of gestation, and weight, considering an energy intake of 30 kcal/kg of ideal weight and a macronutrient distribution of: 55-65% carbohydrates, 10-20% fat, and the remainder as proteins.

Eligibility Criteria

• Age: 18 Years - 42 Years
• Sex: female
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64)
• Sampling Method: Non-Probability Sample

Study Population

Pregnant women attended at the Maternal-Perinatal Hospital "Mónica Pretelini Sáenz".

Sponsors & Collaborators

• Materno-Perinatal Hospital of the State of Mexico: lead

Biospecimen

Retention: SAMPLES WITH DNA

Leukocytes were obtained according to the ACK-lysing buffer (LONZA) protocol. Briefly, a peripheral blood sample was placed in EDTA tube and then centrifuged at 2500 rpm for 10 min. All samples were kept at -80°C until further analysis. DNA was extracted from leukocytes (4000-10,000 cells) in the MagnaPure (Roche) using the MagNAPure LC DNA Isolation Kit 1 (Roche, Germany).

MeSH Terms

Conditions

Body Weight

Condition Hierarchy (Ancestors)

Signs and Symptoms; Pathological Conditions, Signs and Symptoms

Study Officials

• Hugo Mendieta Zerón, PhD.

  Asociación Científica Latina A.C.

  PRINCIPAL INVESTIGATOR

Study Design

• Study Type: observational
• Observational Model: COHORT
• Time Perspective: PROSPECTIVE
• Target Duration: 1 Year
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Médico Internista adscrito a la Unidad de Cuidados Intensivos Obstétricos

Study Record Dates

• First Submitted: March 11, 2015
• First Posted: March 24, 2015
• Study Start: September 1, 2009
• Primary Completion: August 1, 2010
• Study Completion: March 1, 2011
• Last Updated: March 24, 2015

Record last verified: 2015-03