Clinical trial NCT06878781: Postprandial Metabolome and Metabolic Flexibility
Official title: Standardized Meals With Different Macronutrient Ratios on the Postprandial Metabolome and Metabolic Flexibility
Study type: INTERVENTIONAL
Overall status: RECRUITING
Phase: NA
Conditions: Fasting
Postprandial Hyperglycemia
Lipid Metabolism Disorders
Glucose Metabolism Disorders
Insulin Sensitivity
Keywords: metabolic flexibility
postprandial response
mixed meals
Lead sponsor: Instituto Nacional de Medicina Genomica
Sponsor class: OTHER_GOV
Collaborators: name: Universidad Nacional Autonoma de Mexico
First posted: 2025-03-17
Enrollment: 300
Sex: ALL
Minimum age (years): 18
Maximum age (years): 30
Standard ages: ADULT
Accepts healthy volunteers: yes
Allocation: RANDOMIZED
Intervention model: CROSSOVER
Intervention model description: Enrolled subjects are followed every week for one month. At each visit, a questionnaire assesses daily time-activity patterns. Dietary intake is assessed by a food frequency questionnaire and a multi-step 24-hour dietary recall. Subjects will then be randomized to receive the metabolic challenges in a different order. The procedures will be performed before (fasting; 8-10 hours) and after (postprandial) consumption of the metabolic challenges. Capillary blood samples are obtained in the morning after an 8-hour fasting and after test meal consumption. Once the drop of blood is formed, it is placed directly into the CardioCheck Plus® cassette to determine glucose triglyceride, LDL-cholesterol, HDL-cholesterol, and total cholesterol. A second drop of capillary blood shall be placed on a filter paper. Indirect fasting calorimetry is also performed. After the indirect calorimetry, the metabolic and hormonal response to the test meal is performed.
Primary purpose: PREVENTION

Brief summary: Metabolic flexibility is a process in which the body can switch energy substrates in different physiological states. This flexibility plays an important role in an individual's health because losing it increases the risk of obesity, metabolic syndrome, insulin resistance, and type 2 diabetes. Considering that humans spend most of their awakening hours in a postprandial (PP) state, an organism's metabolic flexibility (MF) to respond to a standardized meal's consumption would provide information on the individual's metabolic health. The PP response to glucose following an oral glucose tolerance test or consumption of a high-carbohydrate meal is well described; however, few studies assess the FM and PP metabolome using mixed meals with different macronutrients. The investigators address how metabolic flexibility and metabolome change after consuming standardized meals with different macronutrient ratios. Data collection includes clinical and diet information, indirect calorimetry, and capillary blood sampling during fasting and after consumption of standardized meals. Samples are collected weekly for one month. The data will determine the metabolic flexibility and metabolome after consuming standardized meals with different macronutrient ratios.

Detailed description: Enrolled subjects are followed every week for one month. At each visit, a questionnaire assesses daily time activity patterns relevant to energy expenditure, general health status, including infectious symptoms, and confirmation of basic social and demographic characteristics. Dietary intake is assessed by a food frequency questionnaire and a multi-step 24-hour dietary recall for quantitative analysis. If symptoms of infection are present, participants are treated with ad-hoc broad antibiotics. Anthropometrics are obtained. Subjects will then be randomized to receive the metabolic challenges in a different order. The procedures will be performed before (fasting; 8-10 hours) and after (postprandial) consumption of the metabolic challenges. Capillary blood samples (40µ) are obtained in the morning after an 8-hour fasting and after test meal consumption. To obtain the capillary blood sample, sterilize the ring finger with alcohol and allow it to dry. Then, puncture the area with a sterile 2 mm long lancet. Once the drop of blood is formed, it is placed directly into the CardioCheck Plus® cassette to determine glucose triglyceride, LDL-cholesterol, HDL-cholesterol, and total cholesterol. A second drop of capillary blood shall be placed on a filter paper (S\&S 903) until a circle of filter paper is filled with blood to saturate the paper throughout its thickness. Insulin concentration shall be determined following the protocol for dried blood, which is standardized in the laboratory. Indirect fasting calorimetry is also performed. After the indirect calorimetry, the metabolic and hormonal response to the test meal is performed.

Each challenge should be consumed within 15 minutes. After 5 minutes of rest, indirect calorimetry will be postprandially, lasting 30 minutes. Capillary blood shall be obtained at the following times: 15-30-45-60-90 and 120 min after ingestion of food.

Nutrient composition of standardized meal and meal example.

* High carbohydrate challenge: Energy, 479.6 kcal; Carbohydrate, 83.6%; Lipids, 12.4% and Protein, 4.0% (70 g of hot-Cake, 100 g of mango, 270 ml of peach nectar, and 40 g of strawberry jam)
* High lipid challenge: Energy, 1043.4 kcal; Carbohydrate, 4.9%; Lipids, 86.8%, and Protein, 8.3% (60 g of manchego cheese, 25 g of egg, white, dried, 24 g of bacon, 5 ml of oil, 65 g of cream cheese, 70 g of cream, and 16 g of poblano pepper)
* High protein challenge: Energy, 441.3 kcal; Carbohydrate, 1.6%; Lipids, 5.1%, and Protein, 93.3% (2 scoop Isopure Zero Carb, 180 g of chicken breast, 20 g of lettuce and 24 g of ham turkey breast)

Determination of metabolites in dried blood To determine the concentration of metabolites (amino acids and acyl-carnitines), a circle of 3 mm diameter shall be punched out of the filter paper and placed in a 96-well plate. Add 100 µl of the acyl-carnitine and amino acid standards from the NeoBase PerkinElmer kit. Subsequently, follow the manufacturer's instructions for determining metabolites by liquid chromatography coupled to mass spectrometry (LC-MS).

Determination of the respiratory quotient (RQ) To determine the respiratory quotient (RQ= VmaxCO2/ VmaxO2) and lipid and carbohydrate oxidation examinations, the Cardio Coach CO2 Vmax Encore 29 System calorimeter software (Korr, Inc, UT, USA) will be used according to the instructions of the supplier. Examinations are invariably performed in the morning (8:00-9:00 am) in a thermoneutral environment with controlled pressure, humidity, and temperature, with the patient supine but awake. The investigators examine a maximum of two subjects per day. Oxygen consumption and carbon dioxide production were obtained using a canopy and were monitored continuously for 30 minutes. The initial 10 minutes of the measurement are discarded for the calculation to ensure greater data homogeneity. O2 consumption and CO2 production will be recorded continuously for 30 min. VO2 and VCO2 values will be used in the equation proposed by Weir (Energy Expenditure = \[3.941(VO2) + 1.11(VCO2)\] x 1440 min/day), considered as the standard method \[34\]. Additionally, one day before the test, subjects are instructed to fast for 8 hours and not engage in physical activity or consume caffeine the day before the exam. All participants will be asked to eat the same dinner the night before each test. Dinner provides 15% of the daily energy intake; 20 g of protein, 7 g of lipids, and 34 g of carbohydrates)

Eligibility criteria: Inclusion Criteria:

* be over 18 years of age and under 30 years of age;
* without active drug addictions: smoking, alcoholism, and/or drug addiction;
* in female subjects: regular menstrual cycles;
* give written consent for their inclusion in the study.

Exclusion Criteria:

* subjects with any pathology that requires medication or special treatment, such as type 2 diabetes mellitus, high blood pressure, dyslipidemia, polycystic ovary syn-drome, autoimmune, thyroid, kidney, neurological diseases, and cancer;
* pregnant or lactating women;
* subjects with problems with chewing, salivation, and swallowing.

Elimination criteria:

* subjects who only have samples (blood and calorimetry) from one study period (fasting or postprandial);
* subjects who only have samples (blood and calorimetry) of a metabolic challenge;
* subjects who have an infection at the scheduled appointment;
* subjects who withdraw their informed consent

Primary outcomes:
measure: Metabolic Flexibility Assessed by Respiratory Quotient (RQ) and Substrate Oxidation
timeFrame: Once a week for three weeks
Secondary outcomes:
measure: Postprandial Glucose Levels
timeFrame: Once a week for three weeks
measure: Postprandial Triglyceride Levels
timeFrame: Once a week for three weeks
measure: Postprandial Acyl Carnitines Levels
timeFrame: Once a week for three weeks
measure: Postprandial Amino Acid Levels
timeFrame: Once a week for three weeks
Other outcomes:
Locations:
city: Mexico City
country: Mexico
facility: Berenice Palacios-Gonzalez
Study officials:
affiliation: National Institute of Genomic Medicine
name: Berenice Palacios-Gonzalez, PhD
role: PRINCIPAL_INVESTIGATOR
affiliation: Universidad Nacional Autonoma de Mexico
name: Noemi Meraz-Cruz, PhD
role: PRINCIPAL_INVESTIGATOR
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