Evaluation Of DNA Methylation Pattern In Healthy, Sarcopenic, Obese And Sarcopenic Obese Older Women: A Cross-Sectional Study
September 30, 2024 updated by: Ellen Cristini de Freitas, University of Sao Paulo
INTRODUCTION: Sarcopenic obesity (SO), a functional and clinical condition, is characterized by the coexistence of obesity, marked by excess fat mass and sarcopenia, characterized by reduced strength and muscle mass.
SO is associated with a greater risk of health-related adverse clinical outcomes than older adults with obesity and sarcopenia alone.
Aging is accompanied by numerous changes epigenetic.
These aging-associated epigenetic changes include DNA methylation, histone modification, chromatin remodeling, non-coding RNA (ncRNA) regulation, and RNA modification.
DNA methylation occurs at cytosines in CpG dinucleotides in the genome and undergoes changes with age in various human tissues.
Furthermore, many genes can be hypermethylated or hypomethylated on CpG islands with the aging process.
Soon, a broad exploration of candidate genes may provide insights into the pathogenesis of Sarcopenic obesity.
Therefore, understanding how aging, specifically sarcopenia, obesity and Sarcopenic obesity, is regulated by epigenetic factors, favors the development of new treatment therapies.
Thus, the objective will be to evaluate the epigenetic influence on sarcopenic obesity in older women.
METHODS: This cross-sectional study will include 32 older women who will be classified as healthy, with sarcopenia, obesity and sarcopenic obesity living in the city of Ribeirão Preto - SP.
The older adults will perform total and regional body scan using iDXA, anthropometric assessment, functional capacity tests, peripheral blood collection for analysis of biochemical markers and epigenetics.
For statistical analysis will be used t test, ANOVA, linear regression models and Pearson correlation.
Analyzes of the complete methylome will be performed using bioinformatics tools, including specific software.
EXPECTED RESULTS: It is expected that there will be differences in the patterns of methylation and gene expression in the diseases analyzed.
In addition, it is expected to clarify how epigenetic changes occur throughout this process.
Study Overview
Status
Active, not recruiting
Conditions
Intervention / Treatment
Detailed Description
The sample size was calculated a priori, considering DNA methylation as the primary outcome, on an exploratory basis.
The analyses were conducted using R software (https://www.r-project.org).
For the calculation, an effect size of 0.65, alpha 0.05 and statistical power of 0.80 were used.
Thus, a total of 32 participants will be included in the study (n = 8 in each group).
The degree of methylation will be determined by measuring the amount of marker incorporated for each probe.
The image intensities will be extracted using the Illumina Genome Studio 2011.1,
Methylation module 1.9.0 software.
The methylation score will be represented as a beta value (β) according to the fluorescence intensity ratio.
Beta values can assume any value between 0 (unmethylated) and 1 (completely methylated).
Subsequently, the image intensities will be extracted from the raw data (IDATS) using the Champ package for R (Statistical Computing, VIE, Austria).
All genome-wide methylation data will be analyzed by t-test (within groups), followed by the Benjamini Hochberg test.
Pathway enrichment analysis will be performed using WEBGestalt and Reactome.
The normality of data distribution will be verified by the Shapiro Wilk test, and descriptive statistics will consist of mean and standard deviation values.
ANOVA will be used to compare groups.
Pearson correlations will be used to assess the correlation of DNA methylation patterns with phenotypic variables.
Linear regression models will be applied to analyze the prediction of methylation pattern and selected variables.
The multinomial logistic regression model will be used in the groups for association with telomere length adjusted for BMI, age, ALST).
Statistical significance will be set at 5%, and all analyses will be performed using the Statistical Package for Social Sciences software (SPSS version 17.0 Inc.
Chicago, IL).
Study Type
Observational
Enrollment (Estimated)
32
Contacts and Locations
This section provides the contact details for those conducting the study, and information on where this study is being conducted.
Study Locations
-
-
Sao Paulo
-
Ribeirao Preto, Sao Paulo, Brazil
- University of São Paulo, School of Physical Education and Sports of Ribeirão Preto
-
-
Participation Criteria
Researchers look for people who fit a certain description, called eligibility criteria. Some examples of these criteria are a person's general health condition or prior treatments.
Eligibility Criteria
Ages Eligible for Study
- Adult
- Older Adult
Accepts Healthy Volunteers
Yes
Sampling Method
Non-Probability Sample
Study Population
Participants will be recruited through social media and on the website of the USP School of Physical Education and Sports (EEFERP/USP).
Interested participants must register online and the researcher will then contact them by phone to schedule an interview.
Description
Inclusion Criteria:
- body mass (menor 120 kg);
- sedentary (not practicing physical exercise for at least 3 months);
- not using vitamin or mineral supplements, anxiolytic medications, hypoglycemic agents with activity on cytochrome P450 (CYP450) enzymes;
- classified within the criteria for sarcopenic obesity, sarcopenia and obesity.
Exclusion Criteria:
- alcoholics;
- smokers;
- infectious diseases;
- coronary diseases;
- chronic kidney diseases and
- presenting a score ≤ 13 for the cognitive examination in the Mini-Mental State Examination (MMSE).
Study Plan
This section provides details of the study plan, including how the study is designed and what the study is measuring.
How is the study designed?
Design Details
Cohorts and Interventions
Group / Cohort |
Intervention / Treatment |
|---|---|
|
Healthy older women
Older women who do not have sarcopenia, obesity or sarcopenic obesity.
|
None (Observational Study)
|
|
Older women with sarcopenia
Older women with sarcopenia, i.e. low handgrip strength and low muscle mass index.
|
None (Observational Study)
|
|
Older women with obesity
|
None (Observational Study)
|
|
Older women with sarcopenic obesity
Older women with sarcopenic obesity, i.e. low handgrip strength, low muscle mass index and high body mass index.
|
None (Observational Study)
|
What is the study measuring?
Primary Outcome Measures
Outcome Measure |
Measure Description |
Time Frame |
|---|---|---|
|
Investigate whether the health condition (healthy, sarcopenic, obese or with sarcopenic obesity) promotes changes in DNA methylation
Time Frame: 1 day
|
For this purpose, blood samples will be collected in the morning after 12 hours of fasting in EDTA tubes by experienced nurses.
5 ml of blood will be collected for analysis of biochemical markers and genetic markers (DNA methylation and gene expression).
DNA extraction from peripheral blood will be performed with the AllPrepDNA/RNA/miRNA Universal kit (Qiagen), according to the instructions, using 200 µL of the extracted material.
The quality of the extracted DNA will be assessed with the Greends DNA Quantification Reagent (Invitrogen, Carlsbad, CA, USA).
After extraction, the samples will be stored in a freezer at -80°C for analysis.
DNA will be bisulfite converted using the EZ DNA Methylation-Gold Conversion Kit (Zymo Research, CA, USA) according to the instructions, with conversion of unmethylated cytosine to uracil.
Array-based specific DNA methylation analysis will be performed using the Infinium Human Methylation 850K (EPIC) Beadchip technology.
|
1 day
|
Secondary Outcome Measures
Outcome Measure |
Measure Description |
Time Frame |
|---|---|---|
|
Correlate the epigenetic variables found with biochemical markers, body composition and physical performance.
Time Frame: 1 day
|
To do this, the following assessments will be performed: Six-minute walk test: the total distance covered (meters) in 6 minutes will be measured. Forearm flexion test: the score is obtained by the total number of flexions performed correctly during 30 seconds. Handgrip strength measurement: the handgrip strength test in kg will be performed using a hand dynamometer: The score will be obtained by the total number of complete executions during 30 seconds. Body composition will be performed by total and regional body scanning using iDXA. The exam will provide measurements in grams of total and regional body (right and left lower limbs, trunk, right and left upper limbs and head) of: fat mass (g); percentage of total and regional fat (% G); bone mineral content; appendicular lean soft tissue will be determined by summing the lean soft tissue of the lower and upper limbs on the right and left sides. |
1 day
|
Collaborators and Investigators
This is where you will find people and organizations involved with this study.
Study record dates
These dates track the progress of study record and summary results submissions to ClinicalTrials.gov. Study records and reported results are reviewed by the National Library of Medicine (NLM) to make sure they meet specific quality control standards before being posted on the public website.
Study Major Dates
Study Start (Actual)
February 21, 2024
Primary Completion (Actual)
September 5, 2024
Study Completion (Estimated)
October 5, 2024
Study Registration Dates
First Submitted
September 23, 2024
First Submitted That Met QC Criteria
September 25, 2024
First Posted (Actual)
October 1, 2024
Study Record Updates
Last Update Posted (Actual)
October 2, 2024
Last Update Submitted That Met QC Criteria
September 30, 2024
Last Verified
September 1, 2024
More Information
Terms related to this study
Keywords
Additional Relevant MeSH Terms
Other Study ID Numbers
- 73189623.6.0000.5659
Plan for Individual participant data (IPD)
Plan to Share Individual Participant Data (IPD)?
NO
Drug and device information, study documents
Studies a U.S. FDA-regulated drug product
No
Studies a U.S. FDA-regulated device product
No
This information was retrieved directly from the website clinicaltrials.gov without any changes. If you have any requests to change, remove or update your study details, please contact register@clinicaltrials.gov. As soon as a change is implemented on clinicaltrials.gov, this will be updated automatically on our website as well.