Analyzing the effects of short-term fasting on transcriptome and epigenome signatures compared with food consumption in healthy subjects
| ISRCTN | ISRCTN12651780 |
|---|---|
| DOI | https://doi.org/10.1186/ISRCTN12651780 |
| ClinicalTrials.gov (NCT) | Nil known |
| Clinical Trials Information System (CTIS) | Nil known |
| Protocol serial number | PTRC_1521410 |
| Sponsor | University of Valencia |
| Funders | Conselleria de Innovación, Universidades, Ciencia y Sociedad Digital, Generalitat Valenciana, Universitat de València |
- Submission date
- 06/12/2021
- Registration date
- 09/12/2021
- Last edited
- 03/11/2022
- Recruitment status
- No longer recruiting
- Overall study status
- Completed
- Condition category
- Nutritional, Metabolic, Endocrine
Plain English summary of protocol
Background and study aims
Several studies in animals and some in humans have reported that dietary interventions based on intermittent fasting or on time-restricted feeding lead to promising improvements in the cardiometabolic risk profile (decreasing body weight, reducing blood sugar and fat, etc). However, although some investigations in humans have associated these beneficial effects with epigenetic modifications (changes that modify the activation of certain genes, but not the genetic code sequence of DNA), mainly with DNA methylation and changes in gene expression in key genes related to metabolism, very few studies have analyzed the changes in the transcriptome (gene expression) and in the epigenome (mainly focusing on DNA methylation) at the genome-wide level. Therefore the main aim of this study is to explore the changes in the transcriptome at the genome-wide level in healthy subjects from a Mediterranean population after a short-term fasting period (24 hours) in comparison with frequent food consumption in the same period, and to establish the corresponding transcriptome and methylome signatures that identify the main differentially expressed and or methylated genes depending on the fasting or on the frequent food consumption status in the short-term intervention.
Who can participate?
Men and women (white European) from the general Mediterranean Spanish population (aged 18-50 years)
What does the study involve?
Each participant receives both interventions in a random order. One intervention will consist of a period of 24 hours fasting (only water will be allowed) and the other intervention will consist of a frequent meal pattern (6 times a day, every 3 hours) for the equivalent 24-hour period. In this food consumption intervention, the participant will be provided with typical foods of the Mediterranean diet: fruits, vegetables, legumes, extra virgin olive oil, fish, white meat and nuts). There will be a 2-week break between the interventions.
What are the possible benefits and risks of participating?
There are no direct medical benefits of participating in the study. Participants will be informed that there are no benefits and risks expected.
Where is the study run from?
University of Valencia (Spain)
When is the study starting and how long is it expected to run for?
January 2020 to December 2022
Who is funding the study?
Generalitat Valenciana and University of Valencia (Spain)
Who is the main contact?
Prof. Carolina Ortega-Azorín
Carolina.Ortega@uv.es
Contact information
Scientific
School of Medicine. University of Valencia
Department of Preventive Medicine
Avda Blasco Ibanez, 15
Valencia
46010
Spain
 ORCID ID |
0000-0001-6719-9358 |
|---|---|
| Phone | +34 (0)963864800 |
| carolina.ortega@uv.es |
Study information
| Primary study design | Interventional |
|---|---|
| Study design | Interventional randomized cross over trial |
| Secondary study design | Randomised cross over trial |
| Study type | Participant information sheet |
| Scientific title | Effect of fasting for 24 hours compared to frequent food consumption on anthropometric, biochemical and gene expression and methylation variations in the general population (AYUGEN) |
| Study acronym | AYUGEN |
| Study objectives | The hypothesis is that fasting for 24 hours can change gene expression and DNA-methylation profiles at the genome-wide level in comparison with a pattern of food consumption, allowing us to identify the transcriptome and methylome signatures associated with fasting and frequent food consumption in a Mediterranean population. |
| Ethics approval(s) | Approved 15/01/2021, Ethics Committee on Human Research at the University of Valencia (Avda. Blasco Ibanez 13, Valencia, 46010, Spain; +34 (0) 963864109; vicerec.investigacio@uv.es), ref: CR_1521410 |
| Health condition(s) or problem(s) studied | Short-term fasting |
| Intervention | This is a short-term cross-over intervention trial in 25 participants (including men and women). Two interventions related to food consumption will be carried out. All participants will receive both interventions. Participants will be randomly assigned 1:1 to the order of the two interventions by a computer-generated randomization procedure. The interventions will consist of: 1. Fasting for 24 hours (water allowed) 2. Consumption of foods for the equivalent 24-hour period The frequent food consumption intervention pattern includes eating 6 times a day every 3 hours. In this food consumption intervention, participants will be provided with typical foods of the Mediterranean diet: fruits, vegetables, legumes, extra virgin olive oil, fish, white meat and nuts). A wash-out period of 2 weeks between interventions will be carried out. |
| Intervention type | Behavioural |
| Primary outcome measure(s) |
Comparative genome-wide gene expression and methylation profiles in leukocytes (using DNA and RNA isolated from blood) to identify transcriptome and methylome signatures (detection of differentially expressed genes and DNA methylation sites) of 24-h fasting versus frequent food consumption after both interventions in healthy subjects from a Mediterranean population. Blood samples will be obtained at baseline and after 24 h of the corresponding intervention (fasting or food consumption). RNA and DNA will be isolated by standard procedures and quality control measures will be carried out. Transcriptome-wide human arrays will be used to analyze differential gene expression and RT-PCR will be used for validation. Methylation will be analyzed by arrays and for selected CpG sites by Massarray according to the standard protocols. Differential gene expression and CpG methylation sites will be statistically analyzed and the combination of the most relevant differentially expressed genes and DNA-methylation sites will be combined by computational analyses to build specific signatures for each intervention. |
| Key secondary outcome measure(s) |
1. Plasma glucose and biochemical parameters (lipids) measured by standard procedures at baseline, at 6 h and after the 24 h interventions in both groups |
| Completion date | 31/12/2022 |
Eligibility
| Participant type(s) | Healthy volunteer |
|---|---|
| Age group | Adult |
| Lower age limit | 18 Years |
| Sex | All |
| Target sample size at registration | 25 |
| Total final enrolment | 28 |
| Key inclusion criteria | Current inclusion criteria as of 25/07/2022: 1. White European subjects (men and women) recruited from the Mediterranean population 2. Age ranging from 18 to 50 years old 3. BMI between 21 and 34 kg/m² _____ Previous inclusion criteria: 1. White European subjects (men and women) recruited from the Mediterranean population 2. Age ranging from 18 to 50 years old 3. BMI between 23 and 33 kg/m² |
| Key exclusion criteria | 1. Diabetic subjects 2. Other chronic diseases (cardiovascular, cancer, respiratory diseases, liver diseases, kidney diseases, etc) 3. Subjects with food allergies or food intolerances 4. Alcohol abuse or addiction 5. Immunodeficiency, HIV-positive status, COVID-19 positive status or other acute infections 6. Serious psychiatric disorders: schizophrenia, bipolar disease, eating disorders, depression, etc 7. Any severe co-morbid condition 8. History of major organ transplantation 9. Concurrent therapy with immunosuppressive drugs or cytotoxic agents 10. Current treatment with systemic corticosteroids 11. Current use of weight loss medication 12. Any other condition that may interfere with the completion of the study protocol |
| Date of first enrolment | 20/12/2021 |
| Date of final enrolment | 31/10/2022 |
Locations
Countries of recruitment
- Spain
Study participating centres
Avda. Blasco Ibanez 15
Valencia
46010
Spain
Madrid
28029
Spain
Results and Publications
| Individual participant data (IPD) Intention to share | No |
|---|---|
| IPD sharing plan summary | Not expected to be made available |
| IPD sharing plan | Data will not be available outside the core research group as the informed consent form signed by participants stated that individual-level data will not be publicly available. Researchers who are interested in this study can contact the main investigator (Dr Carolina Ortega; carolina.ortega@uv.es) if they have any questions regarding the data or are interested in further collaborations. |
Study outputs
| Output type | Details | Date created | Date added | Peer reviewed? | Patient-facing? |
|---|---|---|---|---|---|
| Participant information sheet | Participant information sheet | 11/11/2025 | 11/11/2025 | No | Yes |
Editorial Notes
03/11/2022: The total final enrolment was added.
25/07/2022: The following changes have been made:
1. The recruitment end date has been changed from 30/06/2022 to 31/10/2022.
2. The participant inclusion criteria have been changed.
09/12/2021: Trial's existence confirmed by the Ethics Committee on Human Research at the University of Valencia.
