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The Fasting Study - Unraveling the Mechanistic Effects of Prolonged Fasting in Humans.

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ClinicalTrials.gov Identifier: NCT03757767
Recruitment Status : Completed
First Posted : November 29, 2018
Last Update Posted : April 24, 2019
Sponsor:
Information provided by (Responsible Party):
Lydia A. Afman, Wageningen University

Brief Summary:
Elevated levels of plasma triglycerides are increasingly recognized as an important causal risk factor for cardiovascular disease and associated pathologies. Lowering plasma triglycerides may therefore be a therapeutic target to lower cardiovascular disease risk. With this study the investigators want to examine the effects of fasting on adipose tissue metabolism in humans.

Condition or disease Intervention/treatment Phase
Fasting Adipose Tissue Clinical Trial Lipid Metabolism Other: Fasting for 26-hours Not Applicable

Detailed Description:

Rationale: Elevated levels of plasma triglycerides are increasingly recognized as an important causal risk factor for cardiovascular disease and associated pathologies. Lowering plasma triglycerides may therefore be a therapeutic target to lower cardiovascular disease risk. Lipoprotein lipase (LPL) is the enzyme that catalyzes the hydrolysis of plasma triglycerides into fatty acids on the capillary endothelium, thereby lowering plasma triglycerides and stimulating the uptake of these fatty acids into cells. Angiopoietin-like 4 (ANGPTL4) is produced by fat cells and promotes the unfolding and inactivation of LPL, leading to decreased hydrolysis of plasma triglycerides. The role of ANGPTL4 in regulating LPL is especially important during fasting. However, as most studies are performed in animal models, little is known about the effects of fasting on adipose tissue metabolism in humans, and specifically on ANGPTL4 and its relationship with LPL.

Objectives: The key objective of this study is to determine the effect of a prolonged fast on ANGPTL4 gene and protein expression in subcutaneous adipose tissue, the key organ involved in lipid metabolism. Additionally, the effect of a prolonged fast on LPL gene expression, LPL protein expression, and on LPL activity in subcutaneous adipose tissue will be examined. To explore the systemic effects of a prolonged fast, the investigators will also measure ANGPTL4 levels in plasma, and ANGPTL4 and LPL gene expression in circulating immune cells (PBMCs). The secondary objective of this study is to obtain a more holistic view on the changes occurring upon a prolonged fast, by examining the effects of fasting on whole genome expression, post-transcriptional changes, and on markers of metabolic status. The combined results of all measurements will ultimately lead to a better understanding of the mechanistic effects of a prolonged fast and the metabolic functioning in humans.

Study design: The Fasting Study is a single arm intervention study in healthy men and women. The investigators will examine the effects of a fed state, which is 2 hours after the last food intake, versus a prolonged fasted state, which is 26 hours after the last food intake. Adipose tissue biopsies and blood samples will be taken 2 hours after the last meal and 26 hours after the last meal. The 24 hours between sampling points are chosen to prevent effects of the circadian rhythm.

Study population: The study population will consist of 24, 40-70 years old men and women with a BMI of 22-30 kg/m2, selected from the surroundings of Wageningen through the mailing list for potential study research subjects of the division of Human Nutrition and health of Wageningen University. If needed, additional recruitment of research subjects will take place by flyers and posters, or advertisements in local newspapers.

Intervention: All research subjects will start with the consumption of a standardized meal (ad libitum). Directly thereafter the intervention begins, which is a fasting period of twenty-six-hours, during which research subjects are not allowed to eat or drink anything except for water, which the research subjects may consume ad libitum. The investigators will take the first blood samples and adipose tissue biopsies two hours after the consumption of the standardized meal. This is the fed state: the point in time where the body is physiologically fed, based on the average time for plasma free fatty acids to drop. Twenty-four hours after the "fed state", research subjects are in the prolonged fasted state, and the investigators will take the final blood samples and adipose tissue biopsies. The time period of twenty-four-hours between sampling points is chosen to prevent the effects of circadian related changes on our outcome measures.

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Study Type : Interventional  (Clinical Trial)
Actual Enrollment : 24 participants
Allocation: N/A
Intervention Model: Single Group Assignment
Intervention Model Description: Research subjects are measured in the fed state and in the prolonged fasted state.
Masking: None (Open Label)
Primary Purpose: Basic Science
Official Title: The Fasting Study - Unraveling the Mechanistic Effects of Prolonged Fasting in Humans.
Actual Study Start Date : November 12, 2018
Actual Primary Completion Date : January 22, 2019
Actual Study Completion Date : January 22, 2019

Arm Intervention/treatment
Experimental: Intervention arm
Fasting for 26-hours.
Other: Fasting for 26-hours

All research subjects will have to fast for 26 hours in total.

All research subjects start with a standardized meal. Two hours after the standardized meal the samples in the fed state will be taken. Research participants are not allowed to eat anything after the consumption of the standardized meal until the second measurements on day 2, 26-hours later. There are 24 hours between the two measurement points.

Other Name: Prolonged fasting




Primary Outcome Measures :
  1. Change in ANGPTL4 protein and gene expression in adipose tissue [ Time Frame: there are 24 hours between the measurement in de fed and the fasted state ]
    Using Western blotting, the change in ANGPTL4 protein expression in adipose tissue will be measured from fed to a prolonged fasted state. The change in ANGPTL4 gene expression in adipose tissue will be measured from fed to a prolonged fasted state using whole genome affymetrix microarrays and targeted quantitative real time PCR.

  2. Change in LPL expression, both gene and protein, and LPL activity in adipose tissue [ Time Frame: There are 24 hours between the measurement in the fed and the fasted state ]
    Using Western blotting, the change in LPL protein expression in adipose tissue will be measured from fed to a prolonged fasted state. The change in LPL gene expression in adipose tissue will be measured from fed to a prolonged fasted state using whole genome affymetrix microarrays and targeted quantitative real time PCR. The change in LPL activity in subcutaneous adipose tissue will be measured from fed to a prolonged fasted state, using a lipoprotein lipase assay.

  3. Change in ANGPTL4 expression in plasma [ Time Frame: There are 24 hours between the measurement in the fed and the fasted state ]
    Using ELISA, the change in ANGPTL4 expression in plasma will be measured from fed to a prolonged fasted state.

  4. Change in ANGPTL4 and LPL gene expression in PBMCs [ Time Frame: There are 24 hours between the measurement in the fed and the fasted state ]
    The change in ANGPTL4 and LPL gene expression in PBMCs from fed to a prolonged fasted state will be measured using whole genome affymetrix microarrays and targeted quantitative real time PCR.


Secondary Outcome Measures :
  1. Change in whole genome expression [ Time Frame: There are 24 hours between the measurement in the fed and the fasted state ]
    Using whole genome affymetrix micorarrays, the change in whole genome gene expression in both adipose tissue and PBMCs will be measured from fed to a prolonged fasted state.

  2. Post-transcriptional changes [ Time Frame: There are 24 hours between the measurement in the fed and the fasted state ]
    Using microRNAarrays of Thermo Fisher Scientific, targeted digital droplet PCR, and quantitative real time PCR, post-transcriptional changes in adipose tissue and PBMCs will be measured from fed to a prolonged fasted state.

  3. Changes in metabolite profiles [ Time Frame: There are 24 hours between the measurement in the fed and the fasted state ]
    Using untargeted and targeted metabolomic platforms, the changes in metabolite profiles in the adipose tissue and plasma/serum will be measured.

  4. Changes in protein profiles [ Time Frame: There are 24 hours between the measurement in the fed and the fasted state ]
    Using untargeted and targeted proteomic platforms, the changes in protein profiles in the adipose tissue and plasma/serum will be measured.


Other Outcome Measures:
  1. BMI [ Time Frame: At baseline ]
    weight (in kg) and height (in meters) will be measured and combined to report BMI in kg/m^2



Information from the National Library of Medicine

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Ages Eligible for Study:   40 Years to 70 Years   (Adult, Older Adult)
Sexes Eligible for Study:   All
Accepts Healthy Volunteers:   Yes
Criteria

Inclusion Criteria:

  • Apparently healthy man or woman
  • Age 40-70y at the time of recruitment
  • BMI of 22-30 kg/m2
  • Willing to fast for 26 hours
  • Willing to give blood samples
  • Willing to give adipose tissue biopsies
  • Willing to consume the standardized meal on test day 1
  • Signed informed consent
  • Having a general practitioner

Exclusion Criteria:

  • Alcohol on average: more than 2 consumptions/day or more than 14 consumptions/week
  • Tobacco smoker
  • Usage of drugs
  • Following a diet within one month of starting the study (for example a ketogenic diet)
  • Allergic to one or more components of the standardized meal
  • Donated or intend to donate blood from 2 months before the study until the end of the study
  • Unstable body weight (weight gain or loss >5 kg in the past three months)
  • Diagnosed with any long-term medical condition that can interfere with the study outcome (i.e. cardiovascular disease, diabetes mellitus type 1 or 2, liver, pulmonary, or renal disease)
  • Use of prescribed medication
  • Use of supplements or over the counter medication (OTC) known to interfere with glucose or lipid homeostasis in the week before the start of the study, and during the study
  • Being pregnant or lactating
  • Participation in another biomedical study (other than the EetMeetWeet study) during this study
  • Members of the research team, or first degree family members of the research team
  • Working, or doing an internship at the division "Human Nutrition and Health" - Wageningen University

Information from the National Library of Medicine

To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor.

Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT03757767


Locations
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Netherlands
Wageningen University, Division of Human Nutrition
Wageningen, Gelderland, Netherlands, 6700 EV
Sponsors and Collaborators
Wageningen University
Investigators
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Principal Investigator: Lydia Afman, PhD Wageningen University
Publications automatically indexed to this study by ClinicalTrials.gov Identifier (NCT Number):
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Responsible Party: Lydia A. Afman, Principal Investigator, Wageningen University
ClinicalTrials.gov Identifier: NCT03757767    
Other Study ID Numbers: NL-66589.081.18
First Posted: November 29, 2018    Key Record Dates
Last Update Posted: April 24, 2019
Last Verified: April 2019
Individual Participant Data (IPD) Sharing Statement:
Plan to Share IPD: Undecided
Plan Description: Collected data will be coded.

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Studies a U.S. FDA-regulated Drug Product: No
Studies a U.S. FDA-regulated Device Product: No
Keywords provided by Lydia A. Afman, Wageningen University:
Physiology
Humans